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中国北方某污水处理厂中碳青霉烯酶基因的流行情况及归宿

Prevalence and Fate of Carbapenemase Genes in a Wastewater Treatment Plant in Northern China.

作者信息

Yang Fengxia, Mao Daqing, Zhou Hao, Luo Yi

机构信息

School of Environmental Science and Engineering, Tianjin University, Tianjin, China.

College of Environmental Science and Engineering, Ministry of Education Key Laboratory of Pollution Processes and Environmental Criteria, Nankai University, Tianjin, China.

出版信息

PLoS One. 2016 May 26;11(5):e0156383. doi: 10.1371/journal.pone.0156383. eCollection 2016.

Abstract

Carbapenemase-producing strains of bacteria, which were primarily found in the medical field, have increasingly been found in the environment, thus posing potential risks to public health. One possible way for carbapenemase genes to enter the environment is via wastewater. Therefore, the goal of this study was to determine the occurrence and fate of five high-risk carbapenemase genes in a wastewater treatment plant (WWTP) in northern China using real-time qPCR. Results showed that the blaKPC-2, blaGES-1, and blaIMP-1 genes prevailed throughout all processing stages (even in the chlorination disinfection unit) in the WWTP, whereas the blaVIM-2 and blaOXA-48 genes were not detected in all samples. Worryingly, considerable amounts of carbapenemase genes ((1.54 ± 0.61) × 103 copies/mL to (2.14± 0.41) × 105 copies/mL) were detected in WWTP effluent samples, while the majority of the carbapenemase genes were transported to the dewatered sludge with concentrations from (6.51 ± 0.14) × 109 copies/g to (6.18 ± 0.63) × 1010 copies/g dry weight. Furthermore, a total of 97 KPC-2-producing strains, belonging to 8 bacterial genera, were isolated from the WWTP. Sequencing of 16S rRNA revealed that most of KPC-2 producing isolates were opportunistic pathogens, including Klebsiella spp. (10.3%), Enterococcus spp. (11.3%), Acinetobacter spp. (19.6%), Escherichia spp. (12.4%), Shigella spp. (17.5%), Stenotrophomonas spp. (10.3%) and Wautersiella spp. (9.3%). Moreover, blaKPC-2 genes were identified for the first time in Paenibacillus spp. isolates (an indigenous bacteria), indicating an increased risk of horizontal transfer between clinical pathogens and environmental bacteria. Indeed, a conjugation experiment demonstrated transfer of the blaKPC-2 gene to an E.coli J53 strain from a Klebsiella strain isolated from the WWTP. To our knowledge, this is the first study to obtain Paenibacillus spp. isolates carrying the carbapenemase gene and to quantify the abundance of carbapenemase genes in the environment.

摘要

产碳青霉烯酶的细菌菌株最初主要在医学领域被发现,如今在环境中也越来越多地被发现,从而对公众健康构成潜在风险。碳青霉烯酶基因进入环境的一种可能途径是通过废水。因此,本研究的目的是利用实时定量聚合酶链反应(qPCR)来确定中国北方一家污水处理厂(WWTP)中5种高风险碳青霉烯酶基因的存在情况和去向。结果表明,blaKPC - 2、blaGES - 1和blaIMP - 1基因在污水处理厂的所有处理阶段(甚至在氯化消毒单元)都普遍存在,而blaVIM - 2和blaOXA - 48基因在所有样本中均未检测到。令人担忧的是,在污水处理厂的出水样本中检测到了大量的碳青霉烯酶基因((1.54±0.61)×10³拷贝/毫升至(2.14±0.41)×10⁵拷贝/毫升),而大多数碳青霉烯酶基因被转移到脱水污泥中,其浓度为(6.51±0.14)×10⁹拷贝/克至(6.18±0.63)×10¹⁰拷贝/克干重。此外,从该污水处理厂共分离出97株产KPC - 2的菌株,它们属于8个细菌属。16S rRNA测序显示,大多数产KPC - 2的分离株是机会致病菌,包括克雷伯菌属(10.3%)、肠球菌属(11.3%)、不动杆菌属(19.6%)、埃希氏菌属(共12.4%)、志贺氏菌属(17.5%)、嗜麦芽窄食单胞菌属(10.3%)和沃氏菌属(9.3%)。此外,在芽孢杆菌属分离株(一种本土细菌)中首次鉴定出blaKPC - 2基因,这表明临床病原体与环境细菌之间水平转移的风险增加。事实上,一项接合实验证明blaKPC - 2基因从污水处理厂分离出的一株克雷伯菌转移到了大肠杆菌J53菌株中。据我们所知,这是第一项获得携带碳青霉烯酶基因的芽孢杆菌属分离株并对环境中碳青霉烯酶基因丰度进行量化的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9e3d/4882038/0fe1065a9f14/pone.0156383.g001.jpg

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