Study On Drug Resistant Phenotype And Resistance Genes Of Klebsiella Pneumoniae From Wild Animals In Zoos In Fujian Province

Objective:Klebsiella pneumoniae is a common opportunistic pathogen and zoonotic pathogen of wild animals in captivity,it infects humans and mammals such as primates,omnivores,cats,canines,herbivores,and birds,amphibians and reptiles,even aquatic animals.The problem of drug resistance caused by antibiotics and other drugs used by veterinarians is becoming more and more obvious,and multi drug resistant strains are also emerging.In this paper,we investigated the infection of Klebsiella pneumoniae from wild animals in three zoos in Fujian Province,and the drug resistance and gene types of Klebsiella pneumoniae isolated from animals.We studied the correlation between drug resistance phenotype and drug resistance genes,analyzed and compared the infection and drug resistance of Klebsiella pneumoniaeanimals in animals from different species,and the relationship between the drug resistance phenotype and drug resistance genes of the strains,in order to improve the understanding of veterinary and other animal workers in the prevention and control of Klebsiella pneumoniae,provide the basis for the prevention and control of Klebsiella pneumoniae in wild animals and related public health and safety work in the future.Method:The feces of different species of wild animals were collected from three zoos in Fujian Province,including 80 from Fuzhou zoo,80 from a zoo in Sanming City and 90 from a zoo in Nanping City.Klebsiella pneumoniae was isolated and identified from feces.Plate dilution method was used to analyze the drug resistance of cefazolin,cefotaxime,tetracycline,chloramphenicol,polymyxin,imipenem,ciprofloxacin,nitrofurantoin,gentamicin and other drugs.The resistance genes of the drug-resistant strains were detected by conventional PCR.Then analyze the relationship between drug resistance gene and drug resistance phenotype of Klebsiella pneumoniae.Result:1.Bacterial isolation:110 strains of Klebsiella pneumoniae were isolated and identified from 250 animal feces samples from three zoos.The highest isolation rate was 47.8%(43/90)in the zoo in Nanping,and47.5%(38/80)in the zoo in Sanming and 36.3%(29/80)in Fuzhou zoo.The isolation rate of primates was the highest,reaching 60.6%(49/81)in total,the highest was 73.7%(14/19)in Nanping.The isolation rate of Klebsiella pneumoniae in carnivorous animals was 25%(8/32),which was significantly lower than that in omnivorous animals(35%(11/30))and herbivores(57.9%(22/38))(P<0.05).2.Drug resistance test:The drug resistance of 110 strains of Klebsiella pneumoniae isolated from the three zoos to cefazolin,cefotaxime,tetracycline,chloramphenicol,polymyxin,imipenem,ciprofloxacin,nitrofurantoin and gentamicin,nitrofurantoin was the most resistant,accounting for 80.9%(89/110),followed by polymyxin,accounting for 71.8%(79/110).The lowest resistance rate was imipenem,with no resistant strains were detected.In addition,it was resistant to gentamicin,cefazolin,cefotaxime,tetracycline,chloramphenicol and ciprofloxacin,in different degrees.3.Drug resistance gene detection:15 of the 17 drug-resistant genes were detected,bla _SHVwas not detected.The detection rate of oqx A was the highest,with 75.5%,the detection rate of?-lactam resistance gene Amp C was 60%.The resistance rate of polymyxin was 71.8%(79/110),but the resistance gene mcr-1 was not detected this time.Conclusion:(1)The total isolation rate of Klebsiella pneumoniae was 44.0%(110/250),and the highest isolation rate was 47.7%(14/19)in Nanping zoo.The isolation rate of primates is the highest among all kinds of animal samples,which may be due to the close relationship between primates and human,which makes them more susceptible to infection.(2)In the aspect of drug resistance phenotype,Fuzhou zoo had the lowest level of drug resistance;Sanming zoo had higher drug resistance level to the zoo in Nanping in gentamicin,cefazolin,cefotaxime and ciprofloxacin;while tetracycline and chloramphenicol were the opposite.(3)In terms of drug resistance genotypes,the correlation between tetracycline resistance phenotype and resistance gene was the best.However,the resistance phenotypes and genotypes of other antibiotics were not well matched.The high detection rates of oqx A and oqx B were75.5%and 23.6%respectively.oqx A and oqx B not only mediate resistance to fluoroquinolones,but also cause low to moderate resistance to chloramphenicol and nitrofurantoin,which may be an important reason for the poor correlation between resistance phenotypes and resistance genes of chloramphenicol and nitrofurantoin.(4)The detection rate of cephalosporins and quinolones resistant bacteria was low(75.5%and 23.6%),but the detection rate of related resistance genes was high.A large number of intermediate bacteria were detected in the actual drug sensitivity test,which suggested that the zoo should pay attention to rational drug use and strengthen feeding management.