Mammalian birnaviruses identified in pigs infected by classical swine fever virus

Currently, the Birnaviridae family contains four genera with all members identified from birds, fishes, and insects only. The present study reports a novel birnavirus unexpectedly identified from classical swine fever virus-infected pigs by viral metagenomic analysis, which is, therefore, named as porcine birnavirus (PBRV). Follow-up reverse transcription-polymerase chain reaction (RT-PCR) screening of archived tissues of diseased pigs identified 16 PBRV strains from nine provinces/autonomous regions in China spanning 21 years (1998-2019), and the viral loads of PBRV in clinical samples were 10(5.08)-10(7.95) genome copies per 0.1 g tissue, showing the replication of PBRVs in the pigs. Genome-based sequence comparison showed that PBRVs are genetically distant from existing members within the Birnaviridae family with 45.8-61.6 per cent and 46.2-63.2 per cent nucleotide sequence similarities in segments A and B, respectively, and the relatively closed viruses are avibirnavirus strains. In addition, indels of 57, 5, and 18 amino acid residues occurred in 16, 2, and 7 locations of the PBRV polyprotein and VP5 and VP1 proteins, respectively, as compared to the reference avibirnaviruses. Phylogenetic analysis showed that PBRVs formed an independent genotype separated from four other genera, which could be classified into two or three subgenotypes (PBRV-A1-2 and PBRV-B1-3) based on the nucleotide sequences of full preVP2 and VP1 genes, respectively. All results showed that PBRV represents a novel porcine virus species, which constitutes the first mammalian birnavirus taxon, thereby naming as Mambirnavirus genus is proposed.