Evaluation of anthelmintic activity of ethanolic and aqueous leaf extracts of Azadirachta indica on phosphatases in Gastrothylax indicus

Abstract

Because of increasing anthelmintic resistance and the impact of conventional anthelmintics on the environment, it is important to look for alternative strategies against helminth parasite in sheep. The present study was performed to evaluate in vitro anthelmintic activity of plant Azadirachta indica on rumen amphistome of sheep.Anthelmintic activity of ethanolic and aqueous extracts of A. indica leaves in comparison with albendazole (positive control) and PBS (negative control) was evaluated through in vitro studies using worm motility inhibition and tegumental enzyme assays. Gastrothylax indicus was exposed in vitro to ethanolic and aqueous extracts(6.25 to 50 mg) of neem leaf to evaluate its anthelmintic efficacy. Mortality was observed at 2.14± 0.28h and 1.71± 0.21h for ethanolic and aqueous extracts respectively for the parasite at the highest test concentration of the plant extract. Anthelmintic albendazole was observed to cause mortality instantly (0.08±0.01h) at 80 μg/ml. Tegumental enzymes of the parasite ALP, ACP, ATPase and G-6-pase were found to be suppressed both by ethanolic and aqueous extracts. Enzyme kinetic studies showed inhibition to be noncompetitive in case of ALP with both the extracts whereas for ACP it was found to be non-competitive with ethanolic and competitive with aqueous extract. Albendazole showed competitive inhibition in case of ALP and non-competitive with ACP.