Identification and characterization of sarcocystis fusiformis antigen before and after heat treatment

ABSTRACT

Zoonotic sarcocysts are transmitted to human by eating undercooked meat. Cooking is effective for inactivating Sarcocystis in meat. S. fusiformis cysts were processed for heat treatment at 55, 60, 65 and 70°C. The cystozoites antigen was prepared before and after heat treatment. Both antigens were separated by Sodium dodecyl sulphate polyacrylamide gel electrophoresis [SDS-PAGE]. Following separation, the proteins were either stained with Coomassie blue stain or electrophoretically transferred from polyacrylamide gel to immobilon-p transfer membrane for Western blot [WB] technique. The blotted proteins were analyzed against anti Sarcocystis hyperimmune serum. By SDS-PAGE protein bands ranging from 10 kD to 220 kD are detected in the separated pre-heated antigen. By WB several bands are immunogenic 20-27 kD, 43 kD, 90 kD, 160 kD, and 215 kD. By SDS-PAGE after heat treatment, the pattern of detected protein bands became slightly faint at 65°C, while at 70°C most bands disappeared. These bands were previously reported to be immunogenic bands of micronemes, dense granules and pellicle. These organelles are responsible for adhesion and invasion of host cells. This work explains the role of heat on the antigenicity and infectivity of Sarcocystis