Isolation And Identification Of ESBLs Gram Negative Bacteria From Pets In Tai'an City And Analysis Of Drug Resistance And Virulence Genes

Gram negative bacteria widely exist in nature,human and animal intestines.Studies have pointed out that many zoonotic diseases in pets show recessive infection,so that pets can be used as a repository of microorganisms.In recent years,with the increasing affluence of Chinese people's material life,the number of dogs and cats is increasing.Due to the non-standard drug use and human drug pet use in the diagnosis and treatment of clinical diseases of dogs and cats,the drug resistance of Gram-negative bacteria from pets can not be ignored,and dogs and cats,as people's closest partner,may spread some drug-resistant pathogenic or conditional pathogenic Gram-negative bacteria to people.The purpose of this study was to understand the prevalence and drug resistance of ESBLs producing Gram-negative bacteria in pets in pet hospitals in Tai'an area.98 anal swabs and oral swabs from healthy and diarrhea dogs and cats were collected in 3 pet hospitals in Tai'an City for the isolation and identification of ESBLs producing Gram-negative bacteria;The confirmation test of ESBLs producing Gram-negative bacteria was carried out by disk method,and its resistance to 14 common drugs was detected;Detection of 18 drug resistance genes in four categories by PCR(?-Lactams,quinolones,tetracyclines,aminoglycosides)and class I integrons to understand the carrying situation of drug resistance gene box;PCR technology was used to detect 16 virulence genes of 79 strains of Escherichia coli with the highest isolation rate.At the same time,the distribution of 8 virulence genes of 39 strains of Proteus mirabilis was detected to understand the characteristics of virulence genes.A total of 170 Gram-negative bacteria were isolated in this study.The isolation rate of Escherichia coli was the highest,46.47%(79 / 170),followed by Proteus mirabilis,22.94%(39 / 170);The results of ESBLs producing Gram-negative bacteria confirmatory test showed that among the 170 isolated Gram-negative bacteria,64 ESBLs producing strains were mainly Escherichia coli and Proteus mirabilis of Enterobacteriaceae.The results of drug resistance phenotype showed that the resistance rates of 64 ESBLs producing strains to ampicillins,tetracyclines,sulfonamides,macrocyclic lipids and cephalosporins were higher,all of which were more than 45%;The drug resistance rate of 106 non ESBLs producing strains to tetracyclines,ampicillins and macrocyclic lipids was more than 45%,indicating that the antibiotic resistance of ESBLs producing bacteria was serious.The results of drug resistance gene test show that,?-Among lactams,the carrying rate of blactx-m resistance gene was the highest,which was 47.06%.The carrying rates of blashv and blaoxa resistance genes were 27.06% and 2.94% respectively.No blapse and blatem resistance genes were detected.The carrying rates of AAC(6 ')-IB Cr and aac4 in aminoglycoside resistance genes were 45.88% and 42.35% respectively.Aac1,Aac2 and AAC3 resistance genes were not detected.The carrying rates of oqxa,qnra,qnrs,qnrb and qnrd resistance genes in quinolones were 40.0%,22.4%,18.8%,6.5% and 1.8% respectively.Qnrc resistance gene was not detected positive.Tet A and tetb resistance genes were not detected in tetracyclines.The results of virulence gene detection showed that 87.3% of E.coli carried irp2 gene,35.4% of the strains carried EAEA gene,6.3% of the strains carried east1 gene,and only2.5% of the strains carried F17 gene,while all strains did not detect Hly A,stx1,Stx2,K88,K99,987 P,LT,STA,BFPA,F18,F41 and cs31 a genes,None of the virulence genes investigated were detected in 5 strains of Escherichia coli.Among the eight virulence genes of Proteus mirabilis,the detection rates of virulence genes MRPA,ZapA,ATFA,ATFC,ucaa and pmfa were the highest,all 100%,followed by urec,the detection rate of virulence gene was 97.44%,and rsba virulence gene was not detected.Through the isolation and identification of ESBLs producing Gram-negative bacteria,the drug resistance of common drugs,the carrying of class ? integron and drug resistance gene,and the carrying of virulence gene in dog and cat anal swabs and oral swabs from three pet hospitals in Tai'an area,it was found that there was ESBLs producing Gram-negative bacteria pollution in Tai'an area.Gram negative bacteria in this area showed tolerance to a variety of drugs,and the multi drug resistance rate was 68.24%.The resistance rate of ESBLs producing Proteus mirabilis to ampicillins,tetracyclines,sulfonamides,macrocyclic lipids and cephalosporins was more than 45%;The resistance rate of non ESBLs producing strains to four antibiotics was more than 45%,indicating that the resistance of ESBLs producing strains was serious.The aim of this study is to understand the drug resistance of gram negative bacteria from pets,and to provide guidance for standardizing and limiting the use of antibiotics and the clinical treatment of pet infectious diseases in Tai'an.