The Study On The Virom Of Fecal Samples From Guangxi Rhesus Monkeys

BackgroundRecently,emerging infectious diseases have been reported frequently,which has been the most important public health problem worldwide.For example,SARS in 2003,avian influenza in 2005,a pandemic H1N1 influenza in 2009,MERS in the Middle East in 2013,H7N9 avian influenza in China in 2012,and the outbreak of Ebola in West Africa in 2014.Interestingly,the viruses of these diseases were all originated from the animals.With the expansion of human activities,the relationship between human and wild animal is getting closer,which increases the chances of animal viruses transmitting to humans.It is reported that 70%of the emerging infectious diseases were caused by the viruses originated from animals.Therefore,the study of animal viruses is very important for human disease prevention and control.Recently,many highly pathogenic viruses have been found in non-human primates(NHPs),which could act as the important natural reservoirs of these viruses transmitting to hummans,such as human Immunodeficiency virus,Ebola and Marburg virus,etc.Viral metagenomics is an effective method to identify the viruses of clinical and environmental samples,based on random PCR and high-throughput sequencing,which is very helpful to detect and discovery viruses.Rhesus monkey is one of the most common non-human primates in the world,which is widely distributed and close to human activities.Guangxi Longhu Mountain is known as one of the national"four big monkey",located in the subtropical inland.Therefore,it is of great significance to study the viral spectrum of rhesus monkeys to protect animal and human health.MethodThe fecal samples were collected from Longhu Mountain in Long’an county of Guangxi province in China.The samples were processed according to our established methods;Viral DNA library were reconstructed and then conducted by Hiseq high throughputting sequencing;The obtained mass sequences of rhesus monkeys were analyzed by our established biological information analysis platform for Viral Metagenomics;Some of novel viruses were selected to confirm and screen by PCR;Viral screening,genomic amplification,cell culture were further studied to obtain evolution,prevalence and other characteristics of the viruses.ResultsIn the study,a total of 340899838 sequences were obtained by Hiseq sequencing,of which 1126694 sequences were annotated to the viruses.By sequence analysis,these viruses can be divided into 53 families and 104 genera.Vertebrate viruses contain 12 families and 31 genera;invertebrate viruses include 14 families;plant viruses contain 18 plant families,and 8 families of phage.The vertebrate viruses mainly includes picornaviruses(53%),picobirnaviruses(28%),circoviruses(11%),parvoviruses(6%)and other viruses.Some of these viruses can cause human diseases,such as enterovirus,norovirus,bocavirus,astrovirus and adenovirus.The results show there exists novel astroviruses that were similar to human astrovirus MLB and VA,and the amino acid(aa)homology is 75-90%;There is a novel enterovirus sharing 59%aa identity with known enterovirus;There exists a novel picornavirus showing only 24-27%amino acid homology with the known picornaviruses;There are human GII.17 and GI.3 noroviruses with 98%nt identities;There are many novel bocaviruses mostly related to human bocaviruses with 51-100%aa identities.All these results show that rhesus monkeys have a large number of novel significant viruses,which need to be further studied.The novel picornavirus and enterovirus were selected to further study.SEV-gx genome was 7367 nt.In total,10%faecal samples from rhesus macaques were positive for SEV-gx.Genomic and polyprotein analyses showed that it has a standard genomic organization and characteristics typical for EVs,displaying more closely related to EV-J strains(approximately 54.0%,43.0-44.1%,52.3-55.2%,61.1-62.7%and 64.0%aa identity in polyprotein,P1,P2 and P3 and combined 2C/3CD regions,respectively).Phylogenetic analysis of P1,2C and 3CD aa indicated that SEV-gx can be classified as a distinct cluster in the EVs.All of this evidence demonstrates SEV-gx is a novel species(tentatively named EV-K)in the EV genus.The complete genomic sequence of Mobovirus A is 7,367 nt.Sequence analysis showed that Mobovirus A and Falcovirus/Tremovirus of Picornaviridae shared a low homology,with P1<22%,P2<24 and P3<24%identities.Furthermore,P1,2C and 3D aa phylogenetic analysis displayed Mobovirus A formed an independent branch in picornaviruses and most closely related to Falcovirus/Tremovirus/Hepotavirus.5’UTR RNA structure showed the IRES of Mobovirus A and five known types of IRES have no obvious similarity.Genomic analysis showed that Mobovirus A has the typical characteristics with 5’UTR-VP2-VPO-VP3-VP1-2A-2B-2C-3A-3B-3C-3D-3’UTR.There are no L protein in the Mobovirus A and protein VPO can be cleaved into VP2 and VP4.Mobovirus A has the characteristics typical for picornaviruses.According to the viral classification by ICTV,the novel virus in this study should be classified as a novel genus in the Picornaviridae(tentatively named Monkey-borne-virus.Mobovirus).ConclusionThis study fully understood the viral spectrum existing in rhesus macaques from Longhu mountain,Guangxi,in China,and also found a large number of novel viruses,such as picornaviruses(including novel enterovirus),human astrovirus-like viruses,human bocavirus-like viruses,and human norovirus,which may greatly enrich the viral database of wide monkeys.This study reported a novel enterovirus species and a novel genus of picornavirus from the rhesus monkeys of Guangxi Longhu mountain,which will contribute to our understanding of the genetic diversity and evolution of picornaviruses.