Analysis Of Virus Diversity In Rodents And Identification Of A Newly Discovered Arenavirus

Spillover of wildlife-borne pathogens affects human health and animal protection,in order to conduct an in-depth analysis of the species and diversity of pathogens carried by wild animals,Wildlife sample collection was carried out,analysis of virus diversity carried by rodents,isolation and identification of newly discovered pathogens.First of all,some wild animals with close human contact were selected for sample collection.Wildlife in close contact with humans includes zoo ornamental animals,tourist attraction animals,rescue wildlife,village and wild wildlife,and captive economic animals.The sample collection range was in seven provinces: Hubei,Hunan,Henan,Guangxi,Ningxia,Heilongjiang and Liaoning.Sample types include tissue samples,blood samples,swab samples,and environmental samples.The collection lasted for one year,and a total of 12,605 wildlife samples were obtained,covering 217 species in 28 orders.The samples were collected and information collated according to the region and type.In the process of sample collection,we have formulated specifications for wild animal sample collection in combination with host characteristics and scientific research needs,and carried out informatization and classification management of wild animal samples that are in close contact with humans,effective utilization,safe use and visual management of each sample are realized.The large number of samples collected provides a guarantee for subsequent studies on the diversity of viruses carried by wild animals in close contact with humans.In order to carry out research on the diversity of viruses carried by wild animals in close contact with humans,the diversity of viruses carried by wild rodents in Ningxia was given priority.The second-generation sequencing method was used to detect the sample virus by using the metagenomic sequencing strategy.The 83 wild rats collected in Ningxia were divided into 21 groups according to the collection location and species,and the viral sequences carried by wild rodents were obtained through bioinformatics analysis and treatment.The results showed that the number of virus reads obtained after screening was 847,and the virus sequence could be classified into 22 virions,including 15 RNA virus families,4 DNA virus families,2 bacteriophage virus families and 1 fungal virus family,of which the virus abundance of the arenaviruses was the highest.It is proved that the wild rodents in Ningxia carry a variety of viruses and have rich diversity.Based on the results obtained by the second-generation sequencing,the arenaviruses found in it was analyzed intensively.The sequence of arenaviruses was verified and amplified by a combination of first-generation and second-generation sequencing.The full length of the four proteins of arenaviruses was successfully obtained,including RDRP(6693bp),NP(1729bp),GPC(1457bp),and ZP(242bp).The sequence of the four proteins is compared with nucleotides,the results show that only RDRP can align to Ippy virus of arenaviruses,but the sequence coverage is only 2%;through amino acid alignment,the results show that RDRP,GPC,NP are all aligned to Alxa virus,and the homology was 80.60%,85.38%,84.46%,but there was no matching result of ZP protein alignment.Based on the results of homology alignment and phylogenetic analysis,we defined the arenavirus found in wild mice in Ningxia as a novel arenavirus,named ZHW virus,or ZHW virus for short.In order to investigate the prevalence of the virus,For screening of 83 wild rats in Ningxia,10 of them were found to be positive for nucleic acid,including 8 jerboas,1 foot rat and 1 gerbil,with a positive rate of 12%.Successful acquisition of ZHW virus isolates.This strain is susceptible to BHK cell lines and can produce significant cytopathy after 24 hours of infection in a cell culture incubator at 37 °C.The sequence determination confirmed that the ZHW virus isolate was consistent with the original sequence of ZHW virus,and transmission electron microscopy showed typical arenaviruses morphological characteristics.A large number of samples with diverse sources and clear information have been reserved for in-depth research on pathogens carried by wild animals.At the same time,rodents were found to carry a variety of viruses and for the first time isolated a strain of a new arenaviruses family ZHW virus.These research results provide technical support and reference for the study of the diversity of viruses carried by wild animals,and provide a theoretical basis for in-depth understanding of the cross-species spread of viruses.