tetano
Editor, Senior Moderator
Sci Total Environ
. 2020 Oct 1;737:140405.
doi: 10.1016/j.scitotenv.2020.140405. Epub 2020 Jun 20.
First environmental surveillance for the presence of SARS-CoV-2 RNA in wastewater and river water in Japan
Eiji Haramoto[SUP] 1 [/SUP], Bikash Malla[SUP] 2 [/SUP], Ocean Thakali[SUP] 3 [/SUP], Masaaki Kitajima[SUP] 4 [/SUP]
Affiliations
Abstract
Wastewater-based epidemiology is a powerful tool to understand the actual incidence of coronavirus disease 2019 (COVID-19) in a community because severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the etiological agent of COVID-19, can be shed in the feces of infected individuals regardless of their symptoms. The present study aimed to assess the presence of SARS-CoV-2 RNA in wastewater and river water in Yamanashi Prefecture, Japan, using four quantitative and two nested PCR assays. Influent and secondary-treated (before chlorination) wastewater samples and river water samples were collected five times from a wastewater treatment plant and three times from a river, respectively, between March 17 and May 7, 2020. The wastewater and river water samples (200-5000 mL) were processed by using two different methods: the electronegative membrane-vortex (EMV) method and the membrane adsorption-direct RNA extraction method. Based on the observed concentrations of indigenous pepper mild mottle virus RNA, the EMV method was found superior to the membrane adsorption-direct RNA extraction method. SARS-CoV-2 RNA was successfully detected in one of five secondary-treated wastewater samples with a concentration of 2.4 ? 10[SUP]3[/SUP] copies/L by N_Sarbeco qPCR assay following the EMV method, with sequence confirmation of the qPCR product, whereas all the influent samples were tested negative for SARS-CoV-2 RNA. This result could be attributed to higher limit of detection for influent (4.0 ? 10[SUP]3[/SUP]-8.2 ? 10[SUP]4[/SUP] copies/L) with a lower filtration volume (200 mL) compared to that for secondary-treated wastewater (1.4 ? 10[SUP]2[/SUP]-2.5 ? 10[SUP]3[/SUP] copies/L) with a higher filtration volume of 5000 mL. None of the river water samples tested positive for SARS-CoV-2 RNA. Comparison with the reported COVID-19 cases in Yamanashi Prefecture showed that SARS-CoV-2 RNA was detected in the secondary-treated wastewater sample when the cases peaked in the community. This is the first study reporting the detection of SARS-CoV-2 RNA in wastewater in Japan.
Keywords: COVID-19; River water; SARS-CoV-2; Wastewater; Wastewater-based epidemiology.
. 2020 Oct 1;737:140405.
doi: 10.1016/j.scitotenv.2020.140405. Epub 2020 Jun 20.
First environmental surveillance for the presence of SARS-CoV-2 RNA in wastewater and river water in Japan
Eiji Haramoto[SUP] 1 [/SUP], Bikash Malla[SUP] 2 [/SUP], Ocean Thakali[SUP] 3 [/SUP], Masaaki Kitajima[SUP] 4 [/SUP]
Affiliations
- PMID: 32783878
- DOI: 10.1016/j.scitotenv.2020.140405
Abstract
Wastewater-based epidemiology is a powerful tool to understand the actual incidence of coronavirus disease 2019 (COVID-19) in a community because severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the etiological agent of COVID-19, can be shed in the feces of infected individuals regardless of their symptoms. The present study aimed to assess the presence of SARS-CoV-2 RNA in wastewater and river water in Yamanashi Prefecture, Japan, using four quantitative and two nested PCR assays. Influent and secondary-treated (before chlorination) wastewater samples and river water samples were collected five times from a wastewater treatment plant and three times from a river, respectively, between March 17 and May 7, 2020. The wastewater and river water samples (200-5000 mL) were processed by using two different methods: the electronegative membrane-vortex (EMV) method and the membrane adsorption-direct RNA extraction method. Based on the observed concentrations of indigenous pepper mild mottle virus RNA, the EMV method was found superior to the membrane adsorption-direct RNA extraction method. SARS-CoV-2 RNA was successfully detected in one of five secondary-treated wastewater samples with a concentration of 2.4 ? 10[SUP]3[/SUP] copies/L by N_Sarbeco qPCR assay following the EMV method, with sequence confirmation of the qPCR product, whereas all the influent samples were tested negative for SARS-CoV-2 RNA. This result could be attributed to higher limit of detection for influent (4.0 ? 10[SUP]3[/SUP]-8.2 ? 10[SUP]4[/SUP] copies/L) with a lower filtration volume (200 mL) compared to that for secondary-treated wastewater (1.4 ? 10[SUP]2[/SUP]-2.5 ? 10[SUP]3[/SUP] copies/L) with a higher filtration volume of 5000 mL. None of the river water samples tested positive for SARS-CoV-2 RNA. Comparison with the reported COVID-19 cases in Yamanashi Prefecture showed that SARS-CoV-2 RNA was detected in the secondary-treated wastewater sample when the cases peaked in the community. This is the first study reporting the detection of SARS-CoV-2 RNA in wastewater in Japan.
Keywords: COVID-19; River water; SARS-CoV-2; Wastewater; Wastewater-based epidemiology.