Study On Virome Profiling Of Horses,Discovery And Identification Of Zoonotic Viruses

Objective:In order to uncover the types of zoonotic viruses exsiting in horses,reveal the epidemic characteristics of important zoonotic viruses,which provide a basis for the prevention and control of horse-borne zoonosis.Methods:(1)Nasal swabs were collected from horses in Yili,Urumqi and Changji,and the viral metagenomic sequencing was applied to detect the types of zoonotic viruses in horses;(2)The positive rates of the zoonotic viruses in nasal swabs would be determined by PCR,the genetic diversity and evolution characteristics of which will be further characterized through in genome sequencing,and evolutionary analysis;(3)According to sequences of HCMV and HPV-23,we establish the SYBR-green I q PCR method for them.Results:(1)Two metagenomics results generate 73 007 152 and 90 048 090 reads respectively,of which 72 069(0.098%),and 90 048(0.01%),were annotated to mammalian viruses(Papillomavirus,Herpesvirus,Retrovirus,Respiratory Syncytial virus,Wenzhou virus,Picobirnavirus and Bunyavirus,etc);(2)Ten nasal swabs of Yili horses were positive for HCMV,with a positive rate was 7%,In addition,the positive rate of the virus in abortion sample of Yili horses was 32.2%.Further investigation revealed that the horse origin HCMV could transmit to workers in Yili mare production of HCMV-positive farm.Sequences analysis indicated that the homology nucleotides among our HCMV UL126 shared99.6%-100%.and shared 99.6%～100%nucleotide sequence identity with UL126 genes of human origin HCMV reference strains;(3)HPV-23 positive rate was 16.7%,1.9%,and 1%in nasal swab samples of horses in Urumqi,Yili and Changji,respectively.In addition,the positive rate of the virus in abortion sample of Yili horses was 35.5%.Further investigation revealed that the horse origin HPV-23 could transmit to workers in Yili mare production of HPV-23-positive farm.Sequences analysis indicated that the among our HPV-23 E2 shared 98.4%～100%nucleotide sequence identityand and 97.2%～100%amino acid sequence identity.and shared 98.1%～99.1%nucleotide sequence identityand 96.2%～99.1%amino acid sequence identitywith E2 genes of human origin HPV-23 reference strain;(4)SYBR Green I q PCR was established for detection of HCMV and HPV-23 were 1.15x10~3copies/μL and 7.9x10~1copies/μL,respectively,which showed higher sensitivity than ordinary PCR.Conclusion:(1)The results reveal the most complete viromic structure of zoonotic viruses in horses;(2)HCMV and HPV-23 can infect horses,may function as a causative agent for abortions in horses,and infect humans laboring in mare stables;(3)The SYBR Green q PCR methods for detection of HCMV and HPV-23 were established.