World Journal of Innovative Research (WJIR) ISSN: 2454-8236, Volume-5, Issue-4, October 2018 Pages 01-04 1 www.wjir.org Abstract— Aqueous leaf extract of Achyranthes aspera was investigated for inhibitory activity against Bitis arietans venom protease and phospholipase A 2 activity. The elemental analysis and phytochemical screening of the plant extract was carried out. The activities of protease and phospholipase A 2 (V o ) of the crude Bitis arietans venom were determined and the data obtained was used to estimate K M , V max and K cat . Inhibition studies were carried out using the same procedure except that different concentrations of the extracts (5%, 10%, 15% for protease assay and 0.5%, 0.75%, 10%, 1.25% and 1.5% for phospholipase A 2 assay) were added to the reaction mixture. The result showed that the Bitis arietans venom protease had a V max of 0.062 + 0.013 μmol/min,K M of 0.496 + 0.095mg/ml and a K cat of 0.125+ 0.001min -1 . The result also indicates that the Bitis arietans phospholipase A 2 had a V max of 3.27 + 0.030min -1 , K M of 8.358 + 0.050 mg/ml and K cat of 0.391+ 0.002min -1 . The aqueous leaf extract produced significant (P<0.05) decrease in the V max , K M and K cat of the Bitis arietans venom phospholipase A 2 in a dose dependent manner and a statistically significant (P<0.05) increase in the V max , K M and K cat of Bitis arietansprotease in a dose dependent manner. The phytochemical screening revealed the presence of flavonoids, tannins, steroids, saponins and terpenoids in the extract while the elemental analysis revealed the presence of Zn, Cr, Ni, Cd, Mn, Fe and Na. The result suggests that aqueous leaf extract of Achyranthes aspera inhibited the Bitis arietans venom phospholipase A 2 in an uncompetitive manner while the protease activity was stimulated by the extracts. It was observed that the use of the leaf of Achyranthes aspera may be important in the treatment of snake bites. Index Terms— Achyranthes aspera, Bitis arietans, antivenom, protease. phospholipase A 2 . I. INTRODUCTION Highlight Snake bite remains a major public health and agricultural problem throughout the world, particularly Africa. It affects man and his domestic animals causing specific problems (cardiotoxic, neurotoxic) and untimely death (Aguiyi, 2011). Hope Chinyere Nwune, Department of Biochemistry, Faculty of Science, University of Maiduguri, PMB 1069, Maiduguri, Borno State, Nigeria Mohammed Adamu Milala, Department of Biochemistry, Faculty of Science, University of Maiduguri, PMB 1069, Maiduguri, Borno State, Nigeria Hassan Zanna, Department of Biochemistry, Faculty of Science, University of Maiduguri, PMB 1069, Maiduguri, Borno State, Nigeria Ya’u Muhammad,Department of Animal Health and Production, School of Agriculture, Binyaminu Usman Polytechnic, Hadejia Jigawa State, Nigeria Antivenom immunotherapy is the only specific treatment against snake venom envenomation. There are various side effects of antivenom such as anaphylactic shock, pyrogen reaction and serum sickness. In addition to this is expensive (Menatchisundaramet al., 2009). Bitis arietans(Puff adder) belonging to the family viperidae is one of the dangerous snakes in Nigeria, and is one of the commonest causes of envenomation in Northern Nigeria. Snake venoms are composed of complex mixtures of active substances mainly peptides and proteins which are able to interfere with biological processes including thrombosis by affecting platelet aggregation and blood coagulation. Some of these proteins include enzymes like phospholipase A 2 and metalloproteases (Ibrahim et al., 2011). Achyranthes aspera which belongs to the family Amarantheceae is a perennial stiff, erect herb, growing up to 1-2m height and is claimed to be used in the treatment of snake bite in the north eastern parts of Nigeria especially Borno, Gombe and Adamawa. Hence this study intends to scientifically validate this claim by investigating the invitro inhibitory effect of the aqueous leaf extract of Achyranthes aspera on Bitis arietans venom protease and phospholipase A 2 activities. II. MATERIALS AND METHOD Location of the research The study was conducted in the Department of Biochemistry, Faculty of Science, University of Maiduguri, PMB 1069, Maiduguri, Borno State, Nigeria. Chemicals All the chemicals used in this study were of analytical grade and purchased from various sources. Snake venom Freeze dried Bitis arietans,Snake Venom was obtained from the Department of Pharmacognosy and Drug Development, Ahmadu Bello University, Zaria, Nigeria. Plant Material The fresh leaves of Achyranthes aspera were collected from Biu, Borno State, Nigeria. This was authenticated by a plant taxonomist, Prof. S.S. Sanusi in Biological Sciences Department, University of Maiduguri, Borno State, Nigeria. The voucher number was obtained and deposited in the herbarium. The plant materials were washed and shade dried for two weeks to a constant weight. The leaves were pounded to fine powder using mortar and pestle and stored in clean dry Effects of Aqueous Leaf Extract of Achyranthes aspera on Bitis arietans Venom Protease and Phospholipase A 2 Activities Hope Chinyere Nwune, Mohammed Adamu Milala, Hassan Zanna, Ya’u Muhammad