A study of cytopathic rotavirus strains isolated from calves with acute enteritis

Nine cytopathic bovine rotavirus strains were isolated in MA-104 cell cultures from fecal specimens of dairy calves suffering from diarrhea. Isolation of the virus was accomplished from three outbreaks which occurred on dairy farms located in Central and Southern Italy. Fecal suspensions were treated with a high concentration (1000 micrograms/ml) of trypsin, and inoculated into MA-104 cell cultures grown out in Eagle's minimum essential medium (MEM) containing 5 micrograms/ml of the enzyme. Cytopathic effects (CPE), characterized by intracytoplasmic inclusion bodies of different sizes and shapes, were observed on the 1st passage with five of the strains and on the 2nd (2 strains) or the 3rd (2 strains) passage for the others. The presence of trypsin and the use of MA-104 cells appeared to be essential for the occurrence of CPE, inasmuch as no CPE was detected when trypsin was omitted in the MA-104 cell system. Replication failed to occur when primary bovine embryo kidney cell cultures with or without trypsin were used. Electron microscopy revealed the presence of particles with a typical rotavirus morphology. In MA-104 cells, the titre of virus reached its maximum 48 hr after inoculation. Small, clear-cut plaques were produced by the isolates in MA-104 cells under the overlay of MEM containing carboxymethyl cellulose, trypsin and DEAE-dextran. The nine rotavirus strains were antigenically related, whereas the relationship to either the Nebraska or the Compton rotaviruses was quite weak.