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Porcine adenoviruses(PAdVs)are classified into three species,PAdV-A,PAdV-B,and PAdV-C.The genomes of PAdV-A and PAdV-C have been well characterized.However,the genome of PAdV-B has never been completely sequenced,and the epidemiology of PAdV-B remains unclear.In our study,we have identified a novel strain of PAdV-B,named PAdV-B-HNU1,in porcine samples collected in China by viral metagenomic assay and general PCR.The genome of PAdV-B-HNU1 is 31,743 bp in length and highly similar to that of California sea lion adenovirus 1(C.sea lion AdV-1),which contains typical mastadenoviral structures and some unique regions at the carboxy-terminal end.Especially,PAdV-B-HNU1 harbors a dUTPase coding region not clustering with other mastadenoviruses except for C.sea lion AdV-1 and a fiber coding region homologous with galectin 4 and 9 of animals.However,the variance of GC contents between PAdV-B-HNU1(55％)and C.sea lion AdV-1(36％)indicates their differential evolutionary paths.Further epidemiologic study revealed a high positive rate(51.7％)of PAdV-B-HNUl in porcine lymph samples,but low positive rates of 10.2％ and 16.1％ in oral swabs and rectal swabs,respectively.In conclusion,this study characterized a novel representative genome of a lymphotropic PAdV-B with unique evolutionary origin,which contributes to the taxonomical and pathogenic studies of PAdVs.