摘要
      水生环境在临床相关抗生素耐药性基因（ARGs）和病原体的传播中发挥着重要作用。关于临床相关获得性耐药基因在海洋环境中的流行情况，尤其是在挪威，目前的知识有限。本研究的目的是利用绿色荧光蛋白（GFP）标记的大肠杆菌菌株作为受体，通过外源质粒捕获，研究卑尔根港海水中自传抗性质粒的存在和特征。我们从处理的13个样品中的4个样品中获得了对氨苄青霉素和头孢噻肟具有耐药性的跨偶联剂。使用Illumina MiSeq和Oxford Nanopore MinION平台，对基于抗生素敏感性模式选择的9种转偶联物进行测序。在这些转偶联物中检测到10个不同的质粒（范围从35kb到136kb），属于不相容组IncFII/IncFIB/Col156、IncFII、IncI1和IncB/O/K/Z。质粒p1A1（IncFII/IncFIB/Col156135.7kb）携带抗性基因blaTEM-1、dfrA17、sul1、sul2、tet（A）、mph（A），aadA5、aph（3〃）-Ib和aph（6）-Id，赋予对六类不同抗生素的抗性。质粒p1A4携带blaCTX-M-55、lnu（F）、aadA17和aac（3）-IId。在从受当地海洋水族馆废水影响的区域捕获的质粒上检测到头孢菌素酶blaCMY-2。除了ARGs，一些质粒还携带毒力因子，如肠毒素、粘附因子和铁载体。我们的研究证明，卑尔根港海水中存在临床上重要的多药耐药性偶联质粒，这些质粒有可能转移到人类微生物群中。正如世界卫生组织所建议的那样，研究结果强调了监测环境中抗生素耐药性的必要性，尤其是在挪威等低流行环境中。
Abstract
Aquatic environments play important roles in the dissemination of clinically-relevant antibiotic resistance genes (ARGs) and pathogens. Limited knowledge exists about the prevalence of clinically-relevant acquired resistance genes in the marine environment, especially in Norway. The aim of the current study was to investigate the presence of and characterize self-transmissible resistance plasmids from Bergen harbor seawater, with exogenous-plasmid capture, using a green fluorescent protein (GFP)-tagged Escherichia coli strain as a recipient. We obtained transconjugants resistant against ampicillin and cefotaxime from four of the 13 samples processed. Nine transconjugants, selected on the basis of antibiotic sensitivity patterns, were sequenced, using Illumina MiSeq and Oxford Nanopore MinION platforms. Ten different plasmids (ranging from 35 kb to 136 kb) belonging to incompatibility groups IncFII/IncFIB/Col156, IncFII, IncI1 and IncB/O/K/Z were detected among these transconjugants. Plasmid p1A1 (IncFII/IncFIB/Col156, 135.7 kb) carried resistance genes blaTEM-1, dfrA17, sul1, sul2, tet(A), mph(A), aadA5, aph(3″)-Ib and aph(6)-Id, conferring resistance against six different classes of antibiotics. Plasmid p1A4 carried blaCTX-M-55, lnu(F), aadA17 and aac(3)-IId. Cephalosporinase blaCMY-2 was detected on plasmids captured from an area impacted by wastewater from a local marine aquarium. Along with ARGs, some plasmids also carried virulence factors, such as enterotoxins, adhesion factors and siderophores. Our study demonstrates the presence of clinically-important multidrug-resistance conjugative plasmids in seawater from Bergen harbor, which have the potential to be transferred to human microbiota. The results highlight the need for surveillance of antibiotic resistance in the environment, as suggested by the World Health Organization, especially in low prevalence settings like Norway.

https://www.sciencedirect.com/science/article/pii/S1438463922001912