Investigation On Bat Virome And Characterization Of Novel Bat Viruses From Puer Of Yunnan Province Of China

Background: Viruses are one of the richest species and many viruses seriously threaten human health.Recently,new viral infectious diseases breakout frequently,causing global panic due to their high unpredictability,burstiness and high mortality.Viral metagenomics(ViralMeta),which is based on the second generation high-throughput sequencing,could directly detect all the virus nucleic acid sequences in the environment.It has become an efficient tool for investigating the known and unknown viruses,especially for discovering new viruses.Bats are widely distributed and are the natural hosts of many viruses,carrying over 170 viruses including a variety of human zoonotic viruses.Bats can transmit viruses to humans through intermediate hosts,aerosol and bitten or scratched.Yunnan province has important strategic significance since it locates in the southwest border of China and contiguous with Southeast Asian countries.A lot of zoonotic viruses have been found in this area.The purpose of this study is to investigate the diversity of bat viruses in Yunnan and discover new bat viruses,so as to provide information for preventing zoonotic diseases especially bat viruses spreading to human.Methods: Eighty-four healthy bats were collected from Puer,Yunnan province in the condition of no affecting bats population stability from July to September in 2016.The species of bats were identified by PCR.The bat samples were detected by high-throughput sequencing technology.The sequences data were analyzed by ViralMeta to obtain information of viruses carried by bats.Two potential new pathogen were identified by PCR,whole genome amplification,epidemiological investigation and virus isolation to obtain the genomic structure,evolutionary relationship with other viruses and pathogenicity of novel viruses.Results:(1)The 84 bats included 17 Rhinolophus affinis and 67 Hipposideros Pomona.A total of 4,585,938 virus reads were gained from high throughput sequencing data.These viruses were classified into 38 families and 150 genera,in which some zoonotic viruses of vertebrate viruses have potential risk for public health,including hantavirus,bocavirus and rhabdoviridae.(2)A hantavirus,named as DodeHV,was identified in 4 cases of Hipposideros Pomona.The complete ORF of S segment,the most sequence of M and the complete sequence of L segment were obtained.The accession numbers of S,M and L sequences in GenBank were MG637438,MG637437 and MG637436,respectively.It shared the highest homology with the strain XSV found in bat from Phu Province,Vietnamese,but had more differences with known bat hantavirus found in China.(3)A rhabdovirus,named as DPRV,was identified in one case of Rhinolophus affinis.Its complete sequence was obtained and it was a novel rhabdovirus species by sequence comparison.The accession number in GenBank was MH102378.DPRV could infect suckling mice and replicate in mice's brain.Conclusion: The virome carried in bats from Puer further enriches the virus reservoir of different bat species in Yunnan province.It is the first time to discover DodeHV from Hipposideros Pomona in China,which is a different strain of the same genotype with bat hantavirus XSV of Vietnamese.It is the first time to discover a novel rhabdovirus species,DPRV,which has the potential pathogenicity.