Therapeutic evaluation of Terminalia bellirica (Combretaceae) dried fruits against Trypanosoma evansi.
P Shaba, N Pandey, O Sharma, R Rao, R Singh
in vitro and in vivo antitrypanosomal, terminalia belirica dried fruits, trypanosoma evansi
P Shaba, N Pandey, O Sharma, R Rao, R Singh. Therapeutic evaluation of Terminalia bellirica (Combretaceae) dried fruits against Trypanosoma evansi.. The Internet Journal of Veterinary Medicine. 2008 Volume 7 Number 1.
African trypanosomosis has re-emerged in the recent years. Limited classes of chemotherapy for the treatment of the disease is marred by resistance to available trypanocides coupled with trypanosomes resistant-strains. In this study, methanolic plant extract (MPE) of
Trypanosomosis, an important blood protozoan zoonotic disease, is caused by flagellate parasites of the genus
Chemotherapy and chemoprophylaxis are the only means of combating the menace of the disease. Chemotherapy of trypamosomosis is faced with problems such as limited choice of trypanocides in the market, high cost, toxicity, and emergence of drug-resistant trypanosome strains that have been reported (Freiburghaus
This study is a follow-up to the previous reported trypanocidal potential of
Materials and methods
Silica gel-G for thin layer chromatography (TLC), solvents (hexane, chloroform, methanol, acetic acid and ethyl acetate) for extraction of plant material and development /analysis of TLC plates, vanillin for spray and iodine for detection of bioactive constituents These were purchased from
Dried fruits of
Preparation of extract
Twenty grams of
Thin layer chromatography (TLC) plates
This was done according to the method of Stahl, 1969. Aliquots (0.2ml) of extract were applied on TLC plates, dried under room temperature and immersed inside the solvent systems in glass jar listed in the next subsection. This was done to detect, if any, the presence of bioactive constituents in applied extract. After full development of plates in solvent systems, plates were dried at room temperature. Then, one set of plates were immersed in iodine vapors in a glass jar. Second set of plates were sprayed with Vanillin-sulphuric acid spray. Both media used facilitated the detection of bioactive constituents. This was carried out according to the method of Stahl, 1969.
The following solvent systems were tested for a suitable solvent to be used in developing TLC plates according to the method of Stahl, 1969.
Chloroform / hexane / acetic acid (50:50:1)
Chloroform / ethyl acetate / acetic acid (50:50:1)
Methanol and chloroform (20: 80)
Swiss albino mice (20-30 g) of either sex were obtained from Animal Research Laboratory Section of Indian Veterinary Research Institute (IVRI), Izatnagar, Bareilly maintained in standard environmental conditions and fed on a standard diet prepared by the institute with water
Estimation of parasite counts was carried out according to Lumsden et al. (1973).. 10 -15 fields of each drop of a blood or incubated media and parasites in triplicate were counted using glass slide under inverted microscope (400X) and an average means parasites count was taken as number of parasites per field.
In vitro tryponocidal activity
Stock of test MPE of
After incubation for antitrypanosomal activity was completed, contents of wells with reduced and apparently killed parasites from MPE of
Stock of test MPE was solubilized in 1% dimethylsuphoxide (DMSO) The concentration in the experiment had no deleterious effect by it self on host cells or parasites.1% DMSO in distilled water was used as control(Young
In vivo antitrypanosomal activity
This was done according to the method of Freiburhgaus
Results of trypanocidal activity were expressed as mean ± SEM. Statistical significance was determined by Sigma Stat (Jandel), USA.
Methanol was suitable (as previously reported by the same group on different medium during preliminary screening of the
Antitrypanosomal activity were expressed as mean ± SEM with significant difference (P<0.05)
At dose rate of 200 mg/kg body weight, the mice in this group survived for 7 days post on set of parasitaemia. There was degree of significant difference (P ≤ 0.05 ) between treated groups with test material in comparison to negative control that survived for only 3 days
African trypanosomosis (either in animals or humans) has been described as neglected disease in our generation. Antitrypanosomal drugs currently in used were developed before 1950 when stringent laws on drugs development and uses were not in placed as today. These drugs are very toxic to the body system (Denis and Barret, 2001).
In this report, Presence of bioactive constituents on TLC plates and subsequent suitable solvent system used in development of TLC plates is comparable to extraction and development of
It can be concluded from the present study that
Financial contributions by India and Nigeria governments towards the research, invaluable advice/inputs by Dr. A. K Mishra, Principal Scientist, contributions by other scientists and technical staff, Division of Parasitology IVRI, Izatnagar IVRI, Regional station Palampur and IVRI, Regional Station, Mukteswar, India are highly acknowledged.