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In vitro antileishmanial and
antitrypanosomal activities of five
medicinal plants from Burkina Faso
W. R. Sawadogo, G. Le Douaron,
A. Maciuk, C. Bories, P. M. Loiseau,
B. Figadère, I. P. Guissou &
O. G. Nacoulma
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In vitro antileishmanial and antitrypanosomal activities
of five medicinal plants from Burkina Faso
W. R. Sawadogo & G. Le Douaron & A. Maciuk &
C. Bories & P. M. Loiseau & B. Figadère & I. P. Guissou &
O. G. Nacoulma
Received: 10 September 2011 /Accepted: 12 October 2011
# Springer-Verlag 2011
Abstract After ethnobotanical surveys in central and
western regions of Burkina Faso, five plants namely
Lantana ukambensis (Verbenaceae), Xeoderris sthulmannii
(Fabaceae), Parinari curatellifollia (Chrysobalanaceae),
Ozoroa insignis (Anacardiaceae), and Ficus platyphylla
(Moraceae) were selected for their traditional use in the
treatment of parasitic diseases and cancer. Our previous
studies have focused on the phytochemical, genotoxicity,
antioxidant, and antiproliferative activities of these plants.
In this study, the methanol extract of each plant was tested
to reveal probable antileishmanial and antitrypanosomal
activities. Colorimetric and spectrophotometric methods
were used for the detection of antileishmanial and anti-
trypanosomal activities. Leishmania donovani (LV9 WT)
and Trypanosoma brucei brucei GVR 35 were used to test
the antileishmanial and antitrypanosomal activities, respec-
tively. All extracts of tested plants showed a significant
antitrypanosomal activity with minimum lethal concentra-
tions between 1.5 and 25 μg/ml, the L. ukambensis extract
being the most active. In the antileishmanial test, only the
extract from L. ukambensis showed significant activity with
an inhibitory concentration (IC50) of 6.9 μg/ml. The results
of this study contribute to the promotion of traditional
medicine products and are preliminary for the isolation of
new natural molecules for the treatment of leishmaniasis
Leishmaniasis is a major public health problem affecting
millions of people worldwide with two million new cases
each year (Bern et al. 2008; Murray et al. 2005). This
pathology is a parasitic disease caused by protozoal
organisms of the Leishmania genus that are transmitted to
humans by sandfly bites. Infections can vary from simple
cutaneous leishmaniasis to mucocutaneous and fatal viscer-
al leishmaniasis or kala-azar (Kouassi et al. 2007). The
pharmaceutical drugs used for leishmaniasis treatment were
pentavalent antimonials and amphotericin B, including its
an array of undesired side effects alongside large-scale
resistance (Lira et al. 1999; Murray et al. 2005).
African human trypanosomiasis or sleeping sickness is
another parasitic disease with a fatal outcome if untreated,
which still affects hundreds of thousands of people per year
(WHO 1998). The causative agents are Trypanosoma
W. R. Sawadogo (*):I. P. Guissou
Institut de Recherche en Sciences de la Santé (IRSS/CNRST),
03 BP 7192, Ouagadougou 03, Burkina Faso
W. R. Sawadogo:G. Le Douaron:A. Maciuk:B. Figadère
Laboratoire de chimie des substances naturelles,
UMR-8076 BioCis, Labex LERMIT,
Faculté de Pharmacie de Châtenay-Malabry, Univ Paris-Sud,
C. Bories:P. M. Loiseau
Chimiothérapie antiparasitaire/UMR-8076 BioCis,
Labex LERMIT, Faculté de Pharmacie de Châtenay-Malabry,
I. P. Guissou
Département de Sciences Pharmaceutiques appliquées/
UFR/SDS/Université de Ouagadougou,
Ouagadougou, Burkina Faso
O. G. Nacoulma
Laboratoire de Biochimie et Chimie appliquées,
UFR/SVT Université de Ouagadougou,
Ouagadougou, Burkina Faso
Author's personal copy
brucei gambiense in West and Central Africa and Trypa-
nosoma brucei rhodesiense in East Africa. These protozoan
parasites are transmitted by the bite of the tsetse fly. Testse-
infested areas occur in poor rural populations and treatment
with pentamidine or melarsoprol is expensive, long-lasting,
and complicated (Kamanzi et al. 2004). In the case of
animals, the causative agent is Trypanosoma brucei brucei,
and African animal trypanosomiasis is a major problem
for the region of sub-Saharan Africa. Ten million square
kilometers of land that could be lush and fertile is not
productive because of the tsetse-transmitted disease
(Aderbauer et al. 2008).
Leishmaniasis and African trypanosomiasis (sleeping
sickness) are parasitic diseases classified by the WHO as
some of the neglected tropical diseases. These pathologies
affect almost exclusively the poor in developing countries.
Those who are affected with these diseases have symptoms
which often lead to impairments of motor function. This in
turn reduces their productivity and increases their level of
poverty. According to the WHO, over 70% of countries and
territories that report the presence of neglected tropical
diseases have low or middle income.
The high level of poverty makes it difficult for the
population to access to pharmaceutical drugs. The use of
medicinal plants is an alternative for solving the health
problems of populations in these countries (Santos et al.
The WHO estimates that over 80% of the population in
developing countries uses traditional medicine for their
primary health needs (Farnsworth et al. 1985).
The identification of natural products and the wide
variety of separation techniques available supports the
possibility of finding natural molecules which in turn
leads to the development of new drugs (Saklani and
Kutty 2008). Twenty-five percent of modern medicines
are made from plants that were first used traditionally
Despite the current popularity of natural product re-
search, development of herbal medicines for the treatment
of tropical infectious diseases such as leishmaniasis and
African trypanosomiasis still remains rare (Croft et al.
2005; Saklani and Kutty 2008).
After ethnobotanical surveys in central and western
regions of Burkina Faso, five plants were selected for their
traditional use in the treatment of parasitic diseases and
cancer: Lantana ukambensis (Verbenaceae), Xeoderris
sthulmannii (Fabaceae), Parinari curatellifollia (Chrysoba-
lanaceae), Ozoroa insignis (Anacardiaceae), and Ficus
platyphylla (Moraceae) (Table 1).
Our previous studies have shown the phytochemical
composition, the non-genotoxic effect, and antioxidant
and anticancer activities of these plants (Sawadogo et al.
In the present work, the methanol extracts of these plants
were examined for in vitro antileishmanial and antitrypano-
somal activities in order to justify their traditional use.
Materials and methods
Plant materials and extraction
Plants were collected in the forest of Denderesso (Bobo
Dioulasso) and identified by Pr. Millogo J., botanist at
University of Ouagadougou (Laboratory of ecology). A
voucher of each plant (Table 1) was deposited at the
herbarium of Centre National de la Recherche Scientifique
et Technologique (CNRST). Each sample was separately
prepared by air drying, grinding, and then finally being kept
in paper bags. The resulting powder of each plant (100 g)
was extracted by maceration in 500 ml of methanol for 24 h.
The filtrate obtained from this maceration was concentrated
under rotavapor BÜCHI to produce a crude dried extract.
Leishmania donovani (LV9WT) promastigotes were cul-
tured at 26°C in an oven with conditioned air 5% CO2in
bottles containing culture medium M199 plus 10% fetal
calf serum (South American Origin, cat no: 7MB0087,
100 ml, Lonza). A suspension volume of parasites
(109parasites/ml) was subcultured into fresh medium every
3 days to get a culture at 106parasites/ml and thus maintain
the strain. The whole culture was performed in a sterile
environment to avoid contamination.
The strain of T. brucei brucei GVR 35 was grown by
infecting Swiss mice (Outbreed) intraperitoneally, 1 to
2 weeks before the in vitro test. This method induces a
chronic infection that lasts a few weeks if mice remain
untreated (Ngurea et al. 1997).
Some 500 μl of blood were collected from the infected
mice and introduced into 5 ml of culture medium M199 at
4°C. The suspension was centrifuged, and the parasites were
collected for the antitrypanosomial test in a 96-well plate.
Table 1 Plants and voucher specimen
Author's personal copy
ThistestwasperformedinvitroonL. donovani promastigotes
LV9 WT. The colorimetric method for rapid quantification of
living cells was used (Chollet et al. 2008; Fernanda et al.
2007; Filho et al. 2009). The test principle is based on the
reduction of the tetrazolium salt MTT (3-(4,5-dimethylth-
iazol-2-yl)-2,5-diphenyl tetrazolium bromide) in the presence
of mitochondrial succinate dehydrogenase of living cells into
colored formazan. The reduction of this reagent induces a
color change in the culture medium, from light pink to dark
blue. The intensity of staining was evaluated in a spectropho-
tometer at570 nm, and the percentageofliving cells ata fixed
concentration was determined and compared with a control
culture (DMSO) and a reference product (amphotericin B,
tested doses, 20–0.02 μM).
The screening was performed in 96-well plates in a final
volume of 200 μl containing 2.105parasites that were treated
with the different concentrations of the samples. The plate
was incubated in a dark oven and maintained for 72 h at
26°C under air conditioning with 5% CO2. Then, 10 μL of
MTT solution at 5 mg/m1 of PBS was added to each well.
The plate was shaken again and incubated for 4 h under the
same conditions. Some 50 μL of a lysis solution composed
of 400 ml of distilled water, 100 ml of Triton X-100, and
16 ml of concentrated HCl was then added to lyse the cells.
The plate was shaken again, and the absorbance was
measured at 570 nm on plate reader, Multiskan MS.
The percentage of parasite death at a fixed concentration
was calculated using the following formula, % of inhibition =
(AO−AS)×100/AO, where AO = absorbance of negative
control and AS = absorbance of the sample. The 50%
inhibitory concentration (IC50) of the tested samples was
determined from a concentration range, by plotting the
Thistestwas performed invitro onthe slowstrainofT. brucei
brucei GVR 35. The pathogenic stage of T. brucei brucei at
20.103parasites/ml was treated with the different concen-
trations of the samples (plant extracts or reference products)
in 96-well plates (Ngurea et al. 1997; Zagana et al. 2007).
an oven (37°C, 5% CO2). Pentamidine was used as reference
drug. Minimum trypanocidal concentration was determined
by observing the lowest concentration for which no parasite
was observed using an inverted microscope (G×200).
All tests were performed in triplicate, and the results
presented are means±standard deviation. The software
GraphPad Prism 5 was used for the graphical determination
of inhibitory concentrations 50% (IC50). Results were
statistically analyzed by the Student’s t test when comparing
two groups or one-way ANOVA for more than two groups.
P values of <0.05 were considered significant.
Results and discussion
Various plant extracts have been showing antitrypanosomal
and antileishmanial activities, and the bioguided fraction-
ation often led to the identification of the chemical series
involved in these activities. Thus, 15 years ago, diallyl-
sulfide from Allium sativum, currently used in food, was
found to exhibit antitrypanosomal properties (Nok et al.
1996). Among the most recent data, it can be noted that
tannins from Anogeisus leiocarpus and Terminalia avicen-
noides were identified as antileishmanial agents (Shuaibu et
al. 2008). Moreover, naphtoquinones and naphtofuranes
were identified as responsible for the antileishmanial
activities from Chondodendron tomentosum bark and
Cedrela odorata while extracts from Aristoloquia pilosa
were active against T. brucei, followed by those of
Tabebuia serratifolia, Tradescantia zebrina, and Zamia
ulei (González-Coloma et al. 2011). Ghosh et al. (2011)
recently described the antileishmanial activity of roots of
‘Indian Valerian’ Valeriana wallichii (Ghosh et al. 2011). A
new antileishmanial flavanone has also been purified from
the leaves of Baccharis retusa (Grecco et al. 2010).
Essential oil of Cymbopogon citratus exhibited in vitro
antileishmanial activity on Leishmania amazonensis and its
major component, citral, was identified as the main active
principle (Santin et al. 2009).
The results of antileishmanial and antitrypanosomal
activities of extracts from the five plants studied in
Burkina-Faso are presented in Table 2.
All plants tested showed significant antitrypanosomal
activity with MLC between 1.5 and 25 μg/ml. The most
Table 2 Antileishmanial and antitrypanosomal activities of the five
plants and reference
Parasite sampleL. donovani (LV9WT)
T. brucei brucei GVR
35, MLC (μg/mL)
Author's personal copy
active samples were those from L. ukambensis and O.
insignis with a MLC value at 1.5 and 6.2 μg/ml,
respectively. Our previous studies showed the presence of
tannins, triterpenes, and steroids in the plants studied
(Sawadogo et al. 2011). These compounds are well known
for their antimicrobial and antiparasitic activities. The
results of the antitrypanosomal test help to justify the
traditional use of these plants in the treatment of some
parasitic diseases. The triterpenoids, flavonoids, and saponins
of these plants are probably responsible for this activity
(Kamanzi et al. 2004; Prytzyk et al. 2003).
In the antileishmanial test, only L. ukambensis had
demonstrated a significant activity with an inhibitory
concentration (IC50) of 6.9 μg/ml. Several studies have
shown that alkaloids (González-Coloma et al. 2011; Kam et
al. 1999; Mohammad et al. 2003), flavanones (Grecco et al.
2010), chalcones (Kayser and Kiderlen 2001; Liu et al.
2003), di- and triterpenoids (Tan et al. 2002), saponins
(Ridoux et al. 2001), and polyphenols (Kolodziej et al.
2001) are phytochemical compounds with antileishmanial
effect. Our previous study showed that L. ukambensis
contained a high concentration of polyphenols, triterpenes,
and saponins (Sawadogo et al. 2011). The antileishmanial
effect of this plant could be attributed to one of these
groups of compounds.
This study produced original results because the scien-
tific literature has not yet reported the antitrypanosomal
effect of these five plants and particularly the leishmanicidal
activity of L. ukambensis.
The results of this study contribute to the promotion of
traditional medicine products of Burkina Faso and are
preliminary for the isolation of new molecules of interest in
the treatment of leishmaniasis and human or animal African
Francophonie (AUF)forproviding financial support (postdoctoralgrant).
We are grateful to the Agence Universitaire de la
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