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 blaCTX-M, blaOXA48, blaNDM, and blaKPC, 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.

中文翻译：

---

对社区废水中的多种传染病病原体和抗菌素耐药基因进行同步生物量浓缩和后续定量


基于废水的传染病病原体监测 (WBS) 越来越被视为人口健康数据的可靠来源。迄今为止，基于废水的监测工作主要集中在单个病原体上。然而，鉴于废水中含有多种在人群中传播的病原体，更全面的方法可以提高其可用性。我们重点关注同时检测 SARS-CoV-2、沙波病毒、空肠弯曲杆菌、大肠杆菌弯曲杆菌、沙门氏菌、致病性大肠杆菌、隐孢子虫属、贾第虫属。和临床相关的抗菌素耐药基因（ARG）。为了实现这一目标，优化了生物量浓度和核酸提取方法，并使用一套（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情况以及时空趋势，并收集了有关芬兰废水中 ARG 的新信息。此外，使用指示微生物crAssphage研究了不同废水浓缩物（即上清液和离心沉淀物的超滤浓缩物）的使用以及样品处理前冷冻和解冻废水的影响。与分析新鲜样品相比，废水冻融降低了 crAssphage 的基因拷贝数 (p < 0.001)。弯曲杆菌在所研究的四个夏季月份中的两个月份（检出率 30%）以及来自集约化畜牧业地区的废水中最为丰富。然而，40% 的样本中检测到了沙门氏菌，没有任何明显的季节性趋势，并且最大的废水处理厂记录了最高的基因拷贝数。 芬兰人体分离细菌中常见的 β-内酰胺酶抗性基因，即 blaCTX-M、blaOXA48、blaNDM 和 blaKPC，在废水中也经常检测到（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废水的可靠性，并证明了同时对多种病原体进行 WBS 的可能性。