Pharmacological Evaluation of Selected Medicinal Plants Used in...

17
Research Article Pharmacological Evaluation of Selected Medicinal Plants Used in the Management of Oral and Skin Infections in Ebem-Ohafia District, Abia State, Nigeria Blessing O. Oyedemi, 1 Sunday O. Oyedemi , 2 Johnson V. Chibuzor, 1 Ifeoma I. Ijeh, 2 Roger M. Coopoosamy, 3 and Ayobami O. Aiyegoro 4 1 Department of Plant Science and Biotechnology, Michael Okpara University of Agriculture, Umudike, Nigeria 2 Department of Biochemistry, Michael Okpara University of Agriculture, Umudike, Nigeria 3 Department of Nature Conservation and Ethnobotany, Mangosuthu University of Technology, P.O. Box 12363, Jacobs, Durban 4026, South Africa 4 GI Microbiology and Biotechnology Unit, Agricultural Research Council, Animal Production Institute, Irene, Pretoria 0062, South Africa Correspondence should be addressed to Sunday O. Oyedemi; [email protected] Received 24 January 2018; Accepted 26 April 2018; Published 4 July 2018 Academic Editor: Carlos H. G. Martins Copyright © 2018 Blessing O. Oyedemi et al. is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Oral and skin infections contribute significantly to the global health challenges responsible for the current trend of increased morbidity and premature death. e purpose of this study was to document medicinal plants used in the management of oral and skin infections in Ebem-Ohafia Local Government Area (LGA), Abia State, and to characterize the in vitro antioxidant and antibacterial activity. e thin layer chromatography (TLC) profiling of ten of the selected folklore medicine was carried out using a various solvent system of different polarity index. e antioxidant capacity of the plant extracts was evaluated using chemical- based methods, and its antibacterial effect was investigated using disc diffusion and microdilution methods. Sixty-one plant species belonging to 26 families were discovered, and the most frequently cited species are Euphorbiaceae (18.03%), Fabaceae (11.47%), and Asteraceae (11.47%). All the plant extracts showed a promising free radical scavenging activity and efficient ferric reducing antioxidant power in a concentration-dependent manner possibly due to their richness in polyphenol with TLC profiling showing maximum three bands of phytochemicals. Also, the plant extracts exhibited a mild to weak antibacterial activity against our panel of bacterial strains having MIC values ranging from 256 to > 512 g/ mL reflected in their zone of inhibition at 10 g/disc. e data obtained for Breynia nivosa (BN), Eleusine indica (EI), Cassia alata (CA), Chromolaena odorata (CO), and Acalypha hispida (AH) extracts substantiate the traditional use of these herbal remedies in the region and open the possibility for the development of cheaper and affordable drugs in the treatment of oral and skin infections. Further studies are needed to identify active ingredient with strong antibacterial and antioxidant capacities along with their molecular mechanisms. 1. Introduction Oral health is a state of being free from the mouth and facial pain, throat cancer, oral infection, and sores that limit human capacity in biting, chewing, smiling, and speaking as well as psychosocial well-being [1]. Owing to the environmental pollution, the concentration of heavy metals such as Ni, Cr, Zn, Hg, and Pb in most food materials and water is rapidly increasing which severely affects both oral and general health condition. For example, dental caries, periodontitis, and throat cancer have been recognised as the most prevalent noncommunicable diseases associated with heavy metals, globally affecting school-aged children and a huge majority of adults [2]. In Nigeria, the rate of deep periodontal pocket is between 15 and 58% (aged 15 years) while the preva- lence of unmanaged dental caries is high raising alarm for Hindawi e Scientific World Journal Volume 2018, Article ID 4757458, 16 pages https://doi.org/10.1155/2018/4757458

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Research ArticlePharmacological Evaluation of Selected Medicinal Plants Usedin the Management of Oral and Skin Infections in Ebem-OhafiaDistrict Abia State Nigeria

Blessing O Oyedemi1 Sunday O Oyedemi 2 Johnson V Chibuzor1 Ifeoma I Ijeh2

Roger M Coopoosamy3 and Ayobami O Aiyegoro4

1Department of Plant Science and Biotechnology Michael Okpara University of Agriculture Umudike Nigeria2Department of Biochemistry Michael Okpara University of Agriculture Umudike Nigeria3Department of Nature Conservation and Ethnobotany Mangosuthu University of Technology PO Box 12363 JacobsDurban 4026 South Africa

4GI Microbiology and Biotechnology Unit Agricultural Research Council Animal Production Institute IrenePretoria 0062 South Africa

Correspondence should be addressed to Sunday O Oyedemi silvanusdemigmailcom

Received 24 January 2018 Accepted 26 April 2018 Published 4 July 2018

Academic Editor Carlos H G Martins

Copyright copy 2018 Blessing O Oyedemi et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

Oral and skin infections contribute significantly to the global health challenges responsible for the current trend of increasedmorbidity and premature death The purpose of this study was to document medicinal plants used in the management of oraland skin infections in Ebem-Ohafia Local Government Area (LGA) Abia State and to characterize the in vitro antioxidant andantibacterial activity The thin layer chromatography (TLC) profiling of ten of the selected folklore medicine was carried out usinga various solvent system of different polarity index The antioxidant capacity of the plant extracts was evaluated using chemical-based methods and its antibacterial effect was investigated using disc diffusion andmicrodilutionmethods Sixty-one plant speciesbelonging to 26 families were discovered and the most frequently cited species are Euphorbiaceae (1803) Fabaceae (1147)and Asteraceae (1147) All the plant extracts showed a promising free radical scavenging activity and efficient ferric reducingantioxidant power in a concentration-dependentmanner possibly due to their richness in polyphenol with TLC profiling showingmaximum three bands of phytochemicals Also the plant extracts exhibited a mild to weak antibacterial activity against our panelof bacterial strains having MIC values ranging from 256 to gt 512 120583g mL reflected in their zone of inhibition at 10 120583gdisc Thedata obtained for Breynia nivosa (BN) Eleusine indica (EI) Cassia alata (CA) Chromolaena odorata (CO) and Acalypha hispida(AH) extracts substantiate the traditional use of these herbal remedies in the region and open the possibility for the developmentof cheaper and affordable drugs in the treatment of oral and skin infections Further studies are needed to identify active ingredientwith strong antibacterial and antioxidant capacities along with their molecular mechanisms

1 Introduction

Oral health is a state of being free from the mouth and facialpain throat cancer oral infection and sores that limit humancapacity in biting chewing smiling and speaking as wellas psychosocial well-being [1] Owing to the environmentalpollution the concentration of heavy metals such as Ni CrZn Hg and Pb in most food materials and water is rapidly

increasing which severely affects both oral and general healthcondition For example dental caries periodontitis andthroat cancer have been recognised as the most prevalentnoncommunicable diseases associated with heavy metalsglobally affecting school-aged children and a huge majorityof adults [2] In Nigeria the rate of deep periodontal pocketis between 15 and 58 (aged 15 years) while the preva-lence of unmanaged dental caries is high raising alarm for

Hindawie Scientific World JournalVolume 2018 Article ID 4757458 16 pageshttpsdoiorg10115520184757458

2 The Scientific World Journal

odontogenic infection Falsetta and colleagues [3] highlightedthat poor oral hygiene and frequent consumption of sugarscreate bacterial adhesion to tooth surfaces via mineralizationand biofilm formations caused by glucosyl and fructosyltransferases The same authors indicated an adverse effectof higher concentration of fluoride in drinking water insome developing countries that is responsible for the causeof fluorosis and dental caries regardless of age and sex Theoral cavity exposed to carcinogens that may advance into pre-cancerous lesions and malignant disorders like leukoplakiasubmucous fibrosis and dental caries resulting from theimbalance in the levels of free radicals generation and naturalantioxidants in the saliva [4] There is a growing consensusthat administration of natural antioxidants to the oral cavityreduces free radicals oxidative stress and oral inflammationtriggered by nicotine alcohol and other toxins introducedinto the mouth [5]

The skin being the largest organ in the body guards theunderlying muscles bones ligaments and internal organsinterfaces with the environment as well as protecting thebody against pathogenic microbes [6] Notwithstandinginfectious skin diseases such as skin abrasions burns acnedermatitis and sebaceous cysts have become everyday inci-dences and constitute a reoccurring health problem thataffects all human age groups [6] Over the years bacterial skindisorders such as impetigo ecthyma folliculitis eczemas andurticarias are one of the most common causes of hospitalvisits in the southeastern part of Nigeria Overcrowding mal-nutrition and humidity heat food and medication allergiescause someof these skin diseases especially among the schoolchildren in urban and rural settings [2] Microbial invasionof the skin surface forms a biofilm that confers a drugtolerance on the bacteria and capacity to escape host immuneresponse [7] The discovery of antibiotic resistance genesin several microorganisms quorum sensing and horizontalgene transfer has limited treatment of skin and soft tissueinfections and thus could pose a serious health challengein a region where there is limited access to microbiologicallaboratory facilities and antimicrobial agents [8] The globalneed for effective alternative medicines for oral and skininfections with minimal side effects and economical use isincreasing due to the rapid growth of antibiotic resistance andopportunistic infections in immunocompromised patients[9]

Plant-derived products are now gaining recognitionworldwide currently recommended for primary healthcarebut only a few plants have received rigorous scientific investi-gation Medicinal plants are natural products used since timeimmemorial for the treatment of various human diseases Allover the world the citizen and health practitioners are nowstarting to rely on herbal medicines as a substitute for scien-tifically proved therapies [10] Herbal products contained sec-ondary metabolites that are synthesized through the pentosephosphate shikimic acid and phenylpropanoid pathwaysThese compounds play a significant defensive role againstpathogenic bacteria and oxidative stress caused by abioticstress in plant [11] Currently one of the strategies used to treatskin and oral infections is the use of natural products fromplant origin perhaps due to the perception that long-term

use of western medicine induces severe complications Con-sequently many Nigerian patients in particular those livingin the rural areas have resorted to seeking relief via tradi-tional healers and herbalists who administer herbal drugsin the management of human diseases Considering therate at which the vegetation gets depleted and the currentfocus on safe complementary and alternative medicine tosynthetic drugs there is therefore the need to document theindigenous knowledge of these plants and to search for moreplants with therapeutic potential in the treatment of bacterialinfections and oxidative stress

There is a paucity of literature in traditional medicineused in the management of orofacial and skin infectionsin Nigeria Our recent ethnobotanical survey conductedin Ebem-Ohafia LGAs in the southeastern part of Nigeriarevealed sixty-one plant species commonly used in the man-agement of oral and skin infections Despite the acclaimedtraditional use of these botanicals as therapeutic agents thereis little scientific evidence to support their folkloric useTherefore this study aimed at investigating the antioxidantcapacity of these herbal remedies in terms of free radicalscavenging reducing power and total phenolic and flavonoidcontent in consonance with the TLC profiling The antibacte-rial effect on some clinically isolated bacteria associated withoral and skin infections was also investigated

2 Materials and Methods

21 Study Area Ebem-Ohafia is a town in Ohafia LGA ofAbia state in the southeastern part ofNigeria situated between5∘ 381015840N and 7∘ 501015840 East and encompasses over twenty-sixhometowns with a population of over 300 000 from thelast population census in 2006 Ebem community is a partof the tropical rainforest characterized by dry and rainyseason with a total annual rainfall of over 1400 mm and anannual temperature range of 23∘C to 32∘C The people of theregion practice subsistence farming under local governmentagencies policy The inhabitants of Ebem use herbal drugseither alone or in combination with other medicines in thetreatment of human diseases Most of the people in Ebemare rural dwellers hence they use plant-based therapies in themanagement of ailments that include oral and skin infections

22 Ethnobotanical Survey The ethnobotanical survey wasconducted between December and May 2016 using a well-structured questionnaire administered to the participantswith indigenous knowledge of plants utilized in the area Theset questions contained the local diagnosis of oral and skindiseases the name of plant samples used for the treatmentmethods of preparation and duration of treatment sideeffects and mode of administration The people interviewedconsisted of women andmen both married and unmarried atthe age of 30 to 65 with little education qualification

23 PlantCollection and Identification The leaves of Ipomoeainvolucrata (II) Acalypha hispida (AH) Breynia nivosa (BN)Jatropha curcas (JC) Chromolaena odorata (CO) Macrolo-bium macrophyllum (MM) Baphia nitida (BNI) Burkeaafricana (BA) Cassia alata (CA) and the root of Eleusine

The Scientific World Journal 3

indica (EI) were collected after the interview The plants wereidentified by their local names and authenticated by MrOriaku Williams and Professor Gabriel Osuagwu from theDepartment of Plant Science and Biotechnology College ofNatural Sciences Michael Okpara University of AgricultureUmudike in Abia state Nigeria The vouchers specimens(VICJON 1ndash10) were prepared and deposited at the depart-mental herbarium The preparations and mode of intake ofthe herbal remedies are described in Table 1

24 Sample Preparation We collected leaves and roots ofthe ten plants that were frequently mentioned in the surveyand oven-dried at 40∘C for 72 h while the root of EI wasdried longer for seven days The dried plant materials werepulverized to a fine powder using an electric blender andstored in an airtight container for further use Thirty grams(30 g) of dried powdered materials were extracted with 150mL of 100methanol for 48 h on amechanical shaker (StuartScientific Orbital 202 SOSI Essex UK) and the extractswere filtered using Buchner funnel and Whatman Number1 filter paper The filtrate was concentrated using a rotaryevaporated at 40∘C to recover the solvent and air-dried in afume chamber to give a yield ranging from 45 to 83 g

25 Thin Layer Chromatography (TLC) Profile Thin layerchromatography is a simple method for analyzing a complexmixture of compounds based on the distance travelled Theplant extracts (1 mgmL) dissolved in methanol and spottedon the plate coated with silica gel 60 F254 as a stationaryphase The slurry was prepared by dissolving 15 g of silicagel 60 F254 in 30 mL of distilled water and immediatelypoured into the plate The plates were air-dried overnightAbout 10 120583L of the plant extracts was gently loaded onthe base of the plate (5 cm above) using the capillary tubeThe plates were allowed to develop in chromatographictanks consisting of three different solvents (mobile phase)chloroform methanol acetic acid (541) until they reach34th of the TLC plate The TLC plate was removed andallowed to dry the spots were detected by iodine vapour andretention factor (Rf) was calculated using the equation Rf= distance travelled by the componentsdistance travelled bythe solvent

26 Total Phenolic Content (TPC) Assay The total phenoliccontent present in these extracts was quantified by the FolinCiocalteu reagent (FCR) using the method of Ghaffari etal [12] Briefly 02 mL of the plant extract (2 mgmL) wasadded to the reaction mixture consisting of 1 mL of 10 vvFCR and 08 mL of Na2CO3 (0075 mgmL) to give a finalconcentration of 02 mgmL of each extract The resultingmixture was incubated at 45∘C with shaking for 15 min andthe absorbance measured at 765 nm A standard curve wasprepared by mixing methanol solution of gallic acid (02mL 0025 - 0400 mgmL) with 1 mL of 10 vv FCR andsodium carbonate (08 mL 0075 mgmL) The experimentwas carried out in triplicate and the results presented asmeanvalues with standard deviation (plusmn SD) The TPC value wasexpressed as milligrams of gallic acid equivalent (GAE) perg of dried sample It was calculated using the formula T =

CtimesVM where T is the TPC (mgg) of extract in GAE C isthe concentration of gallic acid from the calibration curve Vis the volume of the extract mL M is the dry weight (g) ofthe leaf or root powder from which the extract was obtained

27 Total Flavonoid Content (TFC) Assay The concentra-tion of flavonoids in the plant extracts was determined viaaluminum colorimetric assay method [13] Briefly 1 mL of2 wv AlCl3 was prepared in 100vv methanol added to 1mL of the sample solution A yellow colour formation afterincubation at room temperature for 1 h was measured at420 nm using an AJI-C03 UV VIS spectrophotometer Thestandard curve for TFC was obtained using quercetin as astandard drug under the same procedure described in TPCdetermination The TFC was calculated using the formulaT = CtimesVM where T is the TFC (mgg) of extract in QEC is the concentration of quercetin established from thecalibration curve V is the volume of the extract mL M isthe dry weight (g) of the leaf powder from which the extractwas obtained The TFC was present in the extracts calculatedas mgg of quercetin equivalent (QE)

28 Ferric Reducing Antioxidant Power (FRAP) Assay Theability of plant extracts to reduce ferric (Fe3+) to ferrous(Fe2+) formwas evaluated following themethod described byYen and Chen [14] with slight modification A volume of 03mL of different concentrations (0025 - 2 mgmL) from plantextract BHT ascorbic acid and rutin prepared in distilledwater was mixed with reacting mixture consisting of 25 mLof 02 M phosphate buffer (pH 66) and 25 mL of K3Fe(CN)6(1 wv) The resulting mixture was incubated at 50∘C for20 min followed by addition of 25 mL of TCA (10 wv)After vigorous shaking 25 mL of the solution was mixedwith 25 mL of distilled water and 05 mL of FeCl3 (01 wv)left incubated at room temperature for 5 min and then theabsorbance was measured at 700 nm against a blank sample(without extract)

29 Free Radical Scavenging Activity Assays The free radicalscavenging potential of plant extracts was measured in vitroby the 1 11015840-diphenyl-1-picrylhydrazyl (DPPH) described byTariq et al [15] The assay was experimented by reacting16 mL of 0135 mM DPPH dissolved in 100vv methanolwith 04 mL of various concentrations (0078 - 1 mgmL) ofmethanol crude extracts The reaction mixture was vortexedthoroughly and left in the dark at room temperature for30 min The absorbance of the mixture was measured at517 nm after 2 min The method of Re et al [16] wasadopted to determine ABTS radical scavenging activity ofthe plant extracts The ABTS radical solution was generatedby mixing two stock solutions of 7 mM ABTS and 24mM potassium persulphate in the same ratio and allowingthe solution to react for 12 h at room temperature in thedark The resulting solution was diluted with methanol toobtain an absorbance of 0706 units at 734 nm A volumeof 1 mL of various concentrations (003125 ndash 1 mgmL)of the plant extracts react with 25 mL of ABTS radicalsolution in the dark for 15 min and was later measured

4 The Scientific World Journal

Table1Medicinalplantsused

inthem

anagem

ento

fskinor

oralinfections

insoutheastern

partof

Nigeria

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Macrolobium

macrophyllum

Fabaceae

Antho

notha

macroph

ylla

Leaf

Skin

Treatin

gof

woun

dandcut

Squeezethe

leavea

ndplaceo

nthe

affectedarea

ofthes

kin

Jatro

phacurcas

Euph

orbiaceae

Barbados

nut

Leaf

Skinoral

Treatm

ento

fboils

Boiltheleave

with

waterPlace

theh

otleaveo

nthea

ffected

area

ofthes

kin

Eleusin

eind

icaPo

aceae

Yard

grass

Root

Skin

Treatm

ento

fRingw

orm

Use

ther

ootand

scrubon

affectedarea

ofthes

kin

Cassiaalata

Caesalpiniaceae

Ring

worm

bush

Leaf

Skin

Treatm

ento

fRingw

orm

Scrubtheleave

onaffectedarea

ofthe

skin

Chromolaena

odorata

Aste

raceae

Siam

weed

Leaf

Skin

Tosto

pbleeding

andrashes

Squeezethe

leavea

ndrubon

affectedarea

Psidium

guajava(L)

Myrtaceae

Guava

Leaf

Skin

Treatm

ento

fskinbu

rns

scrapessunb

urn

woun

ds

psoriasiseczem

aand

others

Crushedgu

avaleafo

rboiledleaves

are

applieddirectlyto

thea

ffected

skin

providingreliefandareb

elievedto

improveh

ealin

gBryophyllum

pinn

atum

(Lam

)Oken

Crassulaceae

Mira

cleleaf

Dogrsquosliver

Leaf

Skino

ral

Treatm

ento

fboilwo

und

sore

orcut

Crushleafandapplyjuiceo

nthew

ound

surfa

ce

Ficusa

sperifolia

Miq

Moraceae

False

thistle

Leaflatex

Skin

Treatm

ento

fskinrashes

anditches

Appliedas

poultic

etothes

kinitches

Acanthus

montany

(TA

nderson)

Acanthaceae

Bearrsquosbreeches

Stem

bark

Skin

Treatm

ento

fwou

ndsa

ndskin

diseases

Woo

dbark

iscrushedandsoaked

italcoho

landextractapp

liedto

woun

dsandskin

Alliu

msativ

umL

Liliaceae

Garlic

Bulbjuice

Skin

oral

Usedto

cond

ition

thes

kin

chew

edforc

ough

andsore

throat

Thejuice

ismixed

with

lotio

nsforskin

cond

ition

ingFreshbu

lbischew

edor

boiledin

water

anddrinkdecoctionfor

coug

h

Emiliasonchifolia

(L)

Aste

raceae

Scarlettassel

Leaf

Skin

Treatm

ento

fSkintro

ubles

abscessa

ndbruises

Crushedleaves

were

pressedandused

tocoverthe

woun

dor

bruisedsurfa

ces

Sida

acutaBu

rmf

Malvaceae

Broo

mwe

edLeaf

Skin

Treatm

ento

fjaund

icesore

ulcer

Poultic

esmadefrom

boiledleaves

are

appliedon

thea

ffected

parts

The Scientific World Journal 5

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Ricin

uscommun

isLinn

Eu

phorbiaceae

Castorb

ean

Leafroo

tand

seed

Skin

Treatm

ento

fskininfection

anddetoxification

Applycasto

roilon

thea

ffected

area

torelieve

skin

infection

Citru

slim

on(L)Bu

rmf

(prosp

)Ru

taceae

Limon

Leaffruit

andseed

Skin

Treatm

ento

ffluheadache

cold

andoralinfections

Mixequalquantities

ofLimon

juicea

ndho

lybasilkeepitun

derthe

suntill

becomes

thickapplythison

face

Alliu

mcepa

LLiliaceae

Onion

sLeafandbu

lbSkin

Treatm

ento

fperiodo

ntitis

andcold

Theb

ulbismacerated

andmixed

with

water

andthen

takenorally

Breynianivosa

Euph

orbiaceae

Icep

lant

Leaf

Oral

Treatin

gof

toothache

Boiltheleave

with

waterthentake

the

water

Ipom

oeainvolucrata

Con

volvulaceae

Water

convolvu

lus

Leafandste

mOral

Treatm

ento

foralinfectio

nMixtheleave

with

ugbaga

treea

ndbo

ilwith

water

anddrinkthew

ater

Burkea

afric

ana

Caesalpinioideae

Icheku

Leaf

Oral

Treatm

ento

forald

isease

Boiltheleave

with

water

andtakebut

donrsquot

swallowit

Baphianitid

aPapilio

naceae

Cam

wood

Leafor

stem

Oral

Streng

thensthe

teeth

Chew

thes

tem

orscrubtheleave

onthe

teeth

Gongronem

alatifolium

Asclepiadaceae

Utazi

Leaf

Oral

Treatm

ento

fcou

ghand

diabetes

Crushtheleafw

ithmortara

ndmixwith

ginger

orgarlica

nddrink

Anonasenegalen

sisAnn

onaceae

Ubu

ruocha

Root

Oral

Treatm

ento

fmalariaand

mou

thinfection

Boilther

ootsanddrink

Abrusp

recatoriu

sPapilio

noideae

Crabrsquoseyes

Seed

androot

Oralskin

Treatm

ento

fcou

ghskin

carew

ound

Grin

dther

ootand

applythep

oultice

onthea

ffected

skin

areaA

lsocrushedseeds

canbe

takenorally

Acanthus

montanu

sAc

anthaceae

False

thistle

Leafandroot

Oraland

Skin

Treatm

ento

ffurun

cles

lesio

nsandurinary

infections

Boilther

ootand

applythep

oultice

tothe

affectedpartof

theb

odyFo

rinfectio

ns

boilther

ootand

leaves

anddrink

Piperu

mbellatum

Piperaceae

Njamuja

Leaf

Oral

Treatm

ento

fpeptic

ulcers

Crushtheleavesa

ndsoak

inwarm

orho

twater

anddrink

Euphorbiahirta

Euph

orbiaceae

Asth

maw

eed

Who

leplant

Oral

Treatm

ento

fwou

nds

Leaves

arem

acerated

andappliedto

affectedpart

Ficusb

urkei(Miq)Miq

Moraceae

Sand

paper

Leaf

Oral

Treatm

ento

fhypertension

Leaves

areb

oiledandtakenorally

6 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Gmelina

arborea

Verbenaceae

Gmelina

Stem

andleaf

Oral

Treatm

ento

fgon

orrhea

pilesabdo

minalpains

burningsensationsfever

mou

thwo

undanddiabetes

Snake-biteandscorpion

sting

Leaves

grou

ndinto

paste

with

water

are

appliedto

theforeheadforh

eadachein

feversB

oiledste

msa

retakenorally

Lophira

alata

Ochnaceae

Ekki

Stem

andleaf

Oral

Treatm

ento

ffeverm

outh

woun

dandmalaria

Boiltheleavesa

ndste

mandtake

orally

Nau

clealatifolia

Rubiaceae

Ubu

ru-in

uStem

andleaf

Oral

Itisused

inthetreatment

ofmalariaepilepsymou

thwo

undanxietypainfever

Boilther

ootsanddrink

Annona

murica

taAnn

onaceae

Sour

sop

Leaf

Oral

Treatm

ento

fcancera

ndmou

thwo

undinfections

Crushor

boiltheleavesa

ndtake

orally

Psidium

guajava

Myrtaceae

Guava

Leaf

Oral

Treatm

ento

fperiodo

ntal

diseasebu

rnsscrapes

sunb

urnwo

undspsoria

sis

eczemaandothers

Leaves

arec

rushed

andtakenorally

Astra

galusm

embran

aceus

Fabaceae

Beiq

iStem

andleaf

Oral

Treatm

ento

fcoldup

per

respira

tory

infections

allergiesfib

romyalgia

anem

iaH

IVA

IDS

Leaves

arec

rushed

andtakenorallyand

canalso

beappliedto

skin

Buchholziacoria

cea

Capparaceae

Won

derfu

lkola

Leaf

Oral

Itisused

tradition

allyfor

treatingdiabetes

hypertensio

nrheumatism

coldcou

ghand

catarrh

Boilleaves

andtake

orally

Gnetum

afric

anum

Gnetacea

Afang

Leaf

Oral

Enlarged

spleen

sore-th

roatandcathartic

Leaves

canbe

boiledor

cooked

andtaken

orally

Anoridium

man

niAnn

onaceae

Ewuro-ig

boStem

bark

Oral

Treatm

ento

fDiarrhea

coug

hfeverrheumatism

Theb

arkiscrushedandbo

iledandthen

takenorally

Funtum

iaafric

ana

Apocyn

aceae

Akorie

mbamiri

Leafstem

root

Skin

treatmento

fcon

stipatio

nwo

undsw

eakbladder

jaun

dice

Who

leplantisb

oiledandtakenorally

Also

thec

rushed

leaves

area

ppliedto

the

affectedskin

area

The Scientific World Journal 7

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Commiphora

afric

ana

Burseraceae

Turari

dashi

Rootsfruits

Oral

Treatm

ento

fwho

oping

coug

hbron

chitis

Rootsa

remacerated

andtakenorally

Dacryodesedulis

Burseraceae

Elem

iub

eBa

rkroo

tsfruits

Skin

Treatm

ento

fJiggersSkin

diseasesE

leph

antia

sis

Theleavesa

reeatenrawwith

kolanu

tas

anantie

meticLeaf-s

apisinstilledinto

thee

arfore

ar-tr

oubleanda

leaf-decoctio

nisprepared

asa

vapo

ur-bathforfeverish

stiffn

essw

ithheadache

Combretum

mucronatum

Com

bretaceae

Fararg

eza

Rootsa

ndleaf

Skin

Treatm

ento

fskininfection

Ther

ootand

leafarem

acerated

and

appliedto

affectedskin

area

Cleomec

iliata

Capparaceae

Ekuya

Leafseeds

Skin

Treatm

ento

fCon

vulsion

wo

undssores

Seedsa

recrushedandthep

oultice

appliedto

thea

ffected

areaTh

emacerated

leaves

after

boiledaretaken

orally

Ageratum

conyzoides

Aste

raceae

Urata

Who

leplant

Skin

Treatm

ento

fwou

ndU

lcer

Sleeping

sickn

essEyew

ash

Thep

lant

ismacerated

andappliedto

affectedskin

area

Aspilia

afric

ana

Aste

raceae

Yunyun

Ka

lank

uwa

Leafflow

erSkin

Treatm

ento

fskinrashes

cleaning

sorescorneal

opacities

Thefl

ower

issqueezed

andappliedto

affectedskin

area

Centau

reaperrottetii

Aste

raceae

Danyi

Who

leplant

Skin

Treatm

ento

fskin

infectionsSyphilis

Thefl

ower

issqueezed

andap

oultice

appliedto

affectedskin

area

Chrysanthellu

mindicum

Aste

raceae

Abilereoyigi

Who

leplant

Skin

Treatm

ento

fboils

gono

rrheaJaun

dice

Thep

lant

iscrushedandthep

oultice

applieddirectlyto

thes

kinarea

Elephantopus

scaber

Aste

raceae

Elephant

foot

Leafroo

tOral

Treatm

ento

ffevercou

ghLeafandroot

areb

oiledtogether

and

takenorally

Emiliacoccinea

Aste

raceae

Odu

ndun

Leafsap

and

root

Skin

Treatm

ento

fulcerhernia

measle

sWho

leplantissqu

eezedandsaptaken

orally

Acalypha

godseffi

ana

Euph

orbiaceae

Jinwinini

Leaf

Skin

Treatm

ento

fskin

infectionsantim

icrobials

Leafismacerated

andtakenorallyas

well

asappliedto

affectedskin

area

8 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Brideliafer

rugin

eaEu

phorbiaceae

Irikirni

Leafstem

bark

root

Oral

Treatm

ento

fInsom

nia

mou

thwashgono

rrhea

Who

leplantisb

oiledandtakenorally

Euphorbiaheterophylla

Euph

orbiaceae

Egele

Leafroo

tsSkin

Treatm

ento

fSkindiseases

Leafandroot

arem

acerated

andthe

poultic

eapp

liedto

thea

ffected

skin

area

Euphorbiahirta

Euph

orbiaceae

Non

onku

rciya

odne

Who

leplant

exud

ate

Oral

Treatm

ento

fAsth

ma

coug

hshapeo

fbreasts

Thew

holeplantism

acerated

andtaken

orally

Euphorbialaterifl

ora

Euph

orbiaceae

Orow

ere

Leafexud

ate

Skin

Treatm

ento

fdermatitis

constip

ation

Theleafissqu

eezedandthep

oultice

appliedto

thea

ffected

skin

area

Cajanu

scajan

Fabaceae

Orela

Leafseed

Skin

andoral

Treatm

ento

fSmallpox

and

mou

thwash

Leafissqueezed

andappliedto

the

affectedarea

Dan

iellia

thurifera

Fabaceae

Iya

Stew

-woo

ddu

stSkin

Treatm

ento

fscabies

Theb

arkismacerated

andappliedto

the

affectedskin

area

Dialiu

mguineense

Fabaceae

Tasm

iyar

kumi

Leafbark

fruitand

twig

Oral

Treatm

ento

fBronchitis

coug

handdiuretic

Theleafand

bark

arec

rushed

andbo

iled

togetherTh

emixture

isthen

takenorally

Acacianilotica

Fabaceae

Baanigabaruwa

Fruitbark

exud

ate

Skin

Treatm

ento

fskindiseases

fung

alinfections

Thee

xudatesa

resqueezed

from

theb

ark

andapplieddirectlyto

thea

ffected

skin

area

Napoleona

imperia

lisLecythidaceae

Mabun

giTw

igs

Oral

Treatm

ento

fAsth

ma

coug

hTw

igsa

remacerated

andtakenorally

Alliu

msativ

umLiliaceae

Ayoayuu

Bulb

Oral

Treatm

ento

ffevercou

gh

asthmaantim

icrobial

Theb

ulbismacerated

andtakenorally

Hibiscus

asper

Malvaceae

Isapad

angiraa

Leaf

Oraland

skin

Treatm

ento

fcou

gh

woun

dsanddiuretic

Leafisbo

iledandtakenorally

Hibiscus

rosasin

ensis

Malvaceae

Kekekeireagu

Leafstem

flowe

rSkin

Treatm

ento

finfl

uenza

woun

dsand

diuretic

Who

leplantism

acerated

andtaken

orally

Theleafissqu

eezedandappliedin

affectedskin

area

Hibiscus

sabdariffa

Malvaceae

Sobo

gurgu

zuLeafflow

erOral

Treatm

ento

fDiuretic

scoug

handdressin

gwo

unds

Leafisbo

iledandtakenorally

Ficuseleg

ans

Moraceae

Asoro

Leaf

Skinoral

Treatm

ento

fPile

constip

ationandcraw

-craw

Leafissqueezed

andappliedto

the

affectedarea

The Scientific World Journal 9

the absorbance The percentage inhibition of DPPH orABTS radical scavenging activity by the plant extracts wascalculated as (Abscontrol ndash Abssample)(Abscontrol) times 100where Abscontrol is the absorbance of DPPH or ABTS +methanol Abssample is the absorbance of DPPH or ABTSradical + sample extractstandard Here the concentration ofthe extracts needed to decrease the absorbance of DPPH orABTS radical by 50 was calculated Rutin (Sigma-Aldrichge 94 HPLC grade) at the same working concentrations ofthe plant extracts was used as reference drug

210 Bacterial Strains All bacterial strains were culturedon nutrient agar slopes and were incubated for 24 h at37∘C before the MIC An inoculum turbidity equivalent toa 05 McFarland standard (1times107 CFUmL) was prepared innormal saline for each test organism and then diluted 1100in Mueller-Hinton broth just before injection of the platesStandard strains used in the study included S aureus and Ecoli which were a gift from Dr EO Ekundayo (Departmentof Microbiology Michael Okpara University of Agriculture)and clinically isolated strains of K pneumoniae and Pmirabilis were gotten from Department of MicrobiologyFederal Medical Centre (FMC) Umuahia Abia State

211 Antibacterial Susceptibility Using Disc Diffusion MethodThe microbial growth inhibition potential of the plantextracts was determined by using the agar disc diffusionmethod [17] The plant extracts were dissolved in 10 vvDMSO (Merck UK Ltd) to a final concentration of 2 mgmLThe nutrient agar plate was streaked with the standardizedtest bacterial inoculum over the surface evenly and 2mgdiscof the plant extracts transferred onto sterile filter disc papers(about 6 mm) containing the test compounds was placed onthe agar surface in the Petri dishes 100 120583gdisc of standardantibiotic ciprofloxacin was used as control and 100 120583LDMSOdisc served as negative control The plates were thenincubated at 37∘C for 24 h Generally the antimicrobialextracts diffuse into the agar and inhibit the growth of thetest bacteria All tests were done in duplicate and diame-ter zones of inhibition of growth were measured using ameter rule from the edge of each dish after the incubationperiod

212 Minimum Inhibitory Concentration (MIC) Assays Wefollowed the method of Smith et al [18] to determine theMIC ofmethanol crude extracts of selected botanicals againstclinical isolates of bacteria associated with skin and oralinfection A volumeof 100mLof sterileMueller-Hinton broth(Oxoid Basingstoke UK) containing 20 mgL of Ca2+ and10 mgL of Mg2+ was dispensed into a 96-well microtitreplate (Nunc 03 mL total volume per well) All antibacterialagents were dissolved in dimethyl sulphoxide (DMSO) anddiluted in Mueller-Hinton broth to give a stock solutionThen 100 120583L of the antibacterial agent stock solution (2000mgL) was serially diluted into each well and 100 mL ofthe bacterial inoculum added to each well to give a finalconcentration range of 0024 - 1 mgL All procedures wereperformed in duplicate and the plates were incubated for 18

h at 37∘C Briefly 20 mL of a 5 mgL methanol solution of 3-[45-dimethylthiazol-2-yl]-25-diphenyltetrazolium bromide(MTT) (Sigma-Aldrich Ltd South Africa) was added to eachwell and then incubated for 30 min Blue colouration indi-cated bacterial growth The MIC was recorded as the lowestconcentration at which no colour change was observed

213 Statistical Analysis Data analysis was done inMicrosoftExcel to obtain descriptive statistics Means values wereseparated by the Duncan multiple tests using SAS The dif-ferent levels of significance within the groups were analyzedusing one-way analysis of variance (ANOVA) Values wereconsidered significant at P lt005

3 Results and Discussion

31 Ethnobotanical Survey Modern drugs are expensive andsupply to a remote area may be irregular hence result-ing in overreliance of traditional methods of healing as aprimary source of healthcare [19] The rural communitiesof Ebem-Ohafia region of southeastern Nigeria depend onherbal remedies to meet their domestic and health needsDuring our ethnobotanical survey a total of 30 respondentsincluding the elders herbalists and traditional healers wereinterviewed in the study areaMajority of the people (22)wereaged 30ndash65 years dominated by males (25) and one-third(10) had a primary school education while 15 had secondaryeducation and others informal education A total of 61medic-inal plant species belonging to 26 families were identified inthe treatment of oral and skin diseases as shown in Table 1Twenty-seven of them are used in the management of oralinfections and twenty-six plants for the management of skininfection while eight plants are used in the management ofboth infections Table 2 shows the list of family number of theplant species used and their percentage occurrenceThemostrepresented plant family is the Euphorbiaceae having 1803of the plant species followed by Asteraceae and Fabaceae with1147 each [20] Euphorbiaceae is a large family of floweringplants with 300 genera and about 7500 species found usefulin the curing of some common human diseases Asteraceae isa large family of flowering plants having 32 913 species and1911 genera and has been found to possess diverse biologicaleffects Fabaceae is a family of cosmopolitan distributionwith approximately 730 genera and 19400 species richnessat a global level [20] The leaves (60) recorded the mostcommonly used plant part followed by the root (16) andthen stem bark (12) None of the respondents indicated anypossible side effects caused by any of these plants listed inTable 1 The most common methods of preparation includeinfusion decoction and maceration These herbalists ortraditional healers often collect the plants from the fieldand dry and crush them into powder form before storingin bottles to prevent patients from identifying the plantsused for their treatment Most healers refused to provideinformation without payment whereas others thought toprovide information on the plant use is disrespect of theirtraditional belief Furthermore the majority of the plantsmentioned lack scientific data to substantiate their acclaimedfolkloric use From this work it is evident that medicinal

10 The Scientific World Journal

Table 2 The list of family number of the plant species and theirpercentage occurrence

Family Number of plant species OccurrenceFabaceae 7 1147Euphorbiaceae 11 1803Asteraceae 7 1147Myrtaceae 4 656Moraceae 3 492Malvaceae 4 656Liliaceae 2 328Capparaceae 2 328Annonaceae 3 492Combretaceae 1 164Burseraceae 2 328Apocynaceae 1 164Gnetaceae 1 164Lecythidaceae 1 164Verbenaceae 1 164Ochnaceae 1 164Rubiaceae 1 164Piperaceae 1 164Acanthaceae 2 328Papilionoideae 2 328Asclepiadaceae 1 164Caesalpiniaceae 2 328Convolvulaceae 1 164Rutaceae 1 164Crassulaceae 1 164Poaceae 1 164

plants play a significant role in the primary healthcare ofthe people and conservation of the indigenous knowledge ofthis folklore medicine is crucial for the innovations of futuredrug discovery and possible herbal plants trade in the region[19] On the contrary we observed imprecise dosage lowhygiene standards hiding of healing methods and lack ofpatients record as a significant setback to traditional healthpractitioners in the region

32 Thin Layer Chromatography (TLC) Profiling Thin layerchromatography remains the most common useful techniqueby researchers as a quick and simple method to resolvethe constituents of a crude extract followed by advancedextraction methods to purify the active ingredients Previousstudies reported that specific secondary metabolites in plantexhibit a wide range of antioxidant and antibacterial proper-ties [21] TLC was formed on a glass sheet coated with silicagel After the sample is applied to the plate a solvent system(mobile phase) is drawn up via capillary action on the TLCplate at different mobility rate depending on the chemicalcompositionsThe separated spotswith different colourswerevisualized by the UV light projection onto the glass sheetThe variation of coloured spots detected in the crude extractsindicates the presence of different phytochemicals that may

be responsible for the pharmacological effects of selectedplants use in the management of oral and skin infections asacclaimed by the traditional healers We quantified the resultsby measuring the distance travelled by the plant compoundsdivided by the total distance travelled by the mobile phaseFrom Table 3 it is apparent that mobile phase 1 consistingof chloroform and ethanol in the volume ratio of 64 had abetter separation than mobile phase II containing tolueneethyl acetate and acetic acid in the volume ratio of 541 Themethanol extract of AH in bothmobile phases had the highestnumber of three bands with Rf values of 0667 0833 and0989 for mobile phase I and Rf values of 0381 0648 and0781 for mobile phase IIThe plant extracts MM BA CO JCandCAhad two bands inmobile phase I with clear separationbut had single band in mobile phase II The plant extractshaving only one band in either mobile phase suggest a poorsolvent system for compounds separation The TLC profilesprovide a characteristic fingerprint of these plants that mayalso be useful for their identification

33 Antioxidant Assays

331 TPC and TFC Phenolic compounds are secondarymetabolites distributed in fruits vegetables nuts herbsseeds and beverages [22] They are excellent scavengers offree radical that has been implicated in the pathologies ofvarious human diseases the antioxidant activity is dependenton the structure hydrophobic benzenoid rings and hydrogen-bonding potential of the phenolic hydroxyl groups In TPCassay a phenol in plant extracts loses a proton to formphenolate ion which reduces Folin Ciocalteu reagent andthe resulting changes are measured spectrophotometricallyat 765 nm [13] The content of phenol is reported in mg oftannic acid equivalent (TAE)mL of extract As shown inFigure 1 the total phenolic contents of absolute methanolcrude extract of these botanicals range from 700 to 3600mgTAEg The AH extract had the highest concentrationfollowed by CA CO MM BNI BA EI BN and JC whilethe least concentration was recorded in II extract Moreoverthe Folin Ciocalteu reagent (FCR) has been used to measureother compounds such as ascorbic acid amino acids andsugars hence it may not provide a concise amount of thephenolic compounds present in these plants Flavonoids areincreasingly consumed in a significant amount in daily dietsworldwide due to their various health benefits toman [23 24]In TFC assay aluminum ion in the reacting mixture formscomplexes with the C- 4 keto and either C- 3 or C- 5 hydroxylor with ortho hydroxyl groups in the A or B ring of flavonoidsdetected in the plant extracts [14] As shown in Figure 2the total flavonoid content was recorded in the plant extractsranging from 750 to 1500 mgQEg as extrapolated from thestandard quercetin curve The CA extract had the highestconcentration followed by BNI CO II AH BN MM JCand BA while the lowest concentration was recorded in EIextract TPC is found higher than the TFC in some of theextracts supporting the fact that most flavonoids are alsophenolics According to our results all the plant extractsare rich source of phenolic compounds The composition ofTPC and TFC registered in these plants may be difficult to

The Scientific World Journal 11

Table 3 Thin layer chromatography retention factor using mobile phase CHL ET (mobile phase I) and Tol EA AA (mobile phase II)

SN Plant Extract CHL ET (64) Tol EA AA (541)1 Baphia nitida 1 0742 Macrolobium macrophyllum 0706 0941 06953 Chromolaena odorata 0471 0647 06674 Jatropha curcas 0353 0941 06485 Cassia alata 0118 0882 06866 Ipomoea involucrata 0956 -7 Acalypha hispida 0667 0833 0989 0381 0648 07818 Breynia nivosa 0989 07629 Burkea africana 0644 1 075210 Eleusine indica 0978 0743CHL chloroform ET ethanol Tol toluene EA ethylacetate AA acetic acid

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

500

1000

1500

2000

2500

3000

3500

4000

TPC

(mgT

AE

g)

Figure 1 The total phenolic content (TPC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata TAE tannic acid equivalent

compare with others reported elsewhere due to many factorsincluding differences from species to species maturity stageplant parts and assay methods harvesting time standarddrugs and geographical location

332 Reducing Power (RP) Ferric reducing power is a globalmethod often used to assess the capacity of antioxidantsto donate either an electron or hydrogen atom to unpairedelectron [25] In reducing power assay plant extracts exertantioxidant activities via reduction of Fe3+ferric cyanidecomplex to Fe2+ (ferrous form) resulting in changing thegreen colour to blue and sometimes Perlrsquos Prussian bluecolour depending on the antioxidant potential [10] As shownin Figure 3 the reducing power of the methanol crudeextracts and the standard gallic acid rutin and ascorbic acidincreased with increase in concentration Higher absorbanceat 700 nm can be read after potassium ferricyanide reactionwith ferric chloride At 1000 120583gmL the extracts from AH

0

200

400

600

800

1000

1200

1400

1600

TFC

(mgQ

Eg)

AH BN JC CO MM EI BNI BA CAIIPlant extracts

Figure 2 The total flavonoid content (TFC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata QE Quercetin equivalent

BNI MM CA BA EI and BN had absorbance value of 270086 073 055 036 027 and 027 respectively while that ofAH is higher than the standard rutin (254) but comparablewell with gallic acid (275) and ascorbic acid (269) EI andBN extracts had a similar trend of ferric reducing powerthough significantly lower as compared with the standarddrugs AH extract exhibits remarkable antioxidant capacity(RP) correlating with its richness in phenolic and flavonoidcontents It is reasonable to suggest that methanol selectivelyextracts antioxidant principles from these botanicals basedon their polarities and chemical structures which are reflectedin plant extracts with high TFC and TPC having minimal RP[26] We have shown that some plant extracts in particularAH BNI MM and CA extracts may serve as an excellentcandidate to minimize skin and oral infections caused byoxidative stress as a result of imbalance of free radicalsgeneration and natural antioxidants in the saliva or woundarea According to our results the increasing RP of the plantextracts correlateswith their TPCandTFCThis study at least

12 The Scientific World JournalRe

duci

ng P

ower

7

00nm

625 125 250 500 10003125Concentration (gmL)

0

05

1

15

2

25

3

IIAHBNJCCOMMEI

BNIBACAGARutinVit C

Figure 3 The ferric reducing antioxidative capacity of selectedplant extracts used in the management of oral and skin infectionsin southeast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata GA gallic acid Vit C vitamin C (standard antioxidants)

in part lends credence to the ethno-therapeutic use of someplant as natural antioxidant in themanagement of skin or oralinfections in the study area

333 FRSA of Plant Extracts Skin and oral cavity exposedto an array of oxidants such as smokes from cigaretteschemicals UV light nicotine and alcohol that are directlyor indirectly leading to the production of reactive oxygenspecies (ROS) are known to be responsible for some com-mon human diseases [27] These free radicals in particularhydroxyl and superoxide anions are highly reactivemoleculesderived from the metabolism of oxygen Although they playan important role in the biochemical immunological andphysiological processes it could be deleterious to vital organsif excessively accumulated in the human body [28] Food andpharmaceutical industries discovered that medicinal plantscontaining phenol and flavonoid compounds are essentialin the development of novel drugs against stress-induceddiseases [28] In the present investigation the concentration(IC50) at which 50 of free radical scavenged was calculatedand extrapolated from the plot of percentage inhibition ver-sus plant extract concentration The herbal drugs exhibitedrobust DPPH radical scavenging activity in a concentration-dependent manner having IC50 ranging from 2025 to 710120583gmL As shown in Figure 4 MM BA EI JC AH BNIand CA extracts had a potent DPPH radical scavengingactivity corresponding to their TPC TFC and RP Theplant extracts scavenged ABTS radical by the decrease inits absorbance at 734 nm All the plant extracts exhibitedstrong ABTS radical scavenging capacities in the following

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

200

400

600

800

1000

1200

1400

1600

IC50

(g

mL)

Figure 4 DPPH free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

050

100150200250300350400450500

IC50

(g

mL)

AH BN JC CO MM EI BNI BA CAIIPlant samples

Figure 5 ABTS free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

order BN gt JC gt CA gt MM gt BNI gt EI gt CO gt AH gtII gt BA having IC50 values ranging from 25 to 400 120583gmL(Figure 5) DPPH and ABTS radical scavenging activities ofthese herbal remedies displayed a positive correlation withthe working concentrations It is obvious that most of theplant extracts tested in this work possess the ability to quenchthe free radicals generation via electron or proton donationdemonstrated by the termination of radical chain reaction[28] Although both DPPH and ABTS radicals are electron-transfer based methods however it is apparent that ABTSmethod seems to be more sensitive than DPPH radical assaypossibly due to their solubility and diffusivity as shown inthe IC50 values [10] It is worth noting that BA extract withstrong DPPH radical scavenging effect had a moderate ABTS

The Scientific World Journal 13

Table 4 Zone of bacterial inhibition by the selected plant extracts use in the management of oral and skin infections

Diameter of inhibition (mm)SN Plant samples K pneumoniae P mirabilis S aureus E coli1 Cassia alata 15 plusmn 141 9 plusmn 042 15 plusmn 083 13 plusmn 0802 Breynia nivosa 8 plusmn 052 17 plusmn 100 8 plusmn 008 15 plusmn 1023 Burkea africana NI 11 plusmn 084 13 plusmn 072 NI4 Baphia nitida 8 plusmn 050 13 plusmn 122 8 plusmn 051 NI5 Ipomoea involucrata 8 plusmn 052 NI 8 plusmn 050 13 plusmn 0526 Jatropha curcas NI 9 plusmn 030 9 plusmn 062 13 plusmn 0727 Acalypha hispida 21 plusmn 163 NI 11 plusmn 009 17 plusmn 1328 Chromolaena odorata 8 plusmn 042 8 plusmn 035 9 plusmn 009 15 plusmn1209 Eleusine indica 8 plusmn 042 8 plusmn 035 NI 17 plusmn 10010 Macrolobium macrophyllum NI NI 8 plusmn 004 15 plusmn 12311 Ciprofloxacin - - le16(R) ge17(S) le16(R)ge20(S)NI indicates no zone of inhibition S sensitive R resistance

radical scavenging capacity which can be explained by thepartial ionization of the extract in the test medium [29] SinceROS promotes tissue damage and epithelial inflammationvia exposure to sunlight radiation or smoke as a result theantioxidant capacity demonstrated in this work may supportthe traditional use of these plants against the influence ofinflammatory markers and oxidants in the skin and oralcavity [29] Based on the results of the three antioxidantassays it is obvious that MM AH BNI EI and CA extractshad the best FRSA indicating their importance in convertingfree radicals into more stable products and then terminatethe free radical chain reaction Our study showed that themethanol crude extracts from these plants could protectagainst free radical damage in the mouth such as humangingival periodontal tissues and certain skin infections [5]

334 Antibacterial Activity of Plant Extracts Pathogenicbacteria such as Staphylococcus aureus Escherichia coli Kleb-siella pneumoniae and Proteus mirabilis play a significantrole as the primary cause of skin and oral infections [30]Staphylococcus aureus is a part of normal skin known to bethe most problematic in the cause of skin infections as aresult of their resistance mechanisms to first-line antibiotics[30] Although the presence of this organism as a residentoral floral is controversial nonetheless MSSA and MRSAhave been isolated from oral rinse and tongue swab inpatients with oral infections [31] Escherichia coli are Gram-negative bacteria known to be notorious for their resistantmechanisms towards the existing arsenal of antibiotics Oralinfections such as edematous and erythematous and skininfections like omphalitis surgical site infections burnnecrotizing fasciitis and cellulitis at all age groups are associ-ated with enterobacterium E coli [32] Klebsiella pneumoniaeis a Gram-negative encapsulated facultative anaerobic androd-shaped bacterium found in the normal flora of themouth skin and intestines This bacterium is responsiblefor the prolonged surgical wound biliary tract wound andurinary tract infections and it could pose a serious healthrisk when resistant strains invade the throat mouth or

intestines [33] Villegas et al [34] detected a CTX-M-12 120573-lactamase producing K pneumoniae new plasmid-mediatedESBLs highly resistant to ceftazidime from a patient withhospital-acquired wound infection Proteus mirabilis is oneof the most common enterobacteria after E coli identified asGram-negativemotile and rod-shaped bacterium It is foundin clinical specimen implicated in the skin respiratory tractand bacteraemia as well as burns wounds and urinary tractinfections [35] All these pathogenic multidrug-resistancebacteria are causative agent in a periodontal lips ulcer whitesponge nevus and skin infections [36]

335 Susceptibility Test and MIC Determination The resultof disc diffusion assay showed the diameter of inhibitionof bacteria growth classified as le 15 (resistant) and ge 16(susceptible) according to CLSI guideline [37] As shown inTable 4 K pneumoniae and E coli are more susceptible tothe treatment of Acalypha hispida (AH) having a zone ofinhibition (ZI) 21 and 17mm respectively BN and EI extractsinhibit the growth of P mirabilis and E coli having the samezone of inhibition 17 mm corresponding to the report of Al-Zubairi et al [38] The data obtained for the methanol crudeextracts from AH BN and EI show their selective inhibitionof the test bacteria growth The plant extracts with ZI above13 mm against our panel of bacterial strains were selected forMICdeterminationHere themethanol crude extracts exhibitMIC values of 256 to gt 512 120583gmL against the test bacterialstrains but were less active as compared to the standardciprofloxacin 003 - 006 120583gmL (Table 5) Ciprofloxacin isan active broad-spectrum antibiotic use in the treatmentof urinary tract infections prostatitis and acute cystitis viainterfering with bacterial DNA replication and transcriptionthrough inhibition of DNA gyrase or topoisomerase [39] Allthe plant extracts demonstrated a mild to weak antibacterialactivity based on the classification of Rios and Recio [40] whoconsidered antibacterial activity of the crude plant extractsignificant (MIC lt 100 120583gmL) moderate (100 lt MIC = 512120583gmL) or weak (MIC gt 512 120583gmL) The herbal remediesshowed moderate inhibition of bacterial growth except for K

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

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International Journal of

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The Scientific World Journal

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Submit your manuscripts atwwwhindawicom

Page 2: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

2 The Scientific World Journal

odontogenic infection Falsetta and colleagues [3] highlightedthat poor oral hygiene and frequent consumption of sugarscreate bacterial adhesion to tooth surfaces via mineralizationand biofilm formations caused by glucosyl and fructosyltransferases The same authors indicated an adverse effectof higher concentration of fluoride in drinking water insome developing countries that is responsible for the causeof fluorosis and dental caries regardless of age and sex Theoral cavity exposed to carcinogens that may advance into pre-cancerous lesions and malignant disorders like leukoplakiasubmucous fibrosis and dental caries resulting from theimbalance in the levels of free radicals generation and naturalantioxidants in the saliva [4] There is a growing consensusthat administration of natural antioxidants to the oral cavityreduces free radicals oxidative stress and oral inflammationtriggered by nicotine alcohol and other toxins introducedinto the mouth [5]

The skin being the largest organ in the body guards theunderlying muscles bones ligaments and internal organsinterfaces with the environment as well as protecting thebody against pathogenic microbes [6] Notwithstandinginfectious skin diseases such as skin abrasions burns acnedermatitis and sebaceous cysts have become everyday inci-dences and constitute a reoccurring health problem thataffects all human age groups [6] Over the years bacterial skindisorders such as impetigo ecthyma folliculitis eczemas andurticarias are one of the most common causes of hospitalvisits in the southeastern part of Nigeria Overcrowding mal-nutrition and humidity heat food and medication allergiescause someof these skin diseases especially among the schoolchildren in urban and rural settings [2] Microbial invasionof the skin surface forms a biofilm that confers a drugtolerance on the bacteria and capacity to escape host immuneresponse [7] The discovery of antibiotic resistance genesin several microorganisms quorum sensing and horizontalgene transfer has limited treatment of skin and soft tissueinfections and thus could pose a serious health challengein a region where there is limited access to microbiologicallaboratory facilities and antimicrobial agents [8] The globalneed for effective alternative medicines for oral and skininfections with minimal side effects and economical use isincreasing due to the rapid growth of antibiotic resistance andopportunistic infections in immunocompromised patients[9]

Plant-derived products are now gaining recognitionworldwide currently recommended for primary healthcarebut only a few plants have received rigorous scientific investi-gation Medicinal plants are natural products used since timeimmemorial for the treatment of various human diseases Allover the world the citizen and health practitioners are nowstarting to rely on herbal medicines as a substitute for scien-tifically proved therapies [10] Herbal products contained sec-ondary metabolites that are synthesized through the pentosephosphate shikimic acid and phenylpropanoid pathwaysThese compounds play a significant defensive role againstpathogenic bacteria and oxidative stress caused by abioticstress in plant [11] Currently one of the strategies used to treatskin and oral infections is the use of natural products fromplant origin perhaps due to the perception that long-term

use of western medicine induces severe complications Con-sequently many Nigerian patients in particular those livingin the rural areas have resorted to seeking relief via tradi-tional healers and herbalists who administer herbal drugsin the management of human diseases Considering therate at which the vegetation gets depleted and the currentfocus on safe complementary and alternative medicine tosynthetic drugs there is therefore the need to document theindigenous knowledge of these plants and to search for moreplants with therapeutic potential in the treatment of bacterialinfections and oxidative stress

There is a paucity of literature in traditional medicineused in the management of orofacial and skin infectionsin Nigeria Our recent ethnobotanical survey conductedin Ebem-Ohafia LGAs in the southeastern part of Nigeriarevealed sixty-one plant species commonly used in the man-agement of oral and skin infections Despite the acclaimedtraditional use of these botanicals as therapeutic agents thereis little scientific evidence to support their folkloric useTherefore this study aimed at investigating the antioxidantcapacity of these herbal remedies in terms of free radicalscavenging reducing power and total phenolic and flavonoidcontent in consonance with the TLC profiling The antibacte-rial effect on some clinically isolated bacteria associated withoral and skin infections was also investigated

2 Materials and Methods

21 Study Area Ebem-Ohafia is a town in Ohafia LGA ofAbia state in the southeastern part ofNigeria situated between5∘ 381015840N and 7∘ 501015840 East and encompasses over twenty-sixhometowns with a population of over 300 000 from thelast population census in 2006 Ebem community is a partof the tropical rainforest characterized by dry and rainyseason with a total annual rainfall of over 1400 mm and anannual temperature range of 23∘C to 32∘C The people of theregion practice subsistence farming under local governmentagencies policy The inhabitants of Ebem use herbal drugseither alone or in combination with other medicines in thetreatment of human diseases Most of the people in Ebemare rural dwellers hence they use plant-based therapies in themanagement of ailments that include oral and skin infections

22 Ethnobotanical Survey The ethnobotanical survey wasconducted between December and May 2016 using a well-structured questionnaire administered to the participantswith indigenous knowledge of plants utilized in the area Theset questions contained the local diagnosis of oral and skindiseases the name of plant samples used for the treatmentmethods of preparation and duration of treatment sideeffects and mode of administration The people interviewedconsisted of women andmen both married and unmarried atthe age of 30 to 65 with little education qualification

23 PlantCollection and Identification The leaves of Ipomoeainvolucrata (II) Acalypha hispida (AH) Breynia nivosa (BN)Jatropha curcas (JC) Chromolaena odorata (CO) Macrolo-bium macrophyllum (MM) Baphia nitida (BNI) Burkeaafricana (BA) Cassia alata (CA) and the root of Eleusine

The Scientific World Journal 3

indica (EI) were collected after the interview The plants wereidentified by their local names and authenticated by MrOriaku Williams and Professor Gabriel Osuagwu from theDepartment of Plant Science and Biotechnology College ofNatural Sciences Michael Okpara University of AgricultureUmudike in Abia state Nigeria The vouchers specimens(VICJON 1ndash10) were prepared and deposited at the depart-mental herbarium The preparations and mode of intake ofthe herbal remedies are described in Table 1

24 Sample Preparation We collected leaves and roots ofthe ten plants that were frequently mentioned in the surveyand oven-dried at 40∘C for 72 h while the root of EI wasdried longer for seven days The dried plant materials werepulverized to a fine powder using an electric blender andstored in an airtight container for further use Thirty grams(30 g) of dried powdered materials were extracted with 150mL of 100methanol for 48 h on amechanical shaker (StuartScientific Orbital 202 SOSI Essex UK) and the extractswere filtered using Buchner funnel and Whatman Number1 filter paper The filtrate was concentrated using a rotaryevaporated at 40∘C to recover the solvent and air-dried in afume chamber to give a yield ranging from 45 to 83 g

25 Thin Layer Chromatography (TLC) Profile Thin layerchromatography is a simple method for analyzing a complexmixture of compounds based on the distance travelled Theplant extracts (1 mgmL) dissolved in methanol and spottedon the plate coated with silica gel 60 F254 as a stationaryphase The slurry was prepared by dissolving 15 g of silicagel 60 F254 in 30 mL of distilled water and immediatelypoured into the plate The plates were air-dried overnightAbout 10 120583L of the plant extracts was gently loaded onthe base of the plate (5 cm above) using the capillary tubeThe plates were allowed to develop in chromatographictanks consisting of three different solvents (mobile phase)chloroform methanol acetic acid (541) until they reach34th of the TLC plate The TLC plate was removed andallowed to dry the spots were detected by iodine vapour andretention factor (Rf) was calculated using the equation Rf= distance travelled by the componentsdistance travelled bythe solvent

26 Total Phenolic Content (TPC) Assay The total phenoliccontent present in these extracts was quantified by the FolinCiocalteu reagent (FCR) using the method of Ghaffari etal [12] Briefly 02 mL of the plant extract (2 mgmL) wasadded to the reaction mixture consisting of 1 mL of 10 vvFCR and 08 mL of Na2CO3 (0075 mgmL) to give a finalconcentration of 02 mgmL of each extract The resultingmixture was incubated at 45∘C with shaking for 15 min andthe absorbance measured at 765 nm A standard curve wasprepared by mixing methanol solution of gallic acid (02mL 0025 - 0400 mgmL) with 1 mL of 10 vv FCR andsodium carbonate (08 mL 0075 mgmL) The experimentwas carried out in triplicate and the results presented asmeanvalues with standard deviation (plusmn SD) The TPC value wasexpressed as milligrams of gallic acid equivalent (GAE) perg of dried sample It was calculated using the formula T =

CtimesVM where T is the TPC (mgg) of extract in GAE C isthe concentration of gallic acid from the calibration curve Vis the volume of the extract mL M is the dry weight (g) ofthe leaf or root powder from which the extract was obtained

27 Total Flavonoid Content (TFC) Assay The concentra-tion of flavonoids in the plant extracts was determined viaaluminum colorimetric assay method [13] Briefly 1 mL of2 wv AlCl3 was prepared in 100vv methanol added to 1mL of the sample solution A yellow colour formation afterincubation at room temperature for 1 h was measured at420 nm using an AJI-C03 UV VIS spectrophotometer Thestandard curve for TFC was obtained using quercetin as astandard drug under the same procedure described in TPCdetermination The TFC was calculated using the formulaT = CtimesVM where T is the TFC (mgg) of extract in QEC is the concentration of quercetin established from thecalibration curve V is the volume of the extract mL M isthe dry weight (g) of the leaf powder from which the extractwas obtained The TFC was present in the extracts calculatedas mgg of quercetin equivalent (QE)

28 Ferric Reducing Antioxidant Power (FRAP) Assay Theability of plant extracts to reduce ferric (Fe3+) to ferrous(Fe2+) formwas evaluated following themethod described byYen and Chen [14] with slight modification A volume of 03mL of different concentrations (0025 - 2 mgmL) from plantextract BHT ascorbic acid and rutin prepared in distilledwater was mixed with reacting mixture consisting of 25 mLof 02 M phosphate buffer (pH 66) and 25 mL of K3Fe(CN)6(1 wv) The resulting mixture was incubated at 50∘C for20 min followed by addition of 25 mL of TCA (10 wv)After vigorous shaking 25 mL of the solution was mixedwith 25 mL of distilled water and 05 mL of FeCl3 (01 wv)left incubated at room temperature for 5 min and then theabsorbance was measured at 700 nm against a blank sample(without extract)

29 Free Radical Scavenging Activity Assays The free radicalscavenging potential of plant extracts was measured in vitroby the 1 11015840-diphenyl-1-picrylhydrazyl (DPPH) described byTariq et al [15] The assay was experimented by reacting16 mL of 0135 mM DPPH dissolved in 100vv methanolwith 04 mL of various concentrations (0078 - 1 mgmL) ofmethanol crude extracts The reaction mixture was vortexedthoroughly and left in the dark at room temperature for30 min The absorbance of the mixture was measured at517 nm after 2 min The method of Re et al [16] wasadopted to determine ABTS radical scavenging activity ofthe plant extracts The ABTS radical solution was generatedby mixing two stock solutions of 7 mM ABTS and 24mM potassium persulphate in the same ratio and allowingthe solution to react for 12 h at room temperature in thedark The resulting solution was diluted with methanol toobtain an absorbance of 0706 units at 734 nm A volumeof 1 mL of various concentrations (003125 ndash 1 mgmL)of the plant extracts react with 25 mL of ABTS radicalsolution in the dark for 15 min and was later measured

4 The Scientific World Journal

Table1Medicinalplantsused

inthem

anagem

ento

fskinor

oralinfections

insoutheastern

partof

Nigeria

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Macrolobium

macrophyllum

Fabaceae

Antho

notha

macroph

ylla

Leaf

Skin

Treatin

gof

woun

dandcut

Squeezethe

leavea

ndplaceo

nthe

affectedarea

ofthes

kin

Jatro

phacurcas

Euph

orbiaceae

Barbados

nut

Leaf

Skinoral

Treatm

ento

fboils

Boiltheleave

with

waterPlace

theh

otleaveo

nthea

ffected

area

ofthes

kin

Eleusin

eind

icaPo

aceae

Yard

grass

Root

Skin

Treatm

ento

fRingw

orm

Use

ther

ootand

scrubon

affectedarea

ofthes

kin

Cassiaalata

Caesalpiniaceae

Ring

worm

bush

Leaf

Skin

Treatm

ento

fRingw

orm

Scrubtheleave

onaffectedarea

ofthe

skin

Chromolaena

odorata

Aste

raceae

Siam

weed

Leaf

Skin

Tosto

pbleeding

andrashes

Squeezethe

leavea

ndrubon

affectedarea

Psidium

guajava(L)

Myrtaceae

Guava

Leaf

Skin

Treatm

ento

fskinbu

rns

scrapessunb

urn

woun

ds

psoriasiseczem

aand

others

Crushedgu

avaleafo

rboiledleaves

are

applieddirectlyto

thea

ffected

skin

providingreliefandareb

elievedto

improveh

ealin

gBryophyllum

pinn

atum

(Lam

)Oken

Crassulaceae

Mira

cleleaf

Dogrsquosliver

Leaf

Skino

ral

Treatm

ento

fboilwo

und

sore

orcut

Crushleafandapplyjuiceo

nthew

ound

surfa

ce

Ficusa

sperifolia

Miq

Moraceae

False

thistle

Leaflatex

Skin

Treatm

ento

fskinrashes

anditches

Appliedas

poultic

etothes

kinitches

Acanthus

montany

(TA

nderson)

Acanthaceae

Bearrsquosbreeches

Stem

bark

Skin

Treatm

ento

fwou

ndsa

ndskin

diseases

Woo

dbark

iscrushedandsoaked

italcoho

landextractapp

liedto

woun

dsandskin

Alliu

msativ

umL

Liliaceae

Garlic

Bulbjuice

Skin

oral

Usedto

cond

ition

thes

kin

chew

edforc

ough

andsore

throat

Thejuice

ismixed

with

lotio

nsforskin

cond

ition

ingFreshbu

lbischew

edor

boiledin

water

anddrinkdecoctionfor

coug

h

Emiliasonchifolia

(L)

Aste

raceae

Scarlettassel

Leaf

Skin

Treatm

ento

fSkintro

ubles

abscessa

ndbruises

Crushedleaves

were

pressedandused

tocoverthe

woun

dor

bruisedsurfa

ces

Sida

acutaBu

rmf

Malvaceae

Broo

mwe

edLeaf

Skin

Treatm

ento

fjaund

icesore

ulcer

Poultic

esmadefrom

boiledleaves

are

appliedon

thea

ffected

parts

The Scientific World Journal 5

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Ricin

uscommun

isLinn

Eu

phorbiaceae

Castorb

ean

Leafroo

tand

seed

Skin

Treatm

ento

fskininfection

anddetoxification

Applycasto

roilon

thea

ffected

area

torelieve

skin

infection

Citru

slim

on(L)Bu

rmf

(prosp

)Ru

taceae

Limon

Leaffruit

andseed

Skin

Treatm

ento

ffluheadache

cold

andoralinfections

Mixequalquantities

ofLimon

juicea

ndho

lybasilkeepitun

derthe

suntill

becomes

thickapplythison

face

Alliu

mcepa

LLiliaceae

Onion

sLeafandbu

lbSkin

Treatm

ento

fperiodo

ntitis

andcold

Theb

ulbismacerated

andmixed

with

water

andthen

takenorally

Breynianivosa

Euph

orbiaceae

Icep

lant

Leaf

Oral

Treatin

gof

toothache

Boiltheleave

with

waterthentake

the

water

Ipom

oeainvolucrata

Con

volvulaceae

Water

convolvu

lus

Leafandste

mOral

Treatm

ento

foralinfectio

nMixtheleave

with

ugbaga

treea

ndbo

ilwith

water

anddrinkthew

ater

Burkea

afric

ana

Caesalpinioideae

Icheku

Leaf

Oral

Treatm

ento

forald

isease

Boiltheleave

with

water

andtakebut

donrsquot

swallowit

Baphianitid

aPapilio

naceae

Cam

wood

Leafor

stem

Oral

Streng

thensthe

teeth

Chew

thes

tem

orscrubtheleave

onthe

teeth

Gongronem

alatifolium

Asclepiadaceae

Utazi

Leaf

Oral

Treatm

ento

fcou

ghand

diabetes

Crushtheleafw

ithmortara

ndmixwith

ginger

orgarlica

nddrink

Anonasenegalen

sisAnn

onaceae

Ubu

ruocha

Root

Oral

Treatm

ento

fmalariaand

mou

thinfection

Boilther

ootsanddrink

Abrusp

recatoriu

sPapilio

noideae

Crabrsquoseyes

Seed

androot

Oralskin

Treatm

ento

fcou

ghskin

carew

ound

Grin

dther

ootand

applythep

oultice

onthea

ffected

skin

areaA

lsocrushedseeds

canbe

takenorally

Acanthus

montanu

sAc

anthaceae

False

thistle

Leafandroot

Oraland

Skin

Treatm

ento

ffurun

cles

lesio

nsandurinary

infections

Boilther

ootand

applythep

oultice

tothe

affectedpartof

theb

odyFo

rinfectio

ns

boilther

ootand

leaves

anddrink

Piperu

mbellatum

Piperaceae

Njamuja

Leaf

Oral

Treatm

ento

fpeptic

ulcers

Crushtheleavesa

ndsoak

inwarm

orho

twater

anddrink

Euphorbiahirta

Euph

orbiaceae

Asth

maw

eed

Who

leplant

Oral

Treatm

ento

fwou

nds

Leaves

arem

acerated

andappliedto

affectedpart

Ficusb

urkei(Miq)Miq

Moraceae

Sand

paper

Leaf

Oral

Treatm

ento

fhypertension

Leaves

areb

oiledandtakenorally

6 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Gmelina

arborea

Verbenaceae

Gmelina

Stem

andleaf

Oral

Treatm

ento

fgon

orrhea

pilesabdo

minalpains

burningsensationsfever

mou

thwo

undanddiabetes

Snake-biteandscorpion

sting

Leaves

grou

ndinto

paste

with

water

are

appliedto

theforeheadforh

eadachein

feversB

oiledste

msa

retakenorally

Lophira

alata

Ochnaceae

Ekki

Stem

andleaf

Oral

Treatm

ento

ffeverm

outh

woun

dandmalaria

Boiltheleavesa

ndste

mandtake

orally

Nau

clealatifolia

Rubiaceae

Ubu

ru-in

uStem

andleaf

Oral

Itisused

inthetreatment

ofmalariaepilepsymou

thwo

undanxietypainfever

Boilther

ootsanddrink

Annona

murica

taAnn

onaceae

Sour

sop

Leaf

Oral

Treatm

ento

fcancera

ndmou

thwo

undinfections

Crushor

boiltheleavesa

ndtake

orally

Psidium

guajava

Myrtaceae

Guava

Leaf

Oral

Treatm

ento

fperiodo

ntal

diseasebu

rnsscrapes

sunb

urnwo

undspsoria

sis

eczemaandothers

Leaves

arec

rushed

andtakenorally

Astra

galusm

embran

aceus

Fabaceae

Beiq

iStem

andleaf

Oral

Treatm

ento

fcoldup

per

respira

tory

infections

allergiesfib

romyalgia

anem

iaH

IVA

IDS

Leaves

arec

rushed

andtakenorallyand

canalso

beappliedto

skin

Buchholziacoria

cea

Capparaceae

Won

derfu

lkola

Leaf

Oral

Itisused

tradition

allyfor

treatingdiabetes

hypertensio

nrheumatism

coldcou

ghand

catarrh

Boilleaves

andtake

orally

Gnetum

afric

anum

Gnetacea

Afang

Leaf

Oral

Enlarged

spleen

sore-th

roatandcathartic

Leaves

canbe

boiledor

cooked

andtaken

orally

Anoridium

man

niAnn

onaceae

Ewuro-ig

boStem

bark

Oral

Treatm

ento

fDiarrhea

coug

hfeverrheumatism

Theb

arkiscrushedandbo

iledandthen

takenorally

Funtum

iaafric

ana

Apocyn

aceae

Akorie

mbamiri

Leafstem

root

Skin

treatmento

fcon

stipatio

nwo

undsw

eakbladder

jaun

dice

Who

leplantisb

oiledandtakenorally

Also

thec

rushed

leaves

area

ppliedto

the

affectedskin

area

The Scientific World Journal 7

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Commiphora

afric

ana

Burseraceae

Turari

dashi

Rootsfruits

Oral

Treatm

ento

fwho

oping

coug

hbron

chitis

Rootsa

remacerated

andtakenorally

Dacryodesedulis

Burseraceae

Elem

iub

eBa

rkroo

tsfruits

Skin

Treatm

ento

fJiggersSkin

diseasesE

leph

antia

sis

Theleavesa

reeatenrawwith

kolanu

tas

anantie

meticLeaf-s

apisinstilledinto

thee

arfore

ar-tr

oubleanda

leaf-decoctio

nisprepared

asa

vapo

ur-bathforfeverish

stiffn

essw

ithheadache

Combretum

mucronatum

Com

bretaceae

Fararg

eza

Rootsa

ndleaf

Skin

Treatm

ento

fskininfection

Ther

ootand

leafarem

acerated

and

appliedto

affectedskin

area

Cleomec

iliata

Capparaceae

Ekuya

Leafseeds

Skin

Treatm

ento

fCon

vulsion

wo

undssores

Seedsa

recrushedandthep

oultice

appliedto

thea

ffected

areaTh

emacerated

leaves

after

boiledaretaken

orally

Ageratum

conyzoides

Aste

raceae

Urata

Who

leplant

Skin

Treatm

ento

fwou

ndU

lcer

Sleeping

sickn

essEyew

ash

Thep

lant

ismacerated

andappliedto

affectedskin

area

Aspilia

afric

ana

Aste

raceae

Yunyun

Ka

lank

uwa

Leafflow

erSkin

Treatm

ento

fskinrashes

cleaning

sorescorneal

opacities

Thefl

ower

issqueezed

andappliedto

affectedskin

area

Centau

reaperrottetii

Aste

raceae

Danyi

Who

leplant

Skin

Treatm

ento

fskin

infectionsSyphilis

Thefl

ower

issqueezed

andap

oultice

appliedto

affectedskin

area

Chrysanthellu

mindicum

Aste

raceae

Abilereoyigi

Who

leplant

Skin

Treatm

ento

fboils

gono

rrheaJaun

dice

Thep

lant

iscrushedandthep

oultice

applieddirectlyto

thes

kinarea

Elephantopus

scaber

Aste

raceae

Elephant

foot

Leafroo

tOral

Treatm

ento

ffevercou

ghLeafandroot

areb

oiledtogether

and

takenorally

Emiliacoccinea

Aste

raceae

Odu

ndun

Leafsap

and

root

Skin

Treatm

ento

fulcerhernia

measle

sWho

leplantissqu

eezedandsaptaken

orally

Acalypha

godseffi

ana

Euph

orbiaceae

Jinwinini

Leaf

Skin

Treatm

ento

fskin

infectionsantim

icrobials

Leafismacerated

andtakenorallyas

well

asappliedto

affectedskin

area

8 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Brideliafer

rugin

eaEu

phorbiaceae

Irikirni

Leafstem

bark

root

Oral

Treatm

ento

fInsom

nia

mou

thwashgono

rrhea

Who

leplantisb

oiledandtakenorally

Euphorbiaheterophylla

Euph

orbiaceae

Egele

Leafroo

tsSkin

Treatm

ento

fSkindiseases

Leafandroot

arem

acerated

andthe

poultic

eapp

liedto

thea

ffected

skin

area

Euphorbiahirta

Euph

orbiaceae

Non

onku

rciya

odne

Who

leplant

exud

ate

Oral

Treatm

ento

fAsth

ma

coug

hshapeo

fbreasts

Thew

holeplantism

acerated

andtaken

orally

Euphorbialaterifl

ora

Euph

orbiaceae

Orow

ere

Leafexud

ate

Skin

Treatm

ento

fdermatitis

constip

ation

Theleafissqu

eezedandthep

oultice

appliedto

thea

ffected

skin

area

Cajanu

scajan

Fabaceae

Orela

Leafseed

Skin

andoral

Treatm

ento

fSmallpox

and

mou

thwash

Leafissqueezed

andappliedto

the

affectedarea

Dan

iellia

thurifera

Fabaceae

Iya

Stew

-woo

ddu

stSkin

Treatm

ento

fscabies

Theb

arkismacerated

andappliedto

the

affectedskin

area

Dialiu

mguineense

Fabaceae

Tasm

iyar

kumi

Leafbark

fruitand

twig

Oral

Treatm

ento

fBronchitis

coug

handdiuretic

Theleafand

bark

arec

rushed

andbo

iled

togetherTh

emixture

isthen

takenorally

Acacianilotica

Fabaceae

Baanigabaruwa

Fruitbark

exud

ate

Skin

Treatm

ento

fskindiseases

fung

alinfections

Thee

xudatesa

resqueezed

from

theb

ark

andapplieddirectlyto

thea

ffected

skin

area

Napoleona

imperia

lisLecythidaceae

Mabun

giTw

igs

Oral

Treatm

ento

fAsth

ma

coug

hTw

igsa

remacerated

andtakenorally

Alliu

msativ

umLiliaceae

Ayoayuu

Bulb

Oral

Treatm

ento

ffevercou

gh

asthmaantim

icrobial

Theb

ulbismacerated

andtakenorally

Hibiscus

asper

Malvaceae

Isapad

angiraa

Leaf

Oraland

skin

Treatm

ento

fcou

gh

woun

dsanddiuretic

Leafisbo

iledandtakenorally

Hibiscus

rosasin

ensis

Malvaceae

Kekekeireagu

Leafstem

flowe

rSkin

Treatm

ento

finfl

uenza

woun

dsand

diuretic

Who

leplantism

acerated

andtaken

orally

Theleafissqu

eezedandappliedin

affectedskin

area

Hibiscus

sabdariffa

Malvaceae

Sobo

gurgu

zuLeafflow

erOral

Treatm

ento

fDiuretic

scoug

handdressin

gwo

unds

Leafisbo

iledandtakenorally

Ficuseleg

ans

Moraceae

Asoro

Leaf

Skinoral

Treatm

ento

fPile

constip

ationandcraw

-craw

Leafissqueezed

andappliedto

the

affectedarea

The Scientific World Journal 9

the absorbance The percentage inhibition of DPPH orABTS radical scavenging activity by the plant extracts wascalculated as (Abscontrol ndash Abssample)(Abscontrol) times 100where Abscontrol is the absorbance of DPPH or ABTS +methanol Abssample is the absorbance of DPPH or ABTSradical + sample extractstandard Here the concentration ofthe extracts needed to decrease the absorbance of DPPH orABTS radical by 50 was calculated Rutin (Sigma-Aldrichge 94 HPLC grade) at the same working concentrations ofthe plant extracts was used as reference drug

210 Bacterial Strains All bacterial strains were culturedon nutrient agar slopes and were incubated for 24 h at37∘C before the MIC An inoculum turbidity equivalent toa 05 McFarland standard (1times107 CFUmL) was prepared innormal saline for each test organism and then diluted 1100in Mueller-Hinton broth just before injection of the platesStandard strains used in the study included S aureus and Ecoli which were a gift from Dr EO Ekundayo (Departmentof Microbiology Michael Okpara University of Agriculture)and clinically isolated strains of K pneumoniae and Pmirabilis were gotten from Department of MicrobiologyFederal Medical Centre (FMC) Umuahia Abia State

211 Antibacterial Susceptibility Using Disc Diffusion MethodThe microbial growth inhibition potential of the plantextracts was determined by using the agar disc diffusionmethod [17] The plant extracts were dissolved in 10 vvDMSO (Merck UK Ltd) to a final concentration of 2 mgmLThe nutrient agar plate was streaked with the standardizedtest bacterial inoculum over the surface evenly and 2mgdiscof the plant extracts transferred onto sterile filter disc papers(about 6 mm) containing the test compounds was placed onthe agar surface in the Petri dishes 100 120583gdisc of standardantibiotic ciprofloxacin was used as control and 100 120583LDMSOdisc served as negative control The plates were thenincubated at 37∘C for 24 h Generally the antimicrobialextracts diffuse into the agar and inhibit the growth of thetest bacteria All tests were done in duplicate and diame-ter zones of inhibition of growth were measured using ameter rule from the edge of each dish after the incubationperiod

212 Minimum Inhibitory Concentration (MIC) Assays Wefollowed the method of Smith et al [18] to determine theMIC ofmethanol crude extracts of selected botanicals againstclinical isolates of bacteria associated with skin and oralinfection A volumeof 100mLof sterileMueller-Hinton broth(Oxoid Basingstoke UK) containing 20 mgL of Ca2+ and10 mgL of Mg2+ was dispensed into a 96-well microtitreplate (Nunc 03 mL total volume per well) All antibacterialagents were dissolved in dimethyl sulphoxide (DMSO) anddiluted in Mueller-Hinton broth to give a stock solutionThen 100 120583L of the antibacterial agent stock solution (2000mgL) was serially diluted into each well and 100 mL ofthe bacterial inoculum added to each well to give a finalconcentration range of 0024 - 1 mgL All procedures wereperformed in duplicate and the plates were incubated for 18

h at 37∘C Briefly 20 mL of a 5 mgL methanol solution of 3-[45-dimethylthiazol-2-yl]-25-diphenyltetrazolium bromide(MTT) (Sigma-Aldrich Ltd South Africa) was added to eachwell and then incubated for 30 min Blue colouration indi-cated bacterial growth The MIC was recorded as the lowestconcentration at which no colour change was observed

213 Statistical Analysis Data analysis was done inMicrosoftExcel to obtain descriptive statistics Means values wereseparated by the Duncan multiple tests using SAS The dif-ferent levels of significance within the groups were analyzedusing one-way analysis of variance (ANOVA) Values wereconsidered significant at P lt005

3 Results and Discussion

31 Ethnobotanical Survey Modern drugs are expensive andsupply to a remote area may be irregular hence result-ing in overreliance of traditional methods of healing as aprimary source of healthcare [19] The rural communitiesof Ebem-Ohafia region of southeastern Nigeria depend onherbal remedies to meet their domestic and health needsDuring our ethnobotanical survey a total of 30 respondentsincluding the elders herbalists and traditional healers wereinterviewed in the study areaMajority of the people (22)wereaged 30ndash65 years dominated by males (25) and one-third(10) had a primary school education while 15 had secondaryeducation and others informal education A total of 61medic-inal plant species belonging to 26 families were identified inthe treatment of oral and skin diseases as shown in Table 1Twenty-seven of them are used in the management of oralinfections and twenty-six plants for the management of skininfection while eight plants are used in the management ofboth infections Table 2 shows the list of family number of theplant species used and their percentage occurrenceThemostrepresented plant family is the Euphorbiaceae having 1803of the plant species followed by Asteraceae and Fabaceae with1147 each [20] Euphorbiaceae is a large family of floweringplants with 300 genera and about 7500 species found usefulin the curing of some common human diseases Asteraceae isa large family of flowering plants having 32 913 species and1911 genera and has been found to possess diverse biologicaleffects Fabaceae is a family of cosmopolitan distributionwith approximately 730 genera and 19400 species richnessat a global level [20] The leaves (60) recorded the mostcommonly used plant part followed by the root (16) andthen stem bark (12) None of the respondents indicated anypossible side effects caused by any of these plants listed inTable 1 The most common methods of preparation includeinfusion decoction and maceration These herbalists ortraditional healers often collect the plants from the fieldand dry and crush them into powder form before storingin bottles to prevent patients from identifying the plantsused for their treatment Most healers refused to provideinformation without payment whereas others thought toprovide information on the plant use is disrespect of theirtraditional belief Furthermore the majority of the plantsmentioned lack scientific data to substantiate their acclaimedfolkloric use From this work it is evident that medicinal

10 The Scientific World Journal

Table 2 The list of family number of the plant species and theirpercentage occurrence

Family Number of plant species OccurrenceFabaceae 7 1147Euphorbiaceae 11 1803Asteraceae 7 1147Myrtaceae 4 656Moraceae 3 492Malvaceae 4 656Liliaceae 2 328Capparaceae 2 328Annonaceae 3 492Combretaceae 1 164Burseraceae 2 328Apocynaceae 1 164Gnetaceae 1 164Lecythidaceae 1 164Verbenaceae 1 164Ochnaceae 1 164Rubiaceae 1 164Piperaceae 1 164Acanthaceae 2 328Papilionoideae 2 328Asclepiadaceae 1 164Caesalpiniaceae 2 328Convolvulaceae 1 164Rutaceae 1 164Crassulaceae 1 164Poaceae 1 164

plants play a significant role in the primary healthcare ofthe people and conservation of the indigenous knowledge ofthis folklore medicine is crucial for the innovations of futuredrug discovery and possible herbal plants trade in the region[19] On the contrary we observed imprecise dosage lowhygiene standards hiding of healing methods and lack ofpatients record as a significant setback to traditional healthpractitioners in the region

32 Thin Layer Chromatography (TLC) Profiling Thin layerchromatography remains the most common useful techniqueby researchers as a quick and simple method to resolvethe constituents of a crude extract followed by advancedextraction methods to purify the active ingredients Previousstudies reported that specific secondary metabolites in plantexhibit a wide range of antioxidant and antibacterial proper-ties [21] TLC was formed on a glass sheet coated with silicagel After the sample is applied to the plate a solvent system(mobile phase) is drawn up via capillary action on the TLCplate at different mobility rate depending on the chemicalcompositionsThe separated spotswith different colourswerevisualized by the UV light projection onto the glass sheetThe variation of coloured spots detected in the crude extractsindicates the presence of different phytochemicals that may

be responsible for the pharmacological effects of selectedplants use in the management of oral and skin infections asacclaimed by the traditional healers We quantified the resultsby measuring the distance travelled by the plant compoundsdivided by the total distance travelled by the mobile phaseFrom Table 3 it is apparent that mobile phase 1 consistingof chloroform and ethanol in the volume ratio of 64 had abetter separation than mobile phase II containing tolueneethyl acetate and acetic acid in the volume ratio of 541 Themethanol extract of AH in bothmobile phases had the highestnumber of three bands with Rf values of 0667 0833 and0989 for mobile phase I and Rf values of 0381 0648 and0781 for mobile phase IIThe plant extracts MM BA CO JCandCAhad two bands inmobile phase I with clear separationbut had single band in mobile phase II The plant extractshaving only one band in either mobile phase suggest a poorsolvent system for compounds separation The TLC profilesprovide a characteristic fingerprint of these plants that mayalso be useful for their identification

33 Antioxidant Assays

331 TPC and TFC Phenolic compounds are secondarymetabolites distributed in fruits vegetables nuts herbsseeds and beverages [22] They are excellent scavengers offree radical that has been implicated in the pathologies ofvarious human diseases the antioxidant activity is dependenton the structure hydrophobic benzenoid rings and hydrogen-bonding potential of the phenolic hydroxyl groups In TPCassay a phenol in plant extracts loses a proton to formphenolate ion which reduces Folin Ciocalteu reagent andthe resulting changes are measured spectrophotometricallyat 765 nm [13] The content of phenol is reported in mg oftannic acid equivalent (TAE)mL of extract As shown inFigure 1 the total phenolic contents of absolute methanolcrude extract of these botanicals range from 700 to 3600mgTAEg The AH extract had the highest concentrationfollowed by CA CO MM BNI BA EI BN and JC whilethe least concentration was recorded in II extract Moreoverthe Folin Ciocalteu reagent (FCR) has been used to measureother compounds such as ascorbic acid amino acids andsugars hence it may not provide a concise amount of thephenolic compounds present in these plants Flavonoids areincreasingly consumed in a significant amount in daily dietsworldwide due to their various health benefits toman [23 24]In TFC assay aluminum ion in the reacting mixture formscomplexes with the C- 4 keto and either C- 3 or C- 5 hydroxylor with ortho hydroxyl groups in the A or B ring of flavonoidsdetected in the plant extracts [14] As shown in Figure 2the total flavonoid content was recorded in the plant extractsranging from 750 to 1500 mgQEg as extrapolated from thestandard quercetin curve The CA extract had the highestconcentration followed by BNI CO II AH BN MM JCand BA while the lowest concentration was recorded in EIextract TPC is found higher than the TFC in some of theextracts supporting the fact that most flavonoids are alsophenolics According to our results all the plant extractsare rich source of phenolic compounds The composition ofTPC and TFC registered in these plants may be difficult to

The Scientific World Journal 11

Table 3 Thin layer chromatography retention factor using mobile phase CHL ET (mobile phase I) and Tol EA AA (mobile phase II)

SN Plant Extract CHL ET (64) Tol EA AA (541)1 Baphia nitida 1 0742 Macrolobium macrophyllum 0706 0941 06953 Chromolaena odorata 0471 0647 06674 Jatropha curcas 0353 0941 06485 Cassia alata 0118 0882 06866 Ipomoea involucrata 0956 -7 Acalypha hispida 0667 0833 0989 0381 0648 07818 Breynia nivosa 0989 07629 Burkea africana 0644 1 075210 Eleusine indica 0978 0743CHL chloroform ET ethanol Tol toluene EA ethylacetate AA acetic acid

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

500

1000

1500

2000

2500

3000

3500

4000

TPC

(mgT

AE

g)

Figure 1 The total phenolic content (TPC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata TAE tannic acid equivalent

compare with others reported elsewhere due to many factorsincluding differences from species to species maturity stageplant parts and assay methods harvesting time standarddrugs and geographical location

332 Reducing Power (RP) Ferric reducing power is a globalmethod often used to assess the capacity of antioxidantsto donate either an electron or hydrogen atom to unpairedelectron [25] In reducing power assay plant extracts exertantioxidant activities via reduction of Fe3+ferric cyanidecomplex to Fe2+ (ferrous form) resulting in changing thegreen colour to blue and sometimes Perlrsquos Prussian bluecolour depending on the antioxidant potential [10] As shownin Figure 3 the reducing power of the methanol crudeextracts and the standard gallic acid rutin and ascorbic acidincreased with increase in concentration Higher absorbanceat 700 nm can be read after potassium ferricyanide reactionwith ferric chloride At 1000 120583gmL the extracts from AH

0

200

400

600

800

1000

1200

1400

1600

TFC

(mgQ

Eg)

AH BN JC CO MM EI BNI BA CAIIPlant extracts

Figure 2 The total flavonoid content (TFC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata QE Quercetin equivalent

BNI MM CA BA EI and BN had absorbance value of 270086 073 055 036 027 and 027 respectively while that ofAH is higher than the standard rutin (254) but comparablewell with gallic acid (275) and ascorbic acid (269) EI andBN extracts had a similar trend of ferric reducing powerthough significantly lower as compared with the standarddrugs AH extract exhibits remarkable antioxidant capacity(RP) correlating with its richness in phenolic and flavonoidcontents It is reasonable to suggest that methanol selectivelyextracts antioxidant principles from these botanicals basedon their polarities and chemical structures which are reflectedin plant extracts with high TFC and TPC having minimal RP[26] We have shown that some plant extracts in particularAH BNI MM and CA extracts may serve as an excellentcandidate to minimize skin and oral infections caused byoxidative stress as a result of imbalance of free radicalsgeneration and natural antioxidants in the saliva or woundarea According to our results the increasing RP of the plantextracts correlateswith their TPCandTFCThis study at least

12 The Scientific World JournalRe

duci

ng P

ower

7

00nm

625 125 250 500 10003125Concentration (gmL)

0

05

1

15

2

25

3

IIAHBNJCCOMMEI

BNIBACAGARutinVit C

Figure 3 The ferric reducing antioxidative capacity of selectedplant extracts used in the management of oral and skin infectionsin southeast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata GA gallic acid Vit C vitamin C (standard antioxidants)

in part lends credence to the ethno-therapeutic use of someplant as natural antioxidant in themanagement of skin or oralinfections in the study area

333 FRSA of Plant Extracts Skin and oral cavity exposedto an array of oxidants such as smokes from cigaretteschemicals UV light nicotine and alcohol that are directlyor indirectly leading to the production of reactive oxygenspecies (ROS) are known to be responsible for some com-mon human diseases [27] These free radicals in particularhydroxyl and superoxide anions are highly reactivemoleculesderived from the metabolism of oxygen Although they playan important role in the biochemical immunological andphysiological processes it could be deleterious to vital organsif excessively accumulated in the human body [28] Food andpharmaceutical industries discovered that medicinal plantscontaining phenol and flavonoid compounds are essentialin the development of novel drugs against stress-induceddiseases [28] In the present investigation the concentration(IC50) at which 50 of free radical scavenged was calculatedand extrapolated from the plot of percentage inhibition ver-sus plant extract concentration The herbal drugs exhibitedrobust DPPH radical scavenging activity in a concentration-dependent manner having IC50 ranging from 2025 to 710120583gmL As shown in Figure 4 MM BA EI JC AH BNIand CA extracts had a potent DPPH radical scavengingactivity corresponding to their TPC TFC and RP Theplant extracts scavenged ABTS radical by the decrease inits absorbance at 734 nm All the plant extracts exhibitedstrong ABTS radical scavenging capacities in the following

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

200

400

600

800

1000

1200

1400

1600

IC50

(g

mL)

Figure 4 DPPH free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

050

100150200250300350400450500

IC50

(g

mL)

AH BN JC CO MM EI BNI BA CAIIPlant samples

Figure 5 ABTS free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

order BN gt JC gt CA gt MM gt BNI gt EI gt CO gt AH gtII gt BA having IC50 values ranging from 25 to 400 120583gmL(Figure 5) DPPH and ABTS radical scavenging activities ofthese herbal remedies displayed a positive correlation withthe working concentrations It is obvious that most of theplant extracts tested in this work possess the ability to quenchthe free radicals generation via electron or proton donationdemonstrated by the termination of radical chain reaction[28] Although both DPPH and ABTS radicals are electron-transfer based methods however it is apparent that ABTSmethod seems to be more sensitive than DPPH radical assaypossibly due to their solubility and diffusivity as shown inthe IC50 values [10] It is worth noting that BA extract withstrong DPPH radical scavenging effect had a moderate ABTS

The Scientific World Journal 13

Table 4 Zone of bacterial inhibition by the selected plant extracts use in the management of oral and skin infections

Diameter of inhibition (mm)SN Plant samples K pneumoniae P mirabilis S aureus E coli1 Cassia alata 15 plusmn 141 9 plusmn 042 15 plusmn 083 13 plusmn 0802 Breynia nivosa 8 plusmn 052 17 plusmn 100 8 plusmn 008 15 plusmn 1023 Burkea africana NI 11 plusmn 084 13 plusmn 072 NI4 Baphia nitida 8 plusmn 050 13 plusmn 122 8 plusmn 051 NI5 Ipomoea involucrata 8 plusmn 052 NI 8 plusmn 050 13 plusmn 0526 Jatropha curcas NI 9 plusmn 030 9 plusmn 062 13 plusmn 0727 Acalypha hispida 21 plusmn 163 NI 11 plusmn 009 17 plusmn 1328 Chromolaena odorata 8 plusmn 042 8 plusmn 035 9 plusmn 009 15 plusmn1209 Eleusine indica 8 plusmn 042 8 plusmn 035 NI 17 plusmn 10010 Macrolobium macrophyllum NI NI 8 plusmn 004 15 plusmn 12311 Ciprofloxacin - - le16(R) ge17(S) le16(R)ge20(S)NI indicates no zone of inhibition S sensitive R resistance

radical scavenging capacity which can be explained by thepartial ionization of the extract in the test medium [29] SinceROS promotes tissue damage and epithelial inflammationvia exposure to sunlight radiation or smoke as a result theantioxidant capacity demonstrated in this work may supportthe traditional use of these plants against the influence ofinflammatory markers and oxidants in the skin and oralcavity [29] Based on the results of the three antioxidantassays it is obvious that MM AH BNI EI and CA extractshad the best FRSA indicating their importance in convertingfree radicals into more stable products and then terminatethe free radical chain reaction Our study showed that themethanol crude extracts from these plants could protectagainst free radical damage in the mouth such as humangingival periodontal tissues and certain skin infections [5]

334 Antibacterial Activity of Plant Extracts Pathogenicbacteria such as Staphylococcus aureus Escherichia coli Kleb-siella pneumoniae and Proteus mirabilis play a significantrole as the primary cause of skin and oral infections [30]Staphylococcus aureus is a part of normal skin known to bethe most problematic in the cause of skin infections as aresult of their resistance mechanisms to first-line antibiotics[30] Although the presence of this organism as a residentoral floral is controversial nonetheless MSSA and MRSAhave been isolated from oral rinse and tongue swab inpatients with oral infections [31] Escherichia coli are Gram-negative bacteria known to be notorious for their resistantmechanisms towards the existing arsenal of antibiotics Oralinfections such as edematous and erythematous and skininfections like omphalitis surgical site infections burnnecrotizing fasciitis and cellulitis at all age groups are associ-ated with enterobacterium E coli [32] Klebsiella pneumoniaeis a Gram-negative encapsulated facultative anaerobic androd-shaped bacterium found in the normal flora of themouth skin and intestines This bacterium is responsiblefor the prolonged surgical wound biliary tract wound andurinary tract infections and it could pose a serious healthrisk when resistant strains invade the throat mouth or

intestines [33] Villegas et al [34] detected a CTX-M-12 120573-lactamase producing K pneumoniae new plasmid-mediatedESBLs highly resistant to ceftazidime from a patient withhospital-acquired wound infection Proteus mirabilis is oneof the most common enterobacteria after E coli identified asGram-negativemotile and rod-shaped bacterium It is foundin clinical specimen implicated in the skin respiratory tractand bacteraemia as well as burns wounds and urinary tractinfections [35] All these pathogenic multidrug-resistancebacteria are causative agent in a periodontal lips ulcer whitesponge nevus and skin infections [36]

335 Susceptibility Test and MIC Determination The resultof disc diffusion assay showed the diameter of inhibitionof bacteria growth classified as le 15 (resistant) and ge 16(susceptible) according to CLSI guideline [37] As shown inTable 4 K pneumoniae and E coli are more susceptible tothe treatment of Acalypha hispida (AH) having a zone ofinhibition (ZI) 21 and 17mm respectively BN and EI extractsinhibit the growth of P mirabilis and E coli having the samezone of inhibition 17 mm corresponding to the report of Al-Zubairi et al [38] The data obtained for the methanol crudeextracts from AH BN and EI show their selective inhibitionof the test bacteria growth The plant extracts with ZI above13 mm against our panel of bacterial strains were selected forMICdeterminationHere themethanol crude extracts exhibitMIC values of 256 to gt 512 120583gmL against the test bacterialstrains but were less active as compared to the standardciprofloxacin 003 - 006 120583gmL (Table 5) Ciprofloxacin isan active broad-spectrum antibiotic use in the treatmentof urinary tract infections prostatitis and acute cystitis viainterfering with bacterial DNA replication and transcriptionthrough inhibition of DNA gyrase or topoisomerase [39] Allthe plant extracts demonstrated a mild to weak antibacterialactivity based on the classification of Rios and Recio [40] whoconsidered antibacterial activity of the crude plant extractsignificant (MIC lt 100 120583gmL) moderate (100 lt MIC = 512120583gmL) or weak (MIC gt 512 120583gmL) The herbal remediesshowed moderate inhibition of bacterial growth except for K

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

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The Scientific World Journal

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Submit your manuscripts atwwwhindawicom

Page 3: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

The Scientific World Journal 3

indica (EI) were collected after the interview The plants wereidentified by their local names and authenticated by MrOriaku Williams and Professor Gabriel Osuagwu from theDepartment of Plant Science and Biotechnology College ofNatural Sciences Michael Okpara University of AgricultureUmudike in Abia state Nigeria The vouchers specimens(VICJON 1ndash10) were prepared and deposited at the depart-mental herbarium The preparations and mode of intake ofthe herbal remedies are described in Table 1

24 Sample Preparation We collected leaves and roots ofthe ten plants that were frequently mentioned in the surveyand oven-dried at 40∘C for 72 h while the root of EI wasdried longer for seven days The dried plant materials werepulverized to a fine powder using an electric blender andstored in an airtight container for further use Thirty grams(30 g) of dried powdered materials were extracted with 150mL of 100methanol for 48 h on amechanical shaker (StuartScientific Orbital 202 SOSI Essex UK) and the extractswere filtered using Buchner funnel and Whatman Number1 filter paper The filtrate was concentrated using a rotaryevaporated at 40∘C to recover the solvent and air-dried in afume chamber to give a yield ranging from 45 to 83 g

25 Thin Layer Chromatography (TLC) Profile Thin layerchromatography is a simple method for analyzing a complexmixture of compounds based on the distance travelled Theplant extracts (1 mgmL) dissolved in methanol and spottedon the plate coated with silica gel 60 F254 as a stationaryphase The slurry was prepared by dissolving 15 g of silicagel 60 F254 in 30 mL of distilled water and immediatelypoured into the plate The plates were air-dried overnightAbout 10 120583L of the plant extracts was gently loaded onthe base of the plate (5 cm above) using the capillary tubeThe plates were allowed to develop in chromatographictanks consisting of three different solvents (mobile phase)chloroform methanol acetic acid (541) until they reach34th of the TLC plate The TLC plate was removed andallowed to dry the spots were detected by iodine vapour andretention factor (Rf) was calculated using the equation Rf= distance travelled by the componentsdistance travelled bythe solvent

26 Total Phenolic Content (TPC) Assay The total phenoliccontent present in these extracts was quantified by the FolinCiocalteu reagent (FCR) using the method of Ghaffari etal [12] Briefly 02 mL of the plant extract (2 mgmL) wasadded to the reaction mixture consisting of 1 mL of 10 vvFCR and 08 mL of Na2CO3 (0075 mgmL) to give a finalconcentration of 02 mgmL of each extract The resultingmixture was incubated at 45∘C with shaking for 15 min andthe absorbance measured at 765 nm A standard curve wasprepared by mixing methanol solution of gallic acid (02mL 0025 - 0400 mgmL) with 1 mL of 10 vv FCR andsodium carbonate (08 mL 0075 mgmL) The experimentwas carried out in triplicate and the results presented asmeanvalues with standard deviation (plusmn SD) The TPC value wasexpressed as milligrams of gallic acid equivalent (GAE) perg of dried sample It was calculated using the formula T =

CtimesVM where T is the TPC (mgg) of extract in GAE C isthe concentration of gallic acid from the calibration curve Vis the volume of the extract mL M is the dry weight (g) ofthe leaf or root powder from which the extract was obtained

27 Total Flavonoid Content (TFC) Assay The concentra-tion of flavonoids in the plant extracts was determined viaaluminum colorimetric assay method [13] Briefly 1 mL of2 wv AlCl3 was prepared in 100vv methanol added to 1mL of the sample solution A yellow colour formation afterincubation at room temperature for 1 h was measured at420 nm using an AJI-C03 UV VIS spectrophotometer Thestandard curve for TFC was obtained using quercetin as astandard drug under the same procedure described in TPCdetermination The TFC was calculated using the formulaT = CtimesVM where T is the TFC (mgg) of extract in QEC is the concentration of quercetin established from thecalibration curve V is the volume of the extract mL M isthe dry weight (g) of the leaf powder from which the extractwas obtained The TFC was present in the extracts calculatedas mgg of quercetin equivalent (QE)

28 Ferric Reducing Antioxidant Power (FRAP) Assay Theability of plant extracts to reduce ferric (Fe3+) to ferrous(Fe2+) formwas evaluated following themethod described byYen and Chen [14] with slight modification A volume of 03mL of different concentrations (0025 - 2 mgmL) from plantextract BHT ascorbic acid and rutin prepared in distilledwater was mixed with reacting mixture consisting of 25 mLof 02 M phosphate buffer (pH 66) and 25 mL of K3Fe(CN)6(1 wv) The resulting mixture was incubated at 50∘C for20 min followed by addition of 25 mL of TCA (10 wv)After vigorous shaking 25 mL of the solution was mixedwith 25 mL of distilled water and 05 mL of FeCl3 (01 wv)left incubated at room temperature for 5 min and then theabsorbance was measured at 700 nm against a blank sample(without extract)

29 Free Radical Scavenging Activity Assays The free radicalscavenging potential of plant extracts was measured in vitroby the 1 11015840-diphenyl-1-picrylhydrazyl (DPPH) described byTariq et al [15] The assay was experimented by reacting16 mL of 0135 mM DPPH dissolved in 100vv methanolwith 04 mL of various concentrations (0078 - 1 mgmL) ofmethanol crude extracts The reaction mixture was vortexedthoroughly and left in the dark at room temperature for30 min The absorbance of the mixture was measured at517 nm after 2 min The method of Re et al [16] wasadopted to determine ABTS radical scavenging activity ofthe plant extracts The ABTS radical solution was generatedby mixing two stock solutions of 7 mM ABTS and 24mM potassium persulphate in the same ratio and allowingthe solution to react for 12 h at room temperature in thedark The resulting solution was diluted with methanol toobtain an absorbance of 0706 units at 734 nm A volumeof 1 mL of various concentrations (003125 ndash 1 mgmL)of the plant extracts react with 25 mL of ABTS radicalsolution in the dark for 15 min and was later measured

4 The Scientific World Journal

Table1Medicinalplantsused

inthem

anagem

ento

fskinor

oralinfections

insoutheastern

partof

Nigeria

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Macrolobium

macrophyllum

Fabaceae

Antho

notha

macroph

ylla

Leaf

Skin

Treatin

gof

woun

dandcut

Squeezethe

leavea

ndplaceo

nthe

affectedarea

ofthes

kin

Jatro

phacurcas

Euph

orbiaceae

Barbados

nut

Leaf

Skinoral

Treatm

ento

fboils

Boiltheleave

with

waterPlace

theh

otleaveo

nthea

ffected

area

ofthes

kin

Eleusin

eind

icaPo

aceae

Yard

grass

Root

Skin

Treatm

ento

fRingw

orm

Use

ther

ootand

scrubon

affectedarea

ofthes

kin

Cassiaalata

Caesalpiniaceae

Ring

worm

bush

Leaf

Skin

Treatm

ento

fRingw

orm

Scrubtheleave

onaffectedarea

ofthe

skin

Chromolaena

odorata

Aste

raceae

Siam

weed

Leaf

Skin

Tosto

pbleeding

andrashes

Squeezethe

leavea

ndrubon

affectedarea

Psidium

guajava(L)

Myrtaceae

Guava

Leaf

Skin

Treatm

ento

fskinbu

rns

scrapessunb

urn

woun

ds

psoriasiseczem

aand

others

Crushedgu

avaleafo

rboiledleaves

are

applieddirectlyto

thea

ffected

skin

providingreliefandareb

elievedto

improveh

ealin

gBryophyllum

pinn

atum

(Lam

)Oken

Crassulaceae

Mira

cleleaf

Dogrsquosliver

Leaf

Skino

ral

Treatm

ento

fboilwo

und

sore

orcut

Crushleafandapplyjuiceo

nthew

ound

surfa

ce

Ficusa

sperifolia

Miq

Moraceae

False

thistle

Leaflatex

Skin

Treatm

ento

fskinrashes

anditches

Appliedas

poultic

etothes

kinitches

Acanthus

montany

(TA

nderson)

Acanthaceae

Bearrsquosbreeches

Stem

bark

Skin

Treatm

ento

fwou

ndsa

ndskin

diseases

Woo

dbark

iscrushedandsoaked

italcoho

landextractapp

liedto

woun

dsandskin

Alliu

msativ

umL

Liliaceae

Garlic

Bulbjuice

Skin

oral

Usedto

cond

ition

thes

kin

chew

edforc

ough

andsore

throat

Thejuice

ismixed

with

lotio

nsforskin

cond

ition

ingFreshbu

lbischew

edor

boiledin

water

anddrinkdecoctionfor

coug

h

Emiliasonchifolia

(L)

Aste

raceae

Scarlettassel

Leaf

Skin

Treatm

ento

fSkintro

ubles

abscessa

ndbruises

Crushedleaves

were

pressedandused

tocoverthe

woun

dor

bruisedsurfa

ces

Sida

acutaBu

rmf

Malvaceae

Broo

mwe

edLeaf

Skin

Treatm

ento

fjaund

icesore

ulcer

Poultic

esmadefrom

boiledleaves

are

appliedon

thea

ffected

parts

The Scientific World Journal 5

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Ricin

uscommun

isLinn

Eu

phorbiaceae

Castorb

ean

Leafroo

tand

seed

Skin

Treatm

ento

fskininfection

anddetoxification

Applycasto

roilon

thea

ffected

area

torelieve

skin

infection

Citru

slim

on(L)Bu

rmf

(prosp

)Ru

taceae

Limon

Leaffruit

andseed

Skin

Treatm

ento

ffluheadache

cold

andoralinfections

Mixequalquantities

ofLimon

juicea

ndho

lybasilkeepitun

derthe

suntill

becomes

thickapplythison

face

Alliu

mcepa

LLiliaceae

Onion

sLeafandbu

lbSkin

Treatm

ento

fperiodo

ntitis

andcold

Theb

ulbismacerated

andmixed

with

water

andthen

takenorally

Breynianivosa

Euph

orbiaceae

Icep

lant

Leaf

Oral

Treatin

gof

toothache

Boiltheleave

with

waterthentake

the

water

Ipom

oeainvolucrata

Con

volvulaceae

Water

convolvu

lus

Leafandste

mOral

Treatm

ento

foralinfectio

nMixtheleave

with

ugbaga

treea

ndbo

ilwith

water

anddrinkthew

ater

Burkea

afric

ana

Caesalpinioideae

Icheku

Leaf

Oral

Treatm

ento

forald

isease

Boiltheleave

with

water

andtakebut

donrsquot

swallowit

Baphianitid

aPapilio

naceae

Cam

wood

Leafor

stem

Oral

Streng

thensthe

teeth

Chew

thes

tem

orscrubtheleave

onthe

teeth

Gongronem

alatifolium

Asclepiadaceae

Utazi

Leaf

Oral

Treatm

ento

fcou

ghand

diabetes

Crushtheleafw

ithmortara

ndmixwith

ginger

orgarlica

nddrink

Anonasenegalen

sisAnn

onaceae

Ubu

ruocha

Root

Oral

Treatm

ento

fmalariaand

mou

thinfection

Boilther

ootsanddrink

Abrusp

recatoriu

sPapilio

noideae

Crabrsquoseyes

Seed

androot

Oralskin

Treatm

ento

fcou

ghskin

carew

ound

Grin

dther

ootand

applythep

oultice

onthea

ffected

skin

areaA

lsocrushedseeds

canbe

takenorally

Acanthus

montanu

sAc

anthaceae

False

thistle

Leafandroot

Oraland

Skin

Treatm

ento

ffurun

cles

lesio

nsandurinary

infections

Boilther

ootand

applythep

oultice

tothe

affectedpartof

theb

odyFo

rinfectio

ns

boilther

ootand

leaves

anddrink

Piperu

mbellatum

Piperaceae

Njamuja

Leaf

Oral

Treatm

ento

fpeptic

ulcers

Crushtheleavesa

ndsoak

inwarm

orho

twater

anddrink

Euphorbiahirta

Euph

orbiaceae

Asth

maw

eed

Who

leplant

Oral

Treatm

ento

fwou

nds

Leaves

arem

acerated

andappliedto

affectedpart

Ficusb

urkei(Miq)Miq

Moraceae

Sand

paper

Leaf

Oral

Treatm

ento

fhypertension

Leaves

areb

oiledandtakenorally

6 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Gmelina

arborea

Verbenaceae

Gmelina

Stem

andleaf

Oral

Treatm

ento

fgon

orrhea

pilesabdo

minalpains

burningsensationsfever

mou

thwo

undanddiabetes

Snake-biteandscorpion

sting

Leaves

grou

ndinto

paste

with

water

are

appliedto

theforeheadforh

eadachein

feversB

oiledste

msa

retakenorally

Lophira

alata

Ochnaceae

Ekki

Stem

andleaf

Oral

Treatm

ento

ffeverm

outh

woun

dandmalaria

Boiltheleavesa

ndste

mandtake

orally

Nau

clealatifolia

Rubiaceae

Ubu

ru-in

uStem

andleaf

Oral

Itisused

inthetreatment

ofmalariaepilepsymou

thwo

undanxietypainfever

Boilther

ootsanddrink

Annona

murica

taAnn

onaceae

Sour

sop

Leaf

Oral

Treatm

ento

fcancera

ndmou

thwo

undinfections

Crushor

boiltheleavesa

ndtake

orally

Psidium

guajava

Myrtaceae

Guava

Leaf

Oral

Treatm

ento

fperiodo

ntal

diseasebu

rnsscrapes

sunb

urnwo

undspsoria

sis

eczemaandothers

Leaves

arec

rushed

andtakenorally

Astra

galusm

embran

aceus

Fabaceae

Beiq

iStem

andleaf

Oral

Treatm

ento

fcoldup

per

respira

tory

infections

allergiesfib

romyalgia

anem

iaH

IVA

IDS

Leaves

arec

rushed

andtakenorallyand

canalso

beappliedto

skin

Buchholziacoria

cea

Capparaceae

Won

derfu

lkola

Leaf

Oral

Itisused

tradition

allyfor

treatingdiabetes

hypertensio

nrheumatism

coldcou

ghand

catarrh

Boilleaves

andtake

orally

Gnetum

afric

anum

Gnetacea

Afang

Leaf

Oral

Enlarged

spleen

sore-th

roatandcathartic

Leaves

canbe

boiledor

cooked

andtaken

orally

Anoridium

man

niAnn

onaceae

Ewuro-ig

boStem

bark

Oral

Treatm

ento

fDiarrhea

coug

hfeverrheumatism

Theb

arkiscrushedandbo

iledandthen

takenorally

Funtum

iaafric

ana

Apocyn

aceae

Akorie

mbamiri

Leafstem

root

Skin

treatmento

fcon

stipatio

nwo

undsw

eakbladder

jaun

dice

Who

leplantisb

oiledandtakenorally

Also

thec

rushed

leaves

area

ppliedto

the

affectedskin

area

The Scientific World Journal 7

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Commiphora

afric

ana

Burseraceae

Turari

dashi

Rootsfruits

Oral

Treatm

ento

fwho

oping

coug

hbron

chitis

Rootsa

remacerated

andtakenorally

Dacryodesedulis

Burseraceae

Elem

iub

eBa

rkroo

tsfruits

Skin

Treatm

ento

fJiggersSkin

diseasesE

leph

antia

sis

Theleavesa

reeatenrawwith

kolanu

tas

anantie

meticLeaf-s

apisinstilledinto

thee

arfore

ar-tr

oubleanda

leaf-decoctio

nisprepared

asa

vapo

ur-bathforfeverish

stiffn

essw

ithheadache

Combretum

mucronatum

Com

bretaceae

Fararg

eza

Rootsa

ndleaf

Skin

Treatm

ento

fskininfection

Ther

ootand

leafarem

acerated

and

appliedto

affectedskin

area

Cleomec

iliata

Capparaceae

Ekuya

Leafseeds

Skin

Treatm

ento

fCon

vulsion

wo

undssores

Seedsa

recrushedandthep

oultice

appliedto

thea

ffected

areaTh

emacerated

leaves

after

boiledaretaken

orally

Ageratum

conyzoides

Aste

raceae

Urata

Who

leplant

Skin

Treatm

ento

fwou

ndU

lcer

Sleeping

sickn

essEyew

ash

Thep

lant

ismacerated

andappliedto

affectedskin

area

Aspilia

afric

ana

Aste

raceae

Yunyun

Ka

lank

uwa

Leafflow

erSkin

Treatm

ento

fskinrashes

cleaning

sorescorneal

opacities

Thefl

ower

issqueezed

andappliedto

affectedskin

area

Centau

reaperrottetii

Aste

raceae

Danyi

Who

leplant

Skin

Treatm

ento

fskin

infectionsSyphilis

Thefl

ower

issqueezed

andap

oultice

appliedto

affectedskin

area

Chrysanthellu

mindicum

Aste

raceae

Abilereoyigi

Who

leplant

Skin

Treatm

ento

fboils

gono

rrheaJaun

dice

Thep

lant

iscrushedandthep

oultice

applieddirectlyto

thes

kinarea

Elephantopus

scaber

Aste

raceae

Elephant

foot

Leafroo

tOral

Treatm

ento

ffevercou

ghLeafandroot

areb

oiledtogether

and

takenorally

Emiliacoccinea

Aste

raceae

Odu

ndun

Leafsap

and

root

Skin

Treatm

ento

fulcerhernia

measle

sWho

leplantissqu

eezedandsaptaken

orally

Acalypha

godseffi

ana

Euph

orbiaceae

Jinwinini

Leaf

Skin

Treatm

ento

fskin

infectionsantim

icrobials

Leafismacerated

andtakenorallyas

well

asappliedto

affectedskin

area

8 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Brideliafer

rugin

eaEu

phorbiaceae

Irikirni

Leafstem

bark

root

Oral

Treatm

ento

fInsom

nia

mou

thwashgono

rrhea

Who

leplantisb

oiledandtakenorally

Euphorbiaheterophylla

Euph

orbiaceae

Egele

Leafroo

tsSkin

Treatm

ento

fSkindiseases

Leafandroot

arem

acerated

andthe

poultic

eapp

liedto

thea

ffected

skin

area

Euphorbiahirta

Euph

orbiaceae

Non

onku

rciya

odne

Who

leplant

exud

ate

Oral

Treatm

ento

fAsth

ma

coug

hshapeo

fbreasts

Thew

holeplantism

acerated

andtaken

orally

Euphorbialaterifl

ora

Euph

orbiaceae

Orow

ere

Leafexud

ate

Skin

Treatm

ento

fdermatitis

constip

ation

Theleafissqu

eezedandthep

oultice

appliedto

thea

ffected

skin

area

Cajanu

scajan

Fabaceae

Orela

Leafseed

Skin

andoral

Treatm

ento

fSmallpox

and

mou

thwash

Leafissqueezed

andappliedto

the

affectedarea

Dan

iellia

thurifera

Fabaceae

Iya

Stew

-woo

ddu

stSkin

Treatm

ento

fscabies

Theb

arkismacerated

andappliedto

the

affectedskin

area

Dialiu

mguineense

Fabaceae

Tasm

iyar

kumi

Leafbark

fruitand

twig

Oral

Treatm

ento

fBronchitis

coug

handdiuretic

Theleafand

bark

arec

rushed

andbo

iled

togetherTh

emixture

isthen

takenorally

Acacianilotica

Fabaceae

Baanigabaruwa

Fruitbark

exud

ate

Skin

Treatm

ento

fskindiseases

fung

alinfections

Thee

xudatesa

resqueezed

from

theb

ark

andapplieddirectlyto

thea

ffected

skin

area

Napoleona

imperia

lisLecythidaceae

Mabun

giTw

igs

Oral

Treatm

ento

fAsth

ma

coug

hTw

igsa

remacerated

andtakenorally

Alliu

msativ

umLiliaceae

Ayoayuu

Bulb

Oral

Treatm

ento

ffevercou

gh

asthmaantim

icrobial

Theb

ulbismacerated

andtakenorally

Hibiscus

asper

Malvaceae

Isapad

angiraa

Leaf

Oraland

skin

Treatm

ento

fcou

gh

woun

dsanddiuretic

Leafisbo

iledandtakenorally

Hibiscus

rosasin

ensis

Malvaceae

Kekekeireagu

Leafstem

flowe

rSkin

Treatm

ento

finfl

uenza

woun

dsand

diuretic

Who

leplantism

acerated

andtaken

orally

Theleafissqu

eezedandappliedin

affectedskin

area

Hibiscus

sabdariffa

Malvaceae

Sobo

gurgu

zuLeafflow

erOral

Treatm

ento

fDiuretic

scoug

handdressin

gwo

unds

Leafisbo

iledandtakenorally

Ficuseleg

ans

Moraceae

Asoro

Leaf

Skinoral

Treatm

ento

fPile

constip

ationandcraw

-craw

Leafissqueezed

andappliedto

the

affectedarea

The Scientific World Journal 9

the absorbance The percentage inhibition of DPPH orABTS radical scavenging activity by the plant extracts wascalculated as (Abscontrol ndash Abssample)(Abscontrol) times 100where Abscontrol is the absorbance of DPPH or ABTS +methanol Abssample is the absorbance of DPPH or ABTSradical + sample extractstandard Here the concentration ofthe extracts needed to decrease the absorbance of DPPH orABTS radical by 50 was calculated Rutin (Sigma-Aldrichge 94 HPLC grade) at the same working concentrations ofthe plant extracts was used as reference drug

210 Bacterial Strains All bacterial strains were culturedon nutrient agar slopes and were incubated for 24 h at37∘C before the MIC An inoculum turbidity equivalent toa 05 McFarland standard (1times107 CFUmL) was prepared innormal saline for each test organism and then diluted 1100in Mueller-Hinton broth just before injection of the platesStandard strains used in the study included S aureus and Ecoli which were a gift from Dr EO Ekundayo (Departmentof Microbiology Michael Okpara University of Agriculture)and clinically isolated strains of K pneumoniae and Pmirabilis were gotten from Department of MicrobiologyFederal Medical Centre (FMC) Umuahia Abia State

211 Antibacterial Susceptibility Using Disc Diffusion MethodThe microbial growth inhibition potential of the plantextracts was determined by using the agar disc diffusionmethod [17] The plant extracts were dissolved in 10 vvDMSO (Merck UK Ltd) to a final concentration of 2 mgmLThe nutrient agar plate was streaked with the standardizedtest bacterial inoculum over the surface evenly and 2mgdiscof the plant extracts transferred onto sterile filter disc papers(about 6 mm) containing the test compounds was placed onthe agar surface in the Petri dishes 100 120583gdisc of standardantibiotic ciprofloxacin was used as control and 100 120583LDMSOdisc served as negative control The plates were thenincubated at 37∘C for 24 h Generally the antimicrobialextracts diffuse into the agar and inhibit the growth of thetest bacteria All tests were done in duplicate and diame-ter zones of inhibition of growth were measured using ameter rule from the edge of each dish after the incubationperiod

212 Minimum Inhibitory Concentration (MIC) Assays Wefollowed the method of Smith et al [18] to determine theMIC ofmethanol crude extracts of selected botanicals againstclinical isolates of bacteria associated with skin and oralinfection A volumeof 100mLof sterileMueller-Hinton broth(Oxoid Basingstoke UK) containing 20 mgL of Ca2+ and10 mgL of Mg2+ was dispensed into a 96-well microtitreplate (Nunc 03 mL total volume per well) All antibacterialagents were dissolved in dimethyl sulphoxide (DMSO) anddiluted in Mueller-Hinton broth to give a stock solutionThen 100 120583L of the antibacterial agent stock solution (2000mgL) was serially diluted into each well and 100 mL ofthe bacterial inoculum added to each well to give a finalconcentration range of 0024 - 1 mgL All procedures wereperformed in duplicate and the plates were incubated for 18

h at 37∘C Briefly 20 mL of a 5 mgL methanol solution of 3-[45-dimethylthiazol-2-yl]-25-diphenyltetrazolium bromide(MTT) (Sigma-Aldrich Ltd South Africa) was added to eachwell and then incubated for 30 min Blue colouration indi-cated bacterial growth The MIC was recorded as the lowestconcentration at which no colour change was observed

213 Statistical Analysis Data analysis was done inMicrosoftExcel to obtain descriptive statistics Means values wereseparated by the Duncan multiple tests using SAS The dif-ferent levels of significance within the groups were analyzedusing one-way analysis of variance (ANOVA) Values wereconsidered significant at P lt005

3 Results and Discussion

31 Ethnobotanical Survey Modern drugs are expensive andsupply to a remote area may be irregular hence result-ing in overreliance of traditional methods of healing as aprimary source of healthcare [19] The rural communitiesof Ebem-Ohafia region of southeastern Nigeria depend onherbal remedies to meet their domestic and health needsDuring our ethnobotanical survey a total of 30 respondentsincluding the elders herbalists and traditional healers wereinterviewed in the study areaMajority of the people (22)wereaged 30ndash65 years dominated by males (25) and one-third(10) had a primary school education while 15 had secondaryeducation and others informal education A total of 61medic-inal plant species belonging to 26 families were identified inthe treatment of oral and skin diseases as shown in Table 1Twenty-seven of them are used in the management of oralinfections and twenty-six plants for the management of skininfection while eight plants are used in the management ofboth infections Table 2 shows the list of family number of theplant species used and their percentage occurrenceThemostrepresented plant family is the Euphorbiaceae having 1803of the plant species followed by Asteraceae and Fabaceae with1147 each [20] Euphorbiaceae is a large family of floweringplants with 300 genera and about 7500 species found usefulin the curing of some common human diseases Asteraceae isa large family of flowering plants having 32 913 species and1911 genera and has been found to possess diverse biologicaleffects Fabaceae is a family of cosmopolitan distributionwith approximately 730 genera and 19400 species richnessat a global level [20] The leaves (60) recorded the mostcommonly used plant part followed by the root (16) andthen stem bark (12) None of the respondents indicated anypossible side effects caused by any of these plants listed inTable 1 The most common methods of preparation includeinfusion decoction and maceration These herbalists ortraditional healers often collect the plants from the fieldand dry and crush them into powder form before storingin bottles to prevent patients from identifying the plantsused for their treatment Most healers refused to provideinformation without payment whereas others thought toprovide information on the plant use is disrespect of theirtraditional belief Furthermore the majority of the plantsmentioned lack scientific data to substantiate their acclaimedfolkloric use From this work it is evident that medicinal

10 The Scientific World Journal

Table 2 The list of family number of the plant species and theirpercentage occurrence

Family Number of plant species OccurrenceFabaceae 7 1147Euphorbiaceae 11 1803Asteraceae 7 1147Myrtaceae 4 656Moraceae 3 492Malvaceae 4 656Liliaceae 2 328Capparaceae 2 328Annonaceae 3 492Combretaceae 1 164Burseraceae 2 328Apocynaceae 1 164Gnetaceae 1 164Lecythidaceae 1 164Verbenaceae 1 164Ochnaceae 1 164Rubiaceae 1 164Piperaceae 1 164Acanthaceae 2 328Papilionoideae 2 328Asclepiadaceae 1 164Caesalpiniaceae 2 328Convolvulaceae 1 164Rutaceae 1 164Crassulaceae 1 164Poaceae 1 164

plants play a significant role in the primary healthcare ofthe people and conservation of the indigenous knowledge ofthis folklore medicine is crucial for the innovations of futuredrug discovery and possible herbal plants trade in the region[19] On the contrary we observed imprecise dosage lowhygiene standards hiding of healing methods and lack ofpatients record as a significant setback to traditional healthpractitioners in the region

32 Thin Layer Chromatography (TLC) Profiling Thin layerchromatography remains the most common useful techniqueby researchers as a quick and simple method to resolvethe constituents of a crude extract followed by advancedextraction methods to purify the active ingredients Previousstudies reported that specific secondary metabolites in plantexhibit a wide range of antioxidant and antibacterial proper-ties [21] TLC was formed on a glass sheet coated with silicagel After the sample is applied to the plate a solvent system(mobile phase) is drawn up via capillary action on the TLCplate at different mobility rate depending on the chemicalcompositionsThe separated spotswith different colourswerevisualized by the UV light projection onto the glass sheetThe variation of coloured spots detected in the crude extractsindicates the presence of different phytochemicals that may

be responsible for the pharmacological effects of selectedplants use in the management of oral and skin infections asacclaimed by the traditional healers We quantified the resultsby measuring the distance travelled by the plant compoundsdivided by the total distance travelled by the mobile phaseFrom Table 3 it is apparent that mobile phase 1 consistingof chloroform and ethanol in the volume ratio of 64 had abetter separation than mobile phase II containing tolueneethyl acetate and acetic acid in the volume ratio of 541 Themethanol extract of AH in bothmobile phases had the highestnumber of three bands with Rf values of 0667 0833 and0989 for mobile phase I and Rf values of 0381 0648 and0781 for mobile phase IIThe plant extracts MM BA CO JCandCAhad two bands inmobile phase I with clear separationbut had single band in mobile phase II The plant extractshaving only one band in either mobile phase suggest a poorsolvent system for compounds separation The TLC profilesprovide a characteristic fingerprint of these plants that mayalso be useful for their identification

33 Antioxidant Assays

331 TPC and TFC Phenolic compounds are secondarymetabolites distributed in fruits vegetables nuts herbsseeds and beverages [22] They are excellent scavengers offree radical that has been implicated in the pathologies ofvarious human diseases the antioxidant activity is dependenton the structure hydrophobic benzenoid rings and hydrogen-bonding potential of the phenolic hydroxyl groups In TPCassay a phenol in plant extracts loses a proton to formphenolate ion which reduces Folin Ciocalteu reagent andthe resulting changes are measured spectrophotometricallyat 765 nm [13] The content of phenol is reported in mg oftannic acid equivalent (TAE)mL of extract As shown inFigure 1 the total phenolic contents of absolute methanolcrude extract of these botanicals range from 700 to 3600mgTAEg The AH extract had the highest concentrationfollowed by CA CO MM BNI BA EI BN and JC whilethe least concentration was recorded in II extract Moreoverthe Folin Ciocalteu reagent (FCR) has been used to measureother compounds such as ascorbic acid amino acids andsugars hence it may not provide a concise amount of thephenolic compounds present in these plants Flavonoids areincreasingly consumed in a significant amount in daily dietsworldwide due to their various health benefits toman [23 24]In TFC assay aluminum ion in the reacting mixture formscomplexes with the C- 4 keto and either C- 3 or C- 5 hydroxylor with ortho hydroxyl groups in the A or B ring of flavonoidsdetected in the plant extracts [14] As shown in Figure 2the total flavonoid content was recorded in the plant extractsranging from 750 to 1500 mgQEg as extrapolated from thestandard quercetin curve The CA extract had the highestconcentration followed by BNI CO II AH BN MM JCand BA while the lowest concentration was recorded in EIextract TPC is found higher than the TFC in some of theextracts supporting the fact that most flavonoids are alsophenolics According to our results all the plant extractsare rich source of phenolic compounds The composition ofTPC and TFC registered in these plants may be difficult to

The Scientific World Journal 11

Table 3 Thin layer chromatography retention factor using mobile phase CHL ET (mobile phase I) and Tol EA AA (mobile phase II)

SN Plant Extract CHL ET (64) Tol EA AA (541)1 Baphia nitida 1 0742 Macrolobium macrophyllum 0706 0941 06953 Chromolaena odorata 0471 0647 06674 Jatropha curcas 0353 0941 06485 Cassia alata 0118 0882 06866 Ipomoea involucrata 0956 -7 Acalypha hispida 0667 0833 0989 0381 0648 07818 Breynia nivosa 0989 07629 Burkea africana 0644 1 075210 Eleusine indica 0978 0743CHL chloroform ET ethanol Tol toluene EA ethylacetate AA acetic acid

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

500

1000

1500

2000

2500

3000

3500

4000

TPC

(mgT

AE

g)

Figure 1 The total phenolic content (TPC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata TAE tannic acid equivalent

compare with others reported elsewhere due to many factorsincluding differences from species to species maturity stageplant parts and assay methods harvesting time standarddrugs and geographical location

332 Reducing Power (RP) Ferric reducing power is a globalmethod often used to assess the capacity of antioxidantsto donate either an electron or hydrogen atom to unpairedelectron [25] In reducing power assay plant extracts exertantioxidant activities via reduction of Fe3+ferric cyanidecomplex to Fe2+ (ferrous form) resulting in changing thegreen colour to blue and sometimes Perlrsquos Prussian bluecolour depending on the antioxidant potential [10] As shownin Figure 3 the reducing power of the methanol crudeextracts and the standard gallic acid rutin and ascorbic acidincreased with increase in concentration Higher absorbanceat 700 nm can be read after potassium ferricyanide reactionwith ferric chloride At 1000 120583gmL the extracts from AH

0

200

400

600

800

1000

1200

1400

1600

TFC

(mgQ

Eg)

AH BN JC CO MM EI BNI BA CAIIPlant extracts

Figure 2 The total flavonoid content (TFC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata QE Quercetin equivalent

BNI MM CA BA EI and BN had absorbance value of 270086 073 055 036 027 and 027 respectively while that ofAH is higher than the standard rutin (254) but comparablewell with gallic acid (275) and ascorbic acid (269) EI andBN extracts had a similar trend of ferric reducing powerthough significantly lower as compared with the standarddrugs AH extract exhibits remarkable antioxidant capacity(RP) correlating with its richness in phenolic and flavonoidcontents It is reasonable to suggest that methanol selectivelyextracts antioxidant principles from these botanicals basedon their polarities and chemical structures which are reflectedin plant extracts with high TFC and TPC having minimal RP[26] We have shown that some plant extracts in particularAH BNI MM and CA extracts may serve as an excellentcandidate to minimize skin and oral infections caused byoxidative stress as a result of imbalance of free radicalsgeneration and natural antioxidants in the saliva or woundarea According to our results the increasing RP of the plantextracts correlateswith their TPCandTFCThis study at least

12 The Scientific World JournalRe

duci

ng P

ower

7

00nm

625 125 250 500 10003125Concentration (gmL)

0

05

1

15

2

25

3

IIAHBNJCCOMMEI

BNIBACAGARutinVit C

Figure 3 The ferric reducing antioxidative capacity of selectedplant extracts used in the management of oral and skin infectionsin southeast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata GA gallic acid Vit C vitamin C (standard antioxidants)

in part lends credence to the ethno-therapeutic use of someplant as natural antioxidant in themanagement of skin or oralinfections in the study area

333 FRSA of Plant Extracts Skin and oral cavity exposedto an array of oxidants such as smokes from cigaretteschemicals UV light nicotine and alcohol that are directlyor indirectly leading to the production of reactive oxygenspecies (ROS) are known to be responsible for some com-mon human diseases [27] These free radicals in particularhydroxyl and superoxide anions are highly reactivemoleculesderived from the metabolism of oxygen Although they playan important role in the biochemical immunological andphysiological processes it could be deleterious to vital organsif excessively accumulated in the human body [28] Food andpharmaceutical industries discovered that medicinal plantscontaining phenol and flavonoid compounds are essentialin the development of novel drugs against stress-induceddiseases [28] In the present investigation the concentration(IC50) at which 50 of free radical scavenged was calculatedand extrapolated from the plot of percentage inhibition ver-sus plant extract concentration The herbal drugs exhibitedrobust DPPH radical scavenging activity in a concentration-dependent manner having IC50 ranging from 2025 to 710120583gmL As shown in Figure 4 MM BA EI JC AH BNIand CA extracts had a potent DPPH radical scavengingactivity corresponding to their TPC TFC and RP Theplant extracts scavenged ABTS radical by the decrease inits absorbance at 734 nm All the plant extracts exhibitedstrong ABTS radical scavenging capacities in the following

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

200

400

600

800

1000

1200

1400

1600

IC50

(g

mL)

Figure 4 DPPH free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

050

100150200250300350400450500

IC50

(g

mL)

AH BN JC CO MM EI BNI BA CAIIPlant samples

Figure 5 ABTS free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

order BN gt JC gt CA gt MM gt BNI gt EI gt CO gt AH gtII gt BA having IC50 values ranging from 25 to 400 120583gmL(Figure 5) DPPH and ABTS radical scavenging activities ofthese herbal remedies displayed a positive correlation withthe working concentrations It is obvious that most of theplant extracts tested in this work possess the ability to quenchthe free radicals generation via electron or proton donationdemonstrated by the termination of radical chain reaction[28] Although both DPPH and ABTS radicals are electron-transfer based methods however it is apparent that ABTSmethod seems to be more sensitive than DPPH radical assaypossibly due to their solubility and diffusivity as shown inthe IC50 values [10] It is worth noting that BA extract withstrong DPPH radical scavenging effect had a moderate ABTS

The Scientific World Journal 13

Table 4 Zone of bacterial inhibition by the selected plant extracts use in the management of oral and skin infections

Diameter of inhibition (mm)SN Plant samples K pneumoniae P mirabilis S aureus E coli1 Cassia alata 15 plusmn 141 9 plusmn 042 15 plusmn 083 13 plusmn 0802 Breynia nivosa 8 plusmn 052 17 plusmn 100 8 plusmn 008 15 plusmn 1023 Burkea africana NI 11 plusmn 084 13 plusmn 072 NI4 Baphia nitida 8 plusmn 050 13 plusmn 122 8 plusmn 051 NI5 Ipomoea involucrata 8 plusmn 052 NI 8 plusmn 050 13 plusmn 0526 Jatropha curcas NI 9 plusmn 030 9 plusmn 062 13 plusmn 0727 Acalypha hispida 21 plusmn 163 NI 11 plusmn 009 17 plusmn 1328 Chromolaena odorata 8 plusmn 042 8 plusmn 035 9 plusmn 009 15 plusmn1209 Eleusine indica 8 plusmn 042 8 plusmn 035 NI 17 plusmn 10010 Macrolobium macrophyllum NI NI 8 plusmn 004 15 plusmn 12311 Ciprofloxacin - - le16(R) ge17(S) le16(R)ge20(S)NI indicates no zone of inhibition S sensitive R resistance

radical scavenging capacity which can be explained by thepartial ionization of the extract in the test medium [29] SinceROS promotes tissue damage and epithelial inflammationvia exposure to sunlight radiation or smoke as a result theantioxidant capacity demonstrated in this work may supportthe traditional use of these plants against the influence ofinflammatory markers and oxidants in the skin and oralcavity [29] Based on the results of the three antioxidantassays it is obvious that MM AH BNI EI and CA extractshad the best FRSA indicating their importance in convertingfree radicals into more stable products and then terminatethe free radical chain reaction Our study showed that themethanol crude extracts from these plants could protectagainst free radical damage in the mouth such as humangingival periodontal tissues and certain skin infections [5]

334 Antibacterial Activity of Plant Extracts Pathogenicbacteria such as Staphylococcus aureus Escherichia coli Kleb-siella pneumoniae and Proteus mirabilis play a significantrole as the primary cause of skin and oral infections [30]Staphylococcus aureus is a part of normal skin known to bethe most problematic in the cause of skin infections as aresult of their resistance mechanisms to first-line antibiotics[30] Although the presence of this organism as a residentoral floral is controversial nonetheless MSSA and MRSAhave been isolated from oral rinse and tongue swab inpatients with oral infections [31] Escherichia coli are Gram-negative bacteria known to be notorious for their resistantmechanisms towards the existing arsenal of antibiotics Oralinfections such as edematous and erythematous and skininfections like omphalitis surgical site infections burnnecrotizing fasciitis and cellulitis at all age groups are associ-ated with enterobacterium E coli [32] Klebsiella pneumoniaeis a Gram-negative encapsulated facultative anaerobic androd-shaped bacterium found in the normal flora of themouth skin and intestines This bacterium is responsiblefor the prolonged surgical wound biliary tract wound andurinary tract infections and it could pose a serious healthrisk when resistant strains invade the throat mouth or

intestines [33] Villegas et al [34] detected a CTX-M-12 120573-lactamase producing K pneumoniae new plasmid-mediatedESBLs highly resistant to ceftazidime from a patient withhospital-acquired wound infection Proteus mirabilis is oneof the most common enterobacteria after E coli identified asGram-negativemotile and rod-shaped bacterium It is foundin clinical specimen implicated in the skin respiratory tractand bacteraemia as well as burns wounds and urinary tractinfections [35] All these pathogenic multidrug-resistancebacteria are causative agent in a periodontal lips ulcer whitesponge nevus and skin infections [36]

335 Susceptibility Test and MIC Determination The resultof disc diffusion assay showed the diameter of inhibitionof bacteria growth classified as le 15 (resistant) and ge 16(susceptible) according to CLSI guideline [37] As shown inTable 4 K pneumoniae and E coli are more susceptible tothe treatment of Acalypha hispida (AH) having a zone ofinhibition (ZI) 21 and 17mm respectively BN and EI extractsinhibit the growth of P mirabilis and E coli having the samezone of inhibition 17 mm corresponding to the report of Al-Zubairi et al [38] The data obtained for the methanol crudeextracts from AH BN and EI show their selective inhibitionof the test bacteria growth The plant extracts with ZI above13 mm against our panel of bacterial strains were selected forMICdeterminationHere themethanol crude extracts exhibitMIC values of 256 to gt 512 120583gmL against the test bacterialstrains but were less active as compared to the standardciprofloxacin 003 - 006 120583gmL (Table 5) Ciprofloxacin isan active broad-spectrum antibiotic use in the treatmentof urinary tract infections prostatitis and acute cystitis viainterfering with bacterial DNA replication and transcriptionthrough inhibition of DNA gyrase or topoisomerase [39] Allthe plant extracts demonstrated a mild to weak antibacterialactivity based on the classification of Rios and Recio [40] whoconsidered antibacterial activity of the crude plant extractsignificant (MIC lt 100 120583gmL) moderate (100 lt MIC = 512120583gmL) or weak (MIC gt 512 120583gmL) The herbal remediesshowed moderate inhibition of bacterial growth except for K

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

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Submit your manuscripts atwwwhindawicom

Page 4: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

4 The Scientific World Journal

Table1Medicinalplantsused

inthem

anagem

ento

fskinor

oralinfections

insoutheastern

partof

Nigeria

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Macrolobium

macrophyllum

Fabaceae

Antho

notha

macroph

ylla

Leaf

Skin

Treatin

gof

woun

dandcut

Squeezethe

leavea

ndplaceo

nthe

affectedarea

ofthes

kin

Jatro

phacurcas

Euph

orbiaceae

Barbados

nut

Leaf

Skinoral

Treatm

ento

fboils

Boiltheleave

with

waterPlace

theh

otleaveo

nthea

ffected

area

ofthes

kin

Eleusin

eind

icaPo

aceae

Yard

grass

Root

Skin

Treatm

ento

fRingw

orm

Use

ther

ootand

scrubon

affectedarea

ofthes

kin

Cassiaalata

Caesalpiniaceae

Ring

worm

bush

Leaf

Skin

Treatm

ento

fRingw

orm

Scrubtheleave

onaffectedarea

ofthe

skin

Chromolaena

odorata

Aste

raceae

Siam

weed

Leaf

Skin

Tosto

pbleeding

andrashes

Squeezethe

leavea

ndrubon

affectedarea

Psidium

guajava(L)

Myrtaceae

Guava

Leaf

Skin

Treatm

ento

fskinbu

rns

scrapessunb

urn

woun

ds

psoriasiseczem

aand

others

Crushedgu

avaleafo

rboiledleaves

are

applieddirectlyto

thea

ffected

skin

providingreliefandareb

elievedto

improveh

ealin

gBryophyllum

pinn

atum

(Lam

)Oken

Crassulaceae

Mira

cleleaf

Dogrsquosliver

Leaf

Skino

ral

Treatm

ento

fboilwo

und

sore

orcut

Crushleafandapplyjuiceo

nthew

ound

surfa

ce

Ficusa

sperifolia

Miq

Moraceae

False

thistle

Leaflatex

Skin

Treatm

ento

fskinrashes

anditches

Appliedas

poultic

etothes

kinitches

Acanthus

montany

(TA

nderson)

Acanthaceae

Bearrsquosbreeches

Stem

bark

Skin

Treatm

ento

fwou

ndsa

ndskin

diseases

Woo

dbark

iscrushedandsoaked

italcoho

landextractapp

liedto

woun

dsandskin

Alliu

msativ

umL

Liliaceae

Garlic

Bulbjuice

Skin

oral

Usedto

cond

ition

thes

kin

chew

edforc

ough

andsore

throat

Thejuice

ismixed

with

lotio

nsforskin

cond

ition

ingFreshbu

lbischew

edor

boiledin

water

anddrinkdecoctionfor

coug

h

Emiliasonchifolia

(L)

Aste

raceae

Scarlettassel

Leaf

Skin

Treatm

ento

fSkintro

ubles

abscessa

ndbruises

Crushedleaves

were

pressedandused

tocoverthe

woun

dor

bruisedsurfa

ces

Sida

acutaBu

rmf

Malvaceae

Broo

mwe

edLeaf

Skin

Treatm

ento

fjaund

icesore

ulcer

Poultic

esmadefrom

boiledleaves

are

appliedon

thea

ffected

parts

The Scientific World Journal 5

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Ricin

uscommun

isLinn

Eu

phorbiaceae

Castorb

ean

Leafroo

tand

seed

Skin

Treatm

ento

fskininfection

anddetoxification

Applycasto

roilon

thea

ffected

area

torelieve

skin

infection

Citru

slim

on(L)Bu

rmf

(prosp

)Ru

taceae

Limon

Leaffruit

andseed

Skin

Treatm

ento

ffluheadache

cold

andoralinfections

Mixequalquantities

ofLimon

juicea

ndho

lybasilkeepitun

derthe

suntill

becomes

thickapplythison

face

Alliu

mcepa

LLiliaceae

Onion

sLeafandbu

lbSkin

Treatm

ento

fperiodo

ntitis

andcold

Theb

ulbismacerated

andmixed

with

water

andthen

takenorally

Breynianivosa

Euph

orbiaceae

Icep

lant

Leaf

Oral

Treatin

gof

toothache

Boiltheleave

with

waterthentake

the

water

Ipom

oeainvolucrata

Con

volvulaceae

Water

convolvu

lus

Leafandste

mOral

Treatm

ento

foralinfectio

nMixtheleave

with

ugbaga

treea

ndbo

ilwith

water

anddrinkthew

ater

Burkea

afric

ana

Caesalpinioideae

Icheku

Leaf

Oral

Treatm

ento

forald

isease

Boiltheleave

with

water

andtakebut

donrsquot

swallowit

Baphianitid

aPapilio

naceae

Cam

wood

Leafor

stem

Oral

Streng

thensthe

teeth

Chew

thes

tem

orscrubtheleave

onthe

teeth

Gongronem

alatifolium

Asclepiadaceae

Utazi

Leaf

Oral

Treatm

ento

fcou

ghand

diabetes

Crushtheleafw

ithmortara

ndmixwith

ginger

orgarlica

nddrink

Anonasenegalen

sisAnn

onaceae

Ubu

ruocha

Root

Oral

Treatm

ento

fmalariaand

mou

thinfection

Boilther

ootsanddrink

Abrusp

recatoriu

sPapilio

noideae

Crabrsquoseyes

Seed

androot

Oralskin

Treatm

ento

fcou

ghskin

carew

ound

Grin

dther

ootand

applythep

oultice

onthea

ffected

skin

areaA

lsocrushedseeds

canbe

takenorally

Acanthus

montanu

sAc

anthaceae

False

thistle

Leafandroot

Oraland

Skin

Treatm

ento

ffurun

cles

lesio

nsandurinary

infections

Boilther

ootand

applythep

oultice

tothe

affectedpartof

theb

odyFo

rinfectio

ns

boilther

ootand

leaves

anddrink

Piperu

mbellatum

Piperaceae

Njamuja

Leaf

Oral

Treatm

ento

fpeptic

ulcers

Crushtheleavesa

ndsoak

inwarm

orho

twater

anddrink

Euphorbiahirta

Euph

orbiaceae

Asth

maw

eed

Who

leplant

Oral

Treatm

ento

fwou

nds

Leaves

arem

acerated

andappliedto

affectedpart

Ficusb

urkei(Miq)Miq

Moraceae

Sand

paper

Leaf

Oral

Treatm

ento

fhypertension

Leaves

areb

oiledandtakenorally

6 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Gmelina

arborea

Verbenaceae

Gmelina

Stem

andleaf

Oral

Treatm

ento

fgon

orrhea

pilesabdo

minalpains

burningsensationsfever

mou

thwo

undanddiabetes

Snake-biteandscorpion

sting

Leaves

grou

ndinto

paste

with

water

are

appliedto

theforeheadforh

eadachein

feversB

oiledste

msa

retakenorally

Lophira

alata

Ochnaceae

Ekki

Stem

andleaf

Oral

Treatm

ento

ffeverm

outh

woun

dandmalaria

Boiltheleavesa

ndste

mandtake

orally

Nau

clealatifolia

Rubiaceae

Ubu

ru-in

uStem

andleaf

Oral

Itisused

inthetreatment

ofmalariaepilepsymou

thwo

undanxietypainfever

Boilther

ootsanddrink

Annona

murica

taAnn

onaceae

Sour

sop

Leaf

Oral

Treatm

ento

fcancera

ndmou

thwo

undinfections

Crushor

boiltheleavesa

ndtake

orally

Psidium

guajava

Myrtaceae

Guava

Leaf

Oral

Treatm

ento

fperiodo

ntal

diseasebu

rnsscrapes

sunb

urnwo

undspsoria

sis

eczemaandothers

Leaves

arec

rushed

andtakenorally

Astra

galusm

embran

aceus

Fabaceae

Beiq

iStem

andleaf

Oral

Treatm

ento

fcoldup

per

respira

tory

infections

allergiesfib

romyalgia

anem

iaH

IVA

IDS

Leaves

arec

rushed

andtakenorallyand

canalso

beappliedto

skin

Buchholziacoria

cea

Capparaceae

Won

derfu

lkola

Leaf

Oral

Itisused

tradition

allyfor

treatingdiabetes

hypertensio

nrheumatism

coldcou

ghand

catarrh

Boilleaves

andtake

orally

Gnetum

afric

anum

Gnetacea

Afang

Leaf

Oral

Enlarged

spleen

sore-th

roatandcathartic

Leaves

canbe

boiledor

cooked

andtaken

orally

Anoridium

man

niAnn

onaceae

Ewuro-ig

boStem

bark

Oral

Treatm

ento

fDiarrhea

coug

hfeverrheumatism

Theb

arkiscrushedandbo

iledandthen

takenorally

Funtum

iaafric

ana

Apocyn

aceae

Akorie

mbamiri

Leafstem

root

Skin

treatmento

fcon

stipatio

nwo

undsw

eakbladder

jaun

dice

Who

leplantisb

oiledandtakenorally

Also

thec

rushed

leaves

area

ppliedto

the

affectedskin

area

The Scientific World Journal 7

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Commiphora

afric

ana

Burseraceae

Turari

dashi

Rootsfruits

Oral

Treatm

ento

fwho

oping

coug

hbron

chitis

Rootsa

remacerated

andtakenorally

Dacryodesedulis

Burseraceae

Elem

iub

eBa

rkroo

tsfruits

Skin

Treatm

ento

fJiggersSkin

diseasesE

leph

antia

sis

Theleavesa

reeatenrawwith

kolanu

tas

anantie

meticLeaf-s

apisinstilledinto

thee

arfore

ar-tr

oubleanda

leaf-decoctio

nisprepared

asa

vapo

ur-bathforfeverish

stiffn

essw

ithheadache

Combretum

mucronatum

Com

bretaceae

Fararg

eza

Rootsa

ndleaf

Skin

Treatm

ento

fskininfection

Ther

ootand

leafarem

acerated

and

appliedto

affectedskin

area

Cleomec

iliata

Capparaceae

Ekuya

Leafseeds

Skin

Treatm

ento

fCon

vulsion

wo

undssores

Seedsa

recrushedandthep

oultice

appliedto

thea

ffected

areaTh

emacerated

leaves

after

boiledaretaken

orally

Ageratum

conyzoides

Aste

raceae

Urata

Who

leplant

Skin

Treatm

ento

fwou

ndU

lcer

Sleeping

sickn

essEyew

ash

Thep

lant

ismacerated

andappliedto

affectedskin

area

Aspilia

afric

ana

Aste

raceae

Yunyun

Ka

lank

uwa

Leafflow

erSkin

Treatm

ento

fskinrashes

cleaning

sorescorneal

opacities

Thefl

ower

issqueezed

andappliedto

affectedskin

area

Centau

reaperrottetii

Aste

raceae

Danyi

Who

leplant

Skin

Treatm

ento

fskin

infectionsSyphilis

Thefl

ower

issqueezed

andap

oultice

appliedto

affectedskin

area

Chrysanthellu

mindicum

Aste

raceae

Abilereoyigi

Who

leplant

Skin

Treatm

ento

fboils

gono

rrheaJaun

dice

Thep

lant

iscrushedandthep

oultice

applieddirectlyto

thes

kinarea

Elephantopus

scaber

Aste

raceae

Elephant

foot

Leafroo

tOral

Treatm

ento

ffevercou

ghLeafandroot

areb

oiledtogether

and

takenorally

Emiliacoccinea

Aste

raceae

Odu

ndun

Leafsap

and

root

Skin

Treatm

ento

fulcerhernia

measle

sWho

leplantissqu

eezedandsaptaken

orally

Acalypha

godseffi

ana

Euph

orbiaceae

Jinwinini

Leaf

Skin

Treatm

ento

fskin

infectionsantim

icrobials

Leafismacerated

andtakenorallyas

well

asappliedto

affectedskin

area

8 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Brideliafer

rugin

eaEu

phorbiaceae

Irikirni

Leafstem

bark

root

Oral

Treatm

ento

fInsom

nia

mou

thwashgono

rrhea

Who

leplantisb

oiledandtakenorally

Euphorbiaheterophylla

Euph

orbiaceae

Egele

Leafroo

tsSkin

Treatm

ento

fSkindiseases

Leafandroot

arem

acerated

andthe

poultic

eapp

liedto

thea

ffected

skin

area

Euphorbiahirta

Euph

orbiaceae

Non

onku

rciya

odne

Who

leplant

exud

ate

Oral

Treatm

ento

fAsth

ma

coug

hshapeo

fbreasts

Thew

holeplantism

acerated

andtaken

orally

Euphorbialaterifl

ora

Euph

orbiaceae

Orow

ere

Leafexud

ate

Skin

Treatm

ento

fdermatitis

constip

ation

Theleafissqu

eezedandthep

oultice

appliedto

thea

ffected

skin

area

Cajanu

scajan

Fabaceae

Orela

Leafseed

Skin

andoral

Treatm

ento

fSmallpox

and

mou

thwash

Leafissqueezed

andappliedto

the

affectedarea

Dan

iellia

thurifera

Fabaceae

Iya

Stew

-woo

ddu

stSkin

Treatm

ento

fscabies

Theb

arkismacerated

andappliedto

the

affectedskin

area

Dialiu

mguineense

Fabaceae

Tasm

iyar

kumi

Leafbark

fruitand

twig

Oral

Treatm

ento

fBronchitis

coug

handdiuretic

Theleafand

bark

arec

rushed

andbo

iled

togetherTh

emixture

isthen

takenorally

Acacianilotica

Fabaceae

Baanigabaruwa

Fruitbark

exud

ate

Skin

Treatm

ento

fskindiseases

fung

alinfections

Thee

xudatesa

resqueezed

from

theb

ark

andapplieddirectlyto

thea

ffected

skin

area

Napoleona

imperia

lisLecythidaceae

Mabun

giTw

igs

Oral

Treatm

ento

fAsth

ma

coug

hTw

igsa

remacerated

andtakenorally

Alliu

msativ

umLiliaceae

Ayoayuu

Bulb

Oral

Treatm

ento

ffevercou

gh

asthmaantim

icrobial

Theb

ulbismacerated

andtakenorally

Hibiscus

asper

Malvaceae

Isapad

angiraa

Leaf

Oraland

skin

Treatm

ento

fcou

gh

woun

dsanddiuretic

Leafisbo

iledandtakenorally

Hibiscus

rosasin

ensis

Malvaceae

Kekekeireagu

Leafstem

flowe

rSkin

Treatm

ento

finfl

uenza

woun

dsand

diuretic

Who

leplantism

acerated

andtaken

orally

Theleafissqu

eezedandappliedin

affectedskin

area

Hibiscus

sabdariffa

Malvaceae

Sobo

gurgu

zuLeafflow

erOral

Treatm

ento

fDiuretic

scoug

handdressin

gwo

unds

Leafisbo

iledandtakenorally

Ficuseleg

ans

Moraceae

Asoro

Leaf

Skinoral

Treatm

ento

fPile

constip

ationandcraw

-craw

Leafissqueezed

andappliedto

the

affectedarea

The Scientific World Journal 9

the absorbance The percentage inhibition of DPPH orABTS radical scavenging activity by the plant extracts wascalculated as (Abscontrol ndash Abssample)(Abscontrol) times 100where Abscontrol is the absorbance of DPPH or ABTS +methanol Abssample is the absorbance of DPPH or ABTSradical + sample extractstandard Here the concentration ofthe extracts needed to decrease the absorbance of DPPH orABTS radical by 50 was calculated Rutin (Sigma-Aldrichge 94 HPLC grade) at the same working concentrations ofthe plant extracts was used as reference drug

210 Bacterial Strains All bacterial strains were culturedon nutrient agar slopes and were incubated for 24 h at37∘C before the MIC An inoculum turbidity equivalent toa 05 McFarland standard (1times107 CFUmL) was prepared innormal saline for each test organism and then diluted 1100in Mueller-Hinton broth just before injection of the platesStandard strains used in the study included S aureus and Ecoli which were a gift from Dr EO Ekundayo (Departmentof Microbiology Michael Okpara University of Agriculture)and clinically isolated strains of K pneumoniae and Pmirabilis were gotten from Department of MicrobiologyFederal Medical Centre (FMC) Umuahia Abia State

211 Antibacterial Susceptibility Using Disc Diffusion MethodThe microbial growth inhibition potential of the plantextracts was determined by using the agar disc diffusionmethod [17] The plant extracts were dissolved in 10 vvDMSO (Merck UK Ltd) to a final concentration of 2 mgmLThe nutrient agar plate was streaked with the standardizedtest bacterial inoculum over the surface evenly and 2mgdiscof the plant extracts transferred onto sterile filter disc papers(about 6 mm) containing the test compounds was placed onthe agar surface in the Petri dishes 100 120583gdisc of standardantibiotic ciprofloxacin was used as control and 100 120583LDMSOdisc served as negative control The plates were thenincubated at 37∘C for 24 h Generally the antimicrobialextracts diffuse into the agar and inhibit the growth of thetest bacteria All tests were done in duplicate and diame-ter zones of inhibition of growth were measured using ameter rule from the edge of each dish after the incubationperiod

212 Minimum Inhibitory Concentration (MIC) Assays Wefollowed the method of Smith et al [18] to determine theMIC ofmethanol crude extracts of selected botanicals againstclinical isolates of bacteria associated with skin and oralinfection A volumeof 100mLof sterileMueller-Hinton broth(Oxoid Basingstoke UK) containing 20 mgL of Ca2+ and10 mgL of Mg2+ was dispensed into a 96-well microtitreplate (Nunc 03 mL total volume per well) All antibacterialagents were dissolved in dimethyl sulphoxide (DMSO) anddiluted in Mueller-Hinton broth to give a stock solutionThen 100 120583L of the antibacterial agent stock solution (2000mgL) was serially diluted into each well and 100 mL ofthe bacterial inoculum added to each well to give a finalconcentration range of 0024 - 1 mgL All procedures wereperformed in duplicate and the plates were incubated for 18

h at 37∘C Briefly 20 mL of a 5 mgL methanol solution of 3-[45-dimethylthiazol-2-yl]-25-diphenyltetrazolium bromide(MTT) (Sigma-Aldrich Ltd South Africa) was added to eachwell and then incubated for 30 min Blue colouration indi-cated bacterial growth The MIC was recorded as the lowestconcentration at which no colour change was observed

213 Statistical Analysis Data analysis was done inMicrosoftExcel to obtain descriptive statistics Means values wereseparated by the Duncan multiple tests using SAS The dif-ferent levels of significance within the groups were analyzedusing one-way analysis of variance (ANOVA) Values wereconsidered significant at P lt005

3 Results and Discussion

31 Ethnobotanical Survey Modern drugs are expensive andsupply to a remote area may be irregular hence result-ing in overreliance of traditional methods of healing as aprimary source of healthcare [19] The rural communitiesof Ebem-Ohafia region of southeastern Nigeria depend onherbal remedies to meet their domestic and health needsDuring our ethnobotanical survey a total of 30 respondentsincluding the elders herbalists and traditional healers wereinterviewed in the study areaMajority of the people (22)wereaged 30ndash65 years dominated by males (25) and one-third(10) had a primary school education while 15 had secondaryeducation and others informal education A total of 61medic-inal plant species belonging to 26 families were identified inthe treatment of oral and skin diseases as shown in Table 1Twenty-seven of them are used in the management of oralinfections and twenty-six plants for the management of skininfection while eight plants are used in the management ofboth infections Table 2 shows the list of family number of theplant species used and their percentage occurrenceThemostrepresented plant family is the Euphorbiaceae having 1803of the plant species followed by Asteraceae and Fabaceae with1147 each [20] Euphorbiaceae is a large family of floweringplants with 300 genera and about 7500 species found usefulin the curing of some common human diseases Asteraceae isa large family of flowering plants having 32 913 species and1911 genera and has been found to possess diverse biologicaleffects Fabaceae is a family of cosmopolitan distributionwith approximately 730 genera and 19400 species richnessat a global level [20] The leaves (60) recorded the mostcommonly used plant part followed by the root (16) andthen stem bark (12) None of the respondents indicated anypossible side effects caused by any of these plants listed inTable 1 The most common methods of preparation includeinfusion decoction and maceration These herbalists ortraditional healers often collect the plants from the fieldand dry and crush them into powder form before storingin bottles to prevent patients from identifying the plantsused for their treatment Most healers refused to provideinformation without payment whereas others thought toprovide information on the plant use is disrespect of theirtraditional belief Furthermore the majority of the plantsmentioned lack scientific data to substantiate their acclaimedfolkloric use From this work it is evident that medicinal

10 The Scientific World Journal

Table 2 The list of family number of the plant species and theirpercentage occurrence

Family Number of plant species OccurrenceFabaceae 7 1147Euphorbiaceae 11 1803Asteraceae 7 1147Myrtaceae 4 656Moraceae 3 492Malvaceae 4 656Liliaceae 2 328Capparaceae 2 328Annonaceae 3 492Combretaceae 1 164Burseraceae 2 328Apocynaceae 1 164Gnetaceae 1 164Lecythidaceae 1 164Verbenaceae 1 164Ochnaceae 1 164Rubiaceae 1 164Piperaceae 1 164Acanthaceae 2 328Papilionoideae 2 328Asclepiadaceae 1 164Caesalpiniaceae 2 328Convolvulaceae 1 164Rutaceae 1 164Crassulaceae 1 164Poaceae 1 164

plants play a significant role in the primary healthcare ofthe people and conservation of the indigenous knowledge ofthis folklore medicine is crucial for the innovations of futuredrug discovery and possible herbal plants trade in the region[19] On the contrary we observed imprecise dosage lowhygiene standards hiding of healing methods and lack ofpatients record as a significant setback to traditional healthpractitioners in the region

32 Thin Layer Chromatography (TLC) Profiling Thin layerchromatography remains the most common useful techniqueby researchers as a quick and simple method to resolvethe constituents of a crude extract followed by advancedextraction methods to purify the active ingredients Previousstudies reported that specific secondary metabolites in plantexhibit a wide range of antioxidant and antibacterial proper-ties [21] TLC was formed on a glass sheet coated with silicagel After the sample is applied to the plate a solvent system(mobile phase) is drawn up via capillary action on the TLCplate at different mobility rate depending on the chemicalcompositionsThe separated spotswith different colourswerevisualized by the UV light projection onto the glass sheetThe variation of coloured spots detected in the crude extractsindicates the presence of different phytochemicals that may

be responsible for the pharmacological effects of selectedplants use in the management of oral and skin infections asacclaimed by the traditional healers We quantified the resultsby measuring the distance travelled by the plant compoundsdivided by the total distance travelled by the mobile phaseFrom Table 3 it is apparent that mobile phase 1 consistingof chloroform and ethanol in the volume ratio of 64 had abetter separation than mobile phase II containing tolueneethyl acetate and acetic acid in the volume ratio of 541 Themethanol extract of AH in bothmobile phases had the highestnumber of three bands with Rf values of 0667 0833 and0989 for mobile phase I and Rf values of 0381 0648 and0781 for mobile phase IIThe plant extracts MM BA CO JCandCAhad two bands inmobile phase I with clear separationbut had single band in mobile phase II The plant extractshaving only one band in either mobile phase suggest a poorsolvent system for compounds separation The TLC profilesprovide a characteristic fingerprint of these plants that mayalso be useful for their identification

33 Antioxidant Assays

331 TPC and TFC Phenolic compounds are secondarymetabolites distributed in fruits vegetables nuts herbsseeds and beverages [22] They are excellent scavengers offree radical that has been implicated in the pathologies ofvarious human diseases the antioxidant activity is dependenton the structure hydrophobic benzenoid rings and hydrogen-bonding potential of the phenolic hydroxyl groups In TPCassay a phenol in plant extracts loses a proton to formphenolate ion which reduces Folin Ciocalteu reagent andthe resulting changes are measured spectrophotometricallyat 765 nm [13] The content of phenol is reported in mg oftannic acid equivalent (TAE)mL of extract As shown inFigure 1 the total phenolic contents of absolute methanolcrude extract of these botanicals range from 700 to 3600mgTAEg The AH extract had the highest concentrationfollowed by CA CO MM BNI BA EI BN and JC whilethe least concentration was recorded in II extract Moreoverthe Folin Ciocalteu reagent (FCR) has been used to measureother compounds such as ascorbic acid amino acids andsugars hence it may not provide a concise amount of thephenolic compounds present in these plants Flavonoids areincreasingly consumed in a significant amount in daily dietsworldwide due to their various health benefits toman [23 24]In TFC assay aluminum ion in the reacting mixture formscomplexes with the C- 4 keto and either C- 3 or C- 5 hydroxylor with ortho hydroxyl groups in the A or B ring of flavonoidsdetected in the plant extracts [14] As shown in Figure 2the total flavonoid content was recorded in the plant extractsranging from 750 to 1500 mgQEg as extrapolated from thestandard quercetin curve The CA extract had the highestconcentration followed by BNI CO II AH BN MM JCand BA while the lowest concentration was recorded in EIextract TPC is found higher than the TFC in some of theextracts supporting the fact that most flavonoids are alsophenolics According to our results all the plant extractsare rich source of phenolic compounds The composition ofTPC and TFC registered in these plants may be difficult to

The Scientific World Journal 11

Table 3 Thin layer chromatography retention factor using mobile phase CHL ET (mobile phase I) and Tol EA AA (mobile phase II)

SN Plant Extract CHL ET (64) Tol EA AA (541)1 Baphia nitida 1 0742 Macrolobium macrophyllum 0706 0941 06953 Chromolaena odorata 0471 0647 06674 Jatropha curcas 0353 0941 06485 Cassia alata 0118 0882 06866 Ipomoea involucrata 0956 -7 Acalypha hispida 0667 0833 0989 0381 0648 07818 Breynia nivosa 0989 07629 Burkea africana 0644 1 075210 Eleusine indica 0978 0743CHL chloroform ET ethanol Tol toluene EA ethylacetate AA acetic acid

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

500

1000

1500

2000

2500

3000

3500

4000

TPC

(mgT

AE

g)

Figure 1 The total phenolic content (TPC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata TAE tannic acid equivalent

compare with others reported elsewhere due to many factorsincluding differences from species to species maturity stageplant parts and assay methods harvesting time standarddrugs and geographical location

332 Reducing Power (RP) Ferric reducing power is a globalmethod often used to assess the capacity of antioxidantsto donate either an electron or hydrogen atom to unpairedelectron [25] In reducing power assay plant extracts exertantioxidant activities via reduction of Fe3+ferric cyanidecomplex to Fe2+ (ferrous form) resulting in changing thegreen colour to blue and sometimes Perlrsquos Prussian bluecolour depending on the antioxidant potential [10] As shownin Figure 3 the reducing power of the methanol crudeextracts and the standard gallic acid rutin and ascorbic acidincreased with increase in concentration Higher absorbanceat 700 nm can be read after potassium ferricyanide reactionwith ferric chloride At 1000 120583gmL the extracts from AH

0

200

400

600

800

1000

1200

1400

1600

TFC

(mgQ

Eg)

AH BN JC CO MM EI BNI BA CAIIPlant extracts

Figure 2 The total flavonoid content (TFC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata QE Quercetin equivalent

BNI MM CA BA EI and BN had absorbance value of 270086 073 055 036 027 and 027 respectively while that ofAH is higher than the standard rutin (254) but comparablewell with gallic acid (275) and ascorbic acid (269) EI andBN extracts had a similar trend of ferric reducing powerthough significantly lower as compared with the standarddrugs AH extract exhibits remarkable antioxidant capacity(RP) correlating with its richness in phenolic and flavonoidcontents It is reasonable to suggest that methanol selectivelyextracts antioxidant principles from these botanicals basedon their polarities and chemical structures which are reflectedin plant extracts with high TFC and TPC having minimal RP[26] We have shown that some plant extracts in particularAH BNI MM and CA extracts may serve as an excellentcandidate to minimize skin and oral infections caused byoxidative stress as a result of imbalance of free radicalsgeneration and natural antioxidants in the saliva or woundarea According to our results the increasing RP of the plantextracts correlateswith their TPCandTFCThis study at least

12 The Scientific World JournalRe

duci

ng P

ower

7

00nm

625 125 250 500 10003125Concentration (gmL)

0

05

1

15

2

25

3

IIAHBNJCCOMMEI

BNIBACAGARutinVit C

Figure 3 The ferric reducing antioxidative capacity of selectedplant extracts used in the management of oral and skin infectionsin southeast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata GA gallic acid Vit C vitamin C (standard antioxidants)

in part lends credence to the ethno-therapeutic use of someplant as natural antioxidant in themanagement of skin or oralinfections in the study area

333 FRSA of Plant Extracts Skin and oral cavity exposedto an array of oxidants such as smokes from cigaretteschemicals UV light nicotine and alcohol that are directlyor indirectly leading to the production of reactive oxygenspecies (ROS) are known to be responsible for some com-mon human diseases [27] These free radicals in particularhydroxyl and superoxide anions are highly reactivemoleculesderived from the metabolism of oxygen Although they playan important role in the biochemical immunological andphysiological processes it could be deleterious to vital organsif excessively accumulated in the human body [28] Food andpharmaceutical industries discovered that medicinal plantscontaining phenol and flavonoid compounds are essentialin the development of novel drugs against stress-induceddiseases [28] In the present investigation the concentration(IC50) at which 50 of free radical scavenged was calculatedand extrapolated from the plot of percentage inhibition ver-sus plant extract concentration The herbal drugs exhibitedrobust DPPH radical scavenging activity in a concentration-dependent manner having IC50 ranging from 2025 to 710120583gmL As shown in Figure 4 MM BA EI JC AH BNIand CA extracts had a potent DPPH radical scavengingactivity corresponding to their TPC TFC and RP Theplant extracts scavenged ABTS radical by the decrease inits absorbance at 734 nm All the plant extracts exhibitedstrong ABTS radical scavenging capacities in the following

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

200

400

600

800

1000

1200

1400

1600

IC50

(g

mL)

Figure 4 DPPH free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

050

100150200250300350400450500

IC50

(g

mL)

AH BN JC CO MM EI BNI BA CAIIPlant samples

Figure 5 ABTS free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

order BN gt JC gt CA gt MM gt BNI gt EI gt CO gt AH gtII gt BA having IC50 values ranging from 25 to 400 120583gmL(Figure 5) DPPH and ABTS radical scavenging activities ofthese herbal remedies displayed a positive correlation withthe working concentrations It is obvious that most of theplant extracts tested in this work possess the ability to quenchthe free radicals generation via electron or proton donationdemonstrated by the termination of radical chain reaction[28] Although both DPPH and ABTS radicals are electron-transfer based methods however it is apparent that ABTSmethod seems to be more sensitive than DPPH radical assaypossibly due to their solubility and diffusivity as shown inthe IC50 values [10] It is worth noting that BA extract withstrong DPPH radical scavenging effect had a moderate ABTS

The Scientific World Journal 13

Table 4 Zone of bacterial inhibition by the selected plant extracts use in the management of oral and skin infections

Diameter of inhibition (mm)SN Plant samples K pneumoniae P mirabilis S aureus E coli1 Cassia alata 15 plusmn 141 9 plusmn 042 15 plusmn 083 13 plusmn 0802 Breynia nivosa 8 plusmn 052 17 plusmn 100 8 plusmn 008 15 plusmn 1023 Burkea africana NI 11 plusmn 084 13 plusmn 072 NI4 Baphia nitida 8 plusmn 050 13 plusmn 122 8 plusmn 051 NI5 Ipomoea involucrata 8 plusmn 052 NI 8 plusmn 050 13 plusmn 0526 Jatropha curcas NI 9 plusmn 030 9 plusmn 062 13 plusmn 0727 Acalypha hispida 21 plusmn 163 NI 11 plusmn 009 17 plusmn 1328 Chromolaena odorata 8 plusmn 042 8 plusmn 035 9 plusmn 009 15 plusmn1209 Eleusine indica 8 plusmn 042 8 plusmn 035 NI 17 plusmn 10010 Macrolobium macrophyllum NI NI 8 plusmn 004 15 plusmn 12311 Ciprofloxacin - - le16(R) ge17(S) le16(R)ge20(S)NI indicates no zone of inhibition S sensitive R resistance

radical scavenging capacity which can be explained by thepartial ionization of the extract in the test medium [29] SinceROS promotes tissue damage and epithelial inflammationvia exposure to sunlight radiation or smoke as a result theantioxidant capacity demonstrated in this work may supportthe traditional use of these plants against the influence ofinflammatory markers and oxidants in the skin and oralcavity [29] Based on the results of the three antioxidantassays it is obvious that MM AH BNI EI and CA extractshad the best FRSA indicating their importance in convertingfree radicals into more stable products and then terminatethe free radical chain reaction Our study showed that themethanol crude extracts from these plants could protectagainst free radical damage in the mouth such as humangingival periodontal tissues and certain skin infections [5]

334 Antibacterial Activity of Plant Extracts Pathogenicbacteria such as Staphylococcus aureus Escherichia coli Kleb-siella pneumoniae and Proteus mirabilis play a significantrole as the primary cause of skin and oral infections [30]Staphylococcus aureus is a part of normal skin known to bethe most problematic in the cause of skin infections as aresult of their resistance mechanisms to first-line antibiotics[30] Although the presence of this organism as a residentoral floral is controversial nonetheless MSSA and MRSAhave been isolated from oral rinse and tongue swab inpatients with oral infections [31] Escherichia coli are Gram-negative bacteria known to be notorious for their resistantmechanisms towards the existing arsenal of antibiotics Oralinfections such as edematous and erythematous and skininfections like omphalitis surgical site infections burnnecrotizing fasciitis and cellulitis at all age groups are associ-ated with enterobacterium E coli [32] Klebsiella pneumoniaeis a Gram-negative encapsulated facultative anaerobic androd-shaped bacterium found in the normal flora of themouth skin and intestines This bacterium is responsiblefor the prolonged surgical wound biliary tract wound andurinary tract infections and it could pose a serious healthrisk when resistant strains invade the throat mouth or

intestines [33] Villegas et al [34] detected a CTX-M-12 120573-lactamase producing K pneumoniae new plasmid-mediatedESBLs highly resistant to ceftazidime from a patient withhospital-acquired wound infection Proteus mirabilis is oneof the most common enterobacteria after E coli identified asGram-negativemotile and rod-shaped bacterium It is foundin clinical specimen implicated in the skin respiratory tractand bacteraemia as well as burns wounds and urinary tractinfections [35] All these pathogenic multidrug-resistancebacteria are causative agent in a periodontal lips ulcer whitesponge nevus and skin infections [36]

335 Susceptibility Test and MIC Determination The resultof disc diffusion assay showed the diameter of inhibitionof bacteria growth classified as le 15 (resistant) and ge 16(susceptible) according to CLSI guideline [37] As shown inTable 4 K pneumoniae and E coli are more susceptible tothe treatment of Acalypha hispida (AH) having a zone ofinhibition (ZI) 21 and 17mm respectively BN and EI extractsinhibit the growth of P mirabilis and E coli having the samezone of inhibition 17 mm corresponding to the report of Al-Zubairi et al [38] The data obtained for the methanol crudeextracts from AH BN and EI show their selective inhibitionof the test bacteria growth The plant extracts with ZI above13 mm against our panel of bacterial strains were selected forMICdeterminationHere themethanol crude extracts exhibitMIC values of 256 to gt 512 120583gmL against the test bacterialstrains but were less active as compared to the standardciprofloxacin 003 - 006 120583gmL (Table 5) Ciprofloxacin isan active broad-spectrum antibiotic use in the treatmentof urinary tract infections prostatitis and acute cystitis viainterfering with bacterial DNA replication and transcriptionthrough inhibition of DNA gyrase or topoisomerase [39] Allthe plant extracts demonstrated a mild to weak antibacterialactivity based on the classification of Rios and Recio [40] whoconsidered antibacterial activity of the crude plant extractsignificant (MIC lt 100 120583gmL) moderate (100 lt MIC = 512120583gmL) or weak (MIC gt 512 120583gmL) The herbal remediesshowed moderate inhibition of bacterial growth except for K

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

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The Scientific World Journal

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Submit your manuscripts atwwwhindawicom

Page 5: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

The Scientific World Journal 5

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Ricin

uscommun

isLinn

Eu

phorbiaceae

Castorb

ean

Leafroo

tand

seed

Skin

Treatm

ento

fskininfection

anddetoxification

Applycasto

roilon

thea

ffected

area

torelieve

skin

infection

Citru

slim

on(L)Bu

rmf

(prosp

)Ru

taceae

Limon

Leaffruit

andseed

Skin

Treatm

ento

ffluheadache

cold

andoralinfections

Mixequalquantities

ofLimon

juicea

ndho

lybasilkeepitun

derthe

suntill

becomes

thickapplythison

face

Alliu

mcepa

LLiliaceae

Onion

sLeafandbu

lbSkin

Treatm

ento

fperiodo

ntitis

andcold

Theb

ulbismacerated

andmixed

with

water

andthen

takenorally

Breynianivosa

Euph

orbiaceae

Icep

lant

Leaf

Oral

Treatin

gof

toothache

Boiltheleave

with

waterthentake

the

water

Ipom

oeainvolucrata

Con

volvulaceae

Water

convolvu

lus

Leafandste

mOral

Treatm

ento

foralinfectio

nMixtheleave

with

ugbaga

treea

ndbo

ilwith

water

anddrinkthew

ater

Burkea

afric

ana

Caesalpinioideae

Icheku

Leaf

Oral

Treatm

ento

forald

isease

Boiltheleave

with

water

andtakebut

donrsquot

swallowit

Baphianitid

aPapilio

naceae

Cam

wood

Leafor

stem

Oral

Streng

thensthe

teeth

Chew

thes

tem

orscrubtheleave

onthe

teeth

Gongronem

alatifolium

Asclepiadaceae

Utazi

Leaf

Oral

Treatm

ento

fcou

ghand

diabetes

Crushtheleafw

ithmortara

ndmixwith

ginger

orgarlica

nddrink

Anonasenegalen

sisAnn

onaceae

Ubu

ruocha

Root

Oral

Treatm

ento

fmalariaand

mou

thinfection

Boilther

ootsanddrink

Abrusp

recatoriu

sPapilio

noideae

Crabrsquoseyes

Seed

androot

Oralskin

Treatm

ento

fcou

ghskin

carew

ound

Grin

dther

ootand

applythep

oultice

onthea

ffected

skin

areaA

lsocrushedseeds

canbe

takenorally

Acanthus

montanu

sAc

anthaceae

False

thistle

Leafandroot

Oraland

Skin

Treatm

ento

ffurun

cles

lesio

nsandurinary

infections

Boilther

ootand

applythep

oultice

tothe

affectedpartof

theb

odyFo

rinfectio

ns

boilther

ootand

leaves

anddrink

Piperu

mbellatum

Piperaceae

Njamuja

Leaf

Oral

Treatm

ento

fpeptic

ulcers

Crushtheleavesa

ndsoak

inwarm

orho

twater

anddrink

Euphorbiahirta

Euph

orbiaceae

Asth

maw

eed

Who

leplant

Oral

Treatm

ento

fwou

nds

Leaves

arem

acerated

andappliedto

affectedpart

Ficusb

urkei(Miq)Miq

Moraceae

Sand

paper

Leaf

Oral

Treatm

ento

fhypertension

Leaves

areb

oiledandtakenorally

6 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Gmelina

arborea

Verbenaceae

Gmelina

Stem

andleaf

Oral

Treatm

ento

fgon

orrhea

pilesabdo

minalpains

burningsensationsfever

mou

thwo

undanddiabetes

Snake-biteandscorpion

sting

Leaves

grou

ndinto

paste

with

water

are

appliedto

theforeheadforh

eadachein

feversB

oiledste

msa

retakenorally

Lophira

alata

Ochnaceae

Ekki

Stem

andleaf

Oral

Treatm

ento

ffeverm

outh

woun

dandmalaria

Boiltheleavesa

ndste

mandtake

orally

Nau

clealatifolia

Rubiaceae

Ubu

ru-in

uStem

andleaf

Oral

Itisused

inthetreatment

ofmalariaepilepsymou

thwo

undanxietypainfever

Boilther

ootsanddrink

Annona

murica

taAnn

onaceae

Sour

sop

Leaf

Oral

Treatm

ento

fcancera

ndmou

thwo

undinfections

Crushor

boiltheleavesa

ndtake

orally

Psidium

guajava

Myrtaceae

Guava

Leaf

Oral

Treatm

ento

fperiodo

ntal

diseasebu

rnsscrapes

sunb

urnwo

undspsoria

sis

eczemaandothers

Leaves

arec

rushed

andtakenorally

Astra

galusm

embran

aceus

Fabaceae

Beiq

iStem

andleaf

Oral

Treatm

ento

fcoldup

per

respira

tory

infections

allergiesfib

romyalgia

anem

iaH

IVA

IDS

Leaves

arec

rushed

andtakenorallyand

canalso

beappliedto

skin

Buchholziacoria

cea

Capparaceae

Won

derfu

lkola

Leaf

Oral

Itisused

tradition

allyfor

treatingdiabetes

hypertensio

nrheumatism

coldcou

ghand

catarrh

Boilleaves

andtake

orally

Gnetum

afric

anum

Gnetacea

Afang

Leaf

Oral

Enlarged

spleen

sore-th

roatandcathartic

Leaves

canbe

boiledor

cooked

andtaken

orally

Anoridium

man

niAnn

onaceae

Ewuro-ig

boStem

bark

Oral

Treatm

ento

fDiarrhea

coug

hfeverrheumatism

Theb

arkiscrushedandbo

iledandthen

takenorally

Funtum

iaafric

ana

Apocyn

aceae

Akorie

mbamiri

Leafstem

root

Skin

treatmento

fcon

stipatio

nwo

undsw

eakbladder

jaun

dice

Who

leplantisb

oiledandtakenorally

Also

thec

rushed

leaves

area

ppliedto

the

affectedskin

area

The Scientific World Journal 7

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Commiphora

afric

ana

Burseraceae

Turari

dashi

Rootsfruits

Oral

Treatm

ento

fwho

oping

coug

hbron

chitis

Rootsa

remacerated

andtakenorally

Dacryodesedulis

Burseraceae

Elem

iub

eBa

rkroo

tsfruits

Skin

Treatm

ento

fJiggersSkin

diseasesE

leph

antia

sis

Theleavesa

reeatenrawwith

kolanu

tas

anantie

meticLeaf-s

apisinstilledinto

thee

arfore

ar-tr

oubleanda

leaf-decoctio

nisprepared

asa

vapo

ur-bathforfeverish

stiffn

essw

ithheadache

Combretum

mucronatum

Com

bretaceae

Fararg

eza

Rootsa

ndleaf

Skin

Treatm

ento

fskininfection

Ther

ootand

leafarem

acerated

and

appliedto

affectedskin

area

Cleomec

iliata

Capparaceae

Ekuya

Leafseeds

Skin

Treatm

ento

fCon

vulsion

wo

undssores

Seedsa

recrushedandthep

oultice

appliedto

thea

ffected

areaTh

emacerated

leaves

after

boiledaretaken

orally

Ageratum

conyzoides

Aste

raceae

Urata

Who

leplant

Skin

Treatm

ento

fwou

ndU

lcer

Sleeping

sickn

essEyew

ash

Thep

lant

ismacerated

andappliedto

affectedskin

area

Aspilia

afric

ana

Aste

raceae

Yunyun

Ka

lank

uwa

Leafflow

erSkin

Treatm

ento

fskinrashes

cleaning

sorescorneal

opacities

Thefl

ower

issqueezed

andappliedto

affectedskin

area

Centau

reaperrottetii

Aste

raceae

Danyi

Who

leplant

Skin

Treatm

ento

fskin

infectionsSyphilis

Thefl

ower

issqueezed

andap

oultice

appliedto

affectedskin

area

Chrysanthellu

mindicum

Aste

raceae

Abilereoyigi

Who

leplant

Skin

Treatm

ento

fboils

gono

rrheaJaun

dice

Thep

lant

iscrushedandthep

oultice

applieddirectlyto

thes

kinarea

Elephantopus

scaber

Aste

raceae

Elephant

foot

Leafroo

tOral

Treatm

ento

ffevercou

ghLeafandroot

areb

oiledtogether

and

takenorally

Emiliacoccinea

Aste

raceae

Odu

ndun

Leafsap

and

root

Skin

Treatm

ento

fulcerhernia

measle

sWho

leplantissqu

eezedandsaptaken

orally

Acalypha

godseffi

ana

Euph

orbiaceae

Jinwinini

Leaf

Skin

Treatm

ento

fskin

infectionsantim

icrobials

Leafismacerated

andtakenorallyas

well

asappliedto

affectedskin

area

8 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Brideliafer

rugin

eaEu

phorbiaceae

Irikirni

Leafstem

bark

root

Oral

Treatm

ento

fInsom

nia

mou

thwashgono

rrhea

Who

leplantisb

oiledandtakenorally

Euphorbiaheterophylla

Euph

orbiaceae

Egele

Leafroo

tsSkin

Treatm

ento

fSkindiseases

Leafandroot

arem

acerated

andthe

poultic

eapp

liedto

thea

ffected

skin

area

Euphorbiahirta

Euph

orbiaceae

Non

onku

rciya

odne

Who

leplant

exud

ate

Oral

Treatm

ento

fAsth

ma

coug

hshapeo

fbreasts

Thew

holeplantism

acerated

andtaken

orally

Euphorbialaterifl

ora

Euph

orbiaceae

Orow

ere

Leafexud

ate

Skin

Treatm

ento

fdermatitis

constip

ation

Theleafissqu

eezedandthep

oultice

appliedto

thea

ffected

skin

area

Cajanu

scajan

Fabaceae

Orela

Leafseed

Skin

andoral

Treatm

ento

fSmallpox

and

mou

thwash

Leafissqueezed

andappliedto

the

affectedarea

Dan

iellia

thurifera

Fabaceae

Iya

Stew

-woo

ddu

stSkin

Treatm

ento

fscabies

Theb

arkismacerated

andappliedto

the

affectedskin

area

Dialiu

mguineense

Fabaceae

Tasm

iyar

kumi

Leafbark

fruitand

twig

Oral

Treatm

ento

fBronchitis

coug

handdiuretic

Theleafand

bark

arec

rushed

andbo

iled

togetherTh

emixture

isthen

takenorally

Acacianilotica

Fabaceae

Baanigabaruwa

Fruitbark

exud

ate

Skin

Treatm

ento

fskindiseases

fung

alinfections

Thee

xudatesa

resqueezed

from

theb

ark

andapplieddirectlyto

thea

ffected

skin

area

Napoleona

imperia

lisLecythidaceae

Mabun

giTw

igs

Oral

Treatm

ento

fAsth

ma

coug

hTw

igsa

remacerated

andtakenorally

Alliu

msativ

umLiliaceae

Ayoayuu

Bulb

Oral

Treatm

ento

ffevercou

gh

asthmaantim

icrobial

Theb

ulbismacerated

andtakenorally

Hibiscus

asper

Malvaceae

Isapad

angiraa

Leaf

Oraland

skin

Treatm

ento

fcou

gh

woun

dsanddiuretic

Leafisbo

iledandtakenorally

Hibiscus

rosasin

ensis

Malvaceae

Kekekeireagu

Leafstem

flowe

rSkin

Treatm

ento

finfl

uenza

woun

dsand

diuretic

Who

leplantism

acerated

andtaken

orally

Theleafissqu

eezedandappliedin

affectedskin

area

Hibiscus

sabdariffa

Malvaceae

Sobo

gurgu

zuLeafflow

erOral

Treatm

ento

fDiuretic

scoug

handdressin

gwo

unds

Leafisbo

iledandtakenorally

Ficuseleg

ans

Moraceae

Asoro

Leaf

Skinoral

Treatm

ento

fPile

constip

ationandcraw

-craw

Leafissqueezed

andappliedto

the

affectedarea

The Scientific World Journal 9

the absorbance The percentage inhibition of DPPH orABTS radical scavenging activity by the plant extracts wascalculated as (Abscontrol ndash Abssample)(Abscontrol) times 100where Abscontrol is the absorbance of DPPH or ABTS +methanol Abssample is the absorbance of DPPH or ABTSradical + sample extractstandard Here the concentration ofthe extracts needed to decrease the absorbance of DPPH orABTS radical by 50 was calculated Rutin (Sigma-Aldrichge 94 HPLC grade) at the same working concentrations ofthe plant extracts was used as reference drug

210 Bacterial Strains All bacterial strains were culturedon nutrient agar slopes and were incubated for 24 h at37∘C before the MIC An inoculum turbidity equivalent toa 05 McFarland standard (1times107 CFUmL) was prepared innormal saline for each test organism and then diluted 1100in Mueller-Hinton broth just before injection of the platesStandard strains used in the study included S aureus and Ecoli which were a gift from Dr EO Ekundayo (Departmentof Microbiology Michael Okpara University of Agriculture)and clinically isolated strains of K pneumoniae and Pmirabilis were gotten from Department of MicrobiologyFederal Medical Centre (FMC) Umuahia Abia State

211 Antibacterial Susceptibility Using Disc Diffusion MethodThe microbial growth inhibition potential of the plantextracts was determined by using the agar disc diffusionmethod [17] The plant extracts were dissolved in 10 vvDMSO (Merck UK Ltd) to a final concentration of 2 mgmLThe nutrient agar plate was streaked with the standardizedtest bacterial inoculum over the surface evenly and 2mgdiscof the plant extracts transferred onto sterile filter disc papers(about 6 mm) containing the test compounds was placed onthe agar surface in the Petri dishes 100 120583gdisc of standardantibiotic ciprofloxacin was used as control and 100 120583LDMSOdisc served as negative control The plates were thenincubated at 37∘C for 24 h Generally the antimicrobialextracts diffuse into the agar and inhibit the growth of thetest bacteria All tests were done in duplicate and diame-ter zones of inhibition of growth were measured using ameter rule from the edge of each dish after the incubationperiod

212 Minimum Inhibitory Concentration (MIC) Assays Wefollowed the method of Smith et al [18] to determine theMIC ofmethanol crude extracts of selected botanicals againstclinical isolates of bacteria associated with skin and oralinfection A volumeof 100mLof sterileMueller-Hinton broth(Oxoid Basingstoke UK) containing 20 mgL of Ca2+ and10 mgL of Mg2+ was dispensed into a 96-well microtitreplate (Nunc 03 mL total volume per well) All antibacterialagents were dissolved in dimethyl sulphoxide (DMSO) anddiluted in Mueller-Hinton broth to give a stock solutionThen 100 120583L of the antibacterial agent stock solution (2000mgL) was serially diluted into each well and 100 mL ofthe bacterial inoculum added to each well to give a finalconcentration range of 0024 - 1 mgL All procedures wereperformed in duplicate and the plates were incubated for 18

h at 37∘C Briefly 20 mL of a 5 mgL methanol solution of 3-[45-dimethylthiazol-2-yl]-25-diphenyltetrazolium bromide(MTT) (Sigma-Aldrich Ltd South Africa) was added to eachwell and then incubated for 30 min Blue colouration indi-cated bacterial growth The MIC was recorded as the lowestconcentration at which no colour change was observed

213 Statistical Analysis Data analysis was done inMicrosoftExcel to obtain descriptive statistics Means values wereseparated by the Duncan multiple tests using SAS The dif-ferent levels of significance within the groups were analyzedusing one-way analysis of variance (ANOVA) Values wereconsidered significant at P lt005

3 Results and Discussion

31 Ethnobotanical Survey Modern drugs are expensive andsupply to a remote area may be irregular hence result-ing in overreliance of traditional methods of healing as aprimary source of healthcare [19] The rural communitiesof Ebem-Ohafia region of southeastern Nigeria depend onherbal remedies to meet their domestic and health needsDuring our ethnobotanical survey a total of 30 respondentsincluding the elders herbalists and traditional healers wereinterviewed in the study areaMajority of the people (22)wereaged 30ndash65 years dominated by males (25) and one-third(10) had a primary school education while 15 had secondaryeducation and others informal education A total of 61medic-inal plant species belonging to 26 families were identified inthe treatment of oral and skin diseases as shown in Table 1Twenty-seven of them are used in the management of oralinfections and twenty-six plants for the management of skininfection while eight plants are used in the management ofboth infections Table 2 shows the list of family number of theplant species used and their percentage occurrenceThemostrepresented plant family is the Euphorbiaceae having 1803of the plant species followed by Asteraceae and Fabaceae with1147 each [20] Euphorbiaceae is a large family of floweringplants with 300 genera and about 7500 species found usefulin the curing of some common human diseases Asteraceae isa large family of flowering plants having 32 913 species and1911 genera and has been found to possess diverse biologicaleffects Fabaceae is a family of cosmopolitan distributionwith approximately 730 genera and 19400 species richnessat a global level [20] The leaves (60) recorded the mostcommonly used plant part followed by the root (16) andthen stem bark (12) None of the respondents indicated anypossible side effects caused by any of these plants listed inTable 1 The most common methods of preparation includeinfusion decoction and maceration These herbalists ortraditional healers often collect the plants from the fieldand dry and crush them into powder form before storingin bottles to prevent patients from identifying the plantsused for their treatment Most healers refused to provideinformation without payment whereas others thought toprovide information on the plant use is disrespect of theirtraditional belief Furthermore the majority of the plantsmentioned lack scientific data to substantiate their acclaimedfolkloric use From this work it is evident that medicinal

10 The Scientific World Journal

Table 2 The list of family number of the plant species and theirpercentage occurrence

Family Number of plant species OccurrenceFabaceae 7 1147Euphorbiaceae 11 1803Asteraceae 7 1147Myrtaceae 4 656Moraceae 3 492Malvaceae 4 656Liliaceae 2 328Capparaceae 2 328Annonaceae 3 492Combretaceae 1 164Burseraceae 2 328Apocynaceae 1 164Gnetaceae 1 164Lecythidaceae 1 164Verbenaceae 1 164Ochnaceae 1 164Rubiaceae 1 164Piperaceae 1 164Acanthaceae 2 328Papilionoideae 2 328Asclepiadaceae 1 164Caesalpiniaceae 2 328Convolvulaceae 1 164Rutaceae 1 164Crassulaceae 1 164Poaceae 1 164

plants play a significant role in the primary healthcare ofthe people and conservation of the indigenous knowledge ofthis folklore medicine is crucial for the innovations of futuredrug discovery and possible herbal plants trade in the region[19] On the contrary we observed imprecise dosage lowhygiene standards hiding of healing methods and lack ofpatients record as a significant setback to traditional healthpractitioners in the region

32 Thin Layer Chromatography (TLC) Profiling Thin layerchromatography remains the most common useful techniqueby researchers as a quick and simple method to resolvethe constituents of a crude extract followed by advancedextraction methods to purify the active ingredients Previousstudies reported that specific secondary metabolites in plantexhibit a wide range of antioxidant and antibacterial proper-ties [21] TLC was formed on a glass sheet coated with silicagel After the sample is applied to the plate a solvent system(mobile phase) is drawn up via capillary action on the TLCplate at different mobility rate depending on the chemicalcompositionsThe separated spotswith different colourswerevisualized by the UV light projection onto the glass sheetThe variation of coloured spots detected in the crude extractsindicates the presence of different phytochemicals that may

be responsible for the pharmacological effects of selectedplants use in the management of oral and skin infections asacclaimed by the traditional healers We quantified the resultsby measuring the distance travelled by the plant compoundsdivided by the total distance travelled by the mobile phaseFrom Table 3 it is apparent that mobile phase 1 consistingof chloroform and ethanol in the volume ratio of 64 had abetter separation than mobile phase II containing tolueneethyl acetate and acetic acid in the volume ratio of 541 Themethanol extract of AH in bothmobile phases had the highestnumber of three bands with Rf values of 0667 0833 and0989 for mobile phase I and Rf values of 0381 0648 and0781 for mobile phase IIThe plant extracts MM BA CO JCandCAhad two bands inmobile phase I with clear separationbut had single band in mobile phase II The plant extractshaving only one band in either mobile phase suggest a poorsolvent system for compounds separation The TLC profilesprovide a characteristic fingerprint of these plants that mayalso be useful for their identification

33 Antioxidant Assays

331 TPC and TFC Phenolic compounds are secondarymetabolites distributed in fruits vegetables nuts herbsseeds and beverages [22] They are excellent scavengers offree radical that has been implicated in the pathologies ofvarious human diseases the antioxidant activity is dependenton the structure hydrophobic benzenoid rings and hydrogen-bonding potential of the phenolic hydroxyl groups In TPCassay a phenol in plant extracts loses a proton to formphenolate ion which reduces Folin Ciocalteu reagent andthe resulting changes are measured spectrophotometricallyat 765 nm [13] The content of phenol is reported in mg oftannic acid equivalent (TAE)mL of extract As shown inFigure 1 the total phenolic contents of absolute methanolcrude extract of these botanicals range from 700 to 3600mgTAEg The AH extract had the highest concentrationfollowed by CA CO MM BNI BA EI BN and JC whilethe least concentration was recorded in II extract Moreoverthe Folin Ciocalteu reagent (FCR) has been used to measureother compounds such as ascorbic acid amino acids andsugars hence it may not provide a concise amount of thephenolic compounds present in these plants Flavonoids areincreasingly consumed in a significant amount in daily dietsworldwide due to their various health benefits toman [23 24]In TFC assay aluminum ion in the reacting mixture formscomplexes with the C- 4 keto and either C- 3 or C- 5 hydroxylor with ortho hydroxyl groups in the A or B ring of flavonoidsdetected in the plant extracts [14] As shown in Figure 2the total flavonoid content was recorded in the plant extractsranging from 750 to 1500 mgQEg as extrapolated from thestandard quercetin curve The CA extract had the highestconcentration followed by BNI CO II AH BN MM JCand BA while the lowest concentration was recorded in EIextract TPC is found higher than the TFC in some of theextracts supporting the fact that most flavonoids are alsophenolics According to our results all the plant extractsare rich source of phenolic compounds The composition ofTPC and TFC registered in these plants may be difficult to

The Scientific World Journal 11

Table 3 Thin layer chromatography retention factor using mobile phase CHL ET (mobile phase I) and Tol EA AA (mobile phase II)

SN Plant Extract CHL ET (64) Tol EA AA (541)1 Baphia nitida 1 0742 Macrolobium macrophyllum 0706 0941 06953 Chromolaena odorata 0471 0647 06674 Jatropha curcas 0353 0941 06485 Cassia alata 0118 0882 06866 Ipomoea involucrata 0956 -7 Acalypha hispida 0667 0833 0989 0381 0648 07818 Breynia nivosa 0989 07629 Burkea africana 0644 1 075210 Eleusine indica 0978 0743CHL chloroform ET ethanol Tol toluene EA ethylacetate AA acetic acid

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

500

1000

1500

2000

2500

3000

3500

4000

TPC

(mgT

AE

g)

Figure 1 The total phenolic content (TPC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata TAE tannic acid equivalent

compare with others reported elsewhere due to many factorsincluding differences from species to species maturity stageplant parts and assay methods harvesting time standarddrugs and geographical location

332 Reducing Power (RP) Ferric reducing power is a globalmethod often used to assess the capacity of antioxidantsto donate either an electron or hydrogen atom to unpairedelectron [25] In reducing power assay plant extracts exertantioxidant activities via reduction of Fe3+ferric cyanidecomplex to Fe2+ (ferrous form) resulting in changing thegreen colour to blue and sometimes Perlrsquos Prussian bluecolour depending on the antioxidant potential [10] As shownin Figure 3 the reducing power of the methanol crudeextracts and the standard gallic acid rutin and ascorbic acidincreased with increase in concentration Higher absorbanceat 700 nm can be read after potassium ferricyanide reactionwith ferric chloride At 1000 120583gmL the extracts from AH

0

200

400

600

800

1000

1200

1400

1600

TFC

(mgQ

Eg)

AH BN JC CO MM EI BNI BA CAIIPlant extracts

Figure 2 The total flavonoid content (TFC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata QE Quercetin equivalent

BNI MM CA BA EI and BN had absorbance value of 270086 073 055 036 027 and 027 respectively while that ofAH is higher than the standard rutin (254) but comparablewell with gallic acid (275) and ascorbic acid (269) EI andBN extracts had a similar trend of ferric reducing powerthough significantly lower as compared with the standarddrugs AH extract exhibits remarkable antioxidant capacity(RP) correlating with its richness in phenolic and flavonoidcontents It is reasonable to suggest that methanol selectivelyextracts antioxidant principles from these botanicals basedon their polarities and chemical structures which are reflectedin plant extracts with high TFC and TPC having minimal RP[26] We have shown that some plant extracts in particularAH BNI MM and CA extracts may serve as an excellentcandidate to minimize skin and oral infections caused byoxidative stress as a result of imbalance of free radicalsgeneration and natural antioxidants in the saliva or woundarea According to our results the increasing RP of the plantextracts correlateswith their TPCandTFCThis study at least

12 The Scientific World JournalRe

duci

ng P

ower

7

00nm

625 125 250 500 10003125Concentration (gmL)

0

05

1

15

2

25

3

IIAHBNJCCOMMEI

BNIBACAGARutinVit C

Figure 3 The ferric reducing antioxidative capacity of selectedplant extracts used in the management of oral and skin infectionsin southeast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata GA gallic acid Vit C vitamin C (standard antioxidants)

in part lends credence to the ethno-therapeutic use of someplant as natural antioxidant in themanagement of skin or oralinfections in the study area

333 FRSA of Plant Extracts Skin and oral cavity exposedto an array of oxidants such as smokes from cigaretteschemicals UV light nicotine and alcohol that are directlyor indirectly leading to the production of reactive oxygenspecies (ROS) are known to be responsible for some com-mon human diseases [27] These free radicals in particularhydroxyl and superoxide anions are highly reactivemoleculesderived from the metabolism of oxygen Although they playan important role in the biochemical immunological andphysiological processes it could be deleterious to vital organsif excessively accumulated in the human body [28] Food andpharmaceutical industries discovered that medicinal plantscontaining phenol and flavonoid compounds are essentialin the development of novel drugs against stress-induceddiseases [28] In the present investigation the concentration(IC50) at which 50 of free radical scavenged was calculatedand extrapolated from the plot of percentage inhibition ver-sus plant extract concentration The herbal drugs exhibitedrobust DPPH radical scavenging activity in a concentration-dependent manner having IC50 ranging from 2025 to 710120583gmL As shown in Figure 4 MM BA EI JC AH BNIand CA extracts had a potent DPPH radical scavengingactivity corresponding to their TPC TFC and RP Theplant extracts scavenged ABTS radical by the decrease inits absorbance at 734 nm All the plant extracts exhibitedstrong ABTS radical scavenging capacities in the following

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

200

400

600

800

1000

1200

1400

1600

IC50

(g

mL)

Figure 4 DPPH free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

050

100150200250300350400450500

IC50

(g

mL)

AH BN JC CO MM EI BNI BA CAIIPlant samples

Figure 5 ABTS free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

order BN gt JC gt CA gt MM gt BNI gt EI gt CO gt AH gtII gt BA having IC50 values ranging from 25 to 400 120583gmL(Figure 5) DPPH and ABTS radical scavenging activities ofthese herbal remedies displayed a positive correlation withthe working concentrations It is obvious that most of theplant extracts tested in this work possess the ability to quenchthe free radicals generation via electron or proton donationdemonstrated by the termination of radical chain reaction[28] Although both DPPH and ABTS radicals are electron-transfer based methods however it is apparent that ABTSmethod seems to be more sensitive than DPPH radical assaypossibly due to their solubility and diffusivity as shown inthe IC50 values [10] It is worth noting that BA extract withstrong DPPH radical scavenging effect had a moderate ABTS

The Scientific World Journal 13

Table 4 Zone of bacterial inhibition by the selected plant extracts use in the management of oral and skin infections

Diameter of inhibition (mm)SN Plant samples K pneumoniae P mirabilis S aureus E coli1 Cassia alata 15 plusmn 141 9 plusmn 042 15 plusmn 083 13 plusmn 0802 Breynia nivosa 8 plusmn 052 17 plusmn 100 8 plusmn 008 15 plusmn 1023 Burkea africana NI 11 plusmn 084 13 plusmn 072 NI4 Baphia nitida 8 plusmn 050 13 plusmn 122 8 plusmn 051 NI5 Ipomoea involucrata 8 plusmn 052 NI 8 plusmn 050 13 plusmn 0526 Jatropha curcas NI 9 plusmn 030 9 plusmn 062 13 plusmn 0727 Acalypha hispida 21 plusmn 163 NI 11 plusmn 009 17 plusmn 1328 Chromolaena odorata 8 plusmn 042 8 plusmn 035 9 plusmn 009 15 plusmn1209 Eleusine indica 8 plusmn 042 8 plusmn 035 NI 17 plusmn 10010 Macrolobium macrophyllum NI NI 8 plusmn 004 15 plusmn 12311 Ciprofloxacin - - le16(R) ge17(S) le16(R)ge20(S)NI indicates no zone of inhibition S sensitive R resistance

radical scavenging capacity which can be explained by thepartial ionization of the extract in the test medium [29] SinceROS promotes tissue damage and epithelial inflammationvia exposure to sunlight radiation or smoke as a result theantioxidant capacity demonstrated in this work may supportthe traditional use of these plants against the influence ofinflammatory markers and oxidants in the skin and oralcavity [29] Based on the results of the three antioxidantassays it is obvious that MM AH BNI EI and CA extractshad the best FRSA indicating their importance in convertingfree radicals into more stable products and then terminatethe free radical chain reaction Our study showed that themethanol crude extracts from these plants could protectagainst free radical damage in the mouth such as humangingival periodontal tissues and certain skin infections [5]

334 Antibacterial Activity of Plant Extracts Pathogenicbacteria such as Staphylococcus aureus Escherichia coli Kleb-siella pneumoniae and Proteus mirabilis play a significantrole as the primary cause of skin and oral infections [30]Staphylococcus aureus is a part of normal skin known to bethe most problematic in the cause of skin infections as aresult of their resistance mechanisms to first-line antibiotics[30] Although the presence of this organism as a residentoral floral is controversial nonetheless MSSA and MRSAhave been isolated from oral rinse and tongue swab inpatients with oral infections [31] Escherichia coli are Gram-negative bacteria known to be notorious for their resistantmechanisms towards the existing arsenal of antibiotics Oralinfections such as edematous and erythematous and skininfections like omphalitis surgical site infections burnnecrotizing fasciitis and cellulitis at all age groups are associ-ated with enterobacterium E coli [32] Klebsiella pneumoniaeis a Gram-negative encapsulated facultative anaerobic androd-shaped bacterium found in the normal flora of themouth skin and intestines This bacterium is responsiblefor the prolonged surgical wound biliary tract wound andurinary tract infections and it could pose a serious healthrisk when resistant strains invade the throat mouth or

intestines [33] Villegas et al [34] detected a CTX-M-12 120573-lactamase producing K pneumoniae new plasmid-mediatedESBLs highly resistant to ceftazidime from a patient withhospital-acquired wound infection Proteus mirabilis is oneof the most common enterobacteria after E coli identified asGram-negativemotile and rod-shaped bacterium It is foundin clinical specimen implicated in the skin respiratory tractand bacteraemia as well as burns wounds and urinary tractinfections [35] All these pathogenic multidrug-resistancebacteria are causative agent in a periodontal lips ulcer whitesponge nevus and skin infections [36]

335 Susceptibility Test and MIC Determination The resultof disc diffusion assay showed the diameter of inhibitionof bacteria growth classified as le 15 (resistant) and ge 16(susceptible) according to CLSI guideline [37] As shown inTable 4 K pneumoniae and E coli are more susceptible tothe treatment of Acalypha hispida (AH) having a zone ofinhibition (ZI) 21 and 17mm respectively BN and EI extractsinhibit the growth of P mirabilis and E coli having the samezone of inhibition 17 mm corresponding to the report of Al-Zubairi et al [38] The data obtained for the methanol crudeextracts from AH BN and EI show their selective inhibitionof the test bacteria growth The plant extracts with ZI above13 mm against our panel of bacterial strains were selected forMICdeterminationHere themethanol crude extracts exhibitMIC values of 256 to gt 512 120583gmL against the test bacterialstrains but were less active as compared to the standardciprofloxacin 003 - 006 120583gmL (Table 5) Ciprofloxacin isan active broad-spectrum antibiotic use in the treatmentof urinary tract infections prostatitis and acute cystitis viainterfering with bacterial DNA replication and transcriptionthrough inhibition of DNA gyrase or topoisomerase [39] Allthe plant extracts demonstrated a mild to weak antibacterialactivity based on the classification of Rios and Recio [40] whoconsidered antibacterial activity of the crude plant extractsignificant (MIC lt 100 120583gmL) moderate (100 lt MIC = 512120583gmL) or weak (MIC gt 512 120583gmL) The herbal remediesshowed moderate inhibition of bacterial growth except for K

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

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Submit your manuscripts atwwwhindawicom

Page 6: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

6 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Gmelina

arborea

Verbenaceae

Gmelina

Stem

andleaf

Oral

Treatm

ento

fgon

orrhea

pilesabdo

minalpains

burningsensationsfever

mou

thwo

undanddiabetes

Snake-biteandscorpion

sting

Leaves

grou

ndinto

paste

with

water

are

appliedto

theforeheadforh

eadachein

feversB

oiledste

msa

retakenorally

Lophira

alata

Ochnaceae

Ekki

Stem

andleaf

Oral

Treatm

ento

ffeverm

outh

woun

dandmalaria

Boiltheleavesa

ndste

mandtake

orally

Nau

clealatifolia

Rubiaceae

Ubu

ru-in

uStem

andleaf

Oral

Itisused

inthetreatment

ofmalariaepilepsymou

thwo

undanxietypainfever

Boilther

ootsanddrink

Annona

murica

taAnn

onaceae

Sour

sop

Leaf

Oral

Treatm

ento

fcancera

ndmou

thwo

undinfections

Crushor

boiltheleavesa

ndtake

orally

Psidium

guajava

Myrtaceae

Guava

Leaf

Oral

Treatm

ento

fperiodo

ntal

diseasebu

rnsscrapes

sunb

urnwo

undspsoria

sis

eczemaandothers

Leaves

arec

rushed

andtakenorally

Astra

galusm

embran

aceus

Fabaceae

Beiq

iStem

andleaf

Oral

Treatm

ento

fcoldup

per

respira

tory

infections

allergiesfib

romyalgia

anem

iaH

IVA

IDS

Leaves

arec

rushed

andtakenorallyand

canalso

beappliedto

skin

Buchholziacoria

cea

Capparaceae

Won

derfu

lkola

Leaf

Oral

Itisused

tradition

allyfor

treatingdiabetes

hypertensio

nrheumatism

coldcou

ghand

catarrh

Boilleaves

andtake

orally

Gnetum

afric

anum

Gnetacea

Afang

Leaf

Oral

Enlarged

spleen

sore-th

roatandcathartic

Leaves

canbe

boiledor

cooked

andtaken

orally

Anoridium

man

niAnn

onaceae

Ewuro-ig

boStem

bark

Oral

Treatm

ento

fDiarrhea

coug

hfeverrheumatism

Theb

arkiscrushedandbo

iledandthen

takenorally

Funtum

iaafric

ana

Apocyn

aceae

Akorie

mbamiri

Leafstem

root

Skin

treatmento

fcon

stipatio

nwo

undsw

eakbladder

jaun

dice

Who

leplantisb

oiledandtakenorally

Also

thec

rushed

leaves

area

ppliedto

the

affectedskin

area

The Scientific World Journal 7

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Commiphora

afric

ana

Burseraceae

Turari

dashi

Rootsfruits

Oral

Treatm

ento

fwho

oping

coug

hbron

chitis

Rootsa

remacerated

andtakenorally

Dacryodesedulis

Burseraceae

Elem

iub

eBa

rkroo

tsfruits

Skin

Treatm

ento

fJiggersSkin

diseasesE

leph

antia

sis

Theleavesa

reeatenrawwith

kolanu

tas

anantie

meticLeaf-s

apisinstilledinto

thee

arfore

ar-tr

oubleanda

leaf-decoctio

nisprepared

asa

vapo

ur-bathforfeverish

stiffn

essw

ithheadache

Combretum

mucronatum

Com

bretaceae

Fararg

eza

Rootsa

ndleaf

Skin

Treatm

ento

fskininfection

Ther

ootand

leafarem

acerated

and

appliedto

affectedskin

area

Cleomec

iliata

Capparaceae

Ekuya

Leafseeds

Skin

Treatm

ento

fCon

vulsion

wo

undssores

Seedsa

recrushedandthep

oultice

appliedto

thea

ffected

areaTh

emacerated

leaves

after

boiledaretaken

orally

Ageratum

conyzoides

Aste

raceae

Urata

Who

leplant

Skin

Treatm

ento

fwou

ndU

lcer

Sleeping

sickn

essEyew

ash

Thep

lant

ismacerated

andappliedto

affectedskin

area

Aspilia

afric

ana

Aste

raceae

Yunyun

Ka

lank

uwa

Leafflow

erSkin

Treatm

ento

fskinrashes

cleaning

sorescorneal

opacities

Thefl

ower

issqueezed

andappliedto

affectedskin

area

Centau

reaperrottetii

Aste

raceae

Danyi

Who

leplant

Skin

Treatm

ento

fskin

infectionsSyphilis

Thefl

ower

issqueezed

andap

oultice

appliedto

affectedskin

area

Chrysanthellu

mindicum

Aste

raceae

Abilereoyigi

Who

leplant

Skin

Treatm

ento

fboils

gono

rrheaJaun

dice

Thep

lant

iscrushedandthep

oultice

applieddirectlyto

thes

kinarea

Elephantopus

scaber

Aste

raceae

Elephant

foot

Leafroo

tOral

Treatm

ento

ffevercou

ghLeafandroot

areb

oiledtogether

and

takenorally

Emiliacoccinea

Aste

raceae

Odu

ndun

Leafsap

and

root

Skin

Treatm

ento

fulcerhernia

measle

sWho

leplantissqu

eezedandsaptaken

orally

Acalypha

godseffi

ana

Euph

orbiaceae

Jinwinini

Leaf

Skin

Treatm

ento

fskin

infectionsantim

icrobials

Leafismacerated

andtakenorallyas

well

asappliedto

affectedskin

area

8 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Brideliafer

rugin

eaEu

phorbiaceae

Irikirni

Leafstem

bark

root

Oral

Treatm

ento

fInsom

nia

mou

thwashgono

rrhea

Who

leplantisb

oiledandtakenorally

Euphorbiaheterophylla

Euph

orbiaceae

Egele

Leafroo

tsSkin

Treatm

ento

fSkindiseases

Leafandroot

arem

acerated

andthe

poultic

eapp

liedto

thea

ffected

skin

area

Euphorbiahirta

Euph

orbiaceae

Non

onku

rciya

odne

Who

leplant

exud

ate

Oral

Treatm

ento

fAsth

ma

coug

hshapeo

fbreasts

Thew

holeplantism

acerated

andtaken

orally

Euphorbialaterifl

ora

Euph

orbiaceae

Orow

ere

Leafexud

ate

Skin

Treatm

ento

fdermatitis

constip

ation

Theleafissqu

eezedandthep

oultice

appliedto

thea

ffected

skin

area

Cajanu

scajan

Fabaceae

Orela

Leafseed

Skin

andoral

Treatm

ento

fSmallpox

and

mou

thwash

Leafissqueezed

andappliedto

the

affectedarea

Dan

iellia

thurifera

Fabaceae

Iya

Stew

-woo

ddu

stSkin

Treatm

ento

fscabies

Theb

arkismacerated

andappliedto

the

affectedskin

area

Dialiu

mguineense

Fabaceae

Tasm

iyar

kumi

Leafbark

fruitand

twig

Oral

Treatm

ento

fBronchitis

coug

handdiuretic

Theleafand

bark

arec

rushed

andbo

iled

togetherTh

emixture

isthen

takenorally

Acacianilotica

Fabaceae

Baanigabaruwa

Fruitbark

exud

ate

Skin

Treatm

ento

fskindiseases

fung

alinfections

Thee

xudatesa

resqueezed

from

theb

ark

andapplieddirectlyto

thea

ffected

skin

area

Napoleona

imperia

lisLecythidaceae

Mabun

giTw

igs

Oral

Treatm

ento

fAsth

ma

coug

hTw

igsa

remacerated

andtakenorally

Alliu

msativ

umLiliaceae

Ayoayuu

Bulb

Oral

Treatm

ento

ffevercou

gh

asthmaantim

icrobial

Theb

ulbismacerated

andtakenorally

Hibiscus

asper

Malvaceae

Isapad

angiraa

Leaf

Oraland

skin

Treatm

ento

fcou

gh

woun

dsanddiuretic

Leafisbo

iledandtakenorally

Hibiscus

rosasin

ensis

Malvaceae

Kekekeireagu

Leafstem

flowe

rSkin

Treatm

ento

finfl

uenza

woun

dsand

diuretic

Who

leplantism

acerated

andtaken

orally

Theleafissqu

eezedandappliedin

affectedskin

area

Hibiscus

sabdariffa

Malvaceae

Sobo

gurgu

zuLeafflow

erOral

Treatm

ento

fDiuretic

scoug

handdressin

gwo

unds

Leafisbo

iledandtakenorally

Ficuseleg

ans

Moraceae

Asoro

Leaf

Skinoral

Treatm

ento

fPile

constip

ationandcraw

-craw

Leafissqueezed

andappliedto

the

affectedarea

The Scientific World Journal 9

the absorbance The percentage inhibition of DPPH orABTS radical scavenging activity by the plant extracts wascalculated as (Abscontrol ndash Abssample)(Abscontrol) times 100where Abscontrol is the absorbance of DPPH or ABTS +methanol Abssample is the absorbance of DPPH or ABTSradical + sample extractstandard Here the concentration ofthe extracts needed to decrease the absorbance of DPPH orABTS radical by 50 was calculated Rutin (Sigma-Aldrichge 94 HPLC grade) at the same working concentrations ofthe plant extracts was used as reference drug

210 Bacterial Strains All bacterial strains were culturedon nutrient agar slopes and were incubated for 24 h at37∘C before the MIC An inoculum turbidity equivalent toa 05 McFarland standard (1times107 CFUmL) was prepared innormal saline for each test organism and then diluted 1100in Mueller-Hinton broth just before injection of the platesStandard strains used in the study included S aureus and Ecoli which were a gift from Dr EO Ekundayo (Departmentof Microbiology Michael Okpara University of Agriculture)and clinically isolated strains of K pneumoniae and Pmirabilis were gotten from Department of MicrobiologyFederal Medical Centre (FMC) Umuahia Abia State

211 Antibacterial Susceptibility Using Disc Diffusion MethodThe microbial growth inhibition potential of the plantextracts was determined by using the agar disc diffusionmethod [17] The plant extracts were dissolved in 10 vvDMSO (Merck UK Ltd) to a final concentration of 2 mgmLThe nutrient agar plate was streaked with the standardizedtest bacterial inoculum over the surface evenly and 2mgdiscof the plant extracts transferred onto sterile filter disc papers(about 6 mm) containing the test compounds was placed onthe agar surface in the Petri dishes 100 120583gdisc of standardantibiotic ciprofloxacin was used as control and 100 120583LDMSOdisc served as negative control The plates were thenincubated at 37∘C for 24 h Generally the antimicrobialextracts diffuse into the agar and inhibit the growth of thetest bacteria All tests were done in duplicate and diame-ter zones of inhibition of growth were measured using ameter rule from the edge of each dish after the incubationperiod

212 Minimum Inhibitory Concentration (MIC) Assays Wefollowed the method of Smith et al [18] to determine theMIC ofmethanol crude extracts of selected botanicals againstclinical isolates of bacteria associated with skin and oralinfection A volumeof 100mLof sterileMueller-Hinton broth(Oxoid Basingstoke UK) containing 20 mgL of Ca2+ and10 mgL of Mg2+ was dispensed into a 96-well microtitreplate (Nunc 03 mL total volume per well) All antibacterialagents were dissolved in dimethyl sulphoxide (DMSO) anddiluted in Mueller-Hinton broth to give a stock solutionThen 100 120583L of the antibacterial agent stock solution (2000mgL) was serially diluted into each well and 100 mL ofthe bacterial inoculum added to each well to give a finalconcentration range of 0024 - 1 mgL All procedures wereperformed in duplicate and the plates were incubated for 18

h at 37∘C Briefly 20 mL of a 5 mgL methanol solution of 3-[45-dimethylthiazol-2-yl]-25-diphenyltetrazolium bromide(MTT) (Sigma-Aldrich Ltd South Africa) was added to eachwell and then incubated for 30 min Blue colouration indi-cated bacterial growth The MIC was recorded as the lowestconcentration at which no colour change was observed

213 Statistical Analysis Data analysis was done inMicrosoftExcel to obtain descriptive statistics Means values wereseparated by the Duncan multiple tests using SAS The dif-ferent levels of significance within the groups were analyzedusing one-way analysis of variance (ANOVA) Values wereconsidered significant at P lt005

3 Results and Discussion

31 Ethnobotanical Survey Modern drugs are expensive andsupply to a remote area may be irregular hence result-ing in overreliance of traditional methods of healing as aprimary source of healthcare [19] The rural communitiesof Ebem-Ohafia region of southeastern Nigeria depend onherbal remedies to meet their domestic and health needsDuring our ethnobotanical survey a total of 30 respondentsincluding the elders herbalists and traditional healers wereinterviewed in the study areaMajority of the people (22)wereaged 30ndash65 years dominated by males (25) and one-third(10) had a primary school education while 15 had secondaryeducation and others informal education A total of 61medic-inal plant species belonging to 26 families were identified inthe treatment of oral and skin diseases as shown in Table 1Twenty-seven of them are used in the management of oralinfections and twenty-six plants for the management of skininfection while eight plants are used in the management ofboth infections Table 2 shows the list of family number of theplant species used and their percentage occurrenceThemostrepresented plant family is the Euphorbiaceae having 1803of the plant species followed by Asteraceae and Fabaceae with1147 each [20] Euphorbiaceae is a large family of floweringplants with 300 genera and about 7500 species found usefulin the curing of some common human diseases Asteraceae isa large family of flowering plants having 32 913 species and1911 genera and has been found to possess diverse biologicaleffects Fabaceae is a family of cosmopolitan distributionwith approximately 730 genera and 19400 species richnessat a global level [20] The leaves (60) recorded the mostcommonly used plant part followed by the root (16) andthen stem bark (12) None of the respondents indicated anypossible side effects caused by any of these plants listed inTable 1 The most common methods of preparation includeinfusion decoction and maceration These herbalists ortraditional healers often collect the plants from the fieldand dry and crush them into powder form before storingin bottles to prevent patients from identifying the plantsused for their treatment Most healers refused to provideinformation without payment whereas others thought toprovide information on the plant use is disrespect of theirtraditional belief Furthermore the majority of the plantsmentioned lack scientific data to substantiate their acclaimedfolkloric use From this work it is evident that medicinal

10 The Scientific World Journal

Table 2 The list of family number of the plant species and theirpercentage occurrence

Family Number of plant species OccurrenceFabaceae 7 1147Euphorbiaceae 11 1803Asteraceae 7 1147Myrtaceae 4 656Moraceae 3 492Malvaceae 4 656Liliaceae 2 328Capparaceae 2 328Annonaceae 3 492Combretaceae 1 164Burseraceae 2 328Apocynaceae 1 164Gnetaceae 1 164Lecythidaceae 1 164Verbenaceae 1 164Ochnaceae 1 164Rubiaceae 1 164Piperaceae 1 164Acanthaceae 2 328Papilionoideae 2 328Asclepiadaceae 1 164Caesalpiniaceae 2 328Convolvulaceae 1 164Rutaceae 1 164Crassulaceae 1 164Poaceae 1 164

plants play a significant role in the primary healthcare ofthe people and conservation of the indigenous knowledge ofthis folklore medicine is crucial for the innovations of futuredrug discovery and possible herbal plants trade in the region[19] On the contrary we observed imprecise dosage lowhygiene standards hiding of healing methods and lack ofpatients record as a significant setback to traditional healthpractitioners in the region

32 Thin Layer Chromatography (TLC) Profiling Thin layerchromatography remains the most common useful techniqueby researchers as a quick and simple method to resolvethe constituents of a crude extract followed by advancedextraction methods to purify the active ingredients Previousstudies reported that specific secondary metabolites in plantexhibit a wide range of antioxidant and antibacterial proper-ties [21] TLC was formed on a glass sheet coated with silicagel After the sample is applied to the plate a solvent system(mobile phase) is drawn up via capillary action on the TLCplate at different mobility rate depending on the chemicalcompositionsThe separated spotswith different colourswerevisualized by the UV light projection onto the glass sheetThe variation of coloured spots detected in the crude extractsindicates the presence of different phytochemicals that may

be responsible for the pharmacological effects of selectedplants use in the management of oral and skin infections asacclaimed by the traditional healers We quantified the resultsby measuring the distance travelled by the plant compoundsdivided by the total distance travelled by the mobile phaseFrom Table 3 it is apparent that mobile phase 1 consistingof chloroform and ethanol in the volume ratio of 64 had abetter separation than mobile phase II containing tolueneethyl acetate and acetic acid in the volume ratio of 541 Themethanol extract of AH in bothmobile phases had the highestnumber of three bands with Rf values of 0667 0833 and0989 for mobile phase I and Rf values of 0381 0648 and0781 for mobile phase IIThe plant extracts MM BA CO JCandCAhad two bands inmobile phase I with clear separationbut had single band in mobile phase II The plant extractshaving only one band in either mobile phase suggest a poorsolvent system for compounds separation The TLC profilesprovide a characteristic fingerprint of these plants that mayalso be useful for their identification

33 Antioxidant Assays

331 TPC and TFC Phenolic compounds are secondarymetabolites distributed in fruits vegetables nuts herbsseeds and beverages [22] They are excellent scavengers offree radical that has been implicated in the pathologies ofvarious human diseases the antioxidant activity is dependenton the structure hydrophobic benzenoid rings and hydrogen-bonding potential of the phenolic hydroxyl groups In TPCassay a phenol in plant extracts loses a proton to formphenolate ion which reduces Folin Ciocalteu reagent andthe resulting changes are measured spectrophotometricallyat 765 nm [13] The content of phenol is reported in mg oftannic acid equivalent (TAE)mL of extract As shown inFigure 1 the total phenolic contents of absolute methanolcrude extract of these botanicals range from 700 to 3600mgTAEg The AH extract had the highest concentrationfollowed by CA CO MM BNI BA EI BN and JC whilethe least concentration was recorded in II extract Moreoverthe Folin Ciocalteu reagent (FCR) has been used to measureother compounds such as ascorbic acid amino acids andsugars hence it may not provide a concise amount of thephenolic compounds present in these plants Flavonoids areincreasingly consumed in a significant amount in daily dietsworldwide due to their various health benefits toman [23 24]In TFC assay aluminum ion in the reacting mixture formscomplexes with the C- 4 keto and either C- 3 or C- 5 hydroxylor with ortho hydroxyl groups in the A or B ring of flavonoidsdetected in the plant extracts [14] As shown in Figure 2the total flavonoid content was recorded in the plant extractsranging from 750 to 1500 mgQEg as extrapolated from thestandard quercetin curve The CA extract had the highestconcentration followed by BNI CO II AH BN MM JCand BA while the lowest concentration was recorded in EIextract TPC is found higher than the TFC in some of theextracts supporting the fact that most flavonoids are alsophenolics According to our results all the plant extractsare rich source of phenolic compounds The composition ofTPC and TFC registered in these plants may be difficult to

The Scientific World Journal 11

Table 3 Thin layer chromatography retention factor using mobile phase CHL ET (mobile phase I) and Tol EA AA (mobile phase II)

SN Plant Extract CHL ET (64) Tol EA AA (541)1 Baphia nitida 1 0742 Macrolobium macrophyllum 0706 0941 06953 Chromolaena odorata 0471 0647 06674 Jatropha curcas 0353 0941 06485 Cassia alata 0118 0882 06866 Ipomoea involucrata 0956 -7 Acalypha hispida 0667 0833 0989 0381 0648 07818 Breynia nivosa 0989 07629 Burkea africana 0644 1 075210 Eleusine indica 0978 0743CHL chloroform ET ethanol Tol toluene EA ethylacetate AA acetic acid

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

500

1000

1500

2000

2500

3000

3500

4000

TPC

(mgT

AE

g)

Figure 1 The total phenolic content (TPC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata TAE tannic acid equivalent

compare with others reported elsewhere due to many factorsincluding differences from species to species maturity stageplant parts and assay methods harvesting time standarddrugs and geographical location

332 Reducing Power (RP) Ferric reducing power is a globalmethod often used to assess the capacity of antioxidantsto donate either an electron or hydrogen atom to unpairedelectron [25] In reducing power assay plant extracts exertantioxidant activities via reduction of Fe3+ferric cyanidecomplex to Fe2+ (ferrous form) resulting in changing thegreen colour to blue and sometimes Perlrsquos Prussian bluecolour depending on the antioxidant potential [10] As shownin Figure 3 the reducing power of the methanol crudeextracts and the standard gallic acid rutin and ascorbic acidincreased with increase in concentration Higher absorbanceat 700 nm can be read after potassium ferricyanide reactionwith ferric chloride At 1000 120583gmL the extracts from AH

0

200

400

600

800

1000

1200

1400

1600

TFC

(mgQ

Eg)

AH BN JC CO MM EI BNI BA CAIIPlant extracts

Figure 2 The total flavonoid content (TFC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata QE Quercetin equivalent

BNI MM CA BA EI and BN had absorbance value of 270086 073 055 036 027 and 027 respectively while that ofAH is higher than the standard rutin (254) but comparablewell with gallic acid (275) and ascorbic acid (269) EI andBN extracts had a similar trend of ferric reducing powerthough significantly lower as compared with the standarddrugs AH extract exhibits remarkable antioxidant capacity(RP) correlating with its richness in phenolic and flavonoidcontents It is reasonable to suggest that methanol selectivelyextracts antioxidant principles from these botanicals basedon their polarities and chemical structures which are reflectedin plant extracts with high TFC and TPC having minimal RP[26] We have shown that some plant extracts in particularAH BNI MM and CA extracts may serve as an excellentcandidate to minimize skin and oral infections caused byoxidative stress as a result of imbalance of free radicalsgeneration and natural antioxidants in the saliva or woundarea According to our results the increasing RP of the plantextracts correlateswith their TPCandTFCThis study at least

12 The Scientific World JournalRe

duci

ng P

ower

7

00nm

625 125 250 500 10003125Concentration (gmL)

0

05

1

15

2

25

3

IIAHBNJCCOMMEI

BNIBACAGARutinVit C

Figure 3 The ferric reducing antioxidative capacity of selectedplant extracts used in the management of oral and skin infectionsin southeast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata GA gallic acid Vit C vitamin C (standard antioxidants)

in part lends credence to the ethno-therapeutic use of someplant as natural antioxidant in themanagement of skin or oralinfections in the study area

333 FRSA of Plant Extracts Skin and oral cavity exposedto an array of oxidants such as smokes from cigaretteschemicals UV light nicotine and alcohol that are directlyor indirectly leading to the production of reactive oxygenspecies (ROS) are known to be responsible for some com-mon human diseases [27] These free radicals in particularhydroxyl and superoxide anions are highly reactivemoleculesderived from the metabolism of oxygen Although they playan important role in the biochemical immunological andphysiological processes it could be deleterious to vital organsif excessively accumulated in the human body [28] Food andpharmaceutical industries discovered that medicinal plantscontaining phenol and flavonoid compounds are essentialin the development of novel drugs against stress-induceddiseases [28] In the present investigation the concentration(IC50) at which 50 of free radical scavenged was calculatedand extrapolated from the plot of percentage inhibition ver-sus plant extract concentration The herbal drugs exhibitedrobust DPPH radical scavenging activity in a concentration-dependent manner having IC50 ranging from 2025 to 710120583gmL As shown in Figure 4 MM BA EI JC AH BNIand CA extracts had a potent DPPH radical scavengingactivity corresponding to their TPC TFC and RP Theplant extracts scavenged ABTS radical by the decrease inits absorbance at 734 nm All the plant extracts exhibitedstrong ABTS radical scavenging capacities in the following

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

200

400

600

800

1000

1200

1400

1600

IC50

(g

mL)

Figure 4 DPPH free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

050

100150200250300350400450500

IC50

(g

mL)

AH BN JC CO MM EI BNI BA CAIIPlant samples

Figure 5 ABTS free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

order BN gt JC gt CA gt MM gt BNI gt EI gt CO gt AH gtII gt BA having IC50 values ranging from 25 to 400 120583gmL(Figure 5) DPPH and ABTS radical scavenging activities ofthese herbal remedies displayed a positive correlation withthe working concentrations It is obvious that most of theplant extracts tested in this work possess the ability to quenchthe free radicals generation via electron or proton donationdemonstrated by the termination of radical chain reaction[28] Although both DPPH and ABTS radicals are electron-transfer based methods however it is apparent that ABTSmethod seems to be more sensitive than DPPH radical assaypossibly due to their solubility and diffusivity as shown inthe IC50 values [10] It is worth noting that BA extract withstrong DPPH radical scavenging effect had a moderate ABTS

The Scientific World Journal 13

Table 4 Zone of bacterial inhibition by the selected plant extracts use in the management of oral and skin infections

Diameter of inhibition (mm)SN Plant samples K pneumoniae P mirabilis S aureus E coli1 Cassia alata 15 plusmn 141 9 plusmn 042 15 plusmn 083 13 plusmn 0802 Breynia nivosa 8 plusmn 052 17 plusmn 100 8 plusmn 008 15 plusmn 1023 Burkea africana NI 11 plusmn 084 13 plusmn 072 NI4 Baphia nitida 8 plusmn 050 13 plusmn 122 8 plusmn 051 NI5 Ipomoea involucrata 8 plusmn 052 NI 8 plusmn 050 13 plusmn 0526 Jatropha curcas NI 9 plusmn 030 9 plusmn 062 13 plusmn 0727 Acalypha hispida 21 plusmn 163 NI 11 plusmn 009 17 plusmn 1328 Chromolaena odorata 8 plusmn 042 8 plusmn 035 9 plusmn 009 15 plusmn1209 Eleusine indica 8 plusmn 042 8 plusmn 035 NI 17 plusmn 10010 Macrolobium macrophyllum NI NI 8 plusmn 004 15 plusmn 12311 Ciprofloxacin - - le16(R) ge17(S) le16(R)ge20(S)NI indicates no zone of inhibition S sensitive R resistance

radical scavenging capacity which can be explained by thepartial ionization of the extract in the test medium [29] SinceROS promotes tissue damage and epithelial inflammationvia exposure to sunlight radiation or smoke as a result theantioxidant capacity demonstrated in this work may supportthe traditional use of these plants against the influence ofinflammatory markers and oxidants in the skin and oralcavity [29] Based on the results of the three antioxidantassays it is obvious that MM AH BNI EI and CA extractshad the best FRSA indicating their importance in convertingfree radicals into more stable products and then terminatethe free radical chain reaction Our study showed that themethanol crude extracts from these plants could protectagainst free radical damage in the mouth such as humangingival periodontal tissues and certain skin infections [5]

334 Antibacterial Activity of Plant Extracts Pathogenicbacteria such as Staphylococcus aureus Escherichia coli Kleb-siella pneumoniae and Proteus mirabilis play a significantrole as the primary cause of skin and oral infections [30]Staphylococcus aureus is a part of normal skin known to bethe most problematic in the cause of skin infections as aresult of their resistance mechanisms to first-line antibiotics[30] Although the presence of this organism as a residentoral floral is controversial nonetheless MSSA and MRSAhave been isolated from oral rinse and tongue swab inpatients with oral infections [31] Escherichia coli are Gram-negative bacteria known to be notorious for their resistantmechanisms towards the existing arsenal of antibiotics Oralinfections such as edematous and erythematous and skininfections like omphalitis surgical site infections burnnecrotizing fasciitis and cellulitis at all age groups are associ-ated with enterobacterium E coli [32] Klebsiella pneumoniaeis a Gram-negative encapsulated facultative anaerobic androd-shaped bacterium found in the normal flora of themouth skin and intestines This bacterium is responsiblefor the prolonged surgical wound biliary tract wound andurinary tract infections and it could pose a serious healthrisk when resistant strains invade the throat mouth or

intestines [33] Villegas et al [34] detected a CTX-M-12 120573-lactamase producing K pneumoniae new plasmid-mediatedESBLs highly resistant to ceftazidime from a patient withhospital-acquired wound infection Proteus mirabilis is oneof the most common enterobacteria after E coli identified asGram-negativemotile and rod-shaped bacterium It is foundin clinical specimen implicated in the skin respiratory tractand bacteraemia as well as burns wounds and urinary tractinfections [35] All these pathogenic multidrug-resistancebacteria are causative agent in a periodontal lips ulcer whitesponge nevus and skin infections [36]

335 Susceptibility Test and MIC Determination The resultof disc diffusion assay showed the diameter of inhibitionof bacteria growth classified as le 15 (resistant) and ge 16(susceptible) according to CLSI guideline [37] As shown inTable 4 K pneumoniae and E coli are more susceptible tothe treatment of Acalypha hispida (AH) having a zone ofinhibition (ZI) 21 and 17mm respectively BN and EI extractsinhibit the growth of P mirabilis and E coli having the samezone of inhibition 17 mm corresponding to the report of Al-Zubairi et al [38] The data obtained for the methanol crudeextracts from AH BN and EI show their selective inhibitionof the test bacteria growth The plant extracts with ZI above13 mm against our panel of bacterial strains were selected forMICdeterminationHere themethanol crude extracts exhibitMIC values of 256 to gt 512 120583gmL against the test bacterialstrains but were less active as compared to the standardciprofloxacin 003 - 006 120583gmL (Table 5) Ciprofloxacin isan active broad-spectrum antibiotic use in the treatmentof urinary tract infections prostatitis and acute cystitis viainterfering with bacterial DNA replication and transcriptionthrough inhibition of DNA gyrase or topoisomerase [39] Allthe plant extracts demonstrated a mild to weak antibacterialactivity based on the classification of Rios and Recio [40] whoconsidered antibacterial activity of the crude plant extractsignificant (MIC lt 100 120583gmL) moderate (100 lt MIC = 512120583gmL) or weak (MIC gt 512 120583gmL) The herbal remediesshowed moderate inhibition of bacterial growth except for K

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

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Page 7: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

The Scientific World Journal 7

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Commiphora

afric

ana

Burseraceae

Turari

dashi

Rootsfruits

Oral

Treatm

ento

fwho

oping

coug

hbron

chitis

Rootsa

remacerated

andtakenorally

Dacryodesedulis

Burseraceae

Elem

iub

eBa

rkroo

tsfruits

Skin

Treatm

ento

fJiggersSkin

diseasesE

leph

antia

sis

Theleavesa

reeatenrawwith

kolanu

tas

anantie

meticLeaf-s

apisinstilledinto

thee

arfore

ar-tr

oubleanda

leaf-decoctio

nisprepared

asa

vapo

ur-bathforfeverish

stiffn

essw

ithheadache

Combretum

mucronatum

Com

bretaceae

Fararg

eza

Rootsa

ndleaf

Skin

Treatm

ento

fskininfection

Ther

ootand

leafarem

acerated

and

appliedto

affectedskin

area

Cleomec

iliata

Capparaceae

Ekuya

Leafseeds

Skin

Treatm

ento

fCon

vulsion

wo

undssores

Seedsa

recrushedandthep

oultice

appliedto

thea

ffected

areaTh

emacerated

leaves

after

boiledaretaken

orally

Ageratum

conyzoides

Aste

raceae

Urata

Who

leplant

Skin

Treatm

ento

fwou

ndU

lcer

Sleeping

sickn

essEyew

ash

Thep

lant

ismacerated

andappliedto

affectedskin

area

Aspilia

afric

ana

Aste

raceae

Yunyun

Ka

lank

uwa

Leafflow

erSkin

Treatm

ento

fskinrashes

cleaning

sorescorneal

opacities

Thefl

ower

issqueezed

andappliedto

affectedskin

area

Centau

reaperrottetii

Aste

raceae

Danyi

Who

leplant

Skin

Treatm

ento

fskin

infectionsSyphilis

Thefl

ower

issqueezed

andap

oultice

appliedto

affectedskin

area

Chrysanthellu

mindicum

Aste

raceae

Abilereoyigi

Who

leplant

Skin

Treatm

ento

fboils

gono

rrheaJaun

dice

Thep

lant

iscrushedandthep

oultice

applieddirectlyto

thes

kinarea

Elephantopus

scaber

Aste

raceae

Elephant

foot

Leafroo

tOral

Treatm

ento

ffevercou

ghLeafandroot

areb

oiledtogether

and

takenorally

Emiliacoccinea

Aste

raceae

Odu

ndun

Leafsap

and

root

Skin

Treatm

ento

fulcerhernia

measle

sWho

leplantissqu

eezedandsaptaken

orally

Acalypha

godseffi

ana

Euph

orbiaceae

Jinwinini

Leaf

Skin

Treatm

ento

fskin

infectionsantim

icrobials

Leafismacerated

andtakenorallyas

well

asappliedto

affectedskin

area

8 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Brideliafer

rugin

eaEu

phorbiaceae

Irikirni

Leafstem

bark

root

Oral

Treatm

ento

fInsom

nia

mou

thwashgono

rrhea

Who

leplantisb

oiledandtakenorally

Euphorbiaheterophylla

Euph

orbiaceae

Egele

Leafroo

tsSkin

Treatm

ento

fSkindiseases

Leafandroot

arem

acerated

andthe

poultic

eapp

liedto

thea

ffected

skin

area

Euphorbiahirta

Euph

orbiaceae

Non

onku

rciya

odne

Who

leplant

exud

ate

Oral

Treatm

ento

fAsth

ma

coug

hshapeo

fbreasts

Thew

holeplantism

acerated

andtaken

orally

Euphorbialaterifl

ora

Euph

orbiaceae

Orow

ere

Leafexud

ate

Skin

Treatm

ento

fdermatitis

constip

ation

Theleafissqu

eezedandthep

oultice

appliedto

thea

ffected

skin

area

Cajanu

scajan

Fabaceae

Orela

Leafseed

Skin

andoral

Treatm

ento

fSmallpox

and

mou

thwash

Leafissqueezed

andappliedto

the

affectedarea

Dan

iellia

thurifera

Fabaceae

Iya

Stew

-woo

ddu

stSkin

Treatm

ento

fscabies

Theb

arkismacerated

andappliedto

the

affectedskin

area

Dialiu

mguineense

Fabaceae

Tasm

iyar

kumi

Leafbark

fruitand

twig

Oral

Treatm

ento

fBronchitis

coug

handdiuretic

Theleafand

bark

arec

rushed

andbo

iled

togetherTh

emixture

isthen

takenorally

Acacianilotica

Fabaceae

Baanigabaruwa

Fruitbark

exud

ate

Skin

Treatm

ento

fskindiseases

fung

alinfections

Thee

xudatesa

resqueezed

from

theb

ark

andapplieddirectlyto

thea

ffected

skin

area

Napoleona

imperia

lisLecythidaceae

Mabun

giTw

igs

Oral

Treatm

ento

fAsth

ma

coug

hTw

igsa

remacerated

andtakenorally

Alliu

msativ

umLiliaceae

Ayoayuu

Bulb

Oral

Treatm

ento

ffevercou

gh

asthmaantim

icrobial

Theb

ulbismacerated

andtakenorally

Hibiscus

asper

Malvaceae

Isapad

angiraa

Leaf

Oraland

skin

Treatm

ento

fcou

gh

woun

dsanddiuretic

Leafisbo

iledandtakenorally

Hibiscus

rosasin

ensis

Malvaceae

Kekekeireagu

Leafstem

flowe

rSkin

Treatm

ento

finfl

uenza

woun

dsand

diuretic

Who

leplantism

acerated

andtaken

orally

Theleafissqu

eezedandappliedin

affectedskin

area

Hibiscus

sabdariffa

Malvaceae

Sobo

gurgu

zuLeafflow

erOral

Treatm

ento

fDiuretic

scoug

handdressin

gwo

unds

Leafisbo

iledandtakenorally

Ficuseleg

ans

Moraceae

Asoro

Leaf

Skinoral

Treatm

ento

fPile

constip

ationandcraw

-craw

Leafissqueezed

andappliedto

the

affectedarea

The Scientific World Journal 9

the absorbance The percentage inhibition of DPPH orABTS radical scavenging activity by the plant extracts wascalculated as (Abscontrol ndash Abssample)(Abscontrol) times 100where Abscontrol is the absorbance of DPPH or ABTS +methanol Abssample is the absorbance of DPPH or ABTSradical + sample extractstandard Here the concentration ofthe extracts needed to decrease the absorbance of DPPH orABTS radical by 50 was calculated Rutin (Sigma-Aldrichge 94 HPLC grade) at the same working concentrations ofthe plant extracts was used as reference drug

210 Bacterial Strains All bacterial strains were culturedon nutrient agar slopes and were incubated for 24 h at37∘C before the MIC An inoculum turbidity equivalent toa 05 McFarland standard (1times107 CFUmL) was prepared innormal saline for each test organism and then diluted 1100in Mueller-Hinton broth just before injection of the platesStandard strains used in the study included S aureus and Ecoli which were a gift from Dr EO Ekundayo (Departmentof Microbiology Michael Okpara University of Agriculture)and clinically isolated strains of K pneumoniae and Pmirabilis were gotten from Department of MicrobiologyFederal Medical Centre (FMC) Umuahia Abia State

211 Antibacterial Susceptibility Using Disc Diffusion MethodThe microbial growth inhibition potential of the plantextracts was determined by using the agar disc diffusionmethod [17] The plant extracts were dissolved in 10 vvDMSO (Merck UK Ltd) to a final concentration of 2 mgmLThe nutrient agar plate was streaked with the standardizedtest bacterial inoculum over the surface evenly and 2mgdiscof the plant extracts transferred onto sterile filter disc papers(about 6 mm) containing the test compounds was placed onthe agar surface in the Petri dishes 100 120583gdisc of standardantibiotic ciprofloxacin was used as control and 100 120583LDMSOdisc served as negative control The plates were thenincubated at 37∘C for 24 h Generally the antimicrobialextracts diffuse into the agar and inhibit the growth of thetest bacteria All tests were done in duplicate and diame-ter zones of inhibition of growth were measured using ameter rule from the edge of each dish after the incubationperiod

212 Minimum Inhibitory Concentration (MIC) Assays Wefollowed the method of Smith et al [18] to determine theMIC ofmethanol crude extracts of selected botanicals againstclinical isolates of bacteria associated with skin and oralinfection A volumeof 100mLof sterileMueller-Hinton broth(Oxoid Basingstoke UK) containing 20 mgL of Ca2+ and10 mgL of Mg2+ was dispensed into a 96-well microtitreplate (Nunc 03 mL total volume per well) All antibacterialagents were dissolved in dimethyl sulphoxide (DMSO) anddiluted in Mueller-Hinton broth to give a stock solutionThen 100 120583L of the antibacterial agent stock solution (2000mgL) was serially diluted into each well and 100 mL ofthe bacterial inoculum added to each well to give a finalconcentration range of 0024 - 1 mgL All procedures wereperformed in duplicate and the plates were incubated for 18

h at 37∘C Briefly 20 mL of a 5 mgL methanol solution of 3-[45-dimethylthiazol-2-yl]-25-diphenyltetrazolium bromide(MTT) (Sigma-Aldrich Ltd South Africa) was added to eachwell and then incubated for 30 min Blue colouration indi-cated bacterial growth The MIC was recorded as the lowestconcentration at which no colour change was observed

213 Statistical Analysis Data analysis was done inMicrosoftExcel to obtain descriptive statistics Means values wereseparated by the Duncan multiple tests using SAS The dif-ferent levels of significance within the groups were analyzedusing one-way analysis of variance (ANOVA) Values wereconsidered significant at P lt005

3 Results and Discussion

31 Ethnobotanical Survey Modern drugs are expensive andsupply to a remote area may be irregular hence result-ing in overreliance of traditional methods of healing as aprimary source of healthcare [19] The rural communitiesof Ebem-Ohafia region of southeastern Nigeria depend onherbal remedies to meet their domestic and health needsDuring our ethnobotanical survey a total of 30 respondentsincluding the elders herbalists and traditional healers wereinterviewed in the study areaMajority of the people (22)wereaged 30ndash65 years dominated by males (25) and one-third(10) had a primary school education while 15 had secondaryeducation and others informal education A total of 61medic-inal plant species belonging to 26 families were identified inthe treatment of oral and skin diseases as shown in Table 1Twenty-seven of them are used in the management of oralinfections and twenty-six plants for the management of skininfection while eight plants are used in the management ofboth infections Table 2 shows the list of family number of theplant species used and their percentage occurrenceThemostrepresented plant family is the Euphorbiaceae having 1803of the plant species followed by Asteraceae and Fabaceae with1147 each [20] Euphorbiaceae is a large family of floweringplants with 300 genera and about 7500 species found usefulin the curing of some common human diseases Asteraceae isa large family of flowering plants having 32 913 species and1911 genera and has been found to possess diverse biologicaleffects Fabaceae is a family of cosmopolitan distributionwith approximately 730 genera and 19400 species richnessat a global level [20] The leaves (60) recorded the mostcommonly used plant part followed by the root (16) andthen stem bark (12) None of the respondents indicated anypossible side effects caused by any of these plants listed inTable 1 The most common methods of preparation includeinfusion decoction and maceration These herbalists ortraditional healers often collect the plants from the fieldand dry and crush them into powder form before storingin bottles to prevent patients from identifying the plantsused for their treatment Most healers refused to provideinformation without payment whereas others thought toprovide information on the plant use is disrespect of theirtraditional belief Furthermore the majority of the plantsmentioned lack scientific data to substantiate their acclaimedfolkloric use From this work it is evident that medicinal

10 The Scientific World Journal

Table 2 The list of family number of the plant species and theirpercentage occurrence

Family Number of plant species OccurrenceFabaceae 7 1147Euphorbiaceae 11 1803Asteraceae 7 1147Myrtaceae 4 656Moraceae 3 492Malvaceae 4 656Liliaceae 2 328Capparaceae 2 328Annonaceae 3 492Combretaceae 1 164Burseraceae 2 328Apocynaceae 1 164Gnetaceae 1 164Lecythidaceae 1 164Verbenaceae 1 164Ochnaceae 1 164Rubiaceae 1 164Piperaceae 1 164Acanthaceae 2 328Papilionoideae 2 328Asclepiadaceae 1 164Caesalpiniaceae 2 328Convolvulaceae 1 164Rutaceae 1 164Crassulaceae 1 164Poaceae 1 164

plants play a significant role in the primary healthcare ofthe people and conservation of the indigenous knowledge ofthis folklore medicine is crucial for the innovations of futuredrug discovery and possible herbal plants trade in the region[19] On the contrary we observed imprecise dosage lowhygiene standards hiding of healing methods and lack ofpatients record as a significant setback to traditional healthpractitioners in the region

32 Thin Layer Chromatography (TLC) Profiling Thin layerchromatography remains the most common useful techniqueby researchers as a quick and simple method to resolvethe constituents of a crude extract followed by advancedextraction methods to purify the active ingredients Previousstudies reported that specific secondary metabolites in plantexhibit a wide range of antioxidant and antibacterial proper-ties [21] TLC was formed on a glass sheet coated with silicagel After the sample is applied to the plate a solvent system(mobile phase) is drawn up via capillary action on the TLCplate at different mobility rate depending on the chemicalcompositionsThe separated spotswith different colourswerevisualized by the UV light projection onto the glass sheetThe variation of coloured spots detected in the crude extractsindicates the presence of different phytochemicals that may

be responsible for the pharmacological effects of selectedplants use in the management of oral and skin infections asacclaimed by the traditional healers We quantified the resultsby measuring the distance travelled by the plant compoundsdivided by the total distance travelled by the mobile phaseFrom Table 3 it is apparent that mobile phase 1 consistingof chloroform and ethanol in the volume ratio of 64 had abetter separation than mobile phase II containing tolueneethyl acetate and acetic acid in the volume ratio of 541 Themethanol extract of AH in bothmobile phases had the highestnumber of three bands with Rf values of 0667 0833 and0989 for mobile phase I and Rf values of 0381 0648 and0781 for mobile phase IIThe plant extracts MM BA CO JCandCAhad two bands inmobile phase I with clear separationbut had single band in mobile phase II The plant extractshaving only one band in either mobile phase suggest a poorsolvent system for compounds separation The TLC profilesprovide a characteristic fingerprint of these plants that mayalso be useful for their identification

33 Antioxidant Assays

331 TPC and TFC Phenolic compounds are secondarymetabolites distributed in fruits vegetables nuts herbsseeds and beverages [22] They are excellent scavengers offree radical that has been implicated in the pathologies ofvarious human diseases the antioxidant activity is dependenton the structure hydrophobic benzenoid rings and hydrogen-bonding potential of the phenolic hydroxyl groups In TPCassay a phenol in plant extracts loses a proton to formphenolate ion which reduces Folin Ciocalteu reagent andthe resulting changes are measured spectrophotometricallyat 765 nm [13] The content of phenol is reported in mg oftannic acid equivalent (TAE)mL of extract As shown inFigure 1 the total phenolic contents of absolute methanolcrude extract of these botanicals range from 700 to 3600mgTAEg The AH extract had the highest concentrationfollowed by CA CO MM BNI BA EI BN and JC whilethe least concentration was recorded in II extract Moreoverthe Folin Ciocalteu reagent (FCR) has been used to measureother compounds such as ascorbic acid amino acids andsugars hence it may not provide a concise amount of thephenolic compounds present in these plants Flavonoids areincreasingly consumed in a significant amount in daily dietsworldwide due to their various health benefits toman [23 24]In TFC assay aluminum ion in the reacting mixture formscomplexes with the C- 4 keto and either C- 3 or C- 5 hydroxylor with ortho hydroxyl groups in the A or B ring of flavonoidsdetected in the plant extracts [14] As shown in Figure 2the total flavonoid content was recorded in the plant extractsranging from 750 to 1500 mgQEg as extrapolated from thestandard quercetin curve The CA extract had the highestconcentration followed by BNI CO II AH BN MM JCand BA while the lowest concentration was recorded in EIextract TPC is found higher than the TFC in some of theextracts supporting the fact that most flavonoids are alsophenolics According to our results all the plant extractsare rich source of phenolic compounds The composition ofTPC and TFC registered in these plants may be difficult to

The Scientific World Journal 11

Table 3 Thin layer chromatography retention factor using mobile phase CHL ET (mobile phase I) and Tol EA AA (mobile phase II)

SN Plant Extract CHL ET (64) Tol EA AA (541)1 Baphia nitida 1 0742 Macrolobium macrophyllum 0706 0941 06953 Chromolaena odorata 0471 0647 06674 Jatropha curcas 0353 0941 06485 Cassia alata 0118 0882 06866 Ipomoea involucrata 0956 -7 Acalypha hispida 0667 0833 0989 0381 0648 07818 Breynia nivosa 0989 07629 Burkea africana 0644 1 075210 Eleusine indica 0978 0743CHL chloroform ET ethanol Tol toluene EA ethylacetate AA acetic acid

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

500

1000

1500

2000

2500

3000

3500

4000

TPC

(mgT

AE

g)

Figure 1 The total phenolic content (TPC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata TAE tannic acid equivalent

compare with others reported elsewhere due to many factorsincluding differences from species to species maturity stageplant parts and assay methods harvesting time standarddrugs and geographical location

332 Reducing Power (RP) Ferric reducing power is a globalmethod often used to assess the capacity of antioxidantsto donate either an electron or hydrogen atom to unpairedelectron [25] In reducing power assay plant extracts exertantioxidant activities via reduction of Fe3+ferric cyanidecomplex to Fe2+ (ferrous form) resulting in changing thegreen colour to blue and sometimes Perlrsquos Prussian bluecolour depending on the antioxidant potential [10] As shownin Figure 3 the reducing power of the methanol crudeextracts and the standard gallic acid rutin and ascorbic acidincreased with increase in concentration Higher absorbanceat 700 nm can be read after potassium ferricyanide reactionwith ferric chloride At 1000 120583gmL the extracts from AH

0

200

400

600

800

1000

1200

1400

1600

TFC

(mgQ

Eg)

AH BN JC CO MM EI BNI BA CAIIPlant extracts

Figure 2 The total flavonoid content (TFC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata QE Quercetin equivalent

BNI MM CA BA EI and BN had absorbance value of 270086 073 055 036 027 and 027 respectively while that ofAH is higher than the standard rutin (254) but comparablewell with gallic acid (275) and ascorbic acid (269) EI andBN extracts had a similar trend of ferric reducing powerthough significantly lower as compared with the standarddrugs AH extract exhibits remarkable antioxidant capacity(RP) correlating with its richness in phenolic and flavonoidcontents It is reasonable to suggest that methanol selectivelyextracts antioxidant principles from these botanicals basedon their polarities and chemical structures which are reflectedin plant extracts with high TFC and TPC having minimal RP[26] We have shown that some plant extracts in particularAH BNI MM and CA extracts may serve as an excellentcandidate to minimize skin and oral infections caused byoxidative stress as a result of imbalance of free radicalsgeneration and natural antioxidants in the saliva or woundarea According to our results the increasing RP of the plantextracts correlateswith their TPCandTFCThis study at least

12 The Scientific World JournalRe

duci

ng P

ower

7

00nm

625 125 250 500 10003125Concentration (gmL)

0

05

1

15

2

25

3

IIAHBNJCCOMMEI

BNIBACAGARutinVit C

Figure 3 The ferric reducing antioxidative capacity of selectedplant extracts used in the management of oral and skin infectionsin southeast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata GA gallic acid Vit C vitamin C (standard antioxidants)

in part lends credence to the ethno-therapeutic use of someplant as natural antioxidant in themanagement of skin or oralinfections in the study area

333 FRSA of Plant Extracts Skin and oral cavity exposedto an array of oxidants such as smokes from cigaretteschemicals UV light nicotine and alcohol that are directlyor indirectly leading to the production of reactive oxygenspecies (ROS) are known to be responsible for some com-mon human diseases [27] These free radicals in particularhydroxyl and superoxide anions are highly reactivemoleculesderived from the metabolism of oxygen Although they playan important role in the biochemical immunological andphysiological processes it could be deleterious to vital organsif excessively accumulated in the human body [28] Food andpharmaceutical industries discovered that medicinal plantscontaining phenol and flavonoid compounds are essentialin the development of novel drugs against stress-induceddiseases [28] In the present investigation the concentration(IC50) at which 50 of free radical scavenged was calculatedand extrapolated from the plot of percentage inhibition ver-sus plant extract concentration The herbal drugs exhibitedrobust DPPH radical scavenging activity in a concentration-dependent manner having IC50 ranging from 2025 to 710120583gmL As shown in Figure 4 MM BA EI JC AH BNIand CA extracts had a potent DPPH radical scavengingactivity corresponding to their TPC TFC and RP Theplant extracts scavenged ABTS radical by the decrease inits absorbance at 734 nm All the plant extracts exhibitedstrong ABTS radical scavenging capacities in the following

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

200

400

600

800

1000

1200

1400

1600

IC50

(g

mL)

Figure 4 DPPH free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

050

100150200250300350400450500

IC50

(g

mL)

AH BN JC CO MM EI BNI BA CAIIPlant samples

Figure 5 ABTS free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

order BN gt JC gt CA gt MM gt BNI gt EI gt CO gt AH gtII gt BA having IC50 values ranging from 25 to 400 120583gmL(Figure 5) DPPH and ABTS radical scavenging activities ofthese herbal remedies displayed a positive correlation withthe working concentrations It is obvious that most of theplant extracts tested in this work possess the ability to quenchthe free radicals generation via electron or proton donationdemonstrated by the termination of radical chain reaction[28] Although both DPPH and ABTS radicals are electron-transfer based methods however it is apparent that ABTSmethod seems to be more sensitive than DPPH radical assaypossibly due to their solubility and diffusivity as shown inthe IC50 values [10] It is worth noting that BA extract withstrong DPPH radical scavenging effect had a moderate ABTS

The Scientific World Journal 13

Table 4 Zone of bacterial inhibition by the selected plant extracts use in the management of oral and skin infections

Diameter of inhibition (mm)SN Plant samples K pneumoniae P mirabilis S aureus E coli1 Cassia alata 15 plusmn 141 9 plusmn 042 15 plusmn 083 13 plusmn 0802 Breynia nivosa 8 plusmn 052 17 plusmn 100 8 plusmn 008 15 plusmn 1023 Burkea africana NI 11 plusmn 084 13 plusmn 072 NI4 Baphia nitida 8 plusmn 050 13 plusmn 122 8 plusmn 051 NI5 Ipomoea involucrata 8 plusmn 052 NI 8 plusmn 050 13 plusmn 0526 Jatropha curcas NI 9 plusmn 030 9 plusmn 062 13 plusmn 0727 Acalypha hispida 21 plusmn 163 NI 11 plusmn 009 17 plusmn 1328 Chromolaena odorata 8 plusmn 042 8 plusmn 035 9 plusmn 009 15 plusmn1209 Eleusine indica 8 plusmn 042 8 plusmn 035 NI 17 plusmn 10010 Macrolobium macrophyllum NI NI 8 plusmn 004 15 plusmn 12311 Ciprofloxacin - - le16(R) ge17(S) le16(R)ge20(S)NI indicates no zone of inhibition S sensitive R resistance

radical scavenging capacity which can be explained by thepartial ionization of the extract in the test medium [29] SinceROS promotes tissue damage and epithelial inflammationvia exposure to sunlight radiation or smoke as a result theantioxidant capacity demonstrated in this work may supportthe traditional use of these plants against the influence ofinflammatory markers and oxidants in the skin and oralcavity [29] Based on the results of the three antioxidantassays it is obvious that MM AH BNI EI and CA extractshad the best FRSA indicating their importance in convertingfree radicals into more stable products and then terminatethe free radical chain reaction Our study showed that themethanol crude extracts from these plants could protectagainst free radical damage in the mouth such as humangingival periodontal tissues and certain skin infections [5]

334 Antibacterial Activity of Plant Extracts Pathogenicbacteria such as Staphylococcus aureus Escherichia coli Kleb-siella pneumoniae and Proteus mirabilis play a significantrole as the primary cause of skin and oral infections [30]Staphylococcus aureus is a part of normal skin known to bethe most problematic in the cause of skin infections as aresult of their resistance mechanisms to first-line antibiotics[30] Although the presence of this organism as a residentoral floral is controversial nonetheless MSSA and MRSAhave been isolated from oral rinse and tongue swab inpatients with oral infections [31] Escherichia coli are Gram-negative bacteria known to be notorious for their resistantmechanisms towards the existing arsenal of antibiotics Oralinfections such as edematous and erythematous and skininfections like omphalitis surgical site infections burnnecrotizing fasciitis and cellulitis at all age groups are associ-ated with enterobacterium E coli [32] Klebsiella pneumoniaeis a Gram-negative encapsulated facultative anaerobic androd-shaped bacterium found in the normal flora of themouth skin and intestines This bacterium is responsiblefor the prolonged surgical wound biliary tract wound andurinary tract infections and it could pose a serious healthrisk when resistant strains invade the throat mouth or

intestines [33] Villegas et al [34] detected a CTX-M-12 120573-lactamase producing K pneumoniae new plasmid-mediatedESBLs highly resistant to ceftazidime from a patient withhospital-acquired wound infection Proteus mirabilis is oneof the most common enterobacteria after E coli identified asGram-negativemotile and rod-shaped bacterium It is foundin clinical specimen implicated in the skin respiratory tractand bacteraemia as well as burns wounds and urinary tractinfections [35] All these pathogenic multidrug-resistancebacteria are causative agent in a periodontal lips ulcer whitesponge nevus and skin infections [36]

335 Susceptibility Test and MIC Determination The resultof disc diffusion assay showed the diameter of inhibitionof bacteria growth classified as le 15 (resistant) and ge 16(susceptible) according to CLSI guideline [37] As shown inTable 4 K pneumoniae and E coli are more susceptible tothe treatment of Acalypha hispida (AH) having a zone ofinhibition (ZI) 21 and 17mm respectively BN and EI extractsinhibit the growth of P mirabilis and E coli having the samezone of inhibition 17 mm corresponding to the report of Al-Zubairi et al [38] The data obtained for the methanol crudeextracts from AH BN and EI show their selective inhibitionof the test bacteria growth The plant extracts with ZI above13 mm against our panel of bacterial strains were selected forMICdeterminationHere themethanol crude extracts exhibitMIC values of 256 to gt 512 120583gmL against the test bacterialstrains but were less active as compared to the standardciprofloxacin 003 - 006 120583gmL (Table 5) Ciprofloxacin isan active broad-spectrum antibiotic use in the treatmentof urinary tract infections prostatitis and acute cystitis viainterfering with bacterial DNA replication and transcriptionthrough inhibition of DNA gyrase or topoisomerase [39] Allthe plant extracts demonstrated a mild to weak antibacterialactivity based on the classification of Rios and Recio [40] whoconsidered antibacterial activity of the crude plant extractsignificant (MIC lt 100 120583gmL) moderate (100 lt MIC = 512120583gmL) or weak (MIC gt 512 120583gmL) The herbal remediesshowed moderate inhibition of bacterial growth except for K

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

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International Journal of

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Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

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Submit your manuscripts atwwwhindawicom

Page 8: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

8 The Scientific World Journal

Table1Con

tinued

PlantN

ame

Family

Localn

ame

Partused

Infection

Tradition

aluses

Metho

dof

preparation

Brideliafer

rugin

eaEu

phorbiaceae

Irikirni

Leafstem

bark

root

Oral

Treatm

ento

fInsom

nia

mou

thwashgono

rrhea

Who

leplantisb

oiledandtakenorally

Euphorbiaheterophylla

Euph

orbiaceae

Egele

Leafroo

tsSkin

Treatm

ento

fSkindiseases

Leafandroot

arem

acerated

andthe

poultic

eapp

liedto

thea

ffected

skin

area

Euphorbiahirta

Euph

orbiaceae

Non

onku

rciya

odne

Who

leplant

exud

ate

Oral

Treatm

ento

fAsth

ma

coug

hshapeo

fbreasts

Thew

holeplantism

acerated

andtaken

orally

Euphorbialaterifl

ora

Euph

orbiaceae

Orow

ere

Leafexud

ate

Skin

Treatm

ento

fdermatitis

constip

ation

Theleafissqu

eezedandthep

oultice

appliedto

thea

ffected

skin

area

Cajanu

scajan

Fabaceae

Orela

Leafseed

Skin

andoral

Treatm

ento

fSmallpox

and

mou

thwash

Leafissqueezed

andappliedto

the

affectedarea

Dan

iellia

thurifera

Fabaceae

Iya

Stew

-woo

ddu

stSkin

Treatm

ento

fscabies

Theb

arkismacerated

andappliedto

the

affectedskin

area

Dialiu

mguineense

Fabaceae

Tasm

iyar

kumi

Leafbark

fruitand

twig

Oral

Treatm

ento

fBronchitis

coug

handdiuretic

Theleafand

bark

arec

rushed

andbo

iled

togetherTh

emixture

isthen

takenorally

Acacianilotica

Fabaceae

Baanigabaruwa

Fruitbark

exud

ate

Skin

Treatm

ento

fskindiseases

fung

alinfections

Thee

xudatesa

resqueezed

from

theb

ark

andapplieddirectlyto

thea

ffected

skin

area

Napoleona

imperia

lisLecythidaceae

Mabun

giTw

igs

Oral

Treatm

ento

fAsth

ma

coug

hTw

igsa

remacerated

andtakenorally

Alliu

msativ

umLiliaceae

Ayoayuu

Bulb

Oral

Treatm

ento

ffevercou

gh

asthmaantim

icrobial

Theb

ulbismacerated

andtakenorally

Hibiscus

asper

Malvaceae

Isapad

angiraa

Leaf

Oraland

skin

Treatm

ento

fcou

gh

woun

dsanddiuretic

Leafisbo

iledandtakenorally

Hibiscus

rosasin

ensis

Malvaceae

Kekekeireagu

Leafstem

flowe

rSkin

Treatm

ento

finfl

uenza

woun

dsand

diuretic

Who

leplantism

acerated

andtaken

orally

Theleafissqu

eezedandappliedin

affectedskin

area

Hibiscus

sabdariffa

Malvaceae

Sobo

gurgu

zuLeafflow

erOral

Treatm

ento

fDiuretic

scoug

handdressin

gwo

unds

Leafisbo

iledandtakenorally

Ficuseleg

ans

Moraceae

Asoro

Leaf

Skinoral

Treatm

ento

fPile

constip

ationandcraw

-craw

Leafissqueezed

andappliedto

the

affectedarea

The Scientific World Journal 9

the absorbance The percentage inhibition of DPPH orABTS radical scavenging activity by the plant extracts wascalculated as (Abscontrol ndash Abssample)(Abscontrol) times 100where Abscontrol is the absorbance of DPPH or ABTS +methanol Abssample is the absorbance of DPPH or ABTSradical + sample extractstandard Here the concentration ofthe extracts needed to decrease the absorbance of DPPH orABTS radical by 50 was calculated Rutin (Sigma-Aldrichge 94 HPLC grade) at the same working concentrations ofthe plant extracts was used as reference drug

210 Bacterial Strains All bacterial strains were culturedon nutrient agar slopes and were incubated for 24 h at37∘C before the MIC An inoculum turbidity equivalent toa 05 McFarland standard (1times107 CFUmL) was prepared innormal saline for each test organism and then diluted 1100in Mueller-Hinton broth just before injection of the platesStandard strains used in the study included S aureus and Ecoli which were a gift from Dr EO Ekundayo (Departmentof Microbiology Michael Okpara University of Agriculture)and clinically isolated strains of K pneumoniae and Pmirabilis were gotten from Department of MicrobiologyFederal Medical Centre (FMC) Umuahia Abia State

211 Antibacterial Susceptibility Using Disc Diffusion MethodThe microbial growth inhibition potential of the plantextracts was determined by using the agar disc diffusionmethod [17] The plant extracts were dissolved in 10 vvDMSO (Merck UK Ltd) to a final concentration of 2 mgmLThe nutrient agar plate was streaked with the standardizedtest bacterial inoculum over the surface evenly and 2mgdiscof the plant extracts transferred onto sterile filter disc papers(about 6 mm) containing the test compounds was placed onthe agar surface in the Petri dishes 100 120583gdisc of standardantibiotic ciprofloxacin was used as control and 100 120583LDMSOdisc served as negative control The plates were thenincubated at 37∘C for 24 h Generally the antimicrobialextracts diffuse into the agar and inhibit the growth of thetest bacteria All tests were done in duplicate and diame-ter zones of inhibition of growth were measured using ameter rule from the edge of each dish after the incubationperiod

212 Minimum Inhibitory Concentration (MIC) Assays Wefollowed the method of Smith et al [18] to determine theMIC ofmethanol crude extracts of selected botanicals againstclinical isolates of bacteria associated with skin and oralinfection A volumeof 100mLof sterileMueller-Hinton broth(Oxoid Basingstoke UK) containing 20 mgL of Ca2+ and10 mgL of Mg2+ was dispensed into a 96-well microtitreplate (Nunc 03 mL total volume per well) All antibacterialagents were dissolved in dimethyl sulphoxide (DMSO) anddiluted in Mueller-Hinton broth to give a stock solutionThen 100 120583L of the antibacterial agent stock solution (2000mgL) was serially diluted into each well and 100 mL ofthe bacterial inoculum added to each well to give a finalconcentration range of 0024 - 1 mgL All procedures wereperformed in duplicate and the plates were incubated for 18

h at 37∘C Briefly 20 mL of a 5 mgL methanol solution of 3-[45-dimethylthiazol-2-yl]-25-diphenyltetrazolium bromide(MTT) (Sigma-Aldrich Ltd South Africa) was added to eachwell and then incubated for 30 min Blue colouration indi-cated bacterial growth The MIC was recorded as the lowestconcentration at which no colour change was observed

213 Statistical Analysis Data analysis was done inMicrosoftExcel to obtain descriptive statistics Means values wereseparated by the Duncan multiple tests using SAS The dif-ferent levels of significance within the groups were analyzedusing one-way analysis of variance (ANOVA) Values wereconsidered significant at P lt005

3 Results and Discussion

31 Ethnobotanical Survey Modern drugs are expensive andsupply to a remote area may be irregular hence result-ing in overreliance of traditional methods of healing as aprimary source of healthcare [19] The rural communitiesof Ebem-Ohafia region of southeastern Nigeria depend onherbal remedies to meet their domestic and health needsDuring our ethnobotanical survey a total of 30 respondentsincluding the elders herbalists and traditional healers wereinterviewed in the study areaMajority of the people (22)wereaged 30ndash65 years dominated by males (25) and one-third(10) had a primary school education while 15 had secondaryeducation and others informal education A total of 61medic-inal plant species belonging to 26 families were identified inthe treatment of oral and skin diseases as shown in Table 1Twenty-seven of them are used in the management of oralinfections and twenty-six plants for the management of skininfection while eight plants are used in the management ofboth infections Table 2 shows the list of family number of theplant species used and their percentage occurrenceThemostrepresented plant family is the Euphorbiaceae having 1803of the plant species followed by Asteraceae and Fabaceae with1147 each [20] Euphorbiaceae is a large family of floweringplants with 300 genera and about 7500 species found usefulin the curing of some common human diseases Asteraceae isa large family of flowering plants having 32 913 species and1911 genera and has been found to possess diverse biologicaleffects Fabaceae is a family of cosmopolitan distributionwith approximately 730 genera and 19400 species richnessat a global level [20] The leaves (60) recorded the mostcommonly used plant part followed by the root (16) andthen stem bark (12) None of the respondents indicated anypossible side effects caused by any of these plants listed inTable 1 The most common methods of preparation includeinfusion decoction and maceration These herbalists ortraditional healers often collect the plants from the fieldand dry and crush them into powder form before storingin bottles to prevent patients from identifying the plantsused for their treatment Most healers refused to provideinformation without payment whereas others thought toprovide information on the plant use is disrespect of theirtraditional belief Furthermore the majority of the plantsmentioned lack scientific data to substantiate their acclaimedfolkloric use From this work it is evident that medicinal

10 The Scientific World Journal

Table 2 The list of family number of the plant species and theirpercentage occurrence

Family Number of plant species OccurrenceFabaceae 7 1147Euphorbiaceae 11 1803Asteraceae 7 1147Myrtaceae 4 656Moraceae 3 492Malvaceae 4 656Liliaceae 2 328Capparaceae 2 328Annonaceae 3 492Combretaceae 1 164Burseraceae 2 328Apocynaceae 1 164Gnetaceae 1 164Lecythidaceae 1 164Verbenaceae 1 164Ochnaceae 1 164Rubiaceae 1 164Piperaceae 1 164Acanthaceae 2 328Papilionoideae 2 328Asclepiadaceae 1 164Caesalpiniaceae 2 328Convolvulaceae 1 164Rutaceae 1 164Crassulaceae 1 164Poaceae 1 164

plants play a significant role in the primary healthcare ofthe people and conservation of the indigenous knowledge ofthis folklore medicine is crucial for the innovations of futuredrug discovery and possible herbal plants trade in the region[19] On the contrary we observed imprecise dosage lowhygiene standards hiding of healing methods and lack ofpatients record as a significant setback to traditional healthpractitioners in the region

32 Thin Layer Chromatography (TLC) Profiling Thin layerchromatography remains the most common useful techniqueby researchers as a quick and simple method to resolvethe constituents of a crude extract followed by advancedextraction methods to purify the active ingredients Previousstudies reported that specific secondary metabolites in plantexhibit a wide range of antioxidant and antibacterial proper-ties [21] TLC was formed on a glass sheet coated with silicagel After the sample is applied to the plate a solvent system(mobile phase) is drawn up via capillary action on the TLCplate at different mobility rate depending on the chemicalcompositionsThe separated spotswith different colourswerevisualized by the UV light projection onto the glass sheetThe variation of coloured spots detected in the crude extractsindicates the presence of different phytochemicals that may

be responsible for the pharmacological effects of selectedplants use in the management of oral and skin infections asacclaimed by the traditional healers We quantified the resultsby measuring the distance travelled by the plant compoundsdivided by the total distance travelled by the mobile phaseFrom Table 3 it is apparent that mobile phase 1 consistingof chloroform and ethanol in the volume ratio of 64 had abetter separation than mobile phase II containing tolueneethyl acetate and acetic acid in the volume ratio of 541 Themethanol extract of AH in bothmobile phases had the highestnumber of three bands with Rf values of 0667 0833 and0989 for mobile phase I and Rf values of 0381 0648 and0781 for mobile phase IIThe plant extracts MM BA CO JCandCAhad two bands inmobile phase I with clear separationbut had single band in mobile phase II The plant extractshaving only one band in either mobile phase suggest a poorsolvent system for compounds separation The TLC profilesprovide a characteristic fingerprint of these plants that mayalso be useful for their identification

33 Antioxidant Assays

331 TPC and TFC Phenolic compounds are secondarymetabolites distributed in fruits vegetables nuts herbsseeds and beverages [22] They are excellent scavengers offree radical that has been implicated in the pathologies ofvarious human diseases the antioxidant activity is dependenton the structure hydrophobic benzenoid rings and hydrogen-bonding potential of the phenolic hydroxyl groups In TPCassay a phenol in plant extracts loses a proton to formphenolate ion which reduces Folin Ciocalteu reagent andthe resulting changes are measured spectrophotometricallyat 765 nm [13] The content of phenol is reported in mg oftannic acid equivalent (TAE)mL of extract As shown inFigure 1 the total phenolic contents of absolute methanolcrude extract of these botanicals range from 700 to 3600mgTAEg The AH extract had the highest concentrationfollowed by CA CO MM BNI BA EI BN and JC whilethe least concentration was recorded in II extract Moreoverthe Folin Ciocalteu reagent (FCR) has been used to measureother compounds such as ascorbic acid amino acids andsugars hence it may not provide a concise amount of thephenolic compounds present in these plants Flavonoids areincreasingly consumed in a significant amount in daily dietsworldwide due to their various health benefits toman [23 24]In TFC assay aluminum ion in the reacting mixture formscomplexes with the C- 4 keto and either C- 3 or C- 5 hydroxylor with ortho hydroxyl groups in the A or B ring of flavonoidsdetected in the plant extracts [14] As shown in Figure 2the total flavonoid content was recorded in the plant extractsranging from 750 to 1500 mgQEg as extrapolated from thestandard quercetin curve The CA extract had the highestconcentration followed by BNI CO II AH BN MM JCand BA while the lowest concentration was recorded in EIextract TPC is found higher than the TFC in some of theextracts supporting the fact that most flavonoids are alsophenolics According to our results all the plant extractsare rich source of phenolic compounds The composition ofTPC and TFC registered in these plants may be difficult to

The Scientific World Journal 11

Table 3 Thin layer chromatography retention factor using mobile phase CHL ET (mobile phase I) and Tol EA AA (mobile phase II)

SN Plant Extract CHL ET (64) Tol EA AA (541)1 Baphia nitida 1 0742 Macrolobium macrophyllum 0706 0941 06953 Chromolaena odorata 0471 0647 06674 Jatropha curcas 0353 0941 06485 Cassia alata 0118 0882 06866 Ipomoea involucrata 0956 -7 Acalypha hispida 0667 0833 0989 0381 0648 07818 Breynia nivosa 0989 07629 Burkea africana 0644 1 075210 Eleusine indica 0978 0743CHL chloroform ET ethanol Tol toluene EA ethylacetate AA acetic acid

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

500

1000

1500

2000

2500

3000

3500

4000

TPC

(mgT

AE

g)

Figure 1 The total phenolic content (TPC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata TAE tannic acid equivalent

compare with others reported elsewhere due to many factorsincluding differences from species to species maturity stageplant parts and assay methods harvesting time standarddrugs and geographical location

332 Reducing Power (RP) Ferric reducing power is a globalmethod often used to assess the capacity of antioxidantsto donate either an electron or hydrogen atom to unpairedelectron [25] In reducing power assay plant extracts exertantioxidant activities via reduction of Fe3+ferric cyanidecomplex to Fe2+ (ferrous form) resulting in changing thegreen colour to blue and sometimes Perlrsquos Prussian bluecolour depending on the antioxidant potential [10] As shownin Figure 3 the reducing power of the methanol crudeextracts and the standard gallic acid rutin and ascorbic acidincreased with increase in concentration Higher absorbanceat 700 nm can be read after potassium ferricyanide reactionwith ferric chloride At 1000 120583gmL the extracts from AH

0

200

400

600

800

1000

1200

1400

1600

TFC

(mgQ

Eg)

AH BN JC CO MM EI BNI BA CAIIPlant extracts

Figure 2 The total flavonoid content (TFC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata QE Quercetin equivalent

BNI MM CA BA EI and BN had absorbance value of 270086 073 055 036 027 and 027 respectively while that ofAH is higher than the standard rutin (254) but comparablewell with gallic acid (275) and ascorbic acid (269) EI andBN extracts had a similar trend of ferric reducing powerthough significantly lower as compared with the standarddrugs AH extract exhibits remarkable antioxidant capacity(RP) correlating with its richness in phenolic and flavonoidcontents It is reasonable to suggest that methanol selectivelyextracts antioxidant principles from these botanicals basedon their polarities and chemical structures which are reflectedin plant extracts with high TFC and TPC having minimal RP[26] We have shown that some plant extracts in particularAH BNI MM and CA extracts may serve as an excellentcandidate to minimize skin and oral infections caused byoxidative stress as a result of imbalance of free radicalsgeneration and natural antioxidants in the saliva or woundarea According to our results the increasing RP of the plantextracts correlateswith their TPCandTFCThis study at least

12 The Scientific World JournalRe

duci

ng P

ower

7

00nm

625 125 250 500 10003125Concentration (gmL)

0

05

1

15

2

25

3

IIAHBNJCCOMMEI

BNIBACAGARutinVit C

Figure 3 The ferric reducing antioxidative capacity of selectedplant extracts used in the management of oral and skin infectionsin southeast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata GA gallic acid Vit C vitamin C (standard antioxidants)

in part lends credence to the ethno-therapeutic use of someplant as natural antioxidant in themanagement of skin or oralinfections in the study area

333 FRSA of Plant Extracts Skin and oral cavity exposedto an array of oxidants such as smokes from cigaretteschemicals UV light nicotine and alcohol that are directlyor indirectly leading to the production of reactive oxygenspecies (ROS) are known to be responsible for some com-mon human diseases [27] These free radicals in particularhydroxyl and superoxide anions are highly reactivemoleculesderived from the metabolism of oxygen Although they playan important role in the biochemical immunological andphysiological processes it could be deleterious to vital organsif excessively accumulated in the human body [28] Food andpharmaceutical industries discovered that medicinal plantscontaining phenol and flavonoid compounds are essentialin the development of novel drugs against stress-induceddiseases [28] In the present investigation the concentration(IC50) at which 50 of free radical scavenged was calculatedand extrapolated from the plot of percentage inhibition ver-sus plant extract concentration The herbal drugs exhibitedrobust DPPH radical scavenging activity in a concentration-dependent manner having IC50 ranging from 2025 to 710120583gmL As shown in Figure 4 MM BA EI JC AH BNIand CA extracts had a potent DPPH radical scavengingactivity corresponding to their TPC TFC and RP Theplant extracts scavenged ABTS radical by the decrease inits absorbance at 734 nm All the plant extracts exhibitedstrong ABTS radical scavenging capacities in the following

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

200

400

600

800

1000

1200

1400

1600

IC50

(g

mL)

Figure 4 DPPH free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

050

100150200250300350400450500

IC50

(g

mL)

AH BN JC CO MM EI BNI BA CAIIPlant samples

Figure 5 ABTS free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

order BN gt JC gt CA gt MM gt BNI gt EI gt CO gt AH gtII gt BA having IC50 values ranging from 25 to 400 120583gmL(Figure 5) DPPH and ABTS radical scavenging activities ofthese herbal remedies displayed a positive correlation withthe working concentrations It is obvious that most of theplant extracts tested in this work possess the ability to quenchthe free radicals generation via electron or proton donationdemonstrated by the termination of radical chain reaction[28] Although both DPPH and ABTS radicals are electron-transfer based methods however it is apparent that ABTSmethod seems to be more sensitive than DPPH radical assaypossibly due to their solubility and diffusivity as shown inthe IC50 values [10] It is worth noting that BA extract withstrong DPPH radical scavenging effect had a moderate ABTS

The Scientific World Journal 13

Table 4 Zone of bacterial inhibition by the selected plant extracts use in the management of oral and skin infections

Diameter of inhibition (mm)SN Plant samples K pneumoniae P mirabilis S aureus E coli1 Cassia alata 15 plusmn 141 9 plusmn 042 15 plusmn 083 13 plusmn 0802 Breynia nivosa 8 plusmn 052 17 plusmn 100 8 plusmn 008 15 plusmn 1023 Burkea africana NI 11 plusmn 084 13 plusmn 072 NI4 Baphia nitida 8 plusmn 050 13 plusmn 122 8 plusmn 051 NI5 Ipomoea involucrata 8 plusmn 052 NI 8 plusmn 050 13 plusmn 0526 Jatropha curcas NI 9 plusmn 030 9 plusmn 062 13 plusmn 0727 Acalypha hispida 21 plusmn 163 NI 11 plusmn 009 17 plusmn 1328 Chromolaena odorata 8 plusmn 042 8 plusmn 035 9 plusmn 009 15 plusmn1209 Eleusine indica 8 plusmn 042 8 plusmn 035 NI 17 plusmn 10010 Macrolobium macrophyllum NI NI 8 plusmn 004 15 plusmn 12311 Ciprofloxacin - - le16(R) ge17(S) le16(R)ge20(S)NI indicates no zone of inhibition S sensitive R resistance

radical scavenging capacity which can be explained by thepartial ionization of the extract in the test medium [29] SinceROS promotes tissue damage and epithelial inflammationvia exposure to sunlight radiation or smoke as a result theantioxidant capacity demonstrated in this work may supportthe traditional use of these plants against the influence ofinflammatory markers and oxidants in the skin and oralcavity [29] Based on the results of the three antioxidantassays it is obvious that MM AH BNI EI and CA extractshad the best FRSA indicating their importance in convertingfree radicals into more stable products and then terminatethe free radical chain reaction Our study showed that themethanol crude extracts from these plants could protectagainst free radical damage in the mouth such as humangingival periodontal tissues and certain skin infections [5]

334 Antibacterial Activity of Plant Extracts Pathogenicbacteria such as Staphylococcus aureus Escherichia coli Kleb-siella pneumoniae and Proteus mirabilis play a significantrole as the primary cause of skin and oral infections [30]Staphylococcus aureus is a part of normal skin known to bethe most problematic in the cause of skin infections as aresult of their resistance mechanisms to first-line antibiotics[30] Although the presence of this organism as a residentoral floral is controversial nonetheless MSSA and MRSAhave been isolated from oral rinse and tongue swab inpatients with oral infections [31] Escherichia coli are Gram-negative bacteria known to be notorious for their resistantmechanisms towards the existing arsenal of antibiotics Oralinfections such as edematous and erythematous and skininfections like omphalitis surgical site infections burnnecrotizing fasciitis and cellulitis at all age groups are associ-ated with enterobacterium E coli [32] Klebsiella pneumoniaeis a Gram-negative encapsulated facultative anaerobic androd-shaped bacterium found in the normal flora of themouth skin and intestines This bacterium is responsiblefor the prolonged surgical wound biliary tract wound andurinary tract infections and it could pose a serious healthrisk when resistant strains invade the throat mouth or

intestines [33] Villegas et al [34] detected a CTX-M-12 120573-lactamase producing K pneumoniae new plasmid-mediatedESBLs highly resistant to ceftazidime from a patient withhospital-acquired wound infection Proteus mirabilis is oneof the most common enterobacteria after E coli identified asGram-negativemotile and rod-shaped bacterium It is foundin clinical specimen implicated in the skin respiratory tractand bacteraemia as well as burns wounds and urinary tractinfections [35] All these pathogenic multidrug-resistancebacteria are causative agent in a periodontal lips ulcer whitesponge nevus and skin infections [36]

335 Susceptibility Test and MIC Determination The resultof disc diffusion assay showed the diameter of inhibitionof bacteria growth classified as le 15 (resistant) and ge 16(susceptible) according to CLSI guideline [37] As shown inTable 4 K pneumoniae and E coli are more susceptible tothe treatment of Acalypha hispida (AH) having a zone ofinhibition (ZI) 21 and 17mm respectively BN and EI extractsinhibit the growth of P mirabilis and E coli having the samezone of inhibition 17 mm corresponding to the report of Al-Zubairi et al [38] The data obtained for the methanol crudeextracts from AH BN and EI show their selective inhibitionof the test bacteria growth The plant extracts with ZI above13 mm against our panel of bacterial strains were selected forMICdeterminationHere themethanol crude extracts exhibitMIC values of 256 to gt 512 120583gmL against the test bacterialstrains but were less active as compared to the standardciprofloxacin 003 - 006 120583gmL (Table 5) Ciprofloxacin isan active broad-spectrum antibiotic use in the treatmentof urinary tract infections prostatitis and acute cystitis viainterfering with bacterial DNA replication and transcriptionthrough inhibition of DNA gyrase or topoisomerase [39] Allthe plant extracts demonstrated a mild to weak antibacterialactivity based on the classification of Rios and Recio [40] whoconsidered antibacterial activity of the crude plant extractsignificant (MIC lt 100 120583gmL) moderate (100 lt MIC = 512120583gmL) or weak (MIC gt 512 120583gmL) The herbal remediesshowed moderate inhibition of bacterial growth except for K

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

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Page 9: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

The Scientific World Journal 9

the absorbance The percentage inhibition of DPPH orABTS radical scavenging activity by the plant extracts wascalculated as (Abscontrol ndash Abssample)(Abscontrol) times 100where Abscontrol is the absorbance of DPPH or ABTS +methanol Abssample is the absorbance of DPPH or ABTSradical + sample extractstandard Here the concentration ofthe extracts needed to decrease the absorbance of DPPH orABTS radical by 50 was calculated Rutin (Sigma-Aldrichge 94 HPLC grade) at the same working concentrations ofthe plant extracts was used as reference drug

210 Bacterial Strains All bacterial strains were culturedon nutrient agar slopes and were incubated for 24 h at37∘C before the MIC An inoculum turbidity equivalent toa 05 McFarland standard (1times107 CFUmL) was prepared innormal saline for each test organism and then diluted 1100in Mueller-Hinton broth just before injection of the platesStandard strains used in the study included S aureus and Ecoli which were a gift from Dr EO Ekundayo (Departmentof Microbiology Michael Okpara University of Agriculture)and clinically isolated strains of K pneumoniae and Pmirabilis were gotten from Department of MicrobiologyFederal Medical Centre (FMC) Umuahia Abia State

211 Antibacterial Susceptibility Using Disc Diffusion MethodThe microbial growth inhibition potential of the plantextracts was determined by using the agar disc diffusionmethod [17] The plant extracts were dissolved in 10 vvDMSO (Merck UK Ltd) to a final concentration of 2 mgmLThe nutrient agar plate was streaked with the standardizedtest bacterial inoculum over the surface evenly and 2mgdiscof the plant extracts transferred onto sterile filter disc papers(about 6 mm) containing the test compounds was placed onthe agar surface in the Petri dishes 100 120583gdisc of standardantibiotic ciprofloxacin was used as control and 100 120583LDMSOdisc served as negative control The plates were thenincubated at 37∘C for 24 h Generally the antimicrobialextracts diffuse into the agar and inhibit the growth of thetest bacteria All tests were done in duplicate and diame-ter zones of inhibition of growth were measured using ameter rule from the edge of each dish after the incubationperiod

212 Minimum Inhibitory Concentration (MIC) Assays Wefollowed the method of Smith et al [18] to determine theMIC ofmethanol crude extracts of selected botanicals againstclinical isolates of bacteria associated with skin and oralinfection A volumeof 100mLof sterileMueller-Hinton broth(Oxoid Basingstoke UK) containing 20 mgL of Ca2+ and10 mgL of Mg2+ was dispensed into a 96-well microtitreplate (Nunc 03 mL total volume per well) All antibacterialagents were dissolved in dimethyl sulphoxide (DMSO) anddiluted in Mueller-Hinton broth to give a stock solutionThen 100 120583L of the antibacterial agent stock solution (2000mgL) was serially diluted into each well and 100 mL ofthe bacterial inoculum added to each well to give a finalconcentration range of 0024 - 1 mgL All procedures wereperformed in duplicate and the plates were incubated for 18

h at 37∘C Briefly 20 mL of a 5 mgL methanol solution of 3-[45-dimethylthiazol-2-yl]-25-diphenyltetrazolium bromide(MTT) (Sigma-Aldrich Ltd South Africa) was added to eachwell and then incubated for 30 min Blue colouration indi-cated bacterial growth The MIC was recorded as the lowestconcentration at which no colour change was observed

213 Statistical Analysis Data analysis was done inMicrosoftExcel to obtain descriptive statistics Means values wereseparated by the Duncan multiple tests using SAS The dif-ferent levels of significance within the groups were analyzedusing one-way analysis of variance (ANOVA) Values wereconsidered significant at P lt005

3 Results and Discussion

31 Ethnobotanical Survey Modern drugs are expensive andsupply to a remote area may be irregular hence result-ing in overreliance of traditional methods of healing as aprimary source of healthcare [19] The rural communitiesof Ebem-Ohafia region of southeastern Nigeria depend onherbal remedies to meet their domestic and health needsDuring our ethnobotanical survey a total of 30 respondentsincluding the elders herbalists and traditional healers wereinterviewed in the study areaMajority of the people (22)wereaged 30ndash65 years dominated by males (25) and one-third(10) had a primary school education while 15 had secondaryeducation and others informal education A total of 61medic-inal plant species belonging to 26 families were identified inthe treatment of oral and skin diseases as shown in Table 1Twenty-seven of them are used in the management of oralinfections and twenty-six plants for the management of skininfection while eight plants are used in the management ofboth infections Table 2 shows the list of family number of theplant species used and their percentage occurrenceThemostrepresented plant family is the Euphorbiaceae having 1803of the plant species followed by Asteraceae and Fabaceae with1147 each [20] Euphorbiaceae is a large family of floweringplants with 300 genera and about 7500 species found usefulin the curing of some common human diseases Asteraceae isa large family of flowering plants having 32 913 species and1911 genera and has been found to possess diverse biologicaleffects Fabaceae is a family of cosmopolitan distributionwith approximately 730 genera and 19400 species richnessat a global level [20] The leaves (60) recorded the mostcommonly used plant part followed by the root (16) andthen stem bark (12) None of the respondents indicated anypossible side effects caused by any of these plants listed inTable 1 The most common methods of preparation includeinfusion decoction and maceration These herbalists ortraditional healers often collect the plants from the fieldand dry and crush them into powder form before storingin bottles to prevent patients from identifying the plantsused for their treatment Most healers refused to provideinformation without payment whereas others thought toprovide information on the plant use is disrespect of theirtraditional belief Furthermore the majority of the plantsmentioned lack scientific data to substantiate their acclaimedfolkloric use From this work it is evident that medicinal

10 The Scientific World Journal

Table 2 The list of family number of the plant species and theirpercentage occurrence

Family Number of plant species OccurrenceFabaceae 7 1147Euphorbiaceae 11 1803Asteraceae 7 1147Myrtaceae 4 656Moraceae 3 492Malvaceae 4 656Liliaceae 2 328Capparaceae 2 328Annonaceae 3 492Combretaceae 1 164Burseraceae 2 328Apocynaceae 1 164Gnetaceae 1 164Lecythidaceae 1 164Verbenaceae 1 164Ochnaceae 1 164Rubiaceae 1 164Piperaceae 1 164Acanthaceae 2 328Papilionoideae 2 328Asclepiadaceae 1 164Caesalpiniaceae 2 328Convolvulaceae 1 164Rutaceae 1 164Crassulaceae 1 164Poaceae 1 164

plants play a significant role in the primary healthcare ofthe people and conservation of the indigenous knowledge ofthis folklore medicine is crucial for the innovations of futuredrug discovery and possible herbal plants trade in the region[19] On the contrary we observed imprecise dosage lowhygiene standards hiding of healing methods and lack ofpatients record as a significant setback to traditional healthpractitioners in the region

32 Thin Layer Chromatography (TLC) Profiling Thin layerchromatography remains the most common useful techniqueby researchers as a quick and simple method to resolvethe constituents of a crude extract followed by advancedextraction methods to purify the active ingredients Previousstudies reported that specific secondary metabolites in plantexhibit a wide range of antioxidant and antibacterial proper-ties [21] TLC was formed on a glass sheet coated with silicagel After the sample is applied to the plate a solvent system(mobile phase) is drawn up via capillary action on the TLCplate at different mobility rate depending on the chemicalcompositionsThe separated spotswith different colourswerevisualized by the UV light projection onto the glass sheetThe variation of coloured spots detected in the crude extractsindicates the presence of different phytochemicals that may

be responsible for the pharmacological effects of selectedplants use in the management of oral and skin infections asacclaimed by the traditional healers We quantified the resultsby measuring the distance travelled by the plant compoundsdivided by the total distance travelled by the mobile phaseFrom Table 3 it is apparent that mobile phase 1 consistingof chloroform and ethanol in the volume ratio of 64 had abetter separation than mobile phase II containing tolueneethyl acetate and acetic acid in the volume ratio of 541 Themethanol extract of AH in bothmobile phases had the highestnumber of three bands with Rf values of 0667 0833 and0989 for mobile phase I and Rf values of 0381 0648 and0781 for mobile phase IIThe plant extracts MM BA CO JCandCAhad two bands inmobile phase I with clear separationbut had single band in mobile phase II The plant extractshaving only one band in either mobile phase suggest a poorsolvent system for compounds separation The TLC profilesprovide a characteristic fingerprint of these plants that mayalso be useful for their identification

33 Antioxidant Assays

331 TPC and TFC Phenolic compounds are secondarymetabolites distributed in fruits vegetables nuts herbsseeds and beverages [22] They are excellent scavengers offree radical that has been implicated in the pathologies ofvarious human diseases the antioxidant activity is dependenton the structure hydrophobic benzenoid rings and hydrogen-bonding potential of the phenolic hydroxyl groups In TPCassay a phenol in plant extracts loses a proton to formphenolate ion which reduces Folin Ciocalteu reagent andthe resulting changes are measured spectrophotometricallyat 765 nm [13] The content of phenol is reported in mg oftannic acid equivalent (TAE)mL of extract As shown inFigure 1 the total phenolic contents of absolute methanolcrude extract of these botanicals range from 700 to 3600mgTAEg The AH extract had the highest concentrationfollowed by CA CO MM BNI BA EI BN and JC whilethe least concentration was recorded in II extract Moreoverthe Folin Ciocalteu reagent (FCR) has been used to measureother compounds such as ascorbic acid amino acids andsugars hence it may not provide a concise amount of thephenolic compounds present in these plants Flavonoids areincreasingly consumed in a significant amount in daily dietsworldwide due to their various health benefits toman [23 24]In TFC assay aluminum ion in the reacting mixture formscomplexes with the C- 4 keto and either C- 3 or C- 5 hydroxylor with ortho hydroxyl groups in the A or B ring of flavonoidsdetected in the plant extracts [14] As shown in Figure 2the total flavonoid content was recorded in the plant extractsranging from 750 to 1500 mgQEg as extrapolated from thestandard quercetin curve The CA extract had the highestconcentration followed by BNI CO II AH BN MM JCand BA while the lowest concentration was recorded in EIextract TPC is found higher than the TFC in some of theextracts supporting the fact that most flavonoids are alsophenolics According to our results all the plant extractsare rich source of phenolic compounds The composition ofTPC and TFC registered in these plants may be difficult to

The Scientific World Journal 11

Table 3 Thin layer chromatography retention factor using mobile phase CHL ET (mobile phase I) and Tol EA AA (mobile phase II)

SN Plant Extract CHL ET (64) Tol EA AA (541)1 Baphia nitida 1 0742 Macrolobium macrophyllum 0706 0941 06953 Chromolaena odorata 0471 0647 06674 Jatropha curcas 0353 0941 06485 Cassia alata 0118 0882 06866 Ipomoea involucrata 0956 -7 Acalypha hispida 0667 0833 0989 0381 0648 07818 Breynia nivosa 0989 07629 Burkea africana 0644 1 075210 Eleusine indica 0978 0743CHL chloroform ET ethanol Tol toluene EA ethylacetate AA acetic acid

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

500

1000

1500

2000

2500

3000

3500

4000

TPC

(mgT

AE

g)

Figure 1 The total phenolic content (TPC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata TAE tannic acid equivalent

compare with others reported elsewhere due to many factorsincluding differences from species to species maturity stageplant parts and assay methods harvesting time standarddrugs and geographical location

332 Reducing Power (RP) Ferric reducing power is a globalmethod often used to assess the capacity of antioxidantsto donate either an electron or hydrogen atom to unpairedelectron [25] In reducing power assay plant extracts exertantioxidant activities via reduction of Fe3+ferric cyanidecomplex to Fe2+ (ferrous form) resulting in changing thegreen colour to blue and sometimes Perlrsquos Prussian bluecolour depending on the antioxidant potential [10] As shownin Figure 3 the reducing power of the methanol crudeextracts and the standard gallic acid rutin and ascorbic acidincreased with increase in concentration Higher absorbanceat 700 nm can be read after potassium ferricyanide reactionwith ferric chloride At 1000 120583gmL the extracts from AH

0

200

400

600

800

1000

1200

1400

1600

TFC

(mgQ

Eg)

AH BN JC CO MM EI BNI BA CAIIPlant extracts

Figure 2 The total flavonoid content (TFC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata QE Quercetin equivalent

BNI MM CA BA EI and BN had absorbance value of 270086 073 055 036 027 and 027 respectively while that ofAH is higher than the standard rutin (254) but comparablewell with gallic acid (275) and ascorbic acid (269) EI andBN extracts had a similar trend of ferric reducing powerthough significantly lower as compared with the standarddrugs AH extract exhibits remarkable antioxidant capacity(RP) correlating with its richness in phenolic and flavonoidcontents It is reasonable to suggest that methanol selectivelyextracts antioxidant principles from these botanicals basedon their polarities and chemical structures which are reflectedin plant extracts with high TFC and TPC having minimal RP[26] We have shown that some plant extracts in particularAH BNI MM and CA extracts may serve as an excellentcandidate to minimize skin and oral infections caused byoxidative stress as a result of imbalance of free radicalsgeneration and natural antioxidants in the saliva or woundarea According to our results the increasing RP of the plantextracts correlateswith their TPCandTFCThis study at least

12 The Scientific World JournalRe

duci

ng P

ower

7

00nm

625 125 250 500 10003125Concentration (gmL)

0

05

1

15

2

25

3

IIAHBNJCCOMMEI

BNIBACAGARutinVit C

Figure 3 The ferric reducing antioxidative capacity of selectedplant extracts used in the management of oral and skin infectionsin southeast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata GA gallic acid Vit C vitamin C (standard antioxidants)

in part lends credence to the ethno-therapeutic use of someplant as natural antioxidant in themanagement of skin or oralinfections in the study area

333 FRSA of Plant Extracts Skin and oral cavity exposedto an array of oxidants such as smokes from cigaretteschemicals UV light nicotine and alcohol that are directlyor indirectly leading to the production of reactive oxygenspecies (ROS) are known to be responsible for some com-mon human diseases [27] These free radicals in particularhydroxyl and superoxide anions are highly reactivemoleculesderived from the metabolism of oxygen Although they playan important role in the biochemical immunological andphysiological processes it could be deleterious to vital organsif excessively accumulated in the human body [28] Food andpharmaceutical industries discovered that medicinal plantscontaining phenol and flavonoid compounds are essentialin the development of novel drugs against stress-induceddiseases [28] In the present investigation the concentration(IC50) at which 50 of free radical scavenged was calculatedand extrapolated from the plot of percentage inhibition ver-sus plant extract concentration The herbal drugs exhibitedrobust DPPH radical scavenging activity in a concentration-dependent manner having IC50 ranging from 2025 to 710120583gmL As shown in Figure 4 MM BA EI JC AH BNIand CA extracts had a potent DPPH radical scavengingactivity corresponding to their TPC TFC and RP Theplant extracts scavenged ABTS radical by the decrease inits absorbance at 734 nm All the plant extracts exhibitedstrong ABTS radical scavenging capacities in the following

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

200

400

600

800

1000

1200

1400

1600

IC50

(g

mL)

Figure 4 DPPH free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

050

100150200250300350400450500

IC50

(g

mL)

AH BN JC CO MM EI BNI BA CAIIPlant samples

Figure 5 ABTS free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

order BN gt JC gt CA gt MM gt BNI gt EI gt CO gt AH gtII gt BA having IC50 values ranging from 25 to 400 120583gmL(Figure 5) DPPH and ABTS radical scavenging activities ofthese herbal remedies displayed a positive correlation withthe working concentrations It is obvious that most of theplant extracts tested in this work possess the ability to quenchthe free radicals generation via electron or proton donationdemonstrated by the termination of radical chain reaction[28] Although both DPPH and ABTS radicals are electron-transfer based methods however it is apparent that ABTSmethod seems to be more sensitive than DPPH radical assaypossibly due to their solubility and diffusivity as shown inthe IC50 values [10] It is worth noting that BA extract withstrong DPPH radical scavenging effect had a moderate ABTS

The Scientific World Journal 13

Table 4 Zone of bacterial inhibition by the selected plant extracts use in the management of oral and skin infections

Diameter of inhibition (mm)SN Plant samples K pneumoniae P mirabilis S aureus E coli1 Cassia alata 15 plusmn 141 9 plusmn 042 15 plusmn 083 13 plusmn 0802 Breynia nivosa 8 plusmn 052 17 plusmn 100 8 plusmn 008 15 plusmn 1023 Burkea africana NI 11 plusmn 084 13 plusmn 072 NI4 Baphia nitida 8 plusmn 050 13 plusmn 122 8 plusmn 051 NI5 Ipomoea involucrata 8 plusmn 052 NI 8 plusmn 050 13 plusmn 0526 Jatropha curcas NI 9 plusmn 030 9 plusmn 062 13 plusmn 0727 Acalypha hispida 21 plusmn 163 NI 11 plusmn 009 17 plusmn 1328 Chromolaena odorata 8 plusmn 042 8 plusmn 035 9 plusmn 009 15 plusmn1209 Eleusine indica 8 plusmn 042 8 plusmn 035 NI 17 plusmn 10010 Macrolobium macrophyllum NI NI 8 plusmn 004 15 plusmn 12311 Ciprofloxacin - - le16(R) ge17(S) le16(R)ge20(S)NI indicates no zone of inhibition S sensitive R resistance

radical scavenging capacity which can be explained by thepartial ionization of the extract in the test medium [29] SinceROS promotes tissue damage and epithelial inflammationvia exposure to sunlight radiation or smoke as a result theantioxidant capacity demonstrated in this work may supportthe traditional use of these plants against the influence ofinflammatory markers and oxidants in the skin and oralcavity [29] Based on the results of the three antioxidantassays it is obvious that MM AH BNI EI and CA extractshad the best FRSA indicating their importance in convertingfree radicals into more stable products and then terminatethe free radical chain reaction Our study showed that themethanol crude extracts from these plants could protectagainst free radical damage in the mouth such as humangingival periodontal tissues and certain skin infections [5]

334 Antibacterial Activity of Plant Extracts Pathogenicbacteria such as Staphylococcus aureus Escherichia coli Kleb-siella pneumoniae and Proteus mirabilis play a significantrole as the primary cause of skin and oral infections [30]Staphylococcus aureus is a part of normal skin known to bethe most problematic in the cause of skin infections as aresult of their resistance mechanisms to first-line antibiotics[30] Although the presence of this organism as a residentoral floral is controversial nonetheless MSSA and MRSAhave been isolated from oral rinse and tongue swab inpatients with oral infections [31] Escherichia coli are Gram-negative bacteria known to be notorious for their resistantmechanisms towards the existing arsenal of antibiotics Oralinfections such as edematous and erythematous and skininfections like omphalitis surgical site infections burnnecrotizing fasciitis and cellulitis at all age groups are associ-ated with enterobacterium E coli [32] Klebsiella pneumoniaeis a Gram-negative encapsulated facultative anaerobic androd-shaped bacterium found in the normal flora of themouth skin and intestines This bacterium is responsiblefor the prolonged surgical wound biliary tract wound andurinary tract infections and it could pose a serious healthrisk when resistant strains invade the throat mouth or

intestines [33] Villegas et al [34] detected a CTX-M-12 120573-lactamase producing K pneumoniae new plasmid-mediatedESBLs highly resistant to ceftazidime from a patient withhospital-acquired wound infection Proteus mirabilis is oneof the most common enterobacteria after E coli identified asGram-negativemotile and rod-shaped bacterium It is foundin clinical specimen implicated in the skin respiratory tractand bacteraemia as well as burns wounds and urinary tractinfections [35] All these pathogenic multidrug-resistancebacteria are causative agent in a periodontal lips ulcer whitesponge nevus and skin infections [36]

335 Susceptibility Test and MIC Determination The resultof disc diffusion assay showed the diameter of inhibitionof bacteria growth classified as le 15 (resistant) and ge 16(susceptible) according to CLSI guideline [37] As shown inTable 4 K pneumoniae and E coli are more susceptible tothe treatment of Acalypha hispida (AH) having a zone ofinhibition (ZI) 21 and 17mm respectively BN and EI extractsinhibit the growth of P mirabilis and E coli having the samezone of inhibition 17 mm corresponding to the report of Al-Zubairi et al [38] The data obtained for the methanol crudeextracts from AH BN and EI show their selective inhibitionof the test bacteria growth The plant extracts with ZI above13 mm against our panel of bacterial strains were selected forMICdeterminationHere themethanol crude extracts exhibitMIC values of 256 to gt 512 120583gmL against the test bacterialstrains but were less active as compared to the standardciprofloxacin 003 - 006 120583gmL (Table 5) Ciprofloxacin isan active broad-spectrum antibiotic use in the treatmentof urinary tract infections prostatitis and acute cystitis viainterfering with bacterial DNA replication and transcriptionthrough inhibition of DNA gyrase or topoisomerase [39] Allthe plant extracts demonstrated a mild to weak antibacterialactivity based on the classification of Rios and Recio [40] whoconsidered antibacterial activity of the crude plant extractsignificant (MIC lt 100 120583gmL) moderate (100 lt MIC = 512120583gmL) or weak (MIC gt 512 120583gmL) The herbal remediesshowed moderate inhibition of bacterial growth except for K

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

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Page 10: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

10 The Scientific World Journal

Table 2 The list of family number of the plant species and theirpercentage occurrence

Family Number of plant species OccurrenceFabaceae 7 1147Euphorbiaceae 11 1803Asteraceae 7 1147Myrtaceae 4 656Moraceae 3 492Malvaceae 4 656Liliaceae 2 328Capparaceae 2 328Annonaceae 3 492Combretaceae 1 164Burseraceae 2 328Apocynaceae 1 164Gnetaceae 1 164Lecythidaceae 1 164Verbenaceae 1 164Ochnaceae 1 164Rubiaceae 1 164Piperaceae 1 164Acanthaceae 2 328Papilionoideae 2 328Asclepiadaceae 1 164Caesalpiniaceae 2 328Convolvulaceae 1 164Rutaceae 1 164Crassulaceae 1 164Poaceae 1 164

plants play a significant role in the primary healthcare ofthe people and conservation of the indigenous knowledge ofthis folklore medicine is crucial for the innovations of futuredrug discovery and possible herbal plants trade in the region[19] On the contrary we observed imprecise dosage lowhygiene standards hiding of healing methods and lack ofpatients record as a significant setback to traditional healthpractitioners in the region

32 Thin Layer Chromatography (TLC) Profiling Thin layerchromatography remains the most common useful techniqueby researchers as a quick and simple method to resolvethe constituents of a crude extract followed by advancedextraction methods to purify the active ingredients Previousstudies reported that specific secondary metabolites in plantexhibit a wide range of antioxidant and antibacterial proper-ties [21] TLC was formed on a glass sheet coated with silicagel After the sample is applied to the plate a solvent system(mobile phase) is drawn up via capillary action on the TLCplate at different mobility rate depending on the chemicalcompositionsThe separated spotswith different colourswerevisualized by the UV light projection onto the glass sheetThe variation of coloured spots detected in the crude extractsindicates the presence of different phytochemicals that may

be responsible for the pharmacological effects of selectedplants use in the management of oral and skin infections asacclaimed by the traditional healers We quantified the resultsby measuring the distance travelled by the plant compoundsdivided by the total distance travelled by the mobile phaseFrom Table 3 it is apparent that mobile phase 1 consistingof chloroform and ethanol in the volume ratio of 64 had abetter separation than mobile phase II containing tolueneethyl acetate and acetic acid in the volume ratio of 541 Themethanol extract of AH in bothmobile phases had the highestnumber of three bands with Rf values of 0667 0833 and0989 for mobile phase I and Rf values of 0381 0648 and0781 for mobile phase IIThe plant extracts MM BA CO JCandCAhad two bands inmobile phase I with clear separationbut had single band in mobile phase II The plant extractshaving only one band in either mobile phase suggest a poorsolvent system for compounds separation The TLC profilesprovide a characteristic fingerprint of these plants that mayalso be useful for their identification

33 Antioxidant Assays

331 TPC and TFC Phenolic compounds are secondarymetabolites distributed in fruits vegetables nuts herbsseeds and beverages [22] They are excellent scavengers offree radical that has been implicated in the pathologies ofvarious human diseases the antioxidant activity is dependenton the structure hydrophobic benzenoid rings and hydrogen-bonding potential of the phenolic hydroxyl groups In TPCassay a phenol in plant extracts loses a proton to formphenolate ion which reduces Folin Ciocalteu reagent andthe resulting changes are measured spectrophotometricallyat 765 nm [13] The content of phenol is reported in mg oftannic acid equivalent (TAE)mL of extract As shown inFigure 1 the total phenolic contents of absolute methanolcrude extract of these botanicals range from 700 to 3600mgTAEg The AH extract had the highest concentrationfollowed by CA CO MM BNI BA EI BN and JC whilethe least concentration was recorded in II extract Moreoverthe Folin Ciocalteu reagent (FCR) has been used to measureother compounds such as ascorbic acid amino acids andsugars hence it may not provide a concise amount of thephenolic compounds present in these plants Flavonoids areincreasingly consumed in a significant amount in daily dietsworldwide due to their various health benefits toman [23 24]In TFC assay aluminum ion in the reacting mixture formscomplexes with the C- 4 keto and either C- 3 or C- 5 hydroxylor with ortho hydroxyl groups in the A or B ring of flavonoidsdetected in the plant extracts [14] As shown in Figure 2the total flavonoid content was recorded in the plant extractsranging from 750 to 1500 mgQEg as extrapolated from thestandard quercetin curve The CA extract had the highestconcentration followed by BNI CO II AH BN MM JCand BA while the lowest concentration was recorded in EIextract TPC is found higher than the TFC in some of theextracts supporting the fact that most flavonoids are alsophenolics According to our results all the plant extractsare rich source of phenolic compounds The composition ofTPC and TFC registered in these plants may be difficult to

The Scientific World Journal 11

Table 3 Thin layer chromatography retention factor using mobile phase CHL ET (mobile phase I) and Tol EA AA (mobile phase II)

SN Plant Extract CHL ET (64) Tol EA AA (541)1 Baphia nitida 1 0742 Macrolobium macrophyllum 0706 0941 06953 Chromolaena odorata 0471 0647 06674 Jatropha curcas 0353 0941 06485 Cassia alata 0118 0882 06866 Ipomoea involucrata 0956 -7 Acalypha hispida 0667 0833 0989 0381 0648 07818 Breynia nivosa 0989 07629 Burkea africana 0644 1 075210 Eleusine indica 0978 0743CHL chloroform ET ethanol Tol toluene EA ethylacetate AA acetic acid

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

500

1000

1500

2000

2500

3000

3500

4000

TPC

(mgT

AE

g)

Figure 1 The total phenolic content (TPC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata TAE tannic acid equivalent

compare with others reported elsewhere due to many factorsincluding differences from species to species maturity stageplant parts and assay methods harvesting time standarddrugs and geographical location

332 Reducing Power (RP) Ferric reducing power is a globalmethod often used to assess the capacity of antioxidantsto donate either an electron or hydrogen atom to unpairedelectron [25] In reducing power assay plant extracts exertantioxidant activities via reduction of Fe3+ferric cyanidecomplex to Fe2+ (ferrous form) resulting in changing thegreen colour to blue and sometimes Perlrsquos Prussian bluecolour depending on the antioxidant potential [10] As shownin Figure 3 the reducing power of the methanol crudeextracts and the standard gallic acid rutin and ascorbic acidincreased with increase in concentration Higher absorbanceat 700 nm can be read after potassium ferricyanide reactionwith ferric chloride At 1000 120583gmL the extracts from AH

0

200

400

600

800

1000

1200

1400

1600

TFC

(mgQ

Eg)

AH BN JC CO MM EI BNI BA CAIIPlant extracts

Figure 2 The total flavonoid content (TFC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata QE Quercetin equivalent

BNI MM CA BA EI and BN had absorbance value of 270086 073 055 036 027 and 027 respectively while that ofAH is higher than the standard rutin (254) but comparablewell with gallic acid (275) and ascorbic acid (269) EI andBN extracts had a similar trend of ferric reducing powerthough significantly lower as compared with the standarddrugs AH extract exhibits remarkable antioxidant capacity(RP) correlating with its richness in phenolic and flavonoidcontents It is reasonable to suggest that methanol selectivelyextracts antioxidant principles from these botanicals basedon their polarities and chemical structures which are reflectedin plant extracts with high TFC and TPC having minimal RP[26] We have shown that some plant extracts in particularAH BNI MM and CA extracts may serve as an excellentcandidate to minimize skin and oral infections caused byoxidative stress as a result of imbalance of free radicalsgeneration and natural antioxidants in the saliva or woundarea According to our results the increasing RP of the plantextracts correlateswith their TPCandTFCThis study at least

12 The Scientific World JournalRe

duci

ng P

ower

7

00nm

625 125 250 500 10003125Concentration (gmL)

0

05

1

15

2

25

3

IIAHBNJCCOMMEI

BNIBACAGARutinVit C

Figure 3 The ferric reducing antioxidative capacity of selectedplant extracts used in the management of oral and skin infectionsin southeast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata GA gallic acid Vit C vitamin C (standard antioxidants)

in part lends credence to the ethno-therapeutic use of someplant as natural antioxidant in themanagement of skin or oralinfections in the study area

333 FRSA of Plant Extracts Skin and oral cavity exposedto an array of oxidants such as smokes from cigaretteschemicals UV light nicotine and alcohol that are directlyor indirectly leading to the production of reactive oxygenspecies (ROS) are known to be responsible for some com-mon human diseases [27] These free radicals in particularhydroxyl and superoxide anions are highly reactivemoleculesderived from the metabolism of oxygen Although they playan important role in the biochemical immunological andphysiological processes it could be deleterious to vital organsif excessively accumulated in the human body [28] Food andpharmaceutical industries discovered that medicinal plantscontaining phenol and flavonoid compounds are essentialin the development of novel drugs against stress-induceddiseases [28] In the present investigation the concentration(IC50) at which 50 of free radical scavenged was calculatedand extrapolated from the plot of percentage inhibition ver-sus plant extract concentration The herbal drugs exhibitedrobust DPPH radical scavenging activity in a concentration-dependent manner having IC50 ranging from 2025 to 710120583gmL As shown in Figure 4 MM BA EI JC AH BNIand CA extracts had a potent DPPH radical scavengingactivity corresponding to their TPC TFC and RP Theplant extracts scavenged ABTS radical by the decrease inits absorbance at 734 nm All the plant extracts exhibitedstrong ABTS radical scavenging capacities in the following

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

200

400

600

800

1000

1200

1400

1600

IC50

(g

mL)

Figure 4 DPPH free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

050

100150200250300350400450500

IC50

(g

mL)

AH BN JC CO MM EI BNI BA CAIIPlant samples

Figure 5 ABTS free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

order BN gt JC gt CA gt MM gt BNI gt EI gt CO gt AH gtII gt BA having IC50 values ranging from 25 to 400 120583gmL(Figure 5) DPPH and ABTS radical scavenging activities ofthese herbal remedies displayed a positive correlation withthe working concentrations It is obvious that most of theplant extracts tested in this work possess the ability to quenchthe free radicals generation via electron or proton donationdemonstrated by the termination of radical chain reaction[28] Although both DPPH and ABTS radicals are electron-transfer based methods however it is apparent that ABTSmethod seems to be more sensitive than DPPH radical assaypossibly due to their solubility and diffusivity as shown inthe IC50 values [10] It is worth noting that BA extract withstrong DPPH radical scavenging effect had a moderate ABTS

The Scientific World Journal 13

Table 4 Zone of bacterial inhibition by the selected plant extracts use in the management of oral and skin infections

Diameter of inhibition (mm)SN Plant samples K pneumoniae P mirabilis S aureus E coli1 Cassia alata 15 plusmn 141 9 plusmn 042 15 plusmn 083 13 plusmn 0802 Breynia nivosa 8 plusmn 052 17 plusmn 100 8 plusmn 008 15 plusmn 1023 Burkea africana NI 11 plusmn 084 13 plusmn 072 NI4 Baphia nitida 8 plusmn 050 13 plusmn 122 8 plusmn 051 NI5 Ipomoea involucrata 8 plusmn 052 NI 8 plusmn 050 13 plusmn 0526 Jatropha curcas NI 9 plusmn 030 9 plusmn 062 13 plusmn 0727 Acalypha hispida 21 plusmn 163 NI 11 plusmn 009 17 plusmn 1328 Chromolaena odorata 8 plusmn 042 8 plusmn 035 9 plusmn 009 15 plusmn1209 Eleusine indica 8 plusmn 042 8 plusmn 035 NI 17 plusmn 10010 Macrolobium macrophyllum NI NI 8 plusmn 004 15 plusmn 12311 Ciprofloxacin - - le16(R) ge17(S) le16(R)ge20(S)NI indicates no zone of inhibition S sensitive R resistance

radical scavenging capacity which can be explained by thepartial ionization of the extract in the test medium [29] SinceROS promotes tissue damage and epithelial inflammationvia exposure to sunlight radiation or smoke as a result theantioxidant capacity demonstrated in this work may supportthe traditional use of these plants against the influence ofinflammatory markers and oxidants in the skin and oralcavity [29] Based on the results of the three antioxidantassays it is obvious that MM AH BNI EI and CA extractshad the best FRSA indicating their importance in convertingfree radicals into more stable products and then terminatethe free radical chain reaction Our study showed that themethanol crude extracts from these plants could protectagainst free radical damage in the mouth such as humangingival periodontal tissues and certain skin infections [5]

334 Antibacterial Activity of Plant Extracts Pathogenicbacteria such as Staphylococcus aureus Escherichia coli Kleb-siella pneumoniae and Proteus mirabilis play a significantrole as the primary cause of skin and oral infections [30]Staphylococcus aureus is a part of normal skin known to bethe most problematic in the cause of skin infections as aresult of their resistance mechanisms to first-line antibiotics[30] Although the presence of this organism as a residentoral floral is controversial nonetheless MSSA and MRSAhave been isolated from oral rinse and tongue swab inpatients with oral infections [31] Escherichia coli are Gram-negative bacteria known to be notorious for their resistantmechanisms towards the existing arsenal of antibiotics Oralinfections such as edematous and erythematous and skininfections like omphalitis surgical site infections burnnecrotizing fasciitis and cellulitis at all age groups are associ-ated with enterobacterium E coli [32] Klebsiella pneumoniaeis a Gram-negative encapsulated facultative anaerobic androd-shaped bacterium found in the normal flora of themouth skin and intestines This bacterium is responsiblefor the prolonged surgical wound biliary tract wound andurinary tract infections and it could pose a serious healthrisk when resistant strains invade the throat mouth or

intestines [33] Villegas et al [34] detected a CTX-M-12 120573-lactamase producing K pneumoniae new plasmid-mediatedESBLs highly resistant to ceftazidime from a patient withhospital-acquired wound infection Proteus mirabilis is oneof the most common enterobacteria after E coli identified asGram-negativemotile and rod-shaped bacterium It is foundin clinical specimen implicated in the skin respiratory tractand bacteraemia as well as burns wounds and urinary tractinfections [35] All these pathogenic multidrug-resistancebacteria are causative agent in a periodontal lips ulcer whitesponge nevus and skin infections [36]

335 Susceptibility Test and MIC Determination The resultof disc diffusion assay showed the diameter of inhibitionof bacteria growth classified as le 15 (resistant) and ge 16(susceptible) according to CLSI guideline [37] As shown inTable 4 K pneumoniae and E coli are more susceptible tothe treatment of Acalypha hispida (AH) having a zone ofinhibition (ZI) 21 and 17mm respectively BN and EI extractsinhibit the growth of P mirabilis and E coli having the samezone of inhibition 17 mm corresponding to the report of Al-Zubairi et al [38] The data obtained for the methanol crudeextracts from AH BN and EI show their selective inhibitionof the test bacteria growth The plant extracts with ZI above13 mm against our panel of bacterial strains were selected forMICdeterminationHere themethanol crude extracts exhibitMIC values of 256 to gt 512 120583gmL against the test bacterialstrains but were less active as compared to the standardciprofloxacin 003 - 006 120583gmL (Table 5) Ciprofloxacin isan active broad-spectrum antibiotic use in the treatmentof urinary tract infections prostatitis and acute cystitis viainterfering with bacterial DNA replication and transcriptionthrough inhibition of DNA gyrase or topoisomerase [39] Allthe plant extracts demonstrated a mild to weak antibacterialactivity based on the classification of Rios and Recio [40] whoconsidered antibacterial activity of the crude plant extractsignificant (MIC lt 100 120583gmL) moderate (100 lt MIC = 512120583gmL) or weak (MIC gt 512 120583gmL) The herbal remediesshowed moderate inhibition of bacterial growth except for K

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

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Page 11: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

The Scientific World Journal 11

Table 3 Thin layer chromatography retention factor using mobile phase CHL ET (mobile phase I) and Tol EA AA (mobile phase II)

SN Plant Extract CHL ET (64) Tol EA AA (541)1 Baphia nitida 1 0742 Macrolobium macrophyllum 0706 0941 06953 Chromolaena odorata 0471 0647 06674 Jatropha curcas 0353 0941 06485 Cassia alata 0118 0882 06866 Ipomoea involucrata 0956 -7 Acalypha hispida 0667 0833 0989 0381 0648 07818 Breynia nivosa 0989 07629 Burkea africana 0644 1 075210 Eleusine indica 0978 0743CHL chloroform ET ethanol Tol toluene EA ethylacetate AA acetic acid

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

500

1000

1500

2000

2500

3000

3500

4000

TPC

(mgT

AE

g)

Figure 1 The total phenolic content (TPC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata TAE tannic acid equivalent

compare with others reported elsewhere due to many factorsincluding differences from species to species maturity stageplant parts and assay methods harvesting time standarddrugs and geographical location

332 Reducing Power (RP) Ferric reducing power is a globalmethod often used to assess the capacity of antioxidantsto donate either an electron or hydrogen atom to unpairedelectron [25] In reducing power assay plant extracts exertantioxidant activities via reduction of Fe3+ferric cyanidecomplex to Fe2+ (ferrous form) resulting in changing thegreen colour to blue and sometimes Perlrsquos Prussian bluecolour depending on the antioxidant potential [10] As shownin Figure 3 the reducing power of the methanol crudeextracts and the standard gallic acid rutin and ascorbic acidincreased with increase in concentration Higher absorbanceat 700 nm can be read after potassium ferricyanide reactionwith ferric chloride At 1000 120583gmL the extracts from AH

0

200

400

600

800

1000

1200

1400

1600

TFC

(mgQ

Eg)

AH BN JC CO MM EI BNI BA CAIIPlant extracts

Figure 2 The total flavonoid content (TFC) of selected plants usedin the management of oral and skin infections in southeast NigeriaII Ipomoea involucrata AH Acalypha hispida BN Breynia nivosaJC Jatropha curcas CO Chromolaena odorata MM Macrolobiummacrophyllum EI Eleusine indica BNI Baphia nitida BA Burkeaafricana CA Cassia alata QE Quercetin equivalent

BNI MM CA BA EI and BN had absorbance value of 270086 073 055 036 027 and 027 respectively while that ofAH is higher than the standard rutin (254) but comparablewell with gallic acid (275) and ascorbic acid (269) EI andBN extracts had a similar trend of ferric reducing powerthough significantly lower as compared with the standarddrugs AH extract exhibits remarkable antioxidant capacity(RP) correlating with its richness in phenolic and flavonoidcontents It is reasonable to suggest that methanol selectivelyextracts antioxidant principles from these botanicals basedon their polarities and chemical structures which are reflectedin plant extracts with high TFC and TPC having minimal RP[26] We have shown that some plant extracts in particularAH BNI MM and CA extracts may serve as an excellentcandidate to minimize skin and oral infections caused byoxidative stress as a result of imbalance of free radicalsgeneration and natural antioxidants in the saliva or woundarea According to our results the increasing RP of the plantextracts correlateswith their TPCandTFCThis study at least

12 The Scientific World JournalRe

duci

ng P

ower

7

00nm

625 125 250 500 10003125Concentration (gmL)

0

05

1

15

2

25

3

IIAHBNJCCOMMEI

BNIBACAGARutinVit C

Figure 3 The ferric reducing antioxidative capacity of selectedplant extracts used in the management of oral and skin infectionsin southeast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata GA gallic acid Vit C vitamin C (standard antioxidants)

in part lends credence to the ethno-therapeutic use of someplant as natural antioxidant in themanagement of skin or oralinfections in the study area

333 FRSA of Plant Extracts Skin and oral cavity exposedto an array of oxidants such as smokes from cigaretteschemicals UV light nicotine and alcohol that are directlyor indirectly leading to the production of reactive oxygenspecies (ROS) are known to be responsible for some com-mon human diseases [27] These free radicals in particularhydroxyl and superoxide anions are highly reactivemoleculesderived from the metabolism of oxygen Although they playan important role in the biochemical immunological andphysiological processes it could be deleterious to vital organsif excessively accumulated in the human body [28] Food andpharmaceutical industries discovered that medicinal plantscontaining phenol and flavonoid compounds are essentialin the development of novel drugs against stress-induceddiseases [28] In the present investigation the concentration(IC50) at which 50 of free radical scavenged was calculatedand extrapolated from the plot of percentage inhibition ver-sus plant extract concentration The herbal drugs exhibitedrobust DPPH radical scavenging activity in a concentration-dependent manner having IC50 ranging from 2025 to 710120583gmL As shown in Figure 4 MM BA EI JC AH BNIand CA extracts had a potent DPPH radical scavengingactivity corresponding to their TPC TFC and RP Theplant extracts scavenged ABTS radical by the decrease inits absorbance at 734 nm All the plant extracts exhibitedstrong ABTS radical scavenging capacities in the following

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

200

400

600

800

1000

1200

1400

1600

IC50

(g

mL)

Figure 4 DPPH free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

050

100150200250300350400450500

IC50

(g

mL)

AH BN JC CO MM EI BNI BA CAIIPlant samples

Figure 5 ABTS free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

order BN gt JC gt CA gt MM gt BNI gt EI gt CO gt AH gtII gt BA having IC50 values ranging from 25 to 400 120583gmL(Figure 5) DPPH and ABTS radical scavenging activities ofthese herbal remedies displayed a positive correlation withthe working concentrations It is obvious that most of theplant extracts tested in this work possess the ability to quenchthe free radicals generation via electron or proton donationdemonstrated by the termination of radical chain reaction[28] Although both DPPH and ABTS radicals are electron-transfer based methods however it is apparent that ABTSmethod seems to be more sensitive than DPPH radical assaypossibly due to their solubility and diffusivity as shown inthe IC50 values [10] It is worth noting that BA extract withstrong DPPH radical scavenging effect had a moderate ABTS

The Scientific World Journal 13

Table 4 Zone of bacterial inhibition by the selected plant extracts use in the management of oral and skin infections

Diameter of inhibition (mm)SN Plant samples K pneumoniae P mirabilis S aureus E coli1 Cassia alata 15 plusmn 141 9 plusmn 042 15 plusmn 083 13 plusmn 0802 Breynia nivosa 8 plusmn 052 17 plusmn 100 8 plusmn 008 15 plusmn 1023 Burkea africana NI 11 plusmn 084 13 plusmn 072 NI4 Baphia nitida 8 plusmn 050 13 plusmn 122 8 plusmn 051 NI5 Ipomoea involucrata 8 plusmn 052 NI 8 plusmn 050 13 plusmn 0526 Jatropha curcas NI 9 plusmn 030 9 plusmn 062 13 plusmn 0727 Acalypha hispida 21 plusmn 163 NI 11 plusmn 009 17 plusmn 1328 Chromolaena odorata 8 plusmn 042 8 plusmn 035 9 plusmn 009 15 plusmn1209 Eleusine indica 8 plusmn 042 8 plusmn 035 NI 17 plusmn 10010 Macrolobium macrophyllum NI NI 8 plusmn 004 15 plusmn 12311 Ciprofloxacin - - le16(R) ge17(S) le16(R)ge20(S)NI indicates no zone of inhibition S sensitive R resistance

radical scavenging capacity which can be explained by thepartial ionization of the extract in the test medium [29] SinceROS promotes tissue damage and epithelial inflammationvia exposure to sunlight radiation or smoke as a result theantioxidant capacity demonstrated in this work may supportthe traditional use of these plants against the influence ofinflammatory markers and oxidants in the skin and oralcavity [29] Based on the results of the three antioxidantassays it is obvious that MM AH BNI EI and CA extractshad the best FRSA indicating their importance in convertingfree radicals into more stable products and then terminatethe free radical chain reaction Our study showed that themethanol crude extracts from these plants could protectagainst free radical damage in the mouth such as humangingival periodontal tissues and certain skin infections [5]

334 Antibacterial Activity of Plant Extracts Pathogenicbacteria such as Staphylococcus aureus Escherichia coli Kleb-siella pneumoniae and Proteus mirabilis play a significantrole as the primary cause of skin and oral infections [30]Staphylococcus aureus is a part of normal skin known to bethe most problematic in the cause of skin infections as aresult of their resistance mechanisms to first-line antibiotics[30] Although the presence of this organism as a residentoral floral is controversial nonetheless MSSA and MRSAhave been isolated from oral rinse and tongue swab inpatients with oral infections [31] Escherichia coli are Gram-negative bacteria known to be notorious for their resistantmechanisms towards the existing arsenal of antibiotics Oralinfections such as edematous and erythematous and skininfections like omphalitis surgical site infections burnnecrotizing fasciitis and cellulitis at all age groups are associ-ated with enterobacterium E coli [32] Klebsiella pneumoniaeis a Gram-negative encapsulated facultative anaerobic androd-shaped bacterium found in the normal flora of themouth skin and intestines This bacterium is responsiblefor the prolonged surgical wound biliary tract wound andurinary tract infections and it could pose a serious healthrisk when resistant strains invade the throat mouth or

intestines [33] Villegas et al [34] detected a CTX-M-12 120573-lactamase producing K pneumoniae new plasmid-mediatedESBLs highly resistant to ceftazidime from a patient withhospital-acquired wound infection Proteus mirabilis is oneof the most common enterobacteria after E coli identified asGram-negativemotile and rod-shaped bacterium It is foundin clinical specimen implicated in the skin respiratory tractand bacteraemia as well as burns wounds and urinary tractinfections [35] All these pathogenic multidrug-resistancebacteria are causative agent in a periodontal lips ulcer whitesponge nevus and skin infections [36]

335 Susceptibility Test and MIC Determination The resultof disc diffusion assay showed the diameter of inhibitionof bacteria growth classified as le 15 (resistant) and ge 16(susceptible) according to CLSI guideline [37] As shown inTable 4 K pneumoniae and E coli are more susceptible tothe treatment of Acalypha hispida (AH) having a zone ofinhibition (ZI) 21 and 17mm respectively BN and EI extractsinhibit the growth of P mirabilis and E coli having the samezone of inhibition 17 mm corresponding to the report of Al-Zubairi et al [38] The data obtained for the methanol crudeextracts from AH BN and EI show their selective inhibitionof the test bacteria growth The plant extracts with ZI above13 mm against our panel of bacterial strains were selected forMICdeterminationHere themethanol crude extracts exhibitMIC values of 256 to gt 512 120583gmL against the test bacterialstrains but were less active as compared to the standardciprofloxacin 003 - 006 120583gmL (Table 5) Ciprofloxacin isan active broad-spectrum antibiotic use in the treatmentof urinary tract infections prostatitis and acute cystitis viainterfering with bacterial DNA replication and transcriptionthrough inhibition of DNA gyrase or topoisomerase [39] Allthe plant extracts demonstrated a mild to weak antibacterialactivity based on the classification of Rios and Recio [40] whoconsidered antibacterial activity of the crude plant extractsignificant (MIC lt 100 120583gmL) moderate (100 lt MIC = 512120583gmL) or weak (MIC gt 512 120583gmL) The herbal remediesshowed moderate inhibition of bacterial growth except for K

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

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Submit your manuscripts atwwwhindawicom

Page 12: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

12 The Scientific World JournalRe

duci

ng P

ower

7

00nm

625 125 250 500 10003125Concentration (gmL)

0

05

1

15

2

25

3

IIAHBNJCCOMMEI

BNIBACAGARutinVit C

Figure 3 The ferric reducing antioxidative capacity of selectedplant extracts used in the management of oral and skin infectionsin southeast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata GA gallic acid Vit C vitamin C (standard antioxidants)

in part lends credence to the ethno-therapeutic use of someplant as natural antioxidant in themanagement of skin or oralinfections in the study area

333 FRSA of Plant Extracts Skin and oral cavity exposedto an array of oxidants such as smokes from cigaretteschemicals UV light nicotine and alcohol that are directlyor indirectly leading to the production of reactive oxygenspecies (ROS) are known to be responsible for some com-mon human diseases [27] These free radicals in particularhydroxyl and superoxide anions are highly reactivemoleculesderived from the metabolism of oxygen Although they playan important role in the biochemical immunological andphysiological processes it could be deleterious to vital organsif excessively accumulated in the human body [28] Food andpharmaceutical industries discovered that medicinal plantscontaining phenol and flavonoid compounds are essentialin the development of novel drugs against stress-induceddiseases [28] In the present investigation the concentration(IC50) at which 50 of free radical scavenged was calculatedand extrapolated from the plot of percentage inhibition ver-sus plant extract concentration The herbal drugs exhibitedrobust DPPH radical scavenging activity in a concentration-dependent manner having IC50 ranging from 2025 to 710120583gmL As shown in Figure 4 MM BA EI JC AH BNIand CA extracts had a potent DPPH radical scavengingactivity corresponding to their TPC TFC and RP Theplant extracts scavenged ABTS radical by the decrease inits absorbance at 734 nm All the plant extracts exhibitedstrong ABTS radical scavenging capacities in the following

AH BN JC CO MM EI BNI BA CAIIPlant extracts

0

200

400

600

800

1000

1200

1400

1600

IC50

(g

mL)

Figure 4 DPPH free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

050

100150200250300350400450500

IC50

(g

mL)

AH BN JC CO MM EI BNI BA CAIIPlant samples

Figure 5 ABTS free radical scavenging activity of selected plantextracts used in the management of oral and skin infections insoutheast folklore medicine of Nigeria II Ipomoea involucrataAH Acalypha hispida BN Breynia nivosa JC Jatropha curcasCO Chromolaena odorata MM Macrolobium macrophyllum EIEleusine indica BNI Baphia nitida BA Burkea africana CACassiaalata

order BN gt JC gt CA gt MM gt BNI gt EI gt CO gt AH gtII gt BA having IC50 values ranging from 25 to 400 120583gmL(Figure 5) DPPH and ABTS radical scavenging activities ofthese herbal remedies displayed a positive correlation withthe working concentrations It is obvious that most of theplant extracts tested in this work possess the ability to quenchthe free radicals generation via electron or proton donationdemonstrated by the termination of radical chain reaction[28] Although both DPPH and ABTS radicals are electron-transfer based methods however it is apparent that ABTSmethod seems to be more sensitive than DPPH radical assaypossibly due to their solubility and diffusivity as shown inthe IC50 values [10] It is worth noting that BA extract withstrong DPPH radical scavenging effect had a moderate ABTS

The Scientific World Journal 13

Table 4 Zone of bacterial inhibition by the selected plant extracts use in the management of oral and skin infections

Diameter of inhibition (mm)SN Plant samples K pneumoniae P mirabilis S aureus E coli1 Cassia alata 15 plusmn 141 9 plusmn 042 15 plusmn 083 13 plusmn 0802 Breynia nivosa 8 plusmn 052 17 plusmn 100 8 plusmn 008 15 plusmn 1023 Burkea africana NI 11 plusmn 084 13 plusmn 072 NI4 Baphia nitida 8 plusmn 050 13 plusmn 122 8 plusmn 051 NI5 Ipomoea involucrata 8 plusmn 052 NI 8 plusmn 050 13 plusmn 0526 Jatropha curcas NI 9 plusmn 030 9 plusmn 062 13 plusmn 0727 Acalypha hispida 21 plusmn 163 NI 11 plusmn 009 17 plusmn 1328 Chromolaena odorata 8 plusmn 042 8 plusmn 035 9 plusmn 009 15 plusmn1209 Eleusine indica 8 plusmn 042 8 plusmn 035 NI 17 plusmn 10010 Macrolobium macrophyllum NI NI 8 plusmn 004 15 plusmn 12311 Ciprofloxacin - - le16(R) ge17(S) le16(R)ge20(S)NI indicates no zone of inhibition S sensitive R resistance

radical scavenging capacity which can be explained by thepartial ionization of the extract in the test medium [29] SinceROS promotes tissue damage and epithelial inflammationvia exposure to sunlight radiation or smoke as a result theantioxidant capacity demonstrated in this work may supportthe traditional use of these plants against the influence ofinflammatory markers and oxidants in the skin and oralcavity [29] Based on the results of the three antioxidantassays it is obvious that MM AH BNI EI and CA extractshad the best FRSA indicating their importance in convertingfree radicals into more stable products and then terminatethe free radical chain reaction Our study showed that themethanol crude extracts from these plants could protectagainst free radical damage in the mouth such as humangingival periodontal tissues and certain skin infections [5]

334 Antibacterial Activity of Plant Extracts Pathogenicbacteria such as Staphylococcus aureus Escherichia coli Kleb-siella pneumoniae and Proteus mirabilis play a significantrole as the primary cause of skin and oral infections [30]Staphylococcus aureus is a part of normal skin known to bethe most problematic in the cause of skin infections as aresult of their resistance mechanisms to first-line antibiotics[30] Although the presence of this organism as a residentoral floral is controversial nonetheless MSSA and MRSAhave been isolated from oral rinse and tongue swab inpatients with oral infections [31] Escherichia coli are Gram-negative bacteria known to be notorious for their resistantmechanisms towards the existing arsenal of antibiotics Oralinfections such as edematous and erythematous and skininfections like omphalitis surgical site infections burnnecrotizing fasciitis and cellulitis at all age groups are associ-ated with enterobacterium E coli [32] Klebsiella pneumoniaeis a Gram-negative encapsulated facultative anaerobic androd-shaped bacterium found in the normal flora of themouth skin and intestines This bacterium is responsiblefor the prolonged surgical wound biliary tract wound andurinary tract infections and it could pose a serious healthrisk when resistant strains invade the throat mouth or

intestines [33] Villegas et al [34] detected a CTX-M-12 120573-lactamase producing K pneumoniae new plasmid-mediatedESBLs highly resistant to ceftazidime from a patient withhospital-acquired wound infection Proteus mirabilis is oneof the most common enterobacteria after E coli identified asGram-negativemotile and rod-shaped bacterium It is foundin clinical specimen implicated in the skin respiratory tractand bacteraemia as well as burns wounds and urinary tractinfections [35] All these pathogenic multidrug-resistancebacteria are causative agent in a periodontal lips ulcer whitesponge nevus and skin infections [36]

335 Susceptibility Test and MIC Determination The resultof disc diffusion assay showed the diameter of inhibitionof bacteria growth classified as le 15 (resistant) and ge 16(susceptible) according to CLSI guideline [37] As shown inTable 4 K pneumoniae and E coli are more susceptible tothe treatment of Acalypha hispida (AH) having a zone ofinhibition (ZI) 21 and 17mm respectively BN and EI extractsinhibit the growth of P mirabilis and E coli having the samezone of inhibition 17 mm corresponding to the report of Al-Zubairi et al [38] The data obtained for the methanol crudeextracts from AH BN and EI show their selective inhibitionof the test bacteria growth The plant extracts with ZI above13 mm against our panel of bacterial strains were selected forMICdeterminationHere themethanol crude extracts exhibitMIC values of 256 to gt 512 120583gmL against the test bacterialstrains but were less active as compared to the standardciprofloxacin 003 - 006 120583gmL (Table 5) Ciprofloxacin isan active broad-spectrum antibiotic use in the treatmentof urinary tract infections prostatitis and acute cystitis viainterfering with bacterial DNA replication and transcriptionthrough inhibition of DNA gyrase or topoisomerase [39] Allthe plant extracts demonstrated a mild to weak antibacterialactivity based on the classification of Rios and Recio [40] whoconsidered antibacterial activity of the crude plant extractsignificant (MIC lt 100 120583gmL) moderate (100 lt MIC = 512120583gmL) or weak (MIC gt 512 120583gmL) The herbal remediesshowed moderate inhibition of bacterial growth except for K

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

Hindawiwwwhindawicom

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PeptidesInternational Journal of

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GenomicsInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Hindawiwwwhindawicom Volume 2018

BioinformaticsAdvances in

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Hindawiwwwhindawicom Volume 2018

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Hindawiwwwhindawicom Volume 2018

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Submit your manuscripts atwwwhindawicom

Page 13: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

The Scientific World Journal 13

Table 4 Zone of bacterial inhibition by the selected plant extracts use in the management of oral and skin infections

Diameter of inhibition (mm)SN Plant samples K pneumoniae P mirabilis S aureus E coli1 Cassia alata 15 plusmn 141 9 plusmn 042 15 plusmn 083 13 plusmn 0802 Breynia nivosa 8 plusmn 052 17 plusmn 100 8 plusmn 008 15 plusmn 1023 Burkea africana NI 11 plusmn 084 13 plusmn 072 NI4 Baphia nitida 8 plusmn 050 13 plusmn 122 8 plusmn 051 NI5 Ipomoea involucrata 8 plusmn 052 NI 8 plusmn 050 13 plusmn 0526 Jatropha curcas NI 9 plusmn 030 9 plusmn 062 13 plusmn 0727 Acalypha hispida 21 plusmn 163 NI 11 plusmn 009 17 plusmn 1328 Chromolaena odorata 8 plusmn 042 8 plusmn 035 9 plusmn 009 15 plusmn1209 Eleusine indica 8 plusmn 042 8 plusmn 035 NI 17 plusmn 10010 Macrolobium macrophyllum NI NI 8 plusmn 004 15 plusmn 12311 Ciprofloxacin - - le16(R) ge17(S) le16(R)ge20(S)NI indicates no zone of inhibition S sensitive R resistance

radical scavenging capacity which can be explained by thepartial ionization of the extract in the test medium [29] SinceROS promotes tissue damage and epithelial inflammationvia exposure to sunlight radiation or smoke as a result theantioxidant capacity demonstrated in this work may supportthe traditional use of these plants against the influence ofinflammatory markers and oxidants in the skin and oralcavity [29] Based on the results of the three antioxidantassays it is obvious that MM AH BNI EI and CA extractshad the best FRSA indicating their importance in convertingfree radicals into more stable products and then terminatethe free radical chain reaction Our study showed that themethanol crude extracts from these plants could protectagainst free radical damage in the mouth such as humangingival periodontal tissues and certain skin infections [5]

334 Antibacterial Activity of Plant Extracts Pathogenicbacteria such as Staphylococcus aureus Escherichia coli Kleb-siella pneumoniae and Proteus mirabilis play a significantrole as the primary cause of skin and oral infections [30]Staphylococcus aureus is a part of normal skin known to bethe most problematic in the cause of skin infections as aresult of their resistance mechanisms to first-line antibiotics[30] Although the presence of this organism as a residentoral floral is controversial nonetheless MSSA and MRSAhave been isolated from oral rinse and tongue swab inpatients with oral infections [31] Escherichia coli are Gram-negative bacteria known to be notorious for their resistantmechanisms towards the existing arsenal of antibiotics Oralinfections such as edematous and erythematous and skininfections like omphalitis surgical site infections burnnecrotizing fasciitis and cellulitis at all age groups are associ-ated with enterobacterium E coli [32] Klebsiella pneumoniaeis a Gram-negative encapsulated facultative anaerobic androd-shaped bacterium found in the normal flora of themouth skin and intestines This bacterium is responsiblefor the prolonged surgical wound biliary tract wound andurinary tract infections and it could pose a serious healthrisk when resistant strains invade the throat mouth or

intestines [33] Villegas et al [34] detected a CTX-M-12 120573-lactamase producing K pneumoniae new plasmid-mediatedESBLs highly resistant to ceftazidime from a patient withhospital-acquired wound infection Proteus mirabilis is oneof the most common enterobacteria after E coli identified asGram-negativemotile and rod-shaped bacterium It is foundin clinical specimen implicated in the skin respiratory tractand bacteraemia as well as burns wounds and urinary tractinfections [35] All these pathogenic multidrug-resistancebacteria are causative agent in a periodontal lips ulcer whitesponge nevus and skin infections [36]

335 Susceptibility Test and MIC Determination The resultof disc diffusion assay showed the diameter of inhibitionof bacteria growth classified as le 15 (resistant) and ge 16(susceptible) according to CLSI guideline [37] As shown inTable 4 K pneumoniae and E coli are more susceptible tothe treatment of Acalypha hispida (AH) having a zone ofinhibition (ZI) 21 and 17mm respectively BN and EI extractsinhibit the growth of P mirabilis and E coli having the samezone of inhibition 17 mm corresponding to the report of Al-Zubairi et al [38] The data obtained for the methanol crudeextracts from AH BN and EI show their selective inhibitionof the test bacteria growth The plant extracts with ZI above13 mm against our panel of bacterial strains were selected forMICdeterminationHere themethanol crude extracts exhibitMIC values of 256 to gt 512 120583gmL against the test bacterialstrains but were less active as compared to the standardciprofloxacin 003 - 006 120583gmL (Table 5) Ciprofloxacin isan active broad-spectrum antibiotic use in the treatmentof urinary tract infections prostatitis and acute cystitis viainterfering with bacterial DNA replication and transcriptionthrough inhibition of DNA gyrase or topoisomerase [39] Allthe plant extracts demonstrated a mild to weak antibacterialactivity based on the classification of Rios and Recio [40] whoconsidered antibacterial activity of the crude plant extractsignificant (MIC lt 100 120583gmL) moderate (100 lt MIC = 512120583gmL) or weak (MIC gt 512 120583gmL) The herbal remediesshowed moderate inhibition of bacterial growth except for K

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

Hindawiwwwhindawicom

International Journal of

Volume 2018

Zoology

Hindawiwwwhindawicom Volume 2018

Anatomy Research International

PeptidesInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Journal of Parasitology Research

GenomicsInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Hindawiwwwhindawicom Volume 2018

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Neuroscience Journal

Hindawiwwwhindawicom Volume 2018

BioMed Research International

Cell BiologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Biochemistry Research International

ArchaeaHindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Genetics Research International

Hindawiwwwhindawicom Volume 2018

Advances in

Virolog y Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Enzyme Research

Hindawiwwwhindawicom Volume 2018

International Journal of

MicrobiologyHindawiwwwhindawicom

Nucleic AcidsJournal of

Volume 2018

Submit your manuscripts atwwwhindawicom

Page 14: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

14 The Scientific World Journal

Table 5 Minimum inhibitory concentration (MIC) of selectedmedicinal plants against bacterial strains associated with oral andskin infections

Plants SA KP PM ECMIC (120583gmL)

Acalypha hispida 512 256 gt512 512Cassia alata 512 256 256 256Breynia nivosa 256 gt512 256 512Eleusine indica 256 512 256 512Chromolaena odorata 512 256 256 256Ciprofloxacin 003 003 003 003SA S aureus KP K pneumoniae PM P mirabilis EC E coli

pneumoniae and P mirabilis towards BN and AH extractsrespectively BN and EI extracts were more active againstS aureus and P mirabilis CA and CO extracts were moreactive against all the test bacterial strains except S aureusPrevious work reported by Amadi et al [41] confirmed theweak antibacterial activity exhibited by BN extract againstK pneumoniae The antibacterial activity demonstrated byAH extract against our panel of bacterial strains contradictsthat of Aboaba and Yeye [42] who reported MIC value of100 mgmL against E coli S aureus and K pneumoniaeIt is interesting to note that K pneumoniae E coli and Pmirabilis that confer resistance to the extended spectrumcephalosporin and penicillin [43] are more susceptible tothe treatment with CA and CO extracts Furthermore theobserved antibacterial activity exhibited by EI CA and COextracts against the superbugs of modern day healthcare [38]is worth noting whichmay perhaps attribute to a wide varietyof secondary metabolites especially phenolics and flavonoidsthat have been reported with potent antimicrobial properties[44] Among the three extracts Eleusine indica has IC50 gt30 120583g mL towards MCF-7 HT-29 and CEM-SS humancancer cell lines suggesting that continued exposure to thisherbal remedy may alter human organs [38] The probablemechanism of action of these plants is not known but it couldbe attributed to the leakage of the cell wall or alteration in themembrane permeability or inhibition of enzyme gyraseDNAsynthesis and protein synthesis [45] Further researches areneeded to identify the active ingredients curare effect andthe molecular mechanisms of these plants

4 Conclusion

The results of this study therefore at least in part providescientific evidence for the traditional use of these medicinalplants in ethnomedicine by the dwellers of Ebem-Ohafiacommunities in the treatment of skin and oral infections Thefree radicals scavenging activity moderate to high TPC TFCand themild antibacterial properties demonstrated by certainplant extracts are worth harnessing for proper evaluation andintegration into the primary healthcare system Further frac-tionation to identify the single compound responsible for theobserved activities is highly recommended EncouraginglyBN CA CO and AH extracts with potent antioxidant andantibacterial properties did not exhibit any significant toxic

effect as reported by other researchers and thus substantiateits safe use as complementary and alternative therapy for thetreatment or prevention of human diseases

Data Availability

The data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

Nopotential conflicts of interestwere reported by the authors

Acknowledgments

The authors thank the National Research Foundation (NRF)South Africa Agency for Science and Technology Advance-ment for their financial support

References

[1] WorldHealth Organization ldquoOral healthrdquo Fact sheets 318 2012httpwwwwhointmediacentrefactsheetsfs318enindexhtml

[2] F E Dewhirst T Chen J Izard et al ldquoThe human oralmicrobiomerdquo Journal of Bacteriology vol 192 no 19 pp 5002ndash5017 2010

[3] M L Falsetta M I Klein P M Colonne et al ldquoSymbiotic rela-tionship between Streptococcus mutans and Candida albicanssynergizes virulence of plaque biofilms in vivordquo Infection andImmunity vol 82 no 5 pp 1968ndash1981 2014

[4] A Shinde J Ganu and P Naik ldquoEffect of Free Radicals ampAntioxidants on Oxidative Stress A Reviewrdquo Journal of Dentaland Allied Sciences vol 1 no 2 p 63 2012

[5] O Hershkovich I Shafat and R M Nagler ldquoAge-relatedchanges in salivary antioxidant profile possible implications fororal cancerrdquo Journals of Gerontology Series A Biological Sciencesand Medical Sciences vol 62 no 4 pp 361ndash366 2007

[6] S Jagtap S Yavankar K Pardesi and B Chopade ldquoProductionof bioemulsifier by Acinetobactersp from healthy human skinof tribal populationrdquo Indian Journal of Experimental Biologyvol 48 pp 70ndash76 2010

[7] D H Limoli A B Rockel K M Host et al ldquoCationicAntimicrobial Peptides Promote Microbial Mutagenesis andPathoadaptation inChronic InfectionsrdquoPLoS Pathogens vol 10no 4 Article ID e1004083 2014

[8] A Al-Ahmad H Ameen K Pelz et al ldquoAntibiotic resistanceand capacity for biofilm formation of different bacteria isolatedfrom endodontic infections associated with root-filled teethrdquoJournal of Endodontics vol 40 no 2 pp 223ndash230 2014

[9] F A Badria andO A Zidan ldquoNatural products for dental cariespreventionrdquo Journal of Medicinal Food vol 7 no 3 pp 381ndash3842004

[10] S O Oyedemi B O Oyedemi I I Ijeh P E Ohanyerem R MCoopoosamy and O A Aiyegoro ldquoAlpha-amylase inhibitionand antioxidative capacity of some antidiabetic plants used bythe traditional healers in Southeastern Nigeriardquo The ScientificWorld Journal vol 2017 Article ID 3592491 11 pages 2017

[11] T Vogt ldquoPhenylpropanoid biosynthesisrdquo Molecular Plant vol3 no 1 pp 2ndash20 2010

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

Hindawiwwwhindawicom

International Journal of

Volume 2018

Zoology

Hindawiwwwhindawicom Volume 2018

Anatomy Research International

PeptidesInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Journal of Parasitology Research

GenomicsInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Hindawiwwwhindawicom Volume 2018

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Neuroscience Journal

Hindawiwwwhindawicom Volume 2018

BioMed Research International

Cell BiologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Biochemistry Research International

ArchaeaHindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Genetics Research International

Hindawiwwwhindawicom Volume 2018

Advances in

Virolog y Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Enzyme Research

Hindawiwwwhindawicom Volume 2018

International Journal of

MicrobiologyHindawiwwwhindawicom

Nucleic AcidsJournal of

Volume 2018

Submit your manuscripts atwwwhindawicom

Page 15: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

The Scientific World Journal 15

[12] H Ghaffari B J Ghassam S C Nayaka K Ramachandra Kiniand H S Prakash ldquoAntioxidant and neuroprotective activitiesof Hyptis suaveolens (L) Poit against oxidative stress-inducedneurotoxicityrdquoCellular andMolecularNeurobiology vol 34 no3 pp 323ndash331 2014

[13] C Chang M Yang H Wen and J Chern ldquoEstimation of totalflavonoid content in propolis by two complementary colometricmethodsrdquo Journal of Food and Drug Analysis vol 10 no 3 pp178ndash182 2002

[14] G C Yen and H Y Chen ldquoAntioxidant activity of varioustea extracts in relation to their antimutagenicityrdquo Journal ofAgricultural and Food Chemistry vol 43 no 1 pp 27ndash32 1995

[15] A Tariq M Athar J Ara V Sultana S Ehteshamul-HaqueandM Ahmad ldquoBiochemical evaluation of antioxidant activityin extracts and polysaccharide fractions of seaweedsrdquo GlobalJournal of Environmental Science Management vol 1 no 1 pp47ndash62 2015

[16] R Re N Pellegrini A ProteggenteA Pannala M Yang andCRice-Evans ldquoAntioxidant activity applying an improved ABTSradical cation decolorization assayrdquo Free Radical Biology ampMedicine vol 26 no 9-10 pp 1231ndash1237 1999

[17] National Committee for Clinical Laboratory Standards ldquoPer-formance Standards for antimicrobial susceptibility testingrdquo 8thInformational Supplement M100 S12 National Committee forClinical Laboratory Standards Villanova Pa USA 2002

[18] E E Smith E Williamson M Zloh and S Gibbons ldquoIsopi-maric acid from Pinus nigra shows activity against multidrug-resistant and EMRSA strains of Staphylococcus aureusrdquo Phy-totherapy Research vol 19 no 6 pp 538ndash542 2005

[19] N R Farnsworth ldquoEthnopharmacology and drug develop-mentrdquo Ciba Foundation Symposium vol 185 pp 42ndash59 2007

[20] W S Judd C S Campbell E A Kellogg P F Stevens andM JDonoghue Plant Systematics A Phylogenetic Approach SinauerAssociates Sunderland Mass USA 2002

[21] M Wink Functions of Plant Secondary Metabolites and theirExploitation in Biotechnology vol 39 of Annual Plant ReviewsWiley-Blackwell London UK 2010

[22] V Cheynier ldquoPolyphenols in foods aremore complex than oftenthoughtrdquo The American Journal of Clinical Nutrition vol 81supplement 1 pp 223Sndash229S 2005

[23] P C H Hollman and M B Katan ldquoDietary flavonoidsIntake health effects and bioavailabilityrdquo Food and ChemicalToxicology vol 37 no 9-10 pp 937ndash942 1999

[24] G P Hubbard S Wolffram J A Lovegrove and J M GibbinsldquoIngestion of quercetin inhibits platelet aggregation and essen-tial components of the collagen-stimulated platelet activationpathway in humansrdquo Journal of Thrombosis and Haemostasisvol 2 no 12 pp 2138ndash2145 2004

[25] M-C Kou S-Y Chiou C-Y Weng L Wang C-T Hoand M-J Wu ldquoCurcuminoids distinctly exhibit antioxidantactivities and regulate expression of scavenger receptors andheme oxygenase-1rdquo Molecular Nutrition amp Food Research vol57 no 9 pp 1598ndash1610 2013

[26] M-Y Juan and C-C Chou ldquoEnhancement of antioxidantactivity total phenolic and flavonoid content of black soybeansby solid state fermentation with Bacillus subtilis BCRC 14715rdquoFood Microbiology vol 27 no 5 pp 586ndash591 2010

[27] S O Oyedemi and A J Afolayan ldquoIn vitro and in vivo antioxi-dant activity of aqueous leaves extract of Leonotis leonurus (L)R Brrdquo International Journal of Pharmacology vol 7 no 2 pp248ndash256 2011

[28] A Temraz and W H El-Tantawy ldquoCharacterization of antiox-idant activity of extract from Artemisia vulgarisrdquo PakistanJournal of Pharmaceutical Sciences vol 21 no 4 pp 321ndash3262008

[29] I Fridovich ldquoFundamental aspects of reactive oxygen speciesor whatrsquos the matter with oxygenrdquo Annals of the New YorkAcademy of Sciences vol 893 no 22 pp 13ndash18 1999

[30] D J Wilson R N Gonzalez and H H Das ldquoBovine mastitispathogens in New York and Pennsylvania prevalence andeffects on somatic cell count and milk productionrdquo Journal ofDairy Science vol 80 no 10 pp 2592ndash2598 1997

[31] A J Smith M S Jackson and J Bagg ldquoThe ecology ofstaphylococci in the oral cavity a reviewrdquo Journal of MedicalMicrobiology vol 50 no 11 pp 940ndash946 2001

[32] G J Moet R N Jones D J Biedenbach M G Stilwelland T R Fritsche ldquoContemporary causes of skin and softtissue infections in North America Latin America and Europereport from the SENTRY antimicrobial surveillance program(1998ndash2004)rdquo Diagnostic Microbiological Infectious Diseasesvol 57 no 1 pp 7ndash13 2007

[33] K J Ryan and C G Ray Sharris Medicinal MicrobiologyMcGraw Hill New York NY USA 4th edition 2004

[34] MVVillegasACorrea F Perez et al ldquoCTX-M-12120573-lactamasein aKlebsiella pneumoniae clinical isolate in Colombiardquo Antimi-crobial Agents and Chemotherapy vol 48 no 2 pp 629ndash6312004

[35] R Tibbetts J G Frye J Marschall D Warren and W DunneldquoDetection of KPC-2 in a clinical isolate of Proteus mirabilisand first reported description of carbapenemase resistancecaused by a KPC 120573-lactamase in P mirabilisrdquo Journal of ClinicalMicrobiology vol 46 no 9 pp 3080ndash3083 2008

[36] S M JacobsenD J Stickler H L T Mobley andM E ShirtliffldquoComplicated catheter-associated urinary tract infections dueto Escherichia coli and Proteus mirabilisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 26ndash59 2008

[37] Clinical and Laboratory Standards Institute ldquoMethods fordilution antimicrobial susceptibility tests for bacteria that growaerobically approved standardrdquo CLSI document M7-A7 Clini-cal and Laboratory Standards Institute Wayne Pa USA 2006

[38] A S Al-Zubairi A B Abdul S I Abdelwahab C Y Peng SMohan andMM Elhassan ldquoEleucine indica possesses antiox-idant antibacterial and cytotoxic propertiesrdquo Evidence-BasedComplementary and Alternative Medicine vol 2011 Article ID965370 6 pages 2011

[39] Y A Al-Soud and N A Al-Masoudi ldquoA new class of dihalo-quinolones bearingN1015840- aldehydoglycosylhydrazidesmercapto-124-triazole oxadiazoline and 120572-amino ester precursors syn-thesis and antimicrobial activityrdquo Journal of the BrazilianChem-ical Society vol 14 no 5 pp 790ndash796 2003

[40] J L Rios and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 2 pp 80ndash84 2005

[41] E S Amadi C A Oyeka R A Onyeagba O C Ugboguand I Okoli ldquoAntimicrobial screening of Breynia nivosus andAgeratum conyzoides against dental caries organismsrdquo Journalof Biological Sciences vol 7 no 2 pp 354ndash358 2007

[42] S A Aboaba and E Yeye ldquoStudies on phytochemical screeningantimicrobial and toxicity effect of the shoot system ofAcalyphasegetalis Muell Argrdquo African Journal of Pure and AppliedChemistry vol 8 no 12 pp 191ndash195 2014

[43] A Wahab S Begum A Ayub et al ldquoLuteolin and kaempferolfrom Cassia alata antimicrobial and antioxidant activity of its

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

Hindawiwwwhindawicom

International Journal of

Volume 2018

Zoology

Hindawiwwwhindawicom Volume 2018

Anatomy Research International

PeptidesInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Journal of Parasitology Research

GenomicsInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Hindawiwwwhindawicom Volume 2018

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Neuroscience Journal

Hindawiwwwhindawicom Volume 2018

BioMed Research International

Cell BiologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Biochemistry Research International

ArchaeaHindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Genetics Research International

Hindawiwwwhindawicom Volume 2018

Advances in

Virolog y Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Enzyme Research

Hindawiwwwhindawicom Volume 2018

International Journal of

MicrobiologyHindawiwwwhindawicom

Nucleic AcidsJournal of

Volume 2018

Submit your manuscripts atwwwhindawicom

Page 16: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

16 The Scientific World Journal

methanolic extractsrdquo FUUAST Journal of Biology vol 4 no 1pp 1ndash5 2014

[44] K Lewis and F M Ausubel ldquoProspects for plant-derivedantibacterialsrdquo Nature Biotechnology vol 24 no 12 pp 1504ndash1507 2006

[45] G Kapoor S Saigal and A Elongavan ldquoAction and resistancemechanisms of antibiotics A guide for cliniciansrdquo Journal ofAnaesthesiology Clinical Pharmacology vol 33 no 3 pp 300ndash305 2017

Hindawiwwwhindawicom

International Journal of

Volume 2018

Zoology

Hindawiwwwhindawicom Volume 2018

Anatomy Research International

PeptidesInternational Journal of

Hindawiwwwhindawicom Volume 2018

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GenomicsInternational Journal of

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Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Hindawiwwwhindawicom Volume 2018

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Neuroscience Journal

Hindawiwwwhindawicom Volume 2018

BioMed Research International

Cell BiologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Biochemistry Research International

ArchaeaHindawiwwwhindawicom Volume 2018

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Genetics Research International

Hindawiwwwhindawicom Volume 2018

Advances in

Virolog y Stem Cells International

Hindawiwwwhindawicom Volume 2018

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Enzyme Research

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International Journal of

MicrobiologyHindawiwwwhindawicom

Nucleic AcidsJournal of

Volume 2018

Submit your manuscripts atwwwhindawicom

Page 17: Pharmacological Evaluation of Selected Medicinal Plants Used in …downloads.hindawi.com/journals/tswj/2018/4757458.pdf · 2019. 7. 30. · eScienticWorldJournal T : Continued. PlantName

Hindawiwwwhindawicom

International Journal of

Volume 2018

Zoology

Hindawiwwwhindawicom Volume 2018

Anatomy Research International

PeptidesInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Journal of Parasitology Research

GenomicsInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Hindawiwwwhindawicom Volume 2018

BioinformaticsAdvances in

Marine BiologyJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Neuroscience Journal

Hindawiwwwhindawicom Volume 2018

BioMed Research International

Cell BiologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Biochemistry Research International

ArchaeaHindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Genetics Research International

Hindawiwwwhindawicom Volume 2018

Advances in

Virolog y Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Enzyme Research

Hindawiwwwhindawicom Volume 2018

International Journal of

MicrobiologyHindawiwwwhindawicom

Nucleic AcidsJournal of

Volume 2018

Submit your manuscripts atwwwhindawicom


A Review of Medicinal Uses and Pharmacological Activities ...

A Review of Medicinal Uses and Pharmacological Activities ...

Pharmacological basis for the medicinal use of Rhus ...

Pharmacological basis for the medicinal use of Rhus ...

PHARMACOGNOSTICAL AND PHARMACOLOGICAL …ijirc.com/upload/656df1b304abc41ecb353a7b57d9e904.pdf · Indian medicinal plants have come under scientific scrutiny since the ... English

PHARMACOGNOSTICAL AND PHARMACOLOGICAL …ijirc.com/upload/656df1b304abc41ecb353a7b57d9e904.pdf · Indian medicinal plants have come under scientific scrutiny since the ... English

PHARMACOLOGICAL EVALUATION OF INDIAN MEDICINAL …repository-tnmgrmu.ac.in/1967/1/Gopi Krishnan S.pdf · chapter 4 materials and methods 22 4.1 collection and authentication of plant

PHARMACOLOGICAL EVALUATION OF INDIAN MEDICINAL …repository-tnmgrmu.ac.in/1967/1/Gopi Krishnan S.pdf · chapter 4 materials and methods 22 4.1 collection and authentication of plant

Research Article Evaluating Medicinal Plants for Anticancer Activitydownloads.hindawi.com/journals/tswj/2014/721402.pdf · 2019-07-31 · Research Article Evaluating Medicinal Plants

Research Article Evaluating Medicinal Plants for Anticancer Activitydownloads.hindawi.com/journals/tswj/2014/721402.pdf · 2019-07-31 · Research Article Evaluating Medicinal Plants

A Review of Medicinal Uses and Pharmacological Activities …A Review of Medicinal Uses and Pharmacological Activities of Nigella sativa. Title. A Review of Medicinal Uses and Pharmacological

A Review of Medicinal Uses and Pharmacological Activities …A Review of Medicinal Uses and Pharmacological Activities of Nigella sativa. Title. A Review of Medicinal Uses and Pharmacological

ESSENTIAL OILS: ITS MEDICINAL AND PHARMACOLOGICAL …

ESSENTIAL OILS: ITS MEDICINAL AND PHARMACOLOGICAL …

A NEW HETEROBIMETALLIC PALLADIUM-[60]FULLERENE … · Medicinal chemistry and pharmacological potential of fullerenes and carbon nanotubes 6 Inv.6. Matsuo Y. C60 derivatives having

A NEW HETEROBIMETALLIC PALLADIUM-[60]FULLERENE … · Medicinal chemistry and pharmacological potential of fullerenes and carbon nanotubes 6 Inv.6. Matsuo Y. C60 derivatives having

Medicinal Chemistry and Pharmacological Potential of ...dl.booktolearn.com/ebooks2/science/chemistry/1402068441_medici… · Medicinal Chemistry and Pharmacological Potential of Fullerenes

Medicinal Chemistry and Pharmacological Potential of ...dl.booktolearn.com/ebooks2/science/chemistry/1402068441_medici… · Medicinal Chemistry and Pharmacological Potential of Fullerenes

Chemistry, Pharmacology, and Medicinal Property of Sage · Key words: Pharmacological property, Sage (Salvia species), Traditional remedies Sage tea has been traditionally used for

Chemistry, Pharmacology, and Medicinal Property of Sage · Key words: Pharmacological property, Sage (Salvia species), Traditional remedies Sage tea has been traditionally used for

Pharmacological screening of some medicinal plants - Virginia Tech

Pharmacological screening of some medicinal plants - Virginia Tech

Dementia diagnosis, pharmacological and non-pharmacological interventions

Dementia diagnosis, pharmacological and non-pharmacological interventions

Pharmacological Effects of Selected Medicinal Plants and ...

Pharmacological Effects of Selected Medicinal Plants and ...

Medicinal treatment against Chemical threats · are given only as a pharmacological class (e.g. anticonvulsants, vasopressors), ... The patency of the upper airway is a major concern

Medicinal treatment against Chemical threats · are given only as a pharmacological class (e.g. anticonvulsants, vasopressors), ... The patency of the upper airway is a major concern

Non-Pharmacological and Pharmacological Management of ...

Non-Pharmacological and Pharmacological Management of ...

Medicinal And Aromatic Plants - Australia · Keywords: Australian plants, medicinal plants, aromatic plants, ethnopharmacology, phytochemical, pharmacological screening. Contents

Medicinal And Aromatic Plants - Australia · Keywords: Australian plants, medicinal plants, aromatic plants, ethnopharmacology, phytochemical, pharmacological screening. Contents

Ethanomedical, Chemical, Pharmacological, … but an important medicinal plant. ... Fennel, Fenugreek, Clove, Garlic, ... The transverse section of the leaf denoted

Ethanomedical, Chemical, Pharmacological, … but an important medicinal plant. ... Fennel, Fenugreek, Clove, Garlic, ... The transverse section of the leaf denoted

Research Article Phytochemical and Biological …downloads.hindawi.com/journals/tswj/2014/857363.pdfResearch Article Phytochemical and Biological Activities of Four Wild Medicinal

Research Article Phytochemical and Biological …downloads.hindawi.com/journals/tswj/2014/857363.pdfResearch Article Phytochemical and Biological Activities of Four Wild Medicinal

Garlic: A Review of Its Medicinal Effects and Indicated ...bigcanoe.themountainsvoice.com/wp-content/uploads/...178 (12) on pharmacological effects, Agarwal (13) on cardiovascular

Garlic: A Review of Its Medicinal Effects and Indicated ...bigcanoe.themountainsvoice.com/wp-content/uploads/...178 (12) on pharmacological effects, Agarwal (13) on cardiovascular

Phytochemistry and Pharmacological Studies of Citrus macroptera: A Medicinal …downloads.hindawi.com/journals/ecam/2017/9789802.pdf ·  · 2017-06-27Phytochemistry and Pharmacological

Phytochemistry and Pharmacological Studies of Citrus macroptera: A Medicinal …downloads.hindawi.com/journals/ecam/2017/9789802.pdf ·  · 2017-06-27Phytochemistry and Pharmacological