ISOLATION AND CHARACTERIZATION OF ANTI ECHIS OCELLATUS VENOM PRINCIPLES OF CEIBA PENTANDRA
ISOLATION AND CHARACTERIZATION OF ANTI ECHIS OCELLATUS VENOM PRINCIPLES OF CEIBA PENTANDRA
No Thumbnail Available
Date
2011-03
Authors
SARKIYAYI, SHEHU
Journal Title
Journal ISSN
Volume Title
Publisher
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.
Description
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