Expression of cathepsin L gene of fasciola gigantica in different developmental stages

Abstract

5 Abstract: Fascioliasis is one of the most serious parasitic infestations of livestock. It is caused by Fasciola he patica (Temperate re gions) and Fasciola gi gantica (Tropical re gions).It also has a zoonotic importance. The proteolytic enzyme; ca thepsin-L cysteine proteinase (28-28.5 KDa) h elps the parasite in tissue i nvasion, host immune evasion and nutrients intake. This study aimed to check weather Fasciola gigantica flukes d epend upon cathepsin-L throughout t heir life c ycle in snail and mammalian hosts or n ot. For th at ca thepsin-L gene was inv estigated in adult and other d evelopmental stages o f Fasciola gi gantica using semi-quantitative RT-PCR assay. C athepsin-L protein w as d etected by us ing Sodium dodecyl sulphate- polyacrylamide gel electrophoresis (SDS-PAGE) anal ysis and detection of cathepsin-L cys teine proteinase antigen in examined stages by Enzyme linked immunotransfer blot (EITB) analysis. Stages of Fasciola gi gantica were collected from an experimental l ife cycle that wa s c ompleted in our la boratory. The adult worms a nd eg gs were c ollected f rom bile d ucts and gal l bladders, respectively, of slaughtered naturally infected cattle at El-Shohada abattoir, Minufiya governorate, Egypt. The results obtained revealed that ca thepsin-L gene is expressed in adult and other d evelopmental stag es o f F asciola gigantica wi th a molecular weight of 28-28.5 kDa. These results may be promising for developing an effective and reliable method for e arly diagnosis of Fasciola gi gantica and protection cysteine proteinase enzyme of Fasciola gigantica is present in ad ult and other developmental stages with m olecular weight at 28-28.5 kDa and its gene is fully expressed in adult and other d evelopmental stag es.