Viva Final Prese 2

7/31/2019 Viva Final Prese 2

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Antimicrobial Activity ofEuryale

ferox. Salisb:- A ThreatenedAquatic Plant of Kashmir

Himalaya

ByJavid Ahmad Parray

M.Phil Research scholar

Supervisor: Prof.(Dr.) Azra N. KamiliCo-Supervisor: Dr. Raies A. Qadri

P.G Department of Environmental Science

University of Kashmir


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INTRODUCTION


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Medicinal plants contain chemical substances which produce a definite

physiological action on human body and animal system.

Medicinal Plants

Phenols and

Mucilages

Flavnoids

Essential

oils TanninsGlycosides

Steroids

Chemical substances


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pharmacological

activities

Antimicrobial

Anti cancerous

Stimulatory

Anti-

inflammatory

Sedative

Spasmolytic

Secondary metabolites


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The main objective of the work was to determine theAntibacterial

and Antifungalactivity ofEuryale ferox.


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Study Area


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Source: Department of Geology and Remote Sensing, University of Kashmir.

Study Area- IRSP6-LISS III October Image of Mansabal lake


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PLANT

PROFILE


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Parts StudiedSeeds, Rhizome,

Petiole and Leaves.

Euryale ferox


11/74Euryaleferox


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Domain Eukaryota

Kingdom Plantae

Subkingdom ViridaeplantaePhylum Tracheophyta

Sub-phylum Euphyllophytina

Infra-phylum Radiatopses

Class MagnoliopsidaSub-class Nymphaeidae

Super- order Nymphaeanae

Order Nymphaeales

Family NymphaeaceaeSub-family Euphorbioideae

Tribe Euphorbie

Genus Euryale

Species E. Ferox

Scientific Classification

Source: http://Plants.gov/java/Euryale f erox/Classification Servelet
http://zipcodezoo.com/Key/Plantae/Euphorbieae_Tribe.asphttp://zipcodezoo.com/Key/Plantae/Euphorbieae_Tribe.asphttp://zipcodezoo.com/Key/Plantae/Euphorbieae_Tribe.asp


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Bihar Madhubani

English Fox nut

Kashmir Jewer

Punjabi Juwar

Hindi Makhana

Korean Ga-si-yeon-kot

China Qianshi

Japan Onibasu

Sanskrit Mukhauna, Padma

Vietnamese Khiem Thuc

Source: http: //www. herbalextractplus.com/Euryale.cfm

Different names of

Euryale ferox


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An annual aquatic herb with rhizomatous stems,

rhizome erect or repent and unbranched.

Leaves directly arise from rhizomes and are of 4-

10 cm in length with long petiolete.

Floating leaves are prickly on petioles andalong veins.

Leaf blade is abaxially dark purple and

adaxially green and is 1.3-2.7 mm indiameter and primary veins are prickly on

both surfaces .

Seeds are very hard, remains dormant for year.

Botanical Description


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China, Korea, Japan, Russia, Bangladesh,

India and grows wild in J&K and is an non

endemic threatened plant. (Khan et al.,

2000; Dar et al., 2002).

Distribution


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The plant has been traditionally used though out the

world to cure many diseases including chronic-

diarrhea, kidney problem, leucorrhea and hypo-

function of spleen. (Brown, 1995). used as an analgesic, aphrodisiac, astringent,

oxytonic and tonic in China (Duke et al.,

1985).

Leaves are used in the case of difficult parturition(Duke and Ayensu, 2003).

The plant is internally taken to treat vaginal-

discharge, impotence etc.(Brown, 1995).

Medicinal Uses


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In Indian traditional system of medicines the dry seeds

of the plant are being used as immuno-stimulant for

mothers after child birth with relatively poor immune

status (Puri et al., 2000). E. ferox also has been shown to enhance the activities of

superoxide dismutase, catalase and glutathione

peroxidase in V79-4 cells (Lee et al., 2002)

Seed of this plant have cardio protective properties

which may link with ability of makhana to induce

TRP32 and TrX-1 protein and to scavenge ROS

(Samarajit et al., 2006).


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Euryale ferox


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Seeds ofEuryale

ferox


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METHODOLOGY


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Methodology involved:Selection and collection of the medicinal

plant.

Solvent extraction. Selection of specific bacterial and fungal

strains.

Antimicrobial assay Qualitative phyto-chemical screening

METHODOLOGY


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Identification:Plant was identified at KASH,Department of Botany, University

of Kashmir under Acc. No.1015.

Collection: August -September 2008

Identification andCollection


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The powder form of different plant parts

(leaf, rhizome, petiole and seeds)

collected were subjected to hotextraction (Soxhlet extraction).

Solvents: Petroleum ether, Chloroform,

and Methanol.

Crude extraction of the

sample


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Soxhlet extractor


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Fifty grams of dried powder ofeach plant part (leaf, rhizome,

petiole and seed) were used.

Extracts collected were later driedand weighed and kept for further

process.

Whole plant

extraction

Preparation of Discs


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:

Preparation of mediaNutrient Agar Medium Dissolve 37.0 gm in 1000 ml of

distilled water

Nutrient Broth Suspend 8.0 gm in 1000 ml of

distilled water.

Muller Hinton Agar Dissolve 38.0 gm in 1000 ml of

distilled water

Muller Hinton Broth Suspend 21g of the powder in

1000ml of distilled water.

Blood Agar Suspend 40 g of the blood agar

base in 1000 ml of distilled water.

Sabourand Dextrose agar Dissolve 64 g in 1000ml of

distlled water

Preparation of Discs

Dissolve 200 mg of extract in 25 ml of solvent.

(1l=8g)


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Test Microorganisms

Gram Positive Bacterial Strains

Staphylococcusaureus I

Staphylococcus aureus II

Staphylococcus aureus III

Pnemococcus spp.


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E. Coli I

E. Coli II

Pseudomonas aeruoginosa I

Pseudomonas aeruoginosa II

Proteus vulgaris

Salmonella typhii

Salmonella typhimurium

Klebsiella pneumonia

Acintobactersp

Citrobacter fruendi

Shigella flexneri

Gram Negative Bacterial Strains


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Strains were provided by Bacteriological

and Mycological section ( Department ofMicrobiology, SKIMS Soura. Srinagar.


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Disc diffusion method: (Nutrient Agar , MullerHinton Agar and Saboarnd Dextroser agar,Hi

Media)

Kirby-Bauer (Bauer et al, 2002)

method was followed.

SENSITIVITY TEST FOR ANTIMICROBIAL ASSAY

Antimicrobial susceptibility Tests:

The primary purpose of antimicrobial susceptibility testing isto determine the potency of crude drug.

Sterile discs with antimicrobial agents are poured on microbialcultures and inhibition zone diameters (mm) around the discs are

measured as the potency of crude plant extracts.

Mixed culture of bacteria obtained from the polluted drain (that

was spread on the nutrient agar plate) and twelve extracts ( 3 ofeach plant parts) were tested for antibacterial activity.

Extracts which shows maximum activity ( methanol seed andmethanol leaf extract ) were than tested against specific bacterial and

fungal strains.

Dil i S ibili T


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Minimum inhibitory concentration(MIC)

Minimum bactericidal concentration(MBC).

Total activity (ml)

Dilution Susceptibility Tests


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S. aureusATCC.25923

E. coliATCC.25922

Pseudomonasaeroginosa

ATCC.27853

Proteusvulgaris*

Shigella

flxmeriATCC.12022

Salmonellatyphi*

StandardATCCBacterial Strains

Strains were provided by Bacteriological and Mycological section,

Department of Microbiology, SKIMS Soura. Srinagar.


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Medium

Muller Hinton Broth (Hi

Media ) (NCCLS, 1990)

i f i i f f


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Antimicrobial Conc.

mg/l

Volume of solution (ml) Distilled water (ml) Antimicrobial Conc.

mg/l

Final Conc. at 1:20

dilution(g/ml)

10000 1 9 1000 _

10000 0.1 9.9 100 _100 1 9 10 _

100 0.1 9.9 1 _

10000 1 0 10000 500

10000 0.512 0.488 5120 256

10000 0.256 0.744 2560 128

10000 0.128 0.872 1280 64

1000 0.64 0.36 640 321000 0.32 0.68 320 16

1000 0.16 0.84 160 8

100 0.8 0.2 80 4

100 0.4 0.6 40 2

100 0.2 0.8 20 1

10 1 0 10 0.5

10 0.5 0.5 5 0.25

10 0.25 0.75 2.5 0.125

10 0.125 0.875 1.25 0.06

1 0.625 0.375 0.625 0.03

1 0.313 0.687 0.313 0.015

1 0.156 0.844 0.156 0.008

(As per Philips et al ., 1991)

Preparation of dilutions of extracts for use

in MIC determination

Ph t h i l S i Q lit ti T t


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Phyto-chemical Screening Qualitative Tests

Alkaloids

Dragendorffs

test

Flavonoids

NaOHand conc.

HCl test

Phenols

Ferricchloridetest

Glycosides

Fehlingstest

SteroidsLieberman-

Burchards

test

TanninsFerricchloride test

Saponins

Frothingtest.

Physiochemical standards ofUnani

formulations- 1987


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By

Isolation of Phyto-constituents

Thin Layer

chromatography


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RESULTS

.

A i i bi l i i f E l f i


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GPC= Gram Positive Cocii

GNB= Gram Negative Bacillus

+ = Inhibitory activity

- = No activity

Pt = Petroleum ether extract

C = chloroform extract

M = methanol extract

Antimicrobial activity ofEuryale ferox extracts against

mixed culture of GPC & GNB bacteria.

Strains Inhibition Zone Diameter (mm)

Rhizome Petiole Leaf Seed

Pt C M Pt C M Pt C M Pt C M

Mixed culture of GPC& GNB bacteria.

_ _ _ _ _ _ _ _ + _ _ +


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Strains Inhibition Zone Diameter (mm)

Petroleum ether Chloroform Methanol

Mixed culture of

GPC & GNB

bacteria.

_ _ _

GPC= Gram Positive Cocii

GNB= Gram Negative Bacillus;

+ = Inhibitory activity

- = No activity.

Solvent Control


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GPC=Gram Positive Cocci

GNB= Gram Negative

Bacilli

M=methanol

Pt=Petroleum etherC=chloroform

SM=Seed methanol

SC=Seed chloroform

SP= Seed pet ether

LM=Leaf methanol

LC=Leaf Chloroform

LP=Leaf pet etherPM=Petiole methanol

PC=Seed chloroform

PP= Seed pet ether

RM=Leaf methanol

RC=Leaf Chloroform

RP=Leaf pet ether

FIG. 1a: Solvent extracts

ofE. ferox

FIG. 1b: Solvent Control

Plate1: Mixed culture of GPC & GNB


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Fig 2a :E. coli Fig 2b:Staphylococcus aureus

Plate 2: Effect of different concentrations of methanol seed extracts ofE.ferox on E.coli and Staphylococcus aureus

SI = 5 l and S11 =55 l (seed methanol extract)

Antibacterial activity of methanolic seed extracts of Euryale


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S.

No.

Strains Inhibition Zone Diameter (mm)*

160 g 320 g Antibiotics

01 S. aureus I 120.64 23 0.41 Erythromycin(25)

02 S. aureus II 120.81 18 0.70 Vancomycin (20)

03 S. aureus III 101.63 18 0.81 Cotrimaxozole (24)

04 Pnemococcus spp 151.29 20 0.81 Lenzolid (22)

* Results are mean diameter of zone of inhibition of three replicates followed by standard

deviation; Methanol was taken as negative control and was found resistant in all strains.

Antibacterial activity of methanolic seed extracts ofEuryale

ferox against Gram Positive Bacterial strains.

Antibacterial activity of methanolic leaf extracts of Euryale


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S. No Strains Inhibition Zone Diameter (mm) *

160 g 320 g Antibiotic

01 S. aureus I NA NA Erythromycin (25)

02 S. aureus II 90.00 151.28 Vancomycin (20)

03 S. aureus III 171.28 201.73 Cotrimaxozole (24)

04 Pnemococcus

spp

121.07 160.40 Lenzolid (22)

*Results are mean diameter of zone of inhibition of three replicates followed by standard

deviation; Methanol was taken as negative control and was found resistant in all strains

Antibacterial activity of methanolic leaf extracts ofEuryale

ferox against Gram Positive Bacterial strains.


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Gram Positive Bacterial Strains

Inh

ibitionzonesDiameter(mm)

Fig. 1: Comparative assessment of antibacterial activity of methanolic seed and leaf Extract ofE.

ferox against Gram Positive Bacterial Strains (320g)

Antibacterial activity of methanolic seed extracts of Euryale


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S. No. Strains Inhibition Zone Diameter (mm) *

160 g 320 g Antibiotics01 E .coli I 12 1.15 15 0.00 Ceftazidime (30)

02 E. coli II 16 2.16 22 1.63 Gentamycin (20)

03 Pseudomonas aeroginosa I 18 2.82 28 0.81 Amikacin (15)

04 Pseudomonas aeroginosa II 16 0.00 27 1.29 Gatifloxcin (20)

05 Acintobacterspp 10 2.16 14 1.15 Piperacilin (0)

06 Citrobacter freundii NA NA Vancomycin (22)

07 Proteus vulgaris NA 19 1.41 Cefixime

08 Shigella flexmeri 141.41 231.61 Ciprofloaxcin (29)

09 Klebsiela pneumonia NA NA Gentamycin (25)

10 Salmonella typhi 18 0.81 25 0.57 Imipenin (28)

11 Salmonella typhimurum NA 15 1.28 Gatifloxcin (0)



Antibacterial activity of methanolic seed extracts ofEuryale

ferox against Gram Negative Bacterial strains.

Antimicrobial activity of methanolic leaf extracts of Euryale


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S. No. Strains Inhibition Zone Diameter (mm) *

160 g 320 g Antibiotics01 E. coli I 8 1.22 100.16 Ceftazidime (30)

02 E. coli II 13 0.42 200.41 Gentamycin (20)

03 Pseudomonas aeroginosa I 16 1.10 22 0.24 Amikacin (15)

04 Pseudomonas aeroginosa

II

17 1.24 21 0.81 Gatifloxcin (20)

05 Acintobacter spp 10 0.16 16 0.40 Piperacilin (0)

06 Citrobacter freundii NA NA Vancomycin (22)

07 Proteus vulgaris NA 10 0.00 Cefixime

08 Shigella flxmeri 12 0.74 18 0.21 Ciprofloaxcin (29)

09 Klebsella pneumoniae NA NA Gentamycin (25)10 Salmonella typhi 11 0.08 21 0.14 Imipenin (28)

11 Salmonella typhimurum NA 10 0.16 Gatifloxcin (0)

*Results are mean diameter of zone of inhibition of three replicates followed by Standard


Antimicrobial activity of methanolic leaf extracts ofEuryale

ferox against Gram Negative Bacterial strains.


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Fig. 2: Comparative assessment of antibacterial activity of methanolic seed and leaf extract

ofE. ferox against Gram Negative Bacterial Strains (320 g)

InhibitionZonesD

iameter(mm)

Gram Negative Bacterial Strains


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Plate 3: Antibacterial activity of methanolic seed and leaf extract of

E. ferox against S.aureus I and Pneumococcus spp

Fig. 3a: S. aureus I Fig. 3b:Pneumococcus Spp.

SI=160 g, S2=320 g= Seed methanol extract. L1=160 g,

L2=320 g = Leaf methanol extract, Ab= Antibiotic; M= methanol


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Fig. 4a: S. aureus II Fig. 4b: S. aureus III

Plate: 4 Antibacterial activity of methanolic seed and leaf extract ofE. ferox against

S.aureus II and S.aureus IIISI=160 g, S2=320 g= Seed methanol extract. L1=160 g,

L2=320 g = Leaf methanol extract, Ab= Antibiotic; M= methanol


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Fig. 5a: P. aureoginosa I Fig. 5b:P. aureoginosa II

Plate 5: Antibacterial activity of methanolic seed and leaf extract ofE. ferox againstP.

aureoginosa I and P. aureoginosa II

SI=160 g, S2=320 g = Seed methanol extract. L1=160 g, L2=320 g = Leaf

methanol extract, Ab= Antibiotic; M= methanol


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Fig. 6a: S. typhi Fig. 6b: E. coli I

Fig. 6c:E. coli II

Plate 6: Antibacterial activity of methanolic

seed and leaf extract ofE. ferox against S.

typhi,E. coli IandE. coli II.

SI=160 g, S2=320 g = Seed methanol

extract. L1=160 g, L2=320 g = Leafmethanol extract, Ab=Antibiotic;

M=methanol


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Fig. 7a: Shigella flexnerii

Plate 7: Antibacterial activity of methanolic seed and leaf extract ofE. ferox against Shigella

flexnerii andAcintobacter spp.

SI=160 g, S2=320 g = Seed methanol extract. L1=160 g, L2=320 g = Leaf methanol extract,

Ab= Antibiotic; M=methanol

Fig. 7b: Acintobacterspp.


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Fig. 8a: Citrobacter freundii

Plate 8: Antibacterial activity of methanolic seed and leaf extract ofE. ferox against Citrobacter

freundii and Klebsiella pneumonia

SI=160 g, S2=320 g= Seed methanol extract. L1=160 g,L2=320 g = Leaf methanol extract, Ab= Antibiotic; M= methanol

Fig. 8b: Klebsiella pneumonia


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Plate 9: Antibacterial activity of methanolic seed and leaf extract ofE. ferox against Salmonella

typhimurium and Proteus vulgaris

SI=160 g, S2=320 g= Seed methanol extract. L1=160 g,L2=320 g = Leaf methanol extract, Ab= Antibiotic; M= methanol

Fig. 9b: Proteus vulgaris

Fig. 9a: Salmonella typhimurium

Minimum Inhibitory Concentration (MIC) of methanolic seed extract of


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S. No. Strains MIC MBC MIC

Index

Total

Activity

(ml)

Gt. 10 g/ml

MIC MBC

01 S. aureus ATCC.25923 64 128 2.0 593.75 32 128

02 E. coli ATCC.25922 128 256 2.0 296.87 128 500

03 Pseudomonas aeroginosaATCC. 27853

64 256 4.0 593.75 16 64

04 Shigella flxmeri

ATCC.12022

ND ND _ _ ND ND

05 Proteus vulgaris* 500 ND 76 256 500

06 Salmonella typhi* 256 500 1.95 148.43 ND ND

Methanol was used as negative control and exhibited no activity against tested organisms;

ND = Non detectable within the range tested; * = isolated Strains; Gt. = Gentamicin

MIC index = 4 (bactericidal) and = 4 (bacteriostatic)

Total activity =highest value= highest inhibitory activity

y

Euryale ferox against standard bacterial strains (g/ml)

.

Minimum Inhibitory Concentration (MIC) of methanolic leaf extract of


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S. No. Strains MIC MBC MIC

Index

Total

Activity

(ml)

Gt 10 g/ml

MIC MBC

01 S. aureus ATCC.25923 128 500 3.90 737.5 32 128

02 E. coli ATCC.25922 256 500 1.95 368.75 128 500

03 Pseudomonas aeroginosa

ATCC.27853

256 500 1.95 368.75 16 64

04 Shigella flxmeri

ATCC.12022

500 ND ND 188.8 ND ND

05 Proteus vulgaris* ND ND ND _ 256 500

06 Salmonella typhi* 500 ND 188.8 ND

Methanol was used as negative control and exhibited no activity against tested organisms;

ND = non detectable within the range tested; * = isolated Strains; Gt. = Gentamicin.

MIC index = 4 (bactericidal) and = 4 (bacteriostatic)

Total activity =highest value= highest inhibitory activity

y ( )

Euryale ferox against standard bacterial strains (g/ml).


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Standard Bacterial Strains

MIC(g/ml)

Fig. 3: Comparative assessment of MIC of methanolic seed and leaf extract ofE. ferox

against standard bacterial strains (g/ml)


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Fig. 4: Comparative assessment of MBC of methanolic seed and leaf extract of

E. ferox against standard bacterial Strains (g/ml)


MBC(g/ml)

g/ml)


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Fig. 5: Comparative assessment of Total activity of methanolic seed and leaf Extract ofE. ferox

against standard bacterial Strains (ml)

Totala

ctivity(ml)


Antifungal activity of methanolic leaf and seed extract of Euryale ferox.


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S. No. Fungal Strains Leaf extract Seed extract

200 g 400 g 200 g 400 g

01 Crytococcus neoformans NA NA NA NA

02 Aspergilus fumigates NA NA NA NA

03 Candida albicans 9 0.81 16 0.70 70.3 12 0.81

04 C. kruesie 8 0.48 100.28 70.3 9 0.00

05 C.Stealoidea 10 0.62 150.34 80.2 11 0.5

06 C. tropicals NA 150.34 NA 18 0.14

07 C. parapsilosis NA NA NA NA

08 Rhizopus spp NA NA NA NA

09 Pencillum notatum 10 0.81 150.60 80.2 19 0.84


deviation; Nystatin was used as positive control and Methanol as negative control in all

strains.

Antifungal activity of methanolic leaf and seed extract ofEuryale ferox.


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Fig. 6: Comparative assessment of antifungal activity of methanolic Seed and Leaf

extract ofE. ferox against fungal Strains (400 g).

InhibitionZ

oneDiameter(mm

)

Fungal Strains


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Fig. 10a: Aspergillus fumigates

Plate 10: Antifungal activity of methanolic seed and leaf extract of

E. ferox against Aspergillus fumigates and Cryptoccocus neoformans

SI=200 g, S2=400 g = Seed methanol extract. L1=200 g, L2=400 g = Leaf methanol

extract, Ab= Antibiotic; M=methanol

Fig. 10b: Cryptoccocus neoformans


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Fig. 11a: C. parapsilosis

Plate. 11: Antifungal activity of methanolic seed and leaf extract ofE. ferox against C.parapsilosis andRhizopus spp.



Fig. 11b:Rhizopus spp


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Fig. 12a: C. albicans

Plate 12: Antifungal activity of methanolic seed and leaf extract ofE. ferox against C. albicans and

C. kruesie



Fig. 12b: C. Kruesie


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Fig. 13a: P. notatum

Plate 13: Antifungal activity of

methanolic seed and leaf extract

ofE. ferox against P. notatum

C. stealoidea andC. tropicals .

SI=200g, S2=400 g = Seed methanol

extract.L1=200g,L2=400 g = Leaf

methanol extract, Ab= Antibiotic;

M=methanol

Fig. 13c: C. tropicals

Fig. 13b: C. stealoidea

Ph h i l C i f M h li E f


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Constituents tested for Leaf extract Seed extract

Alkaloids + +

Steroids _ +

Glycosides + +

Phenols + +

Tannins + +

Saponins _ +

Flavonoids + +

- = Not present , + = Present.

Phyto-chemical Constituents of Methanolic Extracts of

Euryale ferox


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A= Petiole methanol extractB= Seed methanol extract

C= Rhizome methanol extract

D= Leaf methanol extract

Solvent front =16.5cm

A B C D

Rf l f M h li f E l f


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Methanolic Extracts Rf. value

Seed 0.176, 0.41, 0.58, 0.63 and 0.84

Leaf 0.176, 0.35, 0.45 and 0.59

Petiole 0.26

Rhizome 0.11

Solvent front = 17 cm

solvents used = Chloroform: Methanol (7:3)

Rf values of Methanolic extracts of Euryale ferox

Extraction yield and macroscopic characteristics of the crude

t t f E l f


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Name of the

plant

Common

Name

Solvent Part used Macroscopic characteristics Extract/

yield (%)

Euryale ferox Jewer

Methanol

Chloroform

Petroleum ether

SeedSeed

Seed

Black crystalline powderBrown crystalline powder

Dark green shiny surface

3.84

3.07

0.3

Methanol

ChloroformPetroleum ether

Leaves

LeavesLeaves


Dark green powderYellowish green powder

9.44

1.740.6

Methanol

Chloroform

Petroleum ether

Rhizome

Rhizome

Rhizome

Dark black powder


A dark brown shiny surface

7.88

0.38

1.76

Methanol

Chloroform

Petroleum ether

Petiole

Petiole

Petiole

Dark green powder

Brown crystalline powder

Yellowish blue powder

10.9

0.98

1.66

extracts ofEuryale ferox


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CONCLUSION

O l th li t t f d d l f hibit d hi h t


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Only methanolic extracts of seed and leaf exhibited highest

antimicrobial activity.

Seed extracts exhibited higher inhibitory activity than leaf

extracts. Gram Positive strains were more susceptible than gram negative

strains.

Among the gram positive strains, S.aureusshows maximum zone of

inhibition i.e highest inhibitory activity. However, IZD of some gram negative strains ( i.e. P. aureoginosa)

were more than gram positive strains.

Among the fungal strains, Candida spp. and Pencillum notatum

were more susceptible to both extracts at higher

concentrations.

However C. neoformans,A. fumigates andRhizopus spp.

C.Parapsilosiswere completely resistant.

The results of MIC and MBC depicted that extracts ofE.


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p

ferox can be bactericidal in action and among

standard strains tested, S. aureus ATCC 25923 was

inhibited by least concentration of extracts used. Phyto-chemical screening of methanolic seed and leaf

extracts revealed the presence of alkaloids, phenols,

flavonoids, steroids and tannins while as steroids

and Saponins were present in only seed extract. The separation of methanolic extracts by thin layer

chromatography (TLC) yields different compounds

with seed extract showed five compounds of Rf

values 0.176, 0.41, 0.58, 0.63 and 0.84 while as

leaf extract yields four compounds with Rf. value of

0.176, 0.35, 0.45 and 0.59.

The substances that can inhibit pathogens and have


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p g

a little toxicity to host cells are considered good

for developing antimicrobial drugs.

Therefore, the antimicrobial activity ofE. feroxextracts on these UTI organisms reveals that it can

be safely used to treat these kinds of infections

after proper drug formulation.

The literature reveals that no prior work has been

done on evaluating antimicrobial activity ofE.

ferox and it is now expected after results that it

will be helpful in obtaining broad spectrum herbal

formulation as well as new antimicrobial

substances.


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