Successful Application of Wastewater-Based Epidemiology in Prediction and Monitoring of the Second Wave of COVID-19 in India with Fragmented Sewerage Systems- A Case Study of Jaipur (India) (preprint)

The present study tracked the city-wide dynamics of severe acute respiratory syndrome-corona virus 2 (SARS-CoV-2) RNA in the wastewater from nine different wastewater treatment plants (WWTPs) in Jaipur during second wave of COVID-19 out-break in India. A total of 164 samples were collected weekly between February 19th and June 8th, 2021. SARS-CoV-2 was detected in 47.2% (52/110) influent samples and 37% (20/54) effluent samples. The increasing percentage of positive influent samples correlated with the citys increasing active clinical cases during the second wave of COVID-19 in Jaipur. Furthermore, WBE based evidence clearly showed early detection of about 20 days (9/9 samples reported positive on April 20th, 2021) prior to the maximum cases & maximum deaths reported in the city on May 8th, 2021. The present study further observed the presence of SARS-CoV-2 RNA in treated effluents at the time window of maximum active cases in the city even after tertiary disinfection treatments of UV & Chlorine. The average genome concentration in the effluents and removal efficacy of six commonly used treatments; Activated Sludge Treatment + Chlorine disinfection (ASP + Cl2), Moving Bed Biofilm Reactor (MBBR) with Ultraviolet radiations disinfection (MBBR + UV), MBBR + Chlorine (Cl2), Sequencing Batch Reactor (SBR) and SBR + Cl2 were compared with removal efficacy of SBR + Cl2 (81.2%)> MBBR + UV (68.8%) > SBR (57.1%) > ASP (50%) > MBBR + Cl2(36.4%). The study observed the trends & prevalence of four genes (E, RdRp, N, and ORF1ab gene) based on two different kits and found that prevalence of N> ORF1ab >RdRp> E gene, suggested that the effective genome concentration should be calculated based on the presence/absence of multiple genes. Hence, it is imperative to say that using a combination of different detection genes (E, N, RdRp & ORF1ab genes) reduce false positives in WBE. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=131 SRC="FIGDIR/small/21263417v1_ufig1.gif" ALT="Figure 1"> View larger version (40K): org.highwire.dtl.DTLVardef@156126corg.highwire.dtl.DTLVardef@1d39404org.highwire.dtl.DTLVardef@19a9dcdorg.highwire.dtl.DTLVardef@1ee3a0c_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LISuccessful application of WBE with prediction of 14-20 days for COVID-19 in Jaipur C_LIO_LIA comparison of SARS-CoV-2 RNA removal efficacy of 9 WWTPs was investigated C_LIO_LISBR showed better performance than MBBR with SARS-CoV-2 RNA removal from wastewater C_LIO_LIPresence of SARS-CoV-2 in effluents even after UV and Chlorine disinfection C_LIO_LIUsing a combination of different detection genes reduce false positives in WBE C_LI

Detection of SARS-CoV-2 RNA in fourteen wastewater treatment systems in Uttarakhand and Rajasthan States of North India (preprint)

We investigated the presence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) RNA at different treatment stages of 15 wastewater treatment plants (WWTPs) in two North Indian states of Rajasthan and Uttarakhand. Untreated (influent), biologically treated, and disinfected wastewater samples were collected from May to August 2020. The qualitative analysis of the wastewater for the presence of SARS-CoV-2 RNA was done using different pre-processing methods. SARS-CoV-2 RNA was detected in 11 out of 39 wastewater samples in Jaipur district and 5 out of 17 wastewater samples in Haridwar District using Reverse-Transcriptase Quantitative Polymerase Chain Reaction (RT-qPCR) for qualitative detection. None of the 56 samples tested for post-secondary or tertiary treatment were found positive for SARS-CoV-2 RNA. The findings indicate that there are no SARS-CoV-2 related risks involved with using the treated effluent for non-potable applications. In contrast, untreated wastewater may be a potential route of viral transmission to the WWTP and sanitation workers. Future studies are imperative to understand the survival rates of these viruses in wastewater.

Sewage surveillance for the presence of SARS-CoV-2 genome as a useful wastewater based epidemiology (WBE) tracking tool in India (preprint)

The infection with SARS-CoV-2 is reported to be accompanied by the shedding of the virus in stool samples of infected patients. Earlier reports have suggested that COVID-19 agents can be present in the fecal and sewage samples and thus it can be a good indication of the pandemic extent in a community. However, no such studies have been reported in the Indian context so far. Since, several factors like local population physiology, the climatic conditions, sewage composition, and processing of samples could possibly affect the detection of the viral genome, it becomes absolutely necessary to check for the presence of the SARS-CoV-2 in the wastewater samples from wastewater treatment plants (WWTPs) serving different localities of Jaipur city, which has been under red zone (pandemic hotspots) since early April 2020. Samples from different local municipal WWTPs and hospital wastewater samples were collected and wastewater based epidemiology (WBE) studies for the presence of SARS-CoV-2 were carried out using the RT-PCR technique to confirm the presence of different COVID-19 target genes namely S gene, E gene, ORF1ab gene, RdRp gene and N gene in the viral load of wastewater samples. In the present study, the untreated wastewater samples from the municipal WWTPs and hospital wastewater samples showed the presence of SARS-CoV-2 viral genome, which was correlated with the increased number of COVID-19 positive patients from the concerned areas, as per reported in the publically available health data. This is the first study that investigated the presence of SARS-CoV-2 viral genome in wastewater, at higher ambient temperature (above 40{degrees}C), further validating WBE as a potential tool in predicting and mitigating outbreaks.