ISOLATION AND CHARACTERIZATION OF ANTI ECHIS OCELLATUS VENOM PRINCIPLES OF CEIBA PENTANDRA

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Date
2011-03
Authors
SARKIYAYI, SHEHU
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Abstract
The leaves extract of Ceiba pentandra plant was tested for anti venom activity against Echis ocellatus snake venom. Among parameters investigated include: LD50 of the Echis ocellatus snake venom, phospholipase A2 activity, percentage hemolysis, plasma recalcification time, Invivo analysis of total protein content, white blood cells, packed cell volume and haemoglobin contents. The toxicological studies of Ceiba pentandra was also investigated. The result reveals that Echis ocellatus has an LD50 of 0.280mg/kg + 0.06. Haemolysis due to venom was drastically reduced by the extract from 66% to 27.4% suggesting that the plant has Protective activity against the lethal action of the snake venom and is effective in reducing haemolysis in mice and prolonging the recalcification time . Phospholipase A2 was purified from the venom of Echis ocellatus with a yield of 77% on Sephadex A-25 column and appeared as band in polyacrylamide gel electrophoresis (SDS-PAGE) with molecular weight of 15KDa. Phospholipase A2 activity was drastically reduced in a dose dependent manner with Km of 0.8M, revealing that the purified phospholipase incubated with the extract had some anti venom effects against the phospholipase A2 activity. The invivo studies reveals that there were significant (P<0.05) decrease in packed cell volume, total protein and haemoglobin contents for the venom group and there was only slight changes in the venom/extract and control groups, suggesting that the extract has some inhibitory effect on the venom activity. The IR analysis reveals that the bioactive compound in Ceiba pentandra leaves is an organic ester with carbonyl functional group. The GC/MS result showed that the organic ester compound (Di –n- octyl phthalate) has some inhibitory effects on PLA2. The result has shown that Ceiba pentandra leaves extract possess potent snake venom neutralizing capacity. The plant leaves extract could be used as an antidote for snake bite envenomation.The toxicity profile of aqueous methanol extract of Ceiba pentandra leaves reveals that the LD50 was found to be over 5000 mg/kg. For chronic toxicity test, the mice were injected with graded doses (250 - 500 mg/kg) of the leaves extract daily for 21 days and the effects on body weight, haematological and serum biochemical parameters were determined. The results revealed that there was no significant change in body weight, meaning that the extract is relatively safe for oral medical use. In fact there were no variable changes in the body weight of the mice from day zero (D0 ) to day twenty one (D21) in all the groups. The parameters determined during chronic toxicity test reveals that the concentrations of urea, chloride ion sodium ion ,potassium ions and bicarbonate were 11.2, 83.2, 131.6, 8.44 and 38 mMol/L respectively for the test animals .The result showed that apart from urea, all other parameters were within physiological acceptable range. The results further indicated that ,there was a decrease in Creatinine (88 uMol/L) and Total Protein (57.4 g/L), while AST (19 IU/L), ALT (10 IU/L) and ALP (204 IU/L) concentrations has significantly increased .While there was no significant (p<0.05) change in PCV and bilirubin concentrations The finding reveals that C. pentandra has very low toxicity profile in all the tested animals and the plant is relatively safe for herbal oral medication. The inhibitor (Di –n- octyl phthalate) isolated during this research might become a highly effective therapeutic agent for the reducing of snake bite envenomation. The relevance of these findings would be of importance in the area of development of anti snake bite drugs that are going effective, cheap, accessible and less allergic plant compound that can be scientifically explored.
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A DISSERTATION SUBMITTED TO THE POSTGRADUATE SCHOOL, AHMADU BELLO UNIVERSITY ZARIA, NIGERIA IN PARTIAL FULFILLMENT FOR THE AWARD OF DOCTOR OF PHILOSOPHY IN BIOCHEMISTRY DEPARTMENT OF BIOCHEMISTRY AHMADU BEILO UNIVERSITY ZARIA, NIGERIA OCTORBER, 2011
Keywords
ISOLATION,, CHARACTERIZATION, OCELLATUS,, VENOM,, PRINCIPLES,, CEIBA PENTANDRA
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