Isolation and serotyping of porcine rotaviruses and antigenic comparison with other rotaviruses

Abstract

Seven rotavirus strains were isolated in cell cultures from the intestinal contents of piglets with diarrhea. MA104 cells with pancreatin in the cell culture medium was the host system of choice for virus isolation and replication. A cell culture immunofluorescence test in which MA104 cells were used in microtiter plates was very effective for detecting and assaying rotaviruses. A plaque reduction neutralization test, cross-protection studies in gnotobiotic pigs, and electrophoresis of rotaviral double-stranded RNA were used for comparing viruses. Three strains produced plaques on initial isolation attempts, replicated well in cell cultures, and were antigenically very similar. We suggest that these three strains be considered porcine rotavirus serotype 1, with The Ohio State University (OSU) strain serving as the prototype. The OSU strain was distinct from bovine, simian, canine, and human (Wa and M) rotaviruses by plaque reduction neutralization. Four strains did not produce plaques on initial isolation attempts, were difficult to adapt to cell cultures, and were related to each other but were distinct from the serotype 1 strains. We suggest that the Gottfried (G) strain be tentatively considered as a prototype for porcine rotavirus serotype 2. The G strain was antigenically closely related to canine and simian rotaviruses and less so to human M rotavirus (human rotavirus serotype 3). Canine, simian, and human M rotaviruses were closely related. All seven porcine rotavirus strains caused diarrhea in gnotobiotic pigs. Cell-cultured vaccines of the OSU and G strains caused only mild or no diarrhea in gnotobiotic pigs, and protection occurred when such pigs were challenged with homologous, bur not heterologous, virulent viruses. A survey indicated that 94% of 274 porcine serum samples and 100% of 75 herds were serologically positive to the porcine OSU rotavirus.