Procedure for recovery of enteroviruses from the Japanese cockle Tapes japonica

Abstract

The most likely shellfish to be harvested if sportfishing is reinstated in San Francisco Bay is the Japanese cockle Tapes japonica. The virus levels present in these shellfish are unknown and need to be evaluated before the shellfish beds are open. Towards this end, a procedure for recovering and concentrating enteric viruses from these clams has been evaluated. Effective elution of poliovirus from clam tissues was found to occur with pH 9.5 glycine-buffered saline rather than with the pH 7.5 fluid utilized by other investigators on oysters. Poliovirus desorption was combined with Cat-Floc clarification to remove cytotoxicity from clam tissue homogenates. For assay purpose, viruses were concentrated by mixing the glycine supernatant with a beef extract solution, lowering the pH, and suspending the resulting floc in a small volume of phosphate buffer. This simple technique successfully recovered an average of 73% of the poliovirus added to clam homogenates at levels of 93 and 660 plaque-forming units per 100 g. Coxsackievirus B2 was isolated from clams exposed to raw sewage.