Pasteur Institute of Iran

Introduction: To determine an indicator for Triclabendazole (TCBZ) efficacy, Alanine aminotransferase (ALT) activity in Fasciola hepatica (Iranian isolate) parasite in presence and absence of TCBZ was evaluated by an in vitro cultivation method. Also, ALT enzyme activity between none and parasitized-infected sheep liver tissues was assessed. Method:  The sheep livers were collected and transferred immediately to the Department of Parasitology. Adult living parasites were recovered, washed and divided into two groups, treatment and control groups with 10 parasites in each. We added 15 μg TCBZ to the treatment group; then incubated both groups for 4 h at 37ºC. The parasites, infected and parasite free liver tissues were ground and homogenized by a Mortar and pestle, centrifuged, and supernatants were recovered. Protein concentration and ALT enzyme activity were measured in the supenatants. Results: The results of ALT enzyme activity assay showed 0.03 U/ml/mg protein for treated F. hepatica and 0.01 U/ml/mg protein for untreated samples, the mean values of difference was not significant (p>0.05). The difference between ALT activity in none and parasitized-infected liver was not significant (p>0.05). However, two-sample T-test analysis showed higher ALT activity in treated and untreated parasite in comparison with none and parasitized-infected liver specimens (p<0.05). In addition to ALT protein band for parasite and liver tissue, Cathepsin enzyme (proteases) was detected for parasite by SDS-PAGE analysis. Conclusion: ALT activity cannot be considered as a useful marker for TCBZ efficacy in F. hepatica treatment. However, ALT enzyme showed comparable activities in parasite and its host liver tissue.