Anthelmintic activity of both ethanolic and aqueous extracts of Calotropis procera flowers, Azadirachta indica leaves and Punica granatum fruit peel in comparison with albendazole was evaluated through in vitro studies by the worm motility inhibition assay. Significant anthelmintic effects (p \ 0.0005) were observed on live Gastrothylax indicus worm as evident from their mortality at 4 h post exposure to both ethanolic and aqueous extracts. Phytochemical analysis of extracts revealed the presence of phenols, alkaloids, saponins, tannins, flavonoids, steroids and triterpenoids. LC-50 values were determined to be 12.05 mg/ml ± 3.24 and 23.52 mg/ ml ± 6.4 for C. procera, 24.37 mg/ml ± 4.11 and 21.02 mg/ml ± 4.6 for A. indica, 18.92 mg/ml ± 4.54 and 24.43 mg/ml ± 6.96 for P. granatum ethanolic and aqueous extracts respectively, whereas it was 29.23 lg/ ml ± 4.51 for albendazole. The mean mortality index (MI) was 1.0 and 0.90 for C. procera, 0.90 for A. indica and 0.73 and 0.80 for P. granatum ethanolic and aqueous extracts respectively whereas for albendazole it was 1.0. Percent mean worm motility inhibition (%WMI) was observed to be between 70 and 100 % for different extracts.Various concentrations (5-5000 lg/ml) of all the plant extracts and albendazole were used to detect their cytotoxic effects against HeLa cell line to determine CC-50 by MTT assay. CC-50 values, of all the plant extracts were determined to be [1000 lg/ml and for albendazole it was found to be [10 lM. All the three plants can be potential sources for novel anthelmintics.

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. Important lipogenic enzymes like glucose-6-phosphate dehydrogenase (G-6-PDH) and malate dehydrogenase (MDH) show subcellular distribution pattern. Activity of G-6-PDH was largely restricted to cytosolic fraction while MDH was found in both cytosolic and mitochondrial fraction in Following in vitro treatment with ethanolic and aqueous extracts of fruit peel and commercial anthelmintic, albendazole G-6-PDH activity was decreased by 19-32 %, whereas MDH was suppressed by 24-41 %, compared to the respective control. Albendazole was quite effective when compared with negative control and both the extracts. The results indicate that phytochemicals of plant may act as potential vermifuge or vermicide.

Abstract:The trematode parasite, Gastrothylax indicus was exposed to ethanolic and aqueous flower extract of Calotropis procera to evaluate the anthelmintic efficacy of the plant. The parasites were incubated in 6. 25, 12.5, 25, 50 was found to be suppressed by 43.890, 30.287,18.970 and 22.842% by ethanolic extract and 62.710, 19.780, 57.554 and 10.035% by aqueous extract whereas albendazole inhibited 41.617, 25.650, 64.797 and 26.611% respectively. Enzyme kinetic studies showed inhibition to be non-competitive in case of ACP with both the extracts and albendazole whereas for ALP it was found to be non-competitive with ethanolic and mixed type with aqueous extract. Albendazole showed competitive inhibition in case of ALP.

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.