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在BfT-GermVet监测研究中收集的猪、马、猫和狗的大肠杆菌分离株中1类和2类整合子及相关基因盒的分析与分布

Analysis and distribution of class 1 and class 2 integrons and associated gene cassettes among Escherichia coli isolates from swine, horses, cats and dogs collected in the BfT-GermVet monitoring study.

作者信息

Kadlec Kristina, Schwarz Stefan

机构信息

Institute of Farm Animal Genetics, Friedrich-Loeffler-Institute, Höltystr. 10, 31535 Neustadt-Mariensee, Germany.

出版信息

J Antimicrob Chemother. 2008 Sep;62(3):469-73. doi: 10.1093/jac/dkn233. Epub 2008 Jun 11.

Abstract

OBJECTIVES

In the BfT-GermVet monitoring study, 417 Escherichia coli isolates collected during 2004-06 in Germany from various disease conditions of pigs (n = 87), horses (n = 102) or cats/dogs (n = 228) were investigated for their susceptibility to 24 antimicrobial agents. This study dealt with the identification of integron-associated resistance genes among these isolates.

METHODS

Class 1 and class 2 integrons were detected by PCR. The variable parts of the integrons were cloned and sequenced. Transformation and conjugation experiments were conducted to confirm a plasmid location of the integrons.

RESULTS

Class 1 and/or class 2 integrons, alone or in different combinations, were detected in 79 of the 417 E. coli isolates. Four trimethoprim resistance genes (dfrA1/12/14/17), five streptomycin/spectinomycin resistance genes (aadA1/2/4/5/6), two streptothricin resistance genes (estX, sat2), one gentamicin/tobramycin/kanamycin resistance gene (aadB) and one chloramphenicol resistance gene (catB3) were detected. Seven different cassette arrangements were identified within class 1 integrons: aadA1 (21 isolates), dfrA1 + aadA1 (18 isolates), dfrA17 + aadA5 (9 isolates), dfrA12 + orfF + aadA2 (8 isolates), aadB + aadA1 (1 isolate), dfrA14 + recombined aadA6 (1 isolate) and dfrA1 + catB3 + aadA4 (1 isolate). Three different cassette arrangements in class 2 integrons, dfrA1 + sat2 + aadA1 (24 isolates), estX + sat2 + aadA1 (6 isolates) and estX + sat2 + DeltaaadA1 (1 isolate), were identified. The plasmid location of class 1 and/or class 2 integrons was confirmed in 37 isolates.

CONCLUSIONS

Class 1 and/or class 2 integrons carrying resistance gene cassettes were detected in 18.9% of the isolates tested. This molecular analysis complements the phenotypic susceptibility testing conducted in the BfT-GermVet monitoring study and helps to explain the persistence of resistance genes even without direct selective pressure.

摘要

目的

在德国联邦风险评估研究所(BfT)-德国兽医监测研究中,对2004年至2006年期间从猪(n = 87)、马(n = 102)或猫/狗(n = 228)的各种疾病状况中收集的417株大肠杆菌分离株进行了24种抗菌药物的敏感性研究。本研究旨在鉴定这些分离株中与整合子相关的耐药基因。

方法

通过聚合酶链反应(PCR)检测1类和2类整合子。对整合子的可变部分进行克隆和测序。进行转化和接合实验以确认整合子的质粒定位。

结果

在417株大肠杆菌分离株中,有79株检测到1类和/或2类整合子,单独或呈不同组合形式存在。检测到4个甲氧苄啶耐药基因(dfrA1/12/14/17)、5个链霉素/壮观霉素耐药基因(aadA1/2/4/5/6)、2个链丝菌素耐药基因(estX、sat2)、1个庆大霉素/妥布霉素/卡那霉素耐药基因(aadB)和1个氯霉素耐药基因(catB3)。在1类整合子中鉴定出7种不同的盒式排列:aadA1(21株)、dfrA1 + aadA1(18株)、dfrA17 + aadA5(9株)、dfrA12 + orfF + aadA2(8株)、aadB + aadA1(1株)、dfrA14 + 重组aadA6(1株)和dfrA1 + catB3 + aadA4(1株)。在2类整合子中鉴定出3种不同的盒式排列:dfrA1 + sat2 + aadA1(24株)、estX + sat2 + aadA1(6株)和estX + sat2 + ΔaadA1(1株)。在37株分离株中确认了1类和/或2类整合子的质粒定位。

结论

在所检测的分离株中,18.9%的菌株检测到携带耐药基因盒的1类和/或2类整合子。这种分子分析补充了德国联邦风险评估研究所-德国兽医监测研究中进行的表型药敏试验,并有助于解释即使在没有直接选择压力的情况下耐药基因的持续存在。

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