Department of Microbiology and Ecology, University of Valencia, Av. Dr. Moliner, 50, Burjassot, 46100, Valencia, Spain; Department of Preservation and Food Safety Technologies, Institute of Agrochemistry and Food Technology, IATA-CSIC, Av. Agustín Escardino 7, Paterna, 46980, Valencia, Spain.
Research Group on Quality, Safety and Bioactivity of Plant Foods, Department of Food Science and Technology, CEBAS-CSIC, Campus Universitario de Espinardo, 25, 30100, Murcia, Spain.
Water Res. 2020 Aug 15;181:115942. doi: 10.1016/j.watres.2020.115942. Epub 2020 May 16.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused more than 200,000 reported COVID-19 cases in Spain resulting in more than 20,800 deaths as of April 21, 2020. Faecal shedding of SARS-CoV-2 RNA from COVID-19 patients has extensively been reported. Therefore, we investigated the occurrence of SARS-CoV-2 RNA in six wastewater treatments plants (WWTPs) serving the major municipalities within the Region of Murcia (Spain), the area with the lowest COVID-19 prevalence within Iberian Peninsula. Firstly, an aluminum hydroxide adsorption-precipitation concentration method was validated using a porcine coronavirus (Porcine Epidemic Diarrhea Virus, PEDV) and mengovirus (MgV). The procedure resulted in average recoveries of 10 ± 3.5% and 10 ± 2.1% in influent water (n = 2) and 3.3 ± 1.6% and 6.2 ± 1.0% in effluent water (n = 2) samples for PEDV and MgV, respectively. Then, the method was used to monitor the occurrence of SARS-CoV-2 from March 12 to April 14, 2020 in influent, secondary and tertiary effluent water samples. By using the real-time RT-PCR (RT-qPCR) Diagnostic Panel validated by US CDC that targets three regions of the virus nucleocapsid (N) gene, we estimated quantification of SARS-CoV-2 RNA titers in untreated wastewater samples of 5.4 ± 0.2 log genomic copies/L on average. Two secondary water samples resulted positive (2 out of 18) and all tertiary water samples tested as negative (0 out 12). This environmental surveillance data were compared to declared COVID-19 cases at municipality level, revealing that members of the community were shedding SARS-CoV-2 RNA in their stool even before the first cases were reported by local or national authorities in many of the cities where wastewaters have been sampled. The detection of SARS-CoV-2 in wastewater in early stages of the spread of COVID-19 highlights the relevance of this strategy as an early indicator of the infection within a specific population. At this point, this environmental surveillance could be implemented by municipalities right away as a tool, designed to help authorities to coordinate the exit strategy to gradually lift its coronavirus lockdown.
截至 2020 年 4 月 21 日,严重急性呼吸系统综合征冠状病毒 2(SARS-CoV-2)已在西班牙造成超过 20 万例 COVID-19 病例,导致超过 20800 人死亡。有大量报道称 COVID-19 患者的粪便中存在 SARS-CoV-2 RNA。因此,我们调查了西班牙穆尔西亚地区(西班牙)主要城市的六个废水处理厂(WWTP)中 SARS-CoV-2 RNA 的发生情况,该地区是伊比利亚半岛 COVID-19 发病率最低的地区。首先,我们使用猪冠状病毒(猪流行性腹泻病毒,PEDV)和髓灰质炎病毒(MgV)验证了铝氢氧化物吸附沉淀浓缩法。该程序导致进水(n=2)和出水(n=2)样品中 PEDV 和 MgV 的平均回收率分别为 10±3.5%和 10±2.1%,以及 3.3±1.6%和 6.2±1.0%。然后,该方法用于监测 2020 年 3 月 12 日至 4 月 14 日期间 SARS-CoV-2 在进水、二级和三级出水样品中的发生情况。使用美国疾病预防控制中心(US CDC)验证的实时 RT-PCR(RT-qPCR)诊断试剂盒,该试剂盒针对病毒核衣壳(N)基因的三个区域,我们估计未经处理的废水样品中 SARS-CoV-2 RNA 滴度的平均为 5.4±0.2 log 基因组拷贝/L。有两个二级水样呈阳性(18 个中的 2 个),所有三级水样均呈阴性(12 个中的 0 个)。这些环境监测数据与市一级报告的 COVID-19 病例进行了比较,结果表明,在许多采集废水的城市中,当地或国家当局报告首例病例之前,社区成员就在粪便中排出了 SARS-CoV-2 RNA。在 COVID-19 传播的早期阶段在废水中检测到 SARS-CoV-2,突出了这种策略作为特定人群感染的早期指标的重要性。在这一点上,各市政府可以立即实施这种环境监测,作为一种工具,旨在帮助当局协调逐步解除冠状病毒封锁的退出策略。