Dataset Information

Detection of SARS-CoV-2 RNA in wastewater, river water, and hospital wastewater of Nepal.

ABSTRACT: The applicability of wastewater-based epidemiology (WBE) has been extensively studied throughout the world with remarkable findings. This study reports the presence and reduction of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at two wastewater treatment plants (WWTPs) of Nepal, along with river water, hospital wastewater (HWW), and wastewater from sewer lines collected between July 2020 and February 2021. SARS-CoV-2 RNA was detected in 50%, 54%, 100%, and 100% of water samples from WWTPs, river hospitals, and sewer lines, respectively, by at least one of four quantitative PCR assays tested (CDC-N1, CDC-N2, NIID_2019-nCOV_N, and N_Sarbeco). The CDC-N2 assay detected SARS-CoV-2 RNA in the highest number of raw influent samples of both WWTPs. The highest concentration was observed for an influent sample of WWTP A (5.5 ± 1.0 log₁₀ genome copies/L) by the N_Sarbeco assay. SARS-CoV-2 was detected in 47% (16/34) of the total treated effluents of WWTPs, indicating that biological treatments installed at the tested WWTPs are not enough to eliminate SARS-CoV-2 RNA. One influent sample was positive for N501Y mutation using the mutation-specific qPCR, highlighting a need for further typing of water samples to detect Variants of Concern. Furthermore, crAssphage-normalized SARS-CoV-2 RNA concentrations in raw wastewater did not show any significant association with the number of new coronavirus disease 2019 (COVID-19) cases in the whole district where the WWTPs were located, suggesting a need for further studies focusing on suitability of viral as well as biochemical markers as a population normalizing factor. Detection of SARS-CoV-2 RNA before, after, and during the peaking in number of COVID-19 cases suggests that WBE is a useful tool for COVID-19 case estimation in developing countries.

SUBMITTER: Tandukar S

PROVIDER: S-EPMC8832950 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature

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Detection of SARS-CoV-2 RNA in wastewater, river water, and hospital wastewater of Nepal.

Tandukar Sarmila S Sthapit Niva N Thakali Ocean O Malla Bikash B Sherchan Samendra P SP Shakya Bijay Man BM Shrestha Laxman P LP Sherchand Jeevan B JB Joshi Dev Raj DR Lama Bhupendra B Haramoto Eiji E

The Science of the total environment 20220211

The applicability of wastewater-based epidemiology (WBE) has been extensively studied throughout the world with remarkable findings. This study reports the presence and reduction of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at two wastewater treatment plants (WWTPs) of Nepal, along with river water, hospital wastewater (HWW), and wastewater from sewer lines collected between July 2020 and February 2021. SARS-CoV-2 RNA was detected in 50%, 54%, 100%, and 100% of water samples f ...[more]

PMID: 35157870

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Project description:There are only a few established methods to determine the concentration of encapsulated viruses, such as SARS-CoV-2, in water matrices, limiting the application of wastewater-based epidemiology (WBE)—an important tool for public health research. The present study compared four methods that are commonly used to concentrate non-encapsulated enteric viruses for determining SARS-CoV-2 concentration in wastewater and wastewater-enriched river water samples. The four methods tested were electronegative membrane with Mg+2 addition, aluminum hydroxide-based precipitation, polyethylene glycol (PEG) 8000 precipitation, and ultrafiltration (with porosity of 10 and 50 kDa). Prior to the concentration step, filtration or centrifugation was performed to remove suspended particles from the samples (pretreatment). To evaluate the recovery efficiency (%), samples of SARS-CoV-2 from nasopharyngeal swabs obtained from RT-qPCR-positive patients were used as spiked samples. The second part of the analysis involved the quantification of the SARS-CoV-2 copy number in analytes without SARS-CoV-2-spiked samples. Among the tested methods, pretreatment via centrifugation followed by ultrafiltration with a 50-kDa cut-off was found the most efficient method for wastewater samples with spiked samples (54.3 or 113.01% efficiency). For the wastewater-enriched river samples with spiked samples, pretreatment via centrifugation followed by filtration using an electronegative membrane was the most efficient method (110.8% and 95.9% for N1 and N2 markers, respectively). However, ultrafiltration of the raw river water samples using 10 or 50 kDa cut-off filters and PEG 8000 precipitation showed the best concentration efficiency based on copy number, regardless of the pretreatment approach or sample type (values ranging from 3 × 105 to 6.7 × 103). The effectiveness of the concentration method can vary depending on the type of sample and concentration method. We consider that this study will contribute to more widespread use of WBE for the environmental surveillance of SARS-CoV-2. Graphical abstract Image 1

Project description: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 × 103 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 × 103-8.2 × 104 copies/L) with a lower filtration volume (200 mL) compared to that for secondary-treated wastewater (1.4 × 102-2.5 × 103 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.

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Project description:Wastewater-based epidemiology is a corroborated environmental surveillance tool in the global fight against SARS-CoV-2. The analysis of wastewater for detection of SARS-CoV-2 RNA may assist policymakers to survey a specific infectious community. Herein, we report on a long-term quantification study in Bahrain to investigate the incidence of the SARS-CoV-2 RNA in wastewater during the COVID-19 pandemic. The ∼260,000 population of Muharraq Island in Bahrain is served by a discrete sewerage catchment, and all wastewater flows to a single large Sewage Treatment Plant (STP) with a capacity of 100,000 m3/day. The catchment is predominately domestic, but also serves several hospitals and Bahrain's international airport. Flow-weighted 24-h composite wastewater samples for the period February 2020 to October 2021 were analyzed for the presence of SARS-CoV-2 N1, N2 and E genes. A Spearman rank correlation demonstrated a moderate correlation between the concentration of SARS-CoV-2 N1, N2 and E genes in the wastewater samples and the number of COVID-19 cases reported on the same day of the sampling. SARS-CoV-2 viral genes were detected in wastewater samples shortly after the first cases of COVID-19 were reported by the health authorities in Bahrain by reverse transcription-polymerase chain reaction (RT-qPCR). The viral genes were detected in 55 of 65 samples (84.62%) during the whole study period and the concentration range was found to be between 0 and 11,508 RNA copies/mL across the viral genes tested (in average N1: 518.4, N2: 366.8 and E: 649.3 copies/mL). Furthermore, wastewater samples from two COVID-19-dedicated quarantine facilities were analysed and detected higher SARS-CoV-2 gene concentrations (range 27-19,105 copies/mL; in average N1: 5044, N2: 4833 and E: 8663 copies/mL). Our results highlight the potential use of RT-qPCR for SARS-CoV-2 detection and quantification in wastewater and present the moderate correlation between concentration of SARS-CoV-2 genes with reported COVID-19 cases for a specified population. Indeed, this study identifies this technique as a mechanism for long term monitoring of SARS-CoV-2 infection levels and hence provides public health and policymakers with a useful environmental surveillance tool during and after the current pandemic.

Dataset Information

Detection of SARS-CoV-2 RNA in wastewater, river water, and hospital wastewater of Nepal.

Publications

Detection of SARS-CoV-2 RNA in wastewater, river water, and hospital wastewater of Nepal.

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