Detection And Dissemination Analysis Of Antimicrobial Resistance Genes In Klebsiella Pneumoniae From Broiler Carcasses

Antibiotics are considered the twenty-first greatest medical discovery in21st century’s, inhibiting and killing bacteria, viruses, parasites. In the livestock industry, it can protect animal from diseases and stimulating growth. However, as the large-scale use of antibiotics even abuse, making animal intestinal microbial habitat destruction, leading to the animal’s ability to resist outside body reduced, causing frequent secondary disease. Antibiotic residues in aquaculture products, will affect food safety directly through the food chain through the steady streams until to human, primary drug resistance induced occurs, threatening the health of human.In recent years, there was an increase in multi-drug resistant of Klebsiella pneumoniae with antibiotics, especially the emergence of the extensive use of β-lactams and quinolones and other broad-spectrum antibiotics. Klebsiella pneumoniae is a typical opportunistic pathogen, subjected to human or animal respiratory tract into the lungs caused by pathological mutation, drug resistance mechanism was resistant to β-lactamase gene, quinolones. Detecting propagation resistance of genotypes and resistance genes at the molecular level studies in order to take scientific and effective prevention and control measures are very important.Currently, research on drug resistance of Klebsiella pneumoniae clinical isolates mostly hospital-based, few studies in produce livestock breeding. In this paper, samples were collected from a poultry breeding in Shandong Province plant breeder, body surface, commodity chicken and slaughterhouse wastewater,75 isolated and purified strains used as an experimental of Klebsiella pneumoniae. Resistance rate according to agar disk susceptibility experiments was:Klebsiella pneumoniae resisitant for clinical cephalosporins ampicillin (AMP) 100%, ceftazidime (CAZ), kanamycin (KAN), chloramphenicol (CHL), ciprofloxacin stars (CIP) also showing strong resistance, above 80%.In 75 Klebsiella pneumoniae DNA PCR amplifications, there were 72 β-lactamase producing strains resistant gene deteced,66 blaSHB positive,55blaTEM positive, and 60 blaCTX-M positive. To analysis on the gene type of blaSHV and blaCTX-M, the majority were SHV-11, SHV-1 and CTX-M-1 strains. Also by detecting the bacterial plasmid-mediated quinolone resistance gene, there were 68 positive strains resistant gene found, four qnrA positive,56 qnrB positive,23 qnrS positive,2 qepA positive, and 31 aac(6’)-Ib-cr positivel, a total of 66 strains of ESBLs carrying with quinolone resistance genes.According to integrons gene sequence intI1, intI2 and intI3, primers were designed for 75 Klebsiella pneumoniae. The results showed 70 integron Ⅰ positive, nine integron Ⅱ but no integron Ⅲ found. There were three kinds of gene cassette arrays, dfrA17-aadA5, dfrA12-orfF-aadA2 and dfrA1-sat2-aadA1 respectively, of which the largest frequency of gene cassette is dfrA12-orfF-aadA2,1989bp,19 times.To understand the mechanism of resistance in gene transferring we performed preliminary study basing on the resistance and distribution of resistance genes of Klebsiella pneumoniae, in order to raise awareness of the abuse of antibiotics in farming conditions, strengthen the monitoring of farms surrounding environment, to provide a scientific basis preventing and controlling the potential spread and development of Klebsiella pneumoniae, slowing bacterial resistance.