Bioassay-Guided Fractionation and Anti-Trypanocidal Effect ...

Copyright © 2013 by Modern Scientific Press Company, Florida, USA

International Journal of Modern Biochemistry, 2013, 2(1): 1-14

International Journal of Modern Biochemistry

Journal homepage: www.ModernScientificPress.com/Journals/IJBioChem.aspx

ISSN: 2169-0928

Florida, USA

Article

Bioassay-Guided Fractionation and Anti-Trypanocidal Effect of

Fractions and Crude Methanol Roots Extracts of Securidaca

Longepedunculata in Mice and Rats

Y. Haruna

1, 3, *, H. O.

Kwanashie

1, J. A.

Anuka

1, S. E.

Atawodi

2, I. M.

Hussaini

1

1Department of Pharmacology and Therapeutics Ahmadu Bello University, Main Campus, P.M.B.

1045, Samaru, Zaria, 810271, Kaduna, Nigeria 2Department of Biochemistry, Ahmadu Bello University, Main Campus, P.M.B. 1045, Samaru, Zaria,

Nigeria

3Department of Medicinal Chemistry, Kebbi State University of Science and Technology, P.M.B. 1144,

Aliero, Birnin Kebbi, Nigeria

* Author to whom correspondence should be addressed; E-Mail: [email protected]

Article history: Received 1 January 2013, Received in revised form 20 January 2013, Accepted 21

January 2013, Published 24 January 2013.

Abstract: An In-Vivo study was carried out in order to determine the anti-trypanosomal

effect of methanol root extracts of Securidaca longepedunculata, and its fractions in

trypanosoma brucei brucei first in Swiss albino mice, then in Wister albino rats. Groups of

infected mice were treated with 5%, 10% and 20% of the extracts LD50 which is equivalent

to (0.14, 0.28 & 0.56 mg/kg) intraperitoneally, and the effects of treatment, parasitaemia,

packed cell volume (PCV), body weight, and lifespan were monitored. The same procedure

was repeated in rats except that they were sacrificed on the 11th

day post infection. A

statistically significant (P<0.05) suppression of parasitaemia, alleviation of anaemia, body

weight recovery, and prolonged lifespan was observed dose dependently with both the

crude methanol roots extracts and the fractions of S. longepedunculata. Phytochemical

analysis of the fractions revealed the presence of flavonoids, saponins, tannins, and

alkaloids; which are implicated in to this activity. It is conclusive that the crude extracts

and fractions of this plant has potentials in combating these parasites that constitute serious

problems to livestock and even people living in the tropics especially if explored further.

Keywords: Anti-trypanosomal effect, Extracts, fractions, Parasitaemia, Securidaca

longepedunculata, Trypanosoma brucei brucei.

Int. J. Modern Biochem. 2013, 2(1): 1-14


2

1. Introduction

Human African trypanosomiasis (HAT) or sleeping sickness is a major public health problem

in 36 sub-Saharan African countries and is caused by Trypanosoma brucei gambiense and T. b.

rhodesiense; while in South America, Trypanosoma cruzi is the causative organism of Chagas disease.

Animal Typanosomiasis (nagana) on the other hand is caused by T. b. brucei, T. congolense and T.

vivax. It threatens over 60 Million lives on daily basis (Steverding et al, 2005). Typanosomiasis of

domestic animals is responsible for the death of about 3 million cattle annually which is accompanied

by 20% decrease in animals and 25% decrease in milk productions respectively (WHO, 1975). These

have restricted land use and prevented some of the otherwise, most favourable agricultural lands of

sub-Saharan Africa from being developed (Barry, 1976). Until now, the current chemotherapy of

human trypanosomiasis relies on six drugs: Suramin, Pentamidine, Melarsoprol, Eflorinithine, Arsobal

and Mel B; five of which were developed more than 30 years ago (Steverding etal, 2005). Furthermore

these drugs display undesirable toxic side effects and drug resistance (Kuzoe, 1993., Steverding etal,

2005 and Perez-Morga, 2007). And the mortality rate of melarsoprol treated patients is 1-5%

(Bouteille et al, 1995 and WHO, 2007). These problems together with the associated expensive nature

of most drugs in developing nations with poor economic status make it pertinent to search for new,

better and cheaper trypanocides.

Securidaca longepedunculata Fresen (Polygalaceae) also called the violet tree is the most

popular of all among the traditional medicinal plants in South Africa and is used for almost every

conceivable ailment (Van Wyk and Gericke, 2000), some vernacular names (Hausa) Northern Nigeria

mean ‘the king of medicines’. It is reported that almost all parts of S. longepedunculata (leaves, twigs,

stem, bark, roots and seeds) are used by man for different purposes such as medicines; the root and the

bark are taken orally either powdered or infusion for treating chest complaints, inflammation, abortion,

ritual suicide, tuberculosis, infertility, venereal diseases etc (Neuwinger, 1996., Atawodi et al, 2003)

reported that In-vitro screening of the methanol root extract of S. longepedunculata suggested potent

anti-trypanosomal activities. This In-vitro claim was further confirmed by Ameh, et al (2006).,

Adebaubaurer, et al, (2008) that the roots extract of S. longepedunculata were able to reduce

parasitaemia in mice experimentally infected with T. brucei by 42 and 48% at a dose of 150 mg/kg

b.wt. (i.p) twice daily for three days. Therefore, this work was designed to further explore the In-vivo

anti-trypanosomal potential and identify the possible classes of phytochemicals present in S.

longepedunculata.



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2. Materials and Methods

2.1. Plant Material

The roots of S. longepedunculata were collected from Zuru, Kebbi State, Northern Nigeria. The

plant was authenticated by a taxonomist at the herbarium section of Biological Sciences Department,

Ahmadu Bello University Zaria, Nigeria. Where a voucher specimen was deposited and it was given a

voucher number: 900149. The roots were thoroughly washed with water, air dried and pounded into

powder using pestle and mortar.

2.1.1. Animals and animal husbandry

Healthy adult Swiss albino mice and Wistar albino rats of both sexes weighing approximately

±21.20 g and ± 180 g respectively were used for this study. The animals were obtained from the animal

house of the Faculty of Pharmaceutical Sciences, Ahmadu Bello University, Zaria, Nigeria. They were

housed in cages at the same Departmental animal house, fed on standard rats feed (vital feeds, Jos,

Nigeria) and allowed access to water ad libitum. The animals were allowed to acclimatize to the

laboratory conditions for at least 14 days before being subjected to the experiments. All the

experiments were carried out in a Conducive laboratory setting that has ambient illumination and a

temperature that is close to that of the animal house, according to OECD guidelines.

2.1.2. Test organisms

Trypanosoma brucei brucei organisms were obtained from National Institute of

Trypanosomiasis and Onchocerciasis Research (NITR) No. 1, Sulame Road, Unguwan rimi, Kaduna,

Kaduna State, Nigeria. They were maintained in the animal house of the Faculty of Pharmaceutical

Sciences, Ahmadu Bello University Zairia, Kaduna State Nigeria by continuous passage into donor

rats intraperitoneally.

2.1.3. Extract preparation

The methanol crude extracts of the roots of S. longepedunculata was obtained by Soxlet

extraction using methanol as a solvent for 48 hours. The extract was concentrated to dryness on a

water bath between 40-45oC. The extracts were stored in sealed bottles at room temperature until

tested/fractionated.

2.1.4. Fractionation (Vacuum Liquid Chromatography)

The methanol root extract of S. longepedunculata (with the highest biological activity) was

fractionated using a short column or a Buchner filter funnel fitted with glass frit (10-20µm, porosity D

or porosity 2). It was dry packed with sorbet (silica gel of TLC grade 10-40µm) e.g. Merck 60H or



4

60G silica gel. The sorbet was allowed to settle by gentle tapping under gravity. Then vacuum was

applied via the three way stop cork and the sorbet compressed a hard layer by pressing with a rubber

stopper and tapping. The vacuum was released, solvent of low polarity (n-hexane) was poured quickly

onto the surface of the adsorbent then vacuum was reapplied, the sample was pre-absorbed on celite.

The column was developed with appropriate solvent mixtures starting with solvent of low polarity and

gradually increasing the polarity, pulling the column dry between each fraction collected. Fractions

were collected in a suitable separating funnel and finally into collection bottles. All the solvents used

were distilled before the commencement of the work. The eluates were dried at room temperature and

kept at 4oC until required.

Figure 1: Schematic representation of Bioassay guided isolation of active compounds from Ethyl

acetate fraction of S. longepedunculata only fractions with Trypanocidal activity are shown.

Sub-Fraction (11 - 14) exhibited the highest biological activity.

2.2. Phytochemical Test of the Fractions of S.longepedunculata

The possible presence of saponins, flavonoids, tannins, carbohydrates, glycosides, and

anthraquinones was determined using the method of Sofowora, (1993).

A Fraction 1 -

13

B Fraction 14 -

22

Ethyl acetate fraction

Vacuum Liquid Chromatography Yielded

C Fraction 23 -

31

D Fraction 32 E Fraction 33 - 34

B1 (1-10) B5 (11 -14) B6 (15-20)

(1151((((((

(((151520

30202222

0)

B8 (21 – 32)

Vacuum Liquid Chromatography

Yielded



5

2.3. Estimation of Trypanosome Parasites in the Blood

The parasites in the blood were estimated according to the “rapid matching” method of Herbert

and Lumsden (1976); as described by Atawodi et al (2003 & 2005). The method involves a matching

technique in which microscopic fields were compared with a range of standard logarithm values.

Counting of parasites per field in pure blood or blood approximately diluted with buffered phosphate

saline (PBS PH 7.2) a drop of blood was obtained on a slide by pinching the tip of the pre-sterilized

tail with a sterile needle, immediately covered with a cover slip and the wet mount observed under the

microscope at X40 magnification. The number of trypanosomes per microscopic field was compared

with the table of logarithmic values. The logarithmic values which matched the microscopic

observation were then converted to antilogarithm, from where the absolute number of trypanosomes

per ml of blood was obtained.

3. Preliminary in vivo Trypanocidal Screening of Methanol Root Extracts of

Securidaca longepedunculata on Mice Infected with T. brucei.

The roots of Securidaca longepedunculata were screened using Methanol in Swiss albino mice

infected with trypanosome brucei.

3.1. Experimental Procedure

A standard protocol was drawn up in accordance with the Good Laboratory Practice (GLP)

regulations of the World Health Organization (WHO Document, 1998).

Thirty healthy Swiss albino mice were randomly selected for this study, and divided weight

dependently into groups of 5 mice each, consisting of three methanol extracts groups of 5%, 10%, and

20% of the extract’s LD50 which is equivalent to 0.14, 0.28, & 0.56 mg/kg of respectively, and also a

standard control drug (diminazene aceturate 3.5 mg/kg), infected and not treated group and no

infection no treatment group. Except the no infection no treatment group, all other groups were

infected with T. brucei. Invariably, each animal received inoculums of about 1.0 x 107

parasites per

gramme body weight through needle passage and produced parasitaemia in the mice. On

commencement of the medications (i.e. at peak parasitaemia) the methanol root extract of S.

longepedunculata was given to the three groups in divided doses as shown below for seven days and

the diminazene aceturate was given to the standard group at a therapeutic dose of 3.5 mg/kg just once,

while the infected not treated and the no infection no treatment received no treatment, all the drugs

were given through intra-peritoneal routes after confirming parasitaemia:

GROUP 1: Is a negative control infected with T. brucei but no treatment.



6

GROUP 2: Is a positive control group, not infected and no treatment.

GROUP 3: Infected with T. brucei and treated with diminazene aceturate 3.5 mg/kg once (therapeutic

dose) (i.p).

GROUP 4: Infected with T. brucei and treated with 5% of extracts LD50 0.14 mg/kg of methanol root

extract of (SLE) daily for 7 days.(i.p)

GROUP 5: Infected with T. brucei and treated with 10% of extracts LD50 0.28 mg/kg of (SLE)

methanol extract daily for 7 days.(i.p)

GROUP 6: Infected with T. brucei and treated with 20% of extracts LD50 0.56 mg/kg of (SEL)

methanol extract daily for 7days (i.p.

3.2. Bio-assay Guided Fractionation of the Sub-fractions Obtained from Vacuum Liquid

Chromatography of Root Extracts of Securidaca longepedunculata on Rats Infected with T.

brucei brucei

3.2.1. Experimental procedures

A standard protocol was drawn up in accordance with the Good Laboratory Practice (GLP)

regulations of the World Health Organization (WHO Document, 1998).

Thirty healthy male Wistar albino rats were randomly selected for this study, and were weight

dependently divided into groups of 5 rats each, consisting of three methanol extracts groups of 5%,

10%, and 20% of the extract’s LD50 which is equivalent to 0.14, 0.28, and 0.56 mg/kg respectfully, and

also a standard control drug (diminazene aceturate 3.5 mg/kg), infected and not treated group and no

infection no treatment group. Except the no infection no treatment group, all other groups were

infected with T. brucei invariably, each animal received inoculums of about 1.0 x 107

parasites per

gramme body weight through needle passage and produced parasitaemia in the rats. On

commencement of the medications (i.e at peak parasitaemia) the methanol root extract of S.

longepedunculata was given to the three groups in divided doses as shown above for seven days and

the diminazene aceturate was given to the standard group at a therapeutic dose of 3.5 mg/kg just once,

while the infected not treated and the no infection no treatment received no treatment, all the drugs

were given through intra-peritoneal routes after confirming parasitaemia for seven days in mice, while

in the rats phase, they were all sacrificed on day eleven post infection.

GROUP 1: Is a negative control group that was infected with T. brucei but no treatment.

GROUP 2: No infection no treatment group.

GROUP 3: Infected with T.brucei and treated with diminazene aceturate once 3.5 mg/kg therapeutic

dose (i.p.)

GROUP 4: Infected with T. brucei and treated with 5% of extracts LD50 0.14 mg/kg of fractions (11 to

14) of S. longepedunculata. (SLE) daily for 7 days i.p



7

GROUP 5: Infected with T. brucei and treated with 10% of extracts LD50 0.28mg/kg of fractions (11 to

14) of S. longepedunculata (SLE) daily for 7 days i.p.

GROUP 6: Infected with T. brucei and treated with 20% of extracts LD50 0.56 mg/kg of fractions (11

to 14) S.longepedunculata (SLE) daily for 7 days i.p.

i.p. = intra-peritoneal., b.w. =body weight. SLE= Securdaca longepedunculata extracts

3.3. Packed Cell Volume Determination

Blood samples were collected into heparinized capillary tubes with one end of the tube sealed

with plasticine or a bunsen flame were spinned at 2000 g for five minutes in a micro-haematocrit

centrifuge. The PCVs were determined with the aid of a micro-haematocrit reader and the values

expressed as percentages.

3.3.1. Body weight changes determination

Body weights of all the animals in each group was taken before the commencement of the

experiment, thereafter when all the inoculated animals received medications, the weight of their bodies

was also taken and compared with the initial weight.

3.4. Statistical Analysis

Data were analyzed using Analysis of variance (ANOVA) and student t-test using SPSS-

computer package. In all cases the level of statistical significance was considered at (P< 0.05).

4. Results

Microscopic examination of this work showed that peak parasitaemia was reached in 5 to 6

days post inoculation in both mice and rats (table 1). Medications in all the infected groups

commenced on the 6th

day post inoculation (peak parasitaemia), similarly microscopic examination

revealed a steady increase of parasitaemia in all animals of the negative control group (infected with

T.bruce but not treated) until all the animals died on day 14 post inoculation due to parasitaemia (table

1). After administration of diminazene aceturate (intraperitoneally) microscopic examination showed

that parasites in the group were completely cleared and the mice/rats remained alive throughout the

period of the preliminary studies.



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Table 1. Results of methanolic root extracts of S. longepedunculata in mice infected with T. brucei brucei.

Average parasite level per days of observation

Treatment

Groups

N Dose

(mg/kg)

D4 D5 D6 D7 D8 D9 D10

Normal saline 5 0 2.000± 0.32 4.66 ± 0.43 7.34 ± 0.50 9.86 ± 0.11 11.52± 0.32 13.45± 0.41 *15.08± 0.36

No infection

no treatment

5

0 0 0 0 0 0 0 0

Diminazine

aceturate

5

3.5 2.18 ± 0.27 4.28 ± 0.32 7.26 ± 0.42 0 0 0 0

SLE 5 0.14 1.84 ± 0.29 4.10 ± 0.17 6.68 ± 0.28 7.92 ± 0.30 8.60 ± 0.30 8.24 ± 0.22 *7.50 ± 0.30

SLE 5 0.28 * 3.84 ± 0.30 7.04 ± 0.52 8.84 ± 0.48 9.10 ± 0.31 8.40 ± 0.26 *6.96 ± 0.12

SLE 5 0.56 2.50 ± 0.24 3.90 ± 0.17 7.24 ± 0.33 8.34 ± 0.38 8.40 ± 0.20 7.12 ± 0.36 *5.40 ± 0.23

Average parasiteamia per days of observation

D11 D12 D13 D14 D15 D16 D17 D18 D20 D25 D30

*15.72± 0.47 *19.72± 0.49 *24.64± 0.66 *35.06± 0.50 0 0 0 0 0 0 0

0 0 0 0 0 0 0 0 0 0 0

0 0 0 0 0 0 0 0 0 0 0

*7.06 ± 0.33 *6.84 ± 0.51 *6.84 ± 0.43 *7.72± 0.10 8.68± 0.19 9.86± 0.12 11.97± 0.28 15.80± 0.41 27.10± 0.60 0 0

*6.04 ± 0.12 *5.02 ± 0.07 *4.86 ±0.15 *5.68± 0.19 6.76 ±0.32 7.74 ± 0.36 9.40 ± 0.24 12.32± 0.57 30.46± 1.45 0 0

*4.60± 0.026 *3.70 ± 0.29 *3.20± 0.13 *3.58± 0.18 4.18± 0.22 4.64± 0.28 6.08± 0.24 8.96± 3.35 20.04±4.20 26.98± 6.77 31.34± 87.78

*= Significant parasitaemia reduction (P<0.05) compared to the control group; SLE = Securidaca longepedunculata extract; N = Number of animals used per group.

4.1. Effect of Extract/Fractions Administration on Lifespan on Trypanosome-infected Mice/Rats

The work equally showed a steady decrease in parasitaemia in the plant treated groups dose

dependently (0.14, 0.28, 0.56 mg/kg) respectively. It also showed that the extracts treated animals

survived for 25 days before they died. Though, relapse parasitaemia occurred, but results showed that

on daily administration of the extract of S. longepedunculata by day 8 post administration there was a

statistically significant reduction (P<0.05) in parasitaemia load due to the effect of the extract to as low

as 3.58 ± 0.18, 5.68 ± 0.19, & 7.72 ± 0.10 of 0.56, 0.28 & 0.14 mg/kg dose respectively when

compared to the negative control group with 35.06 ± 0.50 parasite value of the same day 8 post

medication (table 1), while in rats 3.76 ±0.42, 5.50 ± 0.40,& 5.58 ± 0.32 of 0.56, 0.28, &0.56 mg/kg

respectively (table 2).

4.2. Packed Cell Volume and Parasitaemia in Animals Administered Methanol Root Extracts and

Fractions of S. longepedunculata

Packed cell volume (PCV) and anemia are critical issues in the pathogenesis of African

trypanosomiasis contributing to morbidity and mortality thus curtailing the absence longevity

(Jennings et al 1977). Infected mice given S. longepedunculata root extracts and fractions in this study

showed significantly (P<0.05) higher level in PCV recovery compared to the infected not treated

groups given normal saline only (tables 1 &2).



9

Table 2. Bio-assay guided fractionation of pooled Fractions of S. longepeduncalata in rats infected

with T. brucei brucei.

Average parasite level per days of observation

Treatment

Groups

N Dose

(mg/kg) D4 D5 D6 D7 D8 D9 D10

Infection

not treaedt

5 0 2.64 ± 0.29 6.70 ± 0.58 9.66 ± 0.29 11.24± 0.53 13.18± 0.67 18.12± 2.32 24.38 ± 3.32

Normal

saline

5 0 0 0 0 0 0 0 0

Diminazine

aceturate

5 3.5 2.54 ± 0.34 7.08 ± 0.37 9.58 ± 0.25 0 0 0 0

PF 11-14 5 0.14 2.64 ± 0.23 7.56 ± 0.38 10.00± 0.46 8.82 ± 0.25 7.76 ± 0.19 7.18 ± 0.26 6.30 ± 0.33

PF 11-14 5 0.28 2.34 ± 0.18 7.06 ± 0.28 9.48 ± 0.24 8.80 ± 0.23 7.88 ± 0.14 7.06 ± 0.17 5.94 ± 0.42

PF 11-14 5 0.56 2.78 ± 0.28 6.88 ± 0.29 9.60 ± 0.29 9.28 ± 0.24 8.08 ± 0.30 6.88 ± 0.36 5.62 ± 0.47

*= Significant parasitaemia reduction (P<0.05) compared to the control group. PF = Pooled fractions

N= Number of animals used per group

As the parasitaemia increased with time there was also a corresponding drop in % PCV levels.

This suggests that haemoglobin concentration decreases with increase severity of infection. However,

the extracts treated group helped in recovering the PCV and Hb levels dose dependently (table 3).

*= Significant parasitaemia reduction (P<0.05) compared to the control group.

PF= Pooled fractions; PCV= Packed cell volume; Hb Haemoglobin count and WBC= White blood cell.

In the same vein, as the infection increased with time, there was a drop in the mean weight

of the animals (table 4), probably increase in severity of the infection decreased the appetite in the

animals or resulted due to the disease-induced anaemia, but there was a statistically significant (P<

0.05) recovery by day seven of post treatment (table 5).

Table 3. Effect of Pooled Fractions of Securidaca longepedunculata on Heamatological parameters

in Rats infected with Trypanosoma brucei brucei

Treatment Groups Dose (mg/kg) PCV Hb WBC

No infection no

treatment

0 *31.00 ± 1.64 *9.64 ± 0.34 6.18 ± 0.59

Normal saline 0 48.6 ± 3.93 16.18 ± 1.31 9.76 ± 2.72

Diminazine

aceturate

3.5 *54.00 ± 1.41 *17.96 ± 0.47 6.50 ± 1.09

PF 11-14 0.14 *43.20 ± 5.16 *14.82 ± 1. 58 6.64 ± 1.29

PF 11-14 0.28 *42.00 ± 4.72 *13.52 ± 1.54 7.20 ± 0.91

PF 11-14 0.56 *41.80 ± 4.84 *12.56 ± 1.46 8.08 ± 2.54



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Table 4. Body weights of rats infected with T. brucei treated with fractions of

S. longepedunculata methanol roots extracts.

Dose (mg/kg) N Initial weight (g) Initial weight (g)

0.14 5 132.00 ± 5.60 137.50±3.50

0.28 5 140.17±5.08 144.33±2.06

0.56 5 146.50±9.70 150.50±13.56

Table 5. Phytochemical screening of pooled fractions of ethyl acetate fraction of roots of S.

longepedunculata

General Test Specific Test Fraction

1-13

Fraction

14-22

Fraction

23-31

Fraction

32

Fraction

33-34

Test for flavonoids a) shinoda's for flavonids + + + - -

b) sodium hydroxide test for

flavonoids +

+ + + -

c) lead acetate test for flavonoids + + + + +

d) ferric chloride test for

flavonoids +

+ + + +

Test for cardiac

glycosides

a) Lieberman Burkard test for

triterpenes -

- - - -

b) Salkowskill test for terpenoids - - - - -

c)Keller kiliani test for digitalis

glycosides -

- - - -

Test for

anthraquinone

derivatives

a) Borntrager's test for combined

anthraquinones -

- - - -

b) Borntrager's test for free

anthraquinones -

- - - -

Test for

phlobatannins

a) Hydrochloric acid test for

phlobatannins -

- - - -

Test for tannins a) Ferric chloride test for tannins + + - - -

Test for saponins a) Frothing test for saponins + + + + +

Test for

carbohydrates a) Molisch's test for carbohydrates -

+ - + +

b) Fehling's test for reducing

sugars +

+ - - -

c) Test for combined reducing

sugar +

+ - - -

Test for alkaloids a) Dragendoff's test - - - - -

b) Wagner's test + - - - -

c) Mayer's test + - - - -



11

5. Discussion

The preset study demonstrates that trypanosoma brucei brucei when inoculated into mice/rats

develop parasitaemia within three days and reach peak parasitaemia within 5-6 days. This is in

agreement with the reports of Ajagbonna et al (2005)., Anene et al, (2006) and Ameh et al, (2006).The

observed in vivo anti-trypanosomal activity of the methanol root extract of Securidaca

longepedunculata is not surprising since previous reports (Nok et al, 1993.,Asuzu et al, 1990., and

Owolabi et al, 1990) have clearly demonstrated that plants of different families could possesses anti-

trypanosomal activity and vice versa due to the biotransformation of plant materials that may convert

active therapeutic molecules to in active ones. The study indicates that there is significant trypanocidal

activity of S. longepedunculata root extracts and its fractions as shown by the steady clearance of

parasitaemia by the plant extract (tables 1 and 2/figures 2 & 3), which also supports the studies of

Atawodi (et al, 2003) and Ameh et al (2006). This work also agrees with Adebaubaurer et al (2008),

who reported that the roots of Securidaca logepedunculata Fresen (Polygalaceae) were able to reduce

parasitaemia in mice experimentally infected with trypanosoma brucei brucei by 48 and 42% at the

dose of 150 mg/kg b.w. intraperitoneally two times daily for three days.

Figure 2. Screening of methanol root extract of S. longepedunculata in mice infected with T. brucei.



12

Figure 3. Bioassay guided studies of sub- pooled fractions 11-14 of 0.14, 0.28, 0.56 mg/kg of

S. longepedunculata in mice infected with T. brucei.

The anti-trypanosomal effect exhibited by these extracts of S. longepedunculata might be

attributed to the presence of some phytochemicals in them like; flavonoids, saponins, tannins, alkaloids

etc.. Tables 1 and 2 give the parasitaemia pattern among three groups treated with the extracts and

fractions dose dependently, five days post treatment, a statistically significant (P<0.05) dose dependent

parasite suppression was observed, while the diminazine-treated mice/rats were cleared of their

parasites 24 hours post treatment. The relapse seen immediately on withdrawal of treatment in the

extracts treated group after the parasites were statistically significantly reduced (P<0.05) could be as a

result of resistance put forward by the parasites or some limitations on the part of the extract itself such

as the inaccessibility of the extract to other tissues where the flagellates are known to hide as a way of

evading trypanolytic action of drugs (Anosa, 1998). Therefore, this plant provides a readily available

cost effective alternative trypanocide. However, further investigation may explore more of the plant’s

usefulness to humanity.

6. Conclusion

From the results of this study, it can be concluded that plant Securidaca longepedunculata

popularly called violet tree in English has potentials to combat these parasites which constitute serious

threats to livestock industry and public health at large. Hence, it should be explored further.



13

Acknowledgements

Thanks to Kebbi State University of Science and Technology, Aliero, for sponsoring my

studies and thanks to Ahmadu Bello University Zaria for training me.

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Ameh, I.G.; Ajagbonna, O.P.; Etuk, E.U and Yusuf, H. (2006). Laboratory treatment of trypanosoma

evansi – infected rats with a combination of securidaca longepedunculata and diminazene

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