| The Panthera Tigris Altaica and the Panthera Pardus Fontanierii are two of the most endangered species in the world.They have great protection value.The Escherichia coli in their intestines has an important impact on the health and survival of Panthera Tigris Altaica and Panthera Pardus Fontanierii.Currently,the environment is thought to be a reservoir of resistance E.coli and its genes because of the heavy use of antibiotics,which makes the environment a pool of many man-made sources of pollution.However,at present,there are few reports on resistance E.coli in Panthera Tigris Altaica and Panthera Pardus Fontanierii in China.Therefore,this study takes the E.coli isolates of Panthera Tigris Altaica and Panthera Pardus Fontanierii as the research object,combined with the experimental method,whole genome sequencing and bioinformatics analysis and so on the many kinds of biological technology,first to clarify Panthera Tigris Altaica and Panthera Pardus Fontanierii the prevalence of resistance E.coli in our country,To explore the source of E.coli resistance in Panthera Tigris Altaica and Panthera Pardus Fontanierii.In this study,35 fecal samples of Panthera Tigris Altaica were collected in Changbai Mountain and Wanda mountain area,and 11 fecal samples of Panthera Pardus Fontanierii were collected in Liupanshan,Ningxia Province.E.coli was isolated and identified,and the isolation rates were 90.91%and 82.85%,respectively.Antimicrobial susceptibility was conducted by using K-B paper disk method recommended by CLSI,E.coli had different degrees of resistance to 15 antibiotics,the resistance rate toβ-lactam antibiotics was 100%,followed by the resistance rate to tetracycline antibiotics was about 20%.Bioinformatics analysis results show that:In this study,24 Panthera Tigris Altaica and 10Panthera Pardus Fontanierii were successfully assembled by Illumina whole-genome sequencing.The Res Finder database found that 34 experimental strains carried a total of 9resistance genes:mdf（A）detection rate was 94.12%,sit ABCD detection rate was 23.53%,Oqx B、gyr A,、par C、par E detection rate was 2.94%、2.94%、17.65%、5.88%,tet（A）、tet（B）detection rate was 5.88%、2.94%.bla OXY-4-1 detection rate was 2.94%.There were 8different types of plasmid replicates in 34 strains of experimental bacteria retrieved from the Plasmadfinder database,including Inc FII,Inc X1,Inc FIA,Inc Y,P0111,Inc FIB,Inc FIC and Inc I2.11 O antigen serotypes and 8 H antigen serotypes were detected in 24 strains of the Panthera Tigris Altaica,and 7 O antigen serotypes and 6 H antigen serotypes were detected in nine strains of the Panthera Pardus Fontanierii.Virulence Finder database searched virulence factors whose identity ratio was 100%.10 virulence factors were detected in 24 strains of Panthera Tigris Altaica,and the detection rate of Ipf A was the highest（59.09%）.12 virulence factors were detected in 10 strains of Panthera Pardus Fontanierii,and the detection rate of omp T was the highest（62.50%）.Shiga toxin stx2a and stx2b with cytotoxic potential were detected in a test strain of Panthera Tigris Altaica.MLST database retrieval showed that 34strains were divided into 18 ST types.ST939 was the main clone type（n=7）,followed by ST0（n=5）,ST3246（n=4）,ST58（n=2）,and ST126（n=2）.The maximum SNP distance between each strain was 214,490 SNPs.and a minimum distance of 1 SNP.This study takes the top wild carnivores（Panthera Tigris Altaica and Panthera Pardus Fontanierii）as the indicator species of the ecological environment,investigates the drug resistance of E.coli in Panthera Tigris Altaica and Panthera Pardus Fontanierii at the molecular level through bioinformatics analysis,indirectly evaluates the diffusion of E.coli drug resistance gene in animal micro ecosystem under the pressure of human antibiotic selection,and deeply discusses the impact of China’s ecological environment on Panthera Tigris Altaica and Panthera Pardus Fontanierii,It is of great significance to promote the protection and management of the rare wild animals and the sustainable development of the ecological environment. |
