This study aimed to evaluate the prevalence and genetic characteristics of carbapenemase-producing Enterobacteriaceae (CPE) in hospital sewage and river water in the Philippines, which has a typical tropical maritime climate. We collected 83 water samples from 7 hospital sewage and 10 river water sites. CPE were identified using CHROMagar mSuperCARBA, and Gram-negative strains were identified using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) or 16S rRNA gene sequencing. Resistance genes in Enterobacteriaceae strains were identified using PCR and DNA sequencing, and transferability of carbapenemase genes from the CPE was investigated with conjugation experiments. Genotyping was performed using multilocus sequence typing (MLST) for Escherichia coli and Klebsiella pneumoniae Out of 124 Enterobacteriaceae isolates, we identified 51 strains as CPE and divided these into 7 species, 11 E. coli, 14 Klebsiella spp., 15 Enterobacter spp., and 11 others, including 4 additional species. Conjugation experiments via broth mating and using E. coli J53 revealed that 24 isolates can transfer carbapenemase-encoding plasmids. MLST analysis showed that 6 of 11 E. coli isolates belonged to clonal complex 10 (CC10). Of 11 K. pneumoniae strains, 9 unique sequence types (STs) were identified, including ST147. Five types of carbapenemase genes were identified, with the most prevalent being NDM (n = 39), which is epidemic in clinical settings in the Philippines. E. coli CC10 and K. pneumoniae ST147, which are often detected in clinical settings, were the dominant strains. In summary, our results indicate that hospital sewage and river water are contaminated by CPE strains belonging to clinically important clonal groups.IMPORTANCE Carbapenemase-producing Enterobacteriaceae (CPE) cause severe health care-associated infections, and their increasing prevalence is a serious concern. Recently, natural ecosystems have been recognized as important reservoirs of antibiotic resistance genes. We investigated the prevalence and genetic characteristics of CPE isolated from the environment (hospital sewage and river water) in the Philippines and found several CPE, including Escherichia coli and other species, with different carbapenemases. The most prevalent carbapenemase gene type was NDM, which is endemic in clinical settings. This study revealed that isolates belonging to carbapenemase-producing E. coli CC10 and K. pneumoniae sequence type 147 (ST147), which are often detected in clinical settings, were dominant in the natural environment. Our work here provides a report on the presence and characteristics of CPE in the environment in the Philippines and demonstrates that both hospital sewage and river water are contaminated by CPE strains belonging to clinically important clonal groups.

译文

:这项研究旨在评估菲律宾的医院污水和河水中产生碳青霉烯酶的肠杆菌科(CPE)的流行情况和遗传特征,菲律宾是典型的热带海洋性气候。我们从7个医院污水和10个河水站点收集了83个水样。使用CHROMagar mSuperCARBA鉴定CPE,并使用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)或16S rRNA基因测序鉴定革兰氏阴性菌株。利用PCR和DNA测序鉴定肠杆菌科细菌的抗性基因,并通过缀合实验研究了来自CPE的碳青霉烯酶基因的可转移性。使用多基因座序列分型(MLST)对大肠杆菌和肺炎克雷伯菌进行基因分型,从124株肠杆菌科细菌中分离出51株CPE,并将其分为7种,分别为11种大肠杆菌,14种克雷伯菌,15种肠杆菌,其他11种,包括另外4种。通过肉汤交配和使用大肠杆菌J53进行的缀合实验表明,有24种分离株可以转移编码碳青霉烯酶的质粒。 MLST分析显示11株大肠杆菌中有6株属于克隆复合物10(CC10)。在11株肺炎克雷伯菌中,鉴定出9种独特的序列类型(ST),包括ST147。鉴定出五种碳青霉烯酶基因,其中最流行的是NDM(n = 39),这在菲律宾的临床环境中很流行。在临床环境中经常检测到的大肠杆菌CC10和肺炎克雷伯菌ST147是主要菌株。总而言之,我们的结果表明,医院污水和河水被属于临床上重要的克隆群体的CPE菌株污染。近来,自然生态系统已被认为是抗生素抗性基因的重要储存库。我们调查了菲律宾从环境(医院污水和河水)中分离出的CPE的流行情况和遗传特征,发现了几种CPE,包括大肠杆菌和其他物种,具有不同的碳青霉烯酶。最流行的碳青霉烯酶基因类型是NDM,在临床环境中很流行。这项研究表明,在临床环境中经常发现属于产碳青霉烯酶的大肠杆菌CC10和肺炎克雷伯氏菌序列类型147(ST147)的分离株。我们在这里的工作提供了有关菲律宾环境中CPE的存在和特征的报告,并证明了医院污水和河水均被临床上重要的克隆组的CPE菌株污染。

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