ABSTRACT
Hydatidosis is a zoonosis disease caused by the larva of Echinococcus granulosus parasite in man and animals. The Proteases enzymes are necessary for nutrition, migration and evasion of immunity of parasite into host. The aim of this study was to determine the protease activity of hydatid cyst protoscolices [PSC], healthy and cystic liver tissues in order to compare of this biomarker for host and parasite in hydatid disease. In this study, PSC were collected from sheep liver tissue infected with hydatid cysts at an abattoir and washed 3 times with PBS buffer. PSC samples were freeze-thawed and sonicated while collected liver tissues were homogenized. Extract solution samples were centrifuged and stored at - 20degree C. Protease enzymes activity was measured in the extract solutions of PSC and sheep liver tissue samples [healthy and cystic livers]. Samples protein concentrations and protein bands were detected using Bradford and sodium dodecyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE] methods respectively. To determine significant difference between two groups, statistical t-test was performed. The values of protease enzyme activities in healthy, cystic and PSC were determined 0.0028, 0.0087 and 0.50U/ml/mg respectively. Elevation of protease enzyme activity in cystic liver as compared with healthy was not significant. Statistical T-test showed higher protease enzyme activity for PSC as compared to healthy [p<0.05]. SDS-PAGE confirmed 24 kDa and 54 kDa bands for protease enzyme in PSC samples and 24 kDa band in liver samples. Protease enzyme activity and molecular weight as compared to healthy liver tissues could be concerned as a comparative metabolic biomarker for host and parasite in hydatidosis
ABSTRACT
The aim of this study was to detect the Glutathione S-Transferase [GST] enzyme activity of healthy / cystic liver as a diagnostic biomarker for hydatidosis. In order to compare with liver tissue, the level of the GSTs enzyme activityof parasite was also determined. Parasites were collected from sheep liver tissue with hydatid cysts at a local abattoir and washed with PBS buffer. Collected parasites and liver tissues were sonicated or homogenized respectively. Extract solution samples were centrifuged and stored at - 20 degree C. GSTenzyme activities were measured in the extract of parasite and liver tissue samples [healthy and infected livers]. Protein amounts and protein bands were detected using Bradford and sodium do-decyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE] methods respectively. To determine significant difference between two groups,two-sample/-test was performed. GST specific activities of healthy / infected livers and parasites were estimated 304, 1297 and 146 U/ml/mgrespectively. Significant higherGST specific activities in cystic liver than healthy liver was observed [P<0.05]. T-test analysis showed GST activity of parasite was lower than healthy liver tissue. SDS-PAGE showed GST protein bands with 24 kDa in parasite samples and25 kDa in liver tissues. GST activity incystic liver tissue could be concerned as a biomarker for hydatid cyst diagnosis with other hydatid disease parameters