Bacterial hosts and horizontal transfer characteristics of clinically important tet(X)-variant genes in municipal wastewater treatment plants using epicPCR-directed cultivation strategy

IF 11.4 1区环境科学与生态学 Q1 ENGINEERING, ENVIRONMENTAL

Water Research Pub Date : 2024-10-18 DOI:10.1016/j.watres.2024.122658

Shiting Dai, Ziming Han, Shihai Liu, Yang Wang, Yu Zhang, Min Yang

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Abstract

Mobile tet(X)-variant genes confer resistance to a wide range of tetracyclines, including the antibiotic of last-resort, tigecycline, raising significant concerns regarding their potential spread cross-environmental dissemination. However, the bacterial hosts and environmental spread of these genes remain poorly understood. Herein, a retrospective study unveiled the prevalence of tet(X)-variant genes (ranging from tet(X3) to tet(X6)) in activated sludge samples from five municipal wastewater treatment plants (WWTPs) from 2013 to 2021. Among these variants, tet(X4) exhibited the highest detection frequency (100%) and abundance [(2.48 ± 3.07) × 10⁷ copies/g dry weight] with an increasing trend. An epicPCR-directed cultivation strategy was proposed to facilitate the targeted isolation of tet(X4)-carrying bacterial hosts in activated sludge. This strategy involves the identification of bacterial host profiles using epicPCR and subsequent selective isolating target bacteria. Enterobacteriaceae emerged as the primary bacterial host for tet(X4), alongside previously unreported genera like Providencia, Advenella, and Moheibacter. Subsequent selective isolation of the most abundant Enterobacteriaceae based on the epicPCR-informed host spectrum yielded 39 tet(X4)-carrying Escherichia coli strains from the WWTP. Whole genome sequencing of tet(X4)-positive strains revealed that plasmid-mediated horizontal gene transfer is the primary mechanism driving tet(X4) dissemination. Plasmids including IncFIA(HI1)/IncHI1A/IncHI1B(R27) and IncX1, commonly reported in various clinical and animal settings, were identified as the predominant carriers of tet(X4). E. coli strains harbouring tet(X4) in the WWTP showed substantial genetic similarity to strains from hospital and animal sources, underscoring concerns about the potential risk of across diverse sectors. This study provided the first glimpse of the presence of mobile tet(X)-variants in WWTPs, and highlighted the promise of the epicPCR-directed cultivation strategy for exploring bacterial hosts of clinically important ARGs in different habitats from a One Health perspective.

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利用 epicPCR 引导培养策略研究城市污水处理厂中临床重要的 tet(X) 变异基因的细菌宿主和水平转移特征

可移动的 tet（X）变异基因对多种四环素类药物（包括最后的抗生素替加环素）具有抗药性，这引起了人们对其跨环境传播潜力的极大关注。然而，人们对这些基因的细菌宿主和环境传播仍然知之甚少。在本文中，一项回顾性研究揭示了2013年至2021年期间五个城市污水处理厂（WWTPs）活性污泥样本中的tet(X)变体基因（从tet(X3)到tet(X6)不等）的流行情况。在这些变体中，tet(X4)的检测频率（100%）和丰度[（2.48 ± 3.07）×107拷贝/克干重]最高，且呈上升趋势。为便于有针对性地分离活性污泥中携带 tet(X4) 的细菌宿主，提出了一种 epicPCR 引导的培养策略。该策略包括使用 epicPCR 鉴定细菌宿主特征，然后选择性分离目标细菌。肠杆菌科成为 tet(X4) 的主要细菌宿主，此外还有普罗维登斯属、阿文氏菌属和莫氏菌属等以前未报道过的菌属。随后，根据 epicPCR 信息宿主谱对最丰富的肠杆菌科细菌进行选择性分离，从污水处理厂中分离出 39 株携带 tet(X4) 的大肠埃希菌。对 tet(X4) 阳性菌株的全基因组测序显示，质粒介导的水平基因转移是驱动 tet(X4) 传播的主要机制。包括 IncFIA(HI1)/IncHI1A/IncHI1B(R27) 和 IncX1 在内的质粒是 tet(X4) 的主要携带者，这些质粒通常在各种临床和动物环境中被报道。污水处理厂中携带 tet(X4)的大肠杆菌菌株与医院和动物来源的菌株在基因上有很大的相似性，这突显了对不同行业潜在风险的担忧。这项研究首次揭示了污水处理厂中存在的移动 tet（X）变体，并强调了 epicPCR 引导培养策略的前景，该策略可从 "一体健康 "的角度探索不同生境中临床上重要的 ARGs 的细菌宿主。

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来源期刊

Water Research 环境科学-工程：环境

CiteScore

20.80

自引率

9.40%

发文量

1307

审稿时长

38 days

期刊介绍： Water Research, along with its open access companion journal Water Research X, serves as a platform for publishing original research papers covering various aspects of the science and technology related to the anthropogenic water cycle, water quality, and its management worldwide. The audience targeted by the journal comprises biologists, chemical engineers, chemists, civil engineers, environmental engineers, limnologists, and microbiologists. The scope of the journal include: •Treatment processes for water and wastewaters (municipal, agricultural, industrial, and on-site treatment), including resource recovery and residuals management; •Urban hydrology including sewer systems, stormwater management, and green infrastructure; •Drinking water treatment and distribution; •Potable and non-potable water reuse; •Sanitation, public health, and risk assessment; •Anaerobic digestion, solid and hazardous waste management, including source characterization and the effects and control of leachates and gaseous emissions; •Contaminants (chemical, microbial, anthropogenic particles such as nanoparticles or microplastics) and related water quality sensing, monitoring, fate, and assessment; •Anthropogenic impacts on inland, tidal, coastal and urban waters, focusing on surface and ground waters, and point and non-point sources of pollution; •Environmental restoration, linked to surface water, groundwater and groundwater remediation; •Analysis of the interfaces between sediments and water, and between water and atmosphere, focusing specifically on anthropogenic impacts; •Mathematical modelling, systems analysis, machine learning, and beneficial use of big data related to the anthropogenic water cycle; •Socio-economic, policy, and regulations studies.