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从不同环境基质中回收的肺炎克雷伯菌和产酸克雷伯菌中的整合子及其相关基因盒的流行率、分子特征。

Prevalence, molecular characterization of integrons and its associated gene cassettes in Klebsiella pneumoniae and K. oxytoca recovered from diverse environmental matrices.

机构信息

SAMRC Microbial Water Quality Monitoring Centre, University of Fort Hare, Alice, South Africa.

Applied and Environmental Microbiology Research Group (AEMREG), Department of Biochemistry and Microbiology, University of Fort Hare, Alice, South Africa.

出版信息

Sci Rep. 2023 Sep 1;13(1):14373. doi: 10.1038/s41598-023-41591-7.

Abstract

The high prevalence of infections arising from Klebsiella species is related to their ability to acquire and disseminate exogenous genes associated with mobile genetic elements such as integrons. We assessed the prevalence, diversity, and associated gene cassettes (GCs) of integrons in Klebsiella species. The isolates recovered from wastewater and hospital effluents, rivers, and animal droppings were identified using the conventional Polymerase Chain Reaction (PCR) with primers targeting the gryA, pehX, and 16S-23S genes. The antimicrobial resistance profile and the Extended-Spectrum and Metallo β-lactamases production were carried out using standard microbiological techniques. PCR, DNA sequencing analyses, and Restriction Fragment Length Polymorphism were used to characterize the integrons and their associated GCs. Furthermore, the genotypic relationships between the different isolated K. pneumoniae were determined using Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR. About 98% (51/52) of the confirmed isolates harboured an integrase gene, with 80% intI1, while the remaining 20% concurrently harboured intI1 and intI2, with no intI3 observed. About 78% (40/51) of the bacterial strains were positive for a promoter, the P2R2, investigated, while 80% (41/51) harboured at least one of the qacEΔ1 and sul1. Three different GCs arrangements identified were aac(6')-Ib, aadA1-dfrA1, and dfrA1-sat2. At a similarity index of 60%, the ERIC-PCR fingerprints generated were categorized into nine clusters. Our study is the first to reveal the features of integrons in Klebsiella spp. recovered from environmental sources in the Eastern Cape Province, South Africa. We conclude that the organisms' sources are repositories of integrons harbouring various gene cassettes, which can be readily mobilized to other microorganisms in similar or varied niches.

摘要

产酸克雷伯菌物种中感染的高发率与它们获取和传播与移动遗传元件(如整合子)相关的外源性基因的能力有关。我们评估了产酸克雷伯菌物种中整合子的流行率、多样性和相关基因盒(GCs)。使用针对 gryA、pehX 和 16S-23S 基因的常规聚合酶链反应(PCR),从废水和医院废水、河流和动物粪便中分离出的分离物。使用标准微生物技术进行抗生素耐药谱和广谱和金属β-内酰胺酶的产生。使用 PCR、DNA 测序分析和限制性片段长度多态性来表征整合子及其相关的 GCs。此外,使用肠杆菌重复基因间一致性(ERIC)-PCR 确定不同分离的肺炎克雷伯菌之间的基因型关系。大约 98%(51/52)的确认分离物携带整合酶基因,其中 80%为 intI1,而其余 20%同时携带 intI1 和 intI2,未观察到 intI3。在所研究的 P2R2 中,大约 78%(40/51)的细菌菌株为启动子阳性,而 80%(41/51)携带至少一个 qacEΔ1 和 sul1。鉴定出的三种不同的 GCs 排列是 aac(6')-Ib、aadA1-dfrA1 和 dfrA1-sat2。在相似度指数为 60%时,生成的 ERIC-PCR 指纹图谱分为九个簇。我们的研究首次揭示了南非东开普省环境来源的克雷伯菌属中整合子的特征。我们得出结论,这些生物体的来源是携带各种基因盒的整合子的储存库,这些基因盒可以很容易地转移到类似或不同生态位的其他微生物中。

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