Determination of Diagnostic Antigens in Cattle Amphistomiasis Using Western Blotting

Abstract: background: mixed infection with amphistomes seems common in native cattle of iran. the aim of this study was to determine diagnostic antigens in cattle mixed amphistomiasis.methods: specific antigens of cotylophoron cotylophorum, gastrothylax crumenifer and paramphisto­mum cervi (mixed infection), the most common species, were collected from cattle was deter­mined. adult trematodes were collected from the rumen of naturally infected cattle at meat inspec­tion. after their homogenization and centrifugation, somatic antigens were prepared and ana­lyzed by sds-page. specific antigens were determinated by western blot with homologous and heterolo­gous sera. sds-page of whole worms extract was performed at different concentrations and subse­quent gels staining. immunoblotting analysis using sera from cattle naturally infected with am­phistomes, dicrocoelium dendriticum, fasciola spp. and hydatid cyst was performed.results: electrophorese analysis of somatic antigens revealed the presence of 10 and 21 protein bands at 4 µgr/ml and 8 µgr/ml with molecular weights ranging from 25-120 and 25-150 kda, respectively. the best result was taken at 8 mg/ml concentration. although western blot of these proteins demon­strate 5 major antigenic polypeptides ranging from 50 to 100 kda which were recognized by serum of cat­tle naturally infected with mixed amphistomes.