Investigation On Bat Viruses In Southeast China

Bats,also known as the order Chiroptera,are the second largest order of mammals.Bats are present throughout most of the world,with 19 families and 962 genuses.Bats are highly relevant with a large number of pathogens(137 viruses),especially zoonoses virus(61viruses)including highly pathogenic like SARS-like coronavirus,Ebola virus,Marburg virus,Rabies virus,Dengue virus.Recent studies indicate that some novel viruses are discovered in bats,including Bocavirus,Circovirus,Retrovirus,Astrovirus and Hepatitis Virus.In 2003,SARS resulted in over 8,000 infections and over 800 deaths.In 2014,Ebola epidemic in Africa caused 6,388 deaths and 17942 confirmad or suspected cases.In 2016,dengue virus in Argentina led to 15,000 infections.These pathogens share a common host-bat.Bats are natural reservoirs for a large number of zoonotic pathogens,drawing more and more public concerns.Traditional technologies which rely on conserved sequences of viruses are inefficient to discover novel viruses.Next-generation sequencing-based viral metagenomics has shown great power in studying virogenomics of animals and environment.In this study,healthy-looking bats were collected from the southeastern of China in 2015.High throughput sequencing was employed to acquire bioinformation.The viruses acquired from collected bats were divided into 38 families,including 16 vertebrate virus families,4 plant virus families and 5 insect virus families.We mainly focus on vertebrate viruses,such as Herpesviridae,Coronaviridae,Picornaviridae,Adenoviridae,Retroviridae,African swine fever virus(Astroviridae);Calicividididae;Circoviridae;Hepeviridae;Papillomaviridae;Parvoviridae;Parvoviridae;Parvoviridae;Parvoviridae;Parvoviridae;Parvoviridae;Retroviridae;Flaviviridae;Poxviridae;Togaviridae.Results showed that homology and abundance were varied from different species as well as regions.We had a basic knowledge of the ecology of bat virus in the southeast area,determining the direction for the future work.Degenerate primers for Circovirus,Bocavirus and Ifavirus were designed and specific RT-PCR was used to acquire the carrying and distributing situations of the above mentioned viruses.4 circoviruses named Bt CV-ZS62,Bt CV-DS13,Bt CV-CL6 and Bt CV-LJ22 were detected in bats from Zhoushan,Daishan,Changle and Lianjiang respectively.2 Bocavirus were detected in Danshan bats.2 Ifavirus were detected in Xiamen bats.What's more,full-length sequences of the four circoviruses and nearly full-length sequence of bocavirus were obtained.Bt CV-ZS62,BtCV-DS13,Bt CV-CL6 and BtCV-LJ22 were four novel species of circoviruses.Bt Bov-DS10 is a novel species of bocavirus.In order to understand the function of Cirvirus and Bocavirus,the proteins BtBOVDS10-NP and Bt CV-DS13-Cap were selected for prokaryotic expression and seroepi demiology studies.Bats are present widely in China and highly relevant for public health.Therefore,it is important to learn the natural background of the viruses from bats to prevent emerging infectious disease.This study explored a new method of virus survey : PCR based on the annotation information of the virus metagenomic.PCR method which screens samples based on annotation information is fast and accurate.This new method greatly reduces the blindness of traditional virus discovery.It not only broadens the width of the virus ecological survey,but also increases the depth of the survey.