Co-existence of multiple strains of two novel porcine bocaviruses in the same pig, a previously undescribed phenomenon in members of the family Parvoviridae, and evidence for inter- and intra-host genetic diversity and recombination.

Despite the recent discovery of novel bocaviruses from porcine samples, their genetic evolution and diversity are poorly understood. This study reports the identification and complete genome characterization of two novel parvoviruses, porcine bocavirus 3 (PBoV3) and porcine bocavirus 4 (PBoV4), from various porcine tissues/samples, displaying marked intra- and inter-host genetic diversity, with recombination events. Bocaviruses were detected by PCR among 16.5 % (55/333) of porcine samples (lymph nodes, serum, nasopharyngeal and faecal samples) from healthy, sick or deceased pigs from farms and a slaughterhouse in Hong Kong. As marked nucleotide polymorphisms were observed in the partial VP1 sequences, complete VP1 genes from one nasopharyngeal and three faecal specimens were cloned and sequenced, which suggested the presence of two different bocaviruses and demonstrated significant intra- and inter-host genetic diversity. Complete genome sequences revealed the presence of two bocaviruses, PBoV3 and PBoV4, in a faecal and nasopharyngeal specimen, respectively, with two genotypes, PBoV4-1 and PBoV4-2, in the latter. Their genomes encoded three ORFs, characteristic of bocaviruses. Phylogenetic analysis showed that they were distantly related to other bocaviruses, forming a distinct cluster within the genus. Recombination analysis showed possible recombination events among VP1 sequences of PBoV4 strains from a faecal specimen, with two breakpoints identified (with a 68 and 71 bp region), suggesting that different strains/variants within the same host could have arisen from recombination. This is the first report describing marked sequence diversity and the co-existence of two viruses of the family Parvoviridae within the same host, which may have originated from and, in turn, facilitated recombination.

Identification of novel porcine and bovine parvoviruses closely related to human parvovirus 4.

Human parvovirus 4 (PARV4), a recently discovered parvovirus found exclusively in human plasma and liver tissue, was considered phylogenetically distinct from other parvoviruses. Here, we report the discovery of two novel parvoviruses closely related to PARV4, porcine hokovirus (PHoV) and bovine hokovirus (BHoV), from porcine and bovine samples in Hong Kong. Their nearly full-length sequences were also analysed. PARV4-like viruses were detected by PCR among 44.4 % (148/333) of porcine samples (including lymph nodes, liver, serum, nasopharyngeal and faecal samples), 13 % (4/32) of bovine spleen samples and 2 % (7/362) of human serum samples that were sent for human immunodeficiency virus and hepatitis C virus antibody tests. Three distinct parvoviruses were identified, including two novel parvoviruses, PHoV and BHoV, from porcine and bovine samples and PARV4 from humans, respectively. Analysis of genome sequences from seven PHoV strains, from three BHoV strains and from one PARV4 strain showed that the two animal parvoviruses were most similar to PARV4 with 61.5-63 % nt identities and, together with PARV4 (HHoV), formed a distinct cluster within the family Parvoviridae. The three parvoviruses also differed from other parvoviruses by their relatively large predicted VP1 protein and the presence of a small unique conserved putative protein. Based on these results, we propose a separate genus, Hokovirus, to describe these three parvoviruses. The co-detection of porcine reproductive and respiratory syndrome virus, the agent associated with the recent 'high fever' disease outbreaks in pigs in China, from our porcine samples warrants further investigation.