Hennebique Aurélie, Monge-Ruiz Jostin, Roger-Margueritat Morgane, Morand Patrice, Terreaux-Masson Claire, Maurin Max, Mercier Corinne, Landelle Caroline, Buelow Elena
Univ. Grenoble Alpes, CNRS, UMR 5525, VetAgro Sup, Grenoble INP, TIMC 38000, Grenoble, France; Bacteriology Laboratory, Institute of Biology and Pathology, Grenoble Alpes University Hospital, Grenoble, France.
Univ. Grenoble Alpes, CNRS, UMR 5525, VetAgro Sup, Grenoble INP, TIMC 38000, Grenoble, France.
Water Res. 2025 Jun 1;284:123902. doi: 10.1016/j.watres.2025.123902.
In hospitals, the transmission of antibiotic-resistant bacteria (ARB) may occur via biofilms present in sink drains, which can lead to infections. Despite the potential role of sink drains in the transmission of ARB in nosocomial infections, routine surveillance of these drains is lacking in most hospitals. As a result, there is currently no comprehensive understanding of the transmission of ARB and the dissemination of antimicrobial resistance genes (ARGs) and associated mobile genetic elements (MGEs) via sink drains. This study employed a multifaceted approach to monitor the total aerobic bacteria as well as the presence of carbapenemase-producing Enterobacterales (CPEs), the microbiota and the resistome of sink drain biofilms (SDBs) and hospital wastewater (WW) of two separate intensive care units (ICUs) in the same healthcare facility in France. Samples of SDB and WW were collected on a monthly basis, from January to April 2023, in the neonatal (NICU) and the adult (AICU) ICUs of Grenoble Alpes University Hospital. In the NICU, sink drain disinfection with surfactants was performed routinely. In the AICU, routine disinfection is not carried out. Culturable aerobic bacteria were quantified on non-selective media, and CPEs were screened using two selective agars. Isolates were identified by MALDI-TOF MS, and antibiotic susceptibility testing (AST) was performed on Enterobacterales and P. aeruginosa. The resistome was analyzed by high-throughput qPCR targeting >80 ARGs and MGEs. The overall bacterial microbiota was assessed via full-length 16S rRNA sequencing. No CPEs were isolated from SDBs in either ICU by bacterial culture. Culture-independent approaches revealed an overall distinct microbiota composition of the SDBs in the two ICUs. The AICU SDBs were dominated by pathogens containing Gram-negative bacterial genera including Pseudomonas, Stenotrophomona, Klebsiella, and Gram-positive Staphylococcus, while the NICU SDBs were dominated by the Gram-negative genera Achromobacter, Serratia, and Acidovorax, as well as the Gram-positive genera Weisella and Lactiplantibacillus. In contrast, the resistome of the SDBs exhibited no significant differences between the two ICUs, indicating that the abundance of ARGs and MGEs is independent of microbiota composition and disinfection practices. The AICU WW exhibited more distinct aerobic bacteria than the NICU WW. In addition, the AICU WW yielded 15 CPEs, whereas the NICU WW yielded a single CPE. All the CPEs were characterized at the species level. The microbiota of the NICU and AICU WW samples differed from their respective SDBs and exhibited distinct variations over the four-month period:the AICU WW contained a greater number of genes conferring resistance to quinolones and integron integrase genes, whereas the NICU WW exhibited a higher abundance of streptogramin resistance genes. Our study demonstrated that the resistome of the hospital SDBs in the two ICUs of the investigated healthcare institute is independent of the microbiota, the environment, and the local disinfection measures. However, the prevalence of CPEs in the WW pipes collecting the waste from the investigated drains differed. These findings offer valuable insights into the resilience of resistance genes in SDBs in ICUs, underscoring the necessity for innovative strategies to combat antimicrobial resistance in clinical environments.
在医院中,耐抗生素细菌(ARB)可能通过水槽排水管道中存在的生物膜传播,这可能导致感染。尽管水槽排水管道在医院感染中ARB传播方面具有潜在作用,但大多数医院缺乏对这些排水管道的常规监测。因此,目前对于ARB通过水槽排水管道的传播以及抗菌药物耐药基因(ARGs)和相关移动遗传元件(MGEs)的传播情况尚无全面了解。本研究采用多方面方法,对法国同一医疗机构中两个独立重症监护病房(ICUs)的水槽排水生物膜(SDBs)和医院废水(WW)中的总需氧菌、产碳青霉烯酶肠杆菌科细菌(CPEs)、微生物群和耐药组进行监测。2023年1月至4月,每月从格勒诺布尔阿尔卑斯大学医院的新生儿重症监护病房(NICU)和成人重症监护病房(AICU)采集SDB和WW样本。在NICU,常规使用表面活性剂进行水槽排水管道消毒。在AICU,不进行常规消毒。在非选择性培养基上对可培养需氧菌进行定量,使用两种选择性琼脂筛选CPEs。通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)鉴定分离株,并对肠杆菌科细菌和铜绿假单胞菌进行抗生素敏感性测试(AST)。通过针对80多种ARGs和MGEs的高通量定量聚合酶链反应(qPCR)分析耐药组。通过全长16S rRNA测序评估整体细菌微生物群。通过细菌培养,在两个ICU的SDBs中均未分离出CPEs。非培养方法揭示了两个ICU中SDBs的微生物群组成总体上存在差异。AICU的SDBs以含有革兰氏阴性菌属(包括假单胞菌属、嗜麦芽窄食单胞菌属、克雷伯菌属)和革兰氏阳性葡萄球菌属的病原体为主,而NICU的SDBs以革兰氏阴性菌属无色杆菌属、沙雷氏菌属和嗜酸菌属以及革兰氏阳性菌属魏斯氏菌属和植物乳杆菌属为主。相比之下,两个ICU中SDBs的耐药组没有显著差异,表明ARGs和MGEs的丰度与微生物群组成和消毒措施无关。AICU的WW中需氧菌比NICU的WW更具差异性。此外,AICU的WW中分离出15株CPEs,而NICU的WW中仅分离出1株CPE。所有CPEs均在种水平上进行了鉴定。NICU和AICU的WW样本中的微生物群与其各自的SDBs不同,并且在四个月期间呈现出明显的变化:AICU的WW中含有更多赋予对喹诺酮类耐药性的基因和整合子整合酶基因,而NICU的WW中链阳菌素耐药基因的丰度更高。我们的研究表明,所调查医疗机构的两个ICU中医院SDBs的耐药组与微生物群、环境和当地消毒措施无关。然而,从所调查排水管道收集废水的WW管道中CPEs的流行情况有所不同。这些发现为ICU中SDBs中耐药基因的持久性提供了有价值的见解,强调了在临床环境中对抗抗菌药物耐药性的创新策略的必要性。