Metagenomics Analysis Of Zoonotic Patho-genic Microorganisms Of Sheep In Xinjiang Pastures,China

The respiratory and intestinal of sheep have a fancy and rich microbial community,which play a significant role in sheep's health,productivity,disease prevention and control.To investigate the diversity of bacteria,viruses and potential zoonosis pathogenic bacteria carried by sheep in Xinjiang large-scale pastures and to provide scientific basis for effective prevention and control of zoonoses.In this study,rangeland sheep were selected from the pastures in Kashgar and Shihezi regions of Xinjiang,and both oropharyngeal and anal swab samples of each sheep were collected at the same time to extract DNA.Macrogenomic second-generation sequencing was adopted,and species annotation,similarity and diversity analyses were carried out through comparison with Minikrakenv2 database and bioinformatics methods.Investigation were carried out to find out what kinds of of bacteria and viruses carried in sheep in the region.A total of 303 sheep samples,including 72 sheep oropharyngeal swabs and 231 sheep anal swabs,were collected from sheep in large-scale pastures in in Kashgar region of southern Xinjiang and Shihezi city of northern Xinjiang in August,2019.Among them,in Kashgar region,91 sheep anal swabs and 29 sheep oropharyngeal swabs and,in Shihezi city,140 sheep anal swabs and 43 sheep oropharyngeal swabs were collected,respectively.A group of 6 sheep oropharyngeal samples were mixed and12 groups were obtained;and 7 sheep anal samples were mixed and 33 groups were obtained.The results of macrogenomic analysis showed that in the two kinds of samples,on the phylum classification level of bacteria,the dominant microflora were Proteobacteria,Bacteroides,Firmicutes and Actinobacteria;while on the taxonomic level of the genus,each of them has completely different dominant genera.10 bacterial zoonotic pathogens were identified,including Campylobacter coli,Campylobacter jejuni,Escherichia coli,Pasteurella multocida,Acinetobacter baumannii,Salmonella,Pseudomonas guangdongensis,Bacillus cereus,Trueperella pyogenes and Streptococcus porcineus,most of which were bacterial food-borne pathogens.The results of PCoA and ANOSIM analysis showed that there were statistically significant difference between the bacterial species carried by sheep in the two regions and among the potential zoonotic bacterial species in the sheep(P<0.05).The results of the macrogenomic analysis showed that on the genus level,the predominant viruses in both samples were ?-retroviruses;on the species level,three viruses,sheep lung adenoma virus(JSRV),sheep endemic rhinotumor virus and goat endemic endonasal tumor virus,were predominant in both samples.Viruses associated with Zoonosis were not detected in this study;PCoA analysis yielded similar composition of virus species in sheep samples collected from the two regions;ANOSIM analysis showed no statistically significant differences between the virus species carried by the sheep in the two regions(P>0.05).To sum up,using macrogenome sequencing technology,we initially elaborated the diversity and regional variability of oropharyngeal and anal bacteria and virus species in sheep in Xinjiang,and detected potential zoonotic pathogens,of which most of them are predominantly spread of foodborne pathogen,there exist certain risk of food safety and disease transmission.It provides some basic scientific data for sheep healthy breeding and related disease prevention and control.