Effects of crude methanol extract of Adansonia digitata fruit pulp on Naja nigricollis venom-induce toxicity in Wistar rats.

OBJECTIVES: This study evaluated the effects of Crude Methanol Extract of Adansonia digitata Fruit Pulp on Naja nigricollis Venom-Induce Toxicity in Wistar rats. METHODS: A. digitata was extracted using 70% methanol and median lethal dose (LD50) of both the extract and venom were determined using the up-and-down method. Sixty Wistar rats were randomly assigned into 10 groups of 6 rats each and were administered with normal saline, venom only, venom + antivenom, 125 mg/kg, 250 mg/kg and 500 mg/kg crude methanol extract (CME) pre-envenomation, 125 mg/kg, 250 mg/kg and 500 mg/kg CME post-envenomation, and venom + antivenom + 250 mg/kg CME, respectively. Blood samples were collected 8-h post-envenomation in EDTA and plain sample bottles. Erythrocyte osmotic fragility (EOF) test was carried out on the EDTA blood samples while serum was harvested and used for Malondialdehyde (MDA) and Superoxide Dismutase (SOD) assays. RESULTS: LD50 of the CME and venom was >5,000 mg/kg and 0.889 mg/kg, respectively. N. nigricollis-induced oxidative stress was evident in group B through increased % haemolysis, MDA and lowered SOD activities. The groups treated with antivenom only, 250 mg/kg CME post-envenomation and antivenom +250 mg/kg CME significantly (p<0.05) reduced EOF, MDA values and increased SOD. The CME revealed better ameliorative effect than protective via inhibition of EOF, MDA values and increased SOD activity. CONCLUSIONS: The CME when administered singly showed more ameliorative properties and the combination of CME with antivenom for protection was not as effective as when compared to single administration.

Antitrypanosomal properties of Anogeissus leiocarpa extracts and their inhibitory effect on trypanosome alternative oxidase.

Background: African trypanosomiasis is a protozoan disease with huge socio-economic burden to sub-Saharan African exceeding US$4.6 annual loss. To mitigate the incidence of trypanosomal drug resistance, efforts are geared towards discovery of molecules, especially from natural products, with potential to inhibit important molecular target (trypanosome alternative oxidase, TAO) in trypanosomes that are critical to their survival. Method: Crude methanol extract of Anogeissus leiocarpa was subjected to in vitro bioassay-guided antitrypanosomal assay to identify the most active extract with trypanocidal activity. The most active extract was run on a column chromatography yielding five fractions, F1-F5. The fractions were assayed for inhibitory effect on TAO. The most promising TAO inhibitor was subjected to antitrypanosomal evaluation by trypanosome count, drug incubation infectivity test (DIIT) and in vivo studies. Gas chromatography-mass spectrometry (GC-MS) was used to identify and quantify phytochemical constituents of the potential TAO-inhibiting fraction. Results: Ethyl acetate extract (EtOAc) significantly (p<0.05) produced trypanocidal effect and was the most active extract. Of the five fractions, only F4 significantly (p<0.05) inhibited TAO compared to the control. F4 completely immobilised the trypanosomes up to 0.5 µg/µl, yielding an EC50 of 0.024 µg/µl compared to the 0.502 µg/µl of diminazene aceturate positive control group. The DIIT showed that F4 was significantly (p<0.05) potent up to 0.1 µg/µl. F4 significantly (p<0.05) suppressed parasite multiplication in systemic circulation of the treated rats and significantly (p<0.05) maintained high PCV when compared to the 5% DMSO group. Furthermore, F4 significantly (p<0.05) lowered serum concentrations of malondialdehyde. Phytoconstituents identified by the GC-MS include tetradecene; cetene; 3-(benzylthio) acrylic acid, methyl ester; 1-octadecene; 9-heptadecanone; hexadecanoic acid, methyl ester; dibutyl phthalate; eicosene; octadecenoic acid, methyl ester; oleic acid; 2-methyl-Z,Z-3,13-octadecadienol; 1-docosene; 3-phenylthiane, s-oxide; phenol, 3-methyl; phthalic acid, di(2-propylpentyl) ester and 1,4-benzenedicarboxylic acid, bis (2-ethylhexyl) ester. Conclusion: F4 from EtOAc contains six carbohydrates (9.58%), two free fatty acids (6.48%), five fatty acid esters (27.73%), two aromatic compounds (50.63%) and one organosulphide (5.61%). It inhibited TAO and demonstrated antitrypanosomal effects.

In vitro and in vivo antitrypanosomal efficacy of combination therapy of Anogeissus leiocarpus, Khaya senegalensis and potash.

ETHNOPHARMACOLOGICAL RELEVANCE: Pastoralists in Nigeria mix barks of Anogeissus leiocarpus (AL) Khaya senegalensis (KS) and potash (Pt) to treat animal African trypanosomosis. AIM: To evaluate antitrypanosomal potential of A. leiocarpus, K. senegalensis and potash for insights into the traditional claim of antitrypanosomal combination therapy (ATCT). MATERIALS AND METHODS: Fifty microliter each of six different concentrations of AL, KS, Pt, AL + KS, AL + KS + Pt and diminazene aceturate (DA, positive control) was incubated with 50 µL of parasite-laden blood containing 108Trypanosoma congolense cells in a 96-well microtitre plate. Negative control wells were devoid of the extracts and drug but supplemented with phosphate-buffered saline (PBS). Efficacy of treatment was observed at 1 h interval for complete immobilisation or reduced motility of the parasites. Each incubated mixture was inoculated into mouse at the point of complete immobilisation of parasite motility or at the end of 6-h observation period for concentrations that did not immobilise the parasites completely. For in vivo assessment, thirty-five parasitaemic rats were randomly allocated into seven groups of 5 rats each. Each rat in groups I-V was treated with 500 mg/kg of AL, KS, Pt, AL + KS and AL + KS + Pt, respectively, for 7 days. Rats in groups VI and VII were treated with diminazene aceturate 3.5 mg/kg once and PBS 2 mL/kg (7 days), which served as positive and negative controls, respectively. Daily monitoring of parasitaemia through the tail vein, packed cell volume and malondialdehyde were used to assess efficacy of the treatments. RESULTS: The AL + KS + Pt group significantly (p < 0.05) and dose-dependently reduced parasite motility and completely immobilized the parasites at 10, 5 and 2.5 µg/µL with an IC50 of 9.1×10-4 µg/µL. All the mice with conditions that produced complete cessation of parasite motility did not develop parasitaemia within one month of observation. The AL + KS group significantly (p < 0.05) lowered the level of parasitaemia and MDA, and significantly (p < 0.05) maintained higher PCV than PBS group. CONCLUSION: The combination of A. leiocarpus and K. senegalensis showed better antitrypanosomal effects than single drug treatment and offers prospects for ATCT. Our findings support ethnopharmacological use of combined barks of A. leiocarpus and K. senegalensis by pastoralist in the treatment of animal African trypanosomosis in Nigeria.