Finnish Institute for Health and Welfare, Expert Microbiology Unit, Neulaniementie 4, Kuopio FI-70701, Finland; University of Helsinki, Faculty of Veterinary Medicine, Department of Food Hygiene and Environmental Health, Agnes Sjöbergin katu 2, Helsinki FI-00014, Finland.
Finnish Institute for Health and Welfare, Expert Microbiology Unit, Neulaniementie 4, Kuopio FI-70701, Finland.
Environ Int. 2024 Sep;191:108973. doi: 10.1016/j.envint.2024.108973. Epub 2024 Aug 20.
Wastewater-based surveillance (WBS) of infectious disease agents is increasingly seen as a reliable source of population health data. To date, wastewater-based surveillance efforts have largely focused on individual pathogens. However, given that wastewater contains a broad range of pathogens circulating in the population, a more comprehensive approach could enhance its usability. We focused on the simultaneous detection of SARS-CoV-2, sapovirus, Campylobacter jejuni, Campylobacter coli, Salmonella spp., pathogenic Escherichia coli, Cryptosporidium spp., Giardia spp. and antimicrobial resistance genes (ARGs) of clinical relevance. To achieve this goal, biomass concentration and nucleic acid extraction methods were optimized, and samples were analyzed by using a set of (RT)-qPCR and (HT)-qPCR methods. We determined the prevalence and the spatial and temporal trends of the targeted pathogens and collected novel information on ARGs in Finnish wastewater. In addition, the use of different wastewater concentrates, namely the ultrafiltered concentrate of the supernatant and the centrifuged pellet, and the effect of freezing and thawing wastewater prior to sample processing were investigated with the indicator microbe crAssphage. Freeze-thawing of wastewater decreased the gene copy count of crAssphage in comparison to analyzing fresh samples (p < 0.001). Campylobacters were most abundant in two of the four studied summer months (30 % detection rate) and in wastewaters from regions with intensive animal farming. Salmonella, however, was detected in 40 % of the samples without any clear seasonal trends, and the highest gene copy numbers were recorded from the largest wastewater treatment plants. Beta-lactamase resistance genes that have commonly been detected in bacteria isolated from humans in Finland, namely bla, bla, bla, and bla, were also frequently detected in wastewaters (100, 98, 98, and 70 % detection rates, respectively). These results confirm the reliability of using wastewater in public health surveillance and demonstrate the possibility to simultaneously perform WBS of multiple pathogens.
基于污水的传染病原体监测（WBS）正日益被视为获取人群健康数据的可靠来源。迄今为止，基于污水的监测工作主要集中在个别病原体上。然而，鉴于污水中包含人群中广泛传播的多种病原体，更全面的方法可以提高其可用性。我们专注于同时检测 SARS-CoV-2、诺如病毒、空肠弯曲菌、结肠弯曲菌、沙门氏菌、致病性大肠杆菌、隐孢子虫、贾第虫和具有临床相关性的抗生素耐药基因（ARGs）。为了实现这一目标，我们优化了生物量浓度和核酸提取方法，并使用一组（RT）-qPCR 和（HT）-qPCR 方法对样品进行了分析。我们确定了目标病原体的流行率以及时空趋势，并在芬兰污水中收集了有关 ARGs 的新信息。此外，我们还使用了不同的污水浓缩物，即上清液的超滤浓缩物和离心沉淀，并使用指示微生物 crAssphage 研究了在进行样品处理之前对污水进行冷冻和解冻的效果。与分析新鲜样品相比，污水的冷冻和解冻会降低 crAssphage 的基因拷贝数（p<0.001）。空肠弯曲菌和结肠弯曲菌在四个研究夏季月份中的两个月（30%的检出率）和动物养殖密集地区的污水中最为丰富。沙门氏菌在没有明显季节性趋势的情况下，在 40%的样本中被检出，且在最大的污水处理厂中记录到的基因拷贝数最高。芬兰从人类分离的细菌中常见的β-内酰胺酶耐药基因，即 bla、bla、bla 和 bla，也经常在污水中检出（检出率分别为 100%、98%、98%和 70%）。这些结果证实了在公共卫生监测中使用污水的可靠性，并展示了同时进行多种病原体的 WBS 的可能性。
