Abstract：
Vancomycin-resistant Enterococcus faecium (VREfm) is an opportunistic pathogen among the highest global priorities regarding public and environmental health. Following One Health approach, we determined for the first time the antibiotic resistance and virulence genes, and sequence types (STs) affiliation of VREfm recovered simultaneously from marine beach waters, submarine outfall of a wastewater treatment plant and an offshore discharge of untreated sewage, and compared them with the surveillance VREfm from regional university hospital in Croatia to assess the hazard of their transmission and routes of introduction into the natural environment. Importantly, VREfm recovered from wastewater, coastal bathing waters and hospital shared similar virulence, multidrug resistance, and ST profiles, posing a major public health threat. All isolates carried the vanA gene, while one clinical isolate also possessed the vanC2/C3 gene. The hospital strains largely carried the aminoglycoside-resistance genes aac(6\')-Ie-aph(2″)-Ia, and aph(2″)-Ib and aph(2″)-Id, which were also predominant in the environmental isolates. The hyl gene was the most prevalent virulence gene. The isolates belonged to 10 STs of the clonal complex CC17, a major epidemic lineage associated with hospital infections and outbreaks, with ST117 and ST889 common to waterborne and hospital isolates, pointing to their sewage-driven dissemination. To gain better insight into the diversity of accompanying taxons in the surveyed water matrices, microbiome taxonomic profiling was carried out using Illumina-based 16S rDNA sequencing and their resistome features predicted using the PICRUSt2 bioinformatics tool. An additional 60 pathogenic bacterial genera were identified, among which Arcobacter, Acinetobacter, Escherichia-Shigella, Bacteroides and Pseudomonas were the most abundant and associated with a plethora of antibiotic resistance genes and modules, providing further evidence of the hazardous effects of wastewater discharges, including the treated ones, on the natural aquatic environment that should be adequately addressed from a sanitary and technological perspective.
摘要：
耐万古霉素的屎肠球菌（VREfm）是关于公众和环境健康的全球最高优先事项之一的机会性病原体。遵循一种健康方法，我们首次确定了抗生素抗性和毒力基因，和从海洋海滩水域同时回收的VREfm的序列类型（STs）隶属关系，污水处理厂的海底排污口和未经处理的污水的海上排放，并将它们与克罗地亚地区大学医院的监测VREfm进行比较，以评估其传播的危害和引入自然环境的途径。重要的是,从废水中回收的VREfm，沿海沐浴水域和医院具有相似的毒力，多药耐药,和ST剖面，构成重大公共卫生威胁。所有分离株都携带vanA基因,而一个临床分离株也具有vanC2/C3基因。医院菌株主要携带氨基糖苷抗性基因aac（6'）-Ie-aph（2”）-Ia，和aph(2″)-Ib和aph(2″)-Id，在环境分离物中也占主导地位。hyl基因是最普遍的毒力基因。分离株属于10个STs的克隆复合体CC17，这是一个与医院感染和暴发有关的主要流行谱系，ST117和ST889是水传播和医院分离株常见的，指向他们的污水驱动的传播。为了更好地了解被调查水基质中伴随分类群的多样性，使用基于Illumina的16SrDNA测序进行微生物组分类分析，并使用PICRUSt2生物信息学工具预测其抗性组特征。另外鉴定出60个病原细菌属，其中，杆菌，不动杆菌,大肠杆菌志贺氏菌,拟杆菌和假单胞菌是最丰富的，并且与过多的抗生素抗性基因和模块相关，提供废水排放的有害影响的进一步证据，包括治疗过的，关于自然水生环境，应该从卫生和技术的角度加以充分解决。
