Preparation and characterization of excretory and secretory antigen of Cotylophoron cotylophorum and Gastrothylax crumenifer .

Excretory and secretory antigen of adult Cotylophoron cotylophorum and Gastrothylax crumenifer was prepared and characterized by SDS-PAGE. Adult paramphistomes were collected in PBS from rumen of sheep slaughtered at Perambur and Villivakkam slaughter houses, Chennai, Tamil Nadu. Excretory/Secretory (E/S) antigens of G. crumenifer and C. cotylophorum were prepared by incubating in PBS at 28.3 °C overnight in a shaking incubator and centrifuged. The culture supernatant was used as antigen. The protein concentration of E/S antigens of C. cotylophorum and G. crumenifer ranged from 0.4 to 7.1 mg/ml and 3.3 to 5.9 mg/ml, respectively. SDS-PAGE analysis of E/S antigen of C. cotylophorum revealed two polypeptide bands from the range of 32 and 40 kDa. Whereas, SDS-PAGE analysis of E/S antigen of G. crumenifer revealed only one polypeptide band of 35 kDa. Excretory/secretory antigen may be used for serodiagnosis of paramphistomosis infection in sheep.