Biological properties of bovine viral diarrhea virus quasispecies detected in the RK13 cell line

The rabbit kidney cell line RK13 has been reported to be contaminated with noncytopathogenic (ncp) bovine viral diarrhea virus (BVDV). Persistent infection was confirmed by demonstrating the stability of virus titers (104.6±0.5 TCID50/ml) and BVDV positive cells (71.9 ± 3.12 %), over six successive passages. Based on the “exaltation of Newcastle disease virus” (END) and reverse plaque formation methods, two types of ncp viruses were isolated, END-phenomenon-positive and negative. Isolates, RK13/E+ and RK13/E−, demonstrated (1) differing levels of reproducibility in cell cultures, (2) similar antigenicity against BVDV antisera, (3) identical 5′-UTR region nucleotide sequences, (4) four amino acid differences throughout the genomic open reading frame, and (5) better growth ability in primary rabbit cells than other laboratory strains when inoculated in parallel at an MOI of 0.01. Overall, the BVDV population in RK13 cells consists of at least two different END characteristic quasispecies that are adapted to cultures of rabbit origin, giving rise to naturally attenuated BVDV strains that can be used in vaccine development.