Imprints of Lockdown and Treatment Processes on the Wastewater Surveillance of SARS-CoV-2: A Curious Case of Fourteen Plants in Northern India

Abstract

The present study investigated the detection of severe acute respiratory syndrome–coronavirus 2 (SARS-CoV-2) genomes at each treatment stage of 14 aerobic wastewater treatment plants (WWTPs) serving the major municipalities in two states of Rajasthan and Uttarakhand in Northern India. The untreated, primary, secondary and tertiary treated wastewater samples were collected over a time frame ranging from under-lockdown to post-lockdown conditions. The results showed that SARS-CoV-2 RNA was detected in 13 out of 40 wastewater samples in Jaipur district, Rajasthan and in 5 out of 14 wastewater samples in the Haridwar District, Uttarakhand with the E gene predominantly observed as compared to the N and RdRp target genes in later time-points of sampling. The Ct values of genes present in wastewater samples were correlated with the incidence of patient and community cases of COVID-19. This study further indicates that the viral RNA could be detected after the primary treatment but was not present in secondary or tertiary treated samples. This study implies that aerobic biological wastewater treatment systems such as moving bed biofilm reactor (MBBR) technology and sequencing batch reactor (SBR) are effective in virus removal from the wastewater. This work might present a new indication that there is little to no risk in relation to SARS-CoV-2 while reusing the treated wastewater for non-potable applications. In contrast, untreated wastewater might present a potential route of viral transmission through WWTPs to sanitation workers and the public. However, there is a need to investigate the survival and infection rates of SARS-CoV-2 in wastewater.

Successful application of wastewater-based epidemiology in prediction and monitoring of the second wave of COVID-19 with fragmented sewerage systems–a case study of Jaipur (India)

“…Concerned personnel wore standard personal protective equipment (PPE) during the entire sampling process. The collected samples were transported to the laboratory at ambient temperatures of the city during the collection months, as adopted by (Arora et al, 2020 , 2021 ). Figure 1 shows the geographical locations of the sampling sites used in the study.…”

“…Figure 2 describes the methodology involved during the study. The samples for RNA isolation were prepared with slight modifications from the protocol described previously (Arora et al, 2020 , 2021 ). The wastewater samples were surface sterilized using UV treatment for 30 min followed by manual mixing.…”

“…The wastewater samples were surface sterilized using UV treatment for 30 min followed by manual mixing. Furthermore, 1-mL sample was aliquoted and centrifuged at 7000 rpm for 30 min (for removal of debris & unwanted materials) and the supernatant was then processed for RNA extraction as described in (Arora et al, 2021 ).

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“…There are very few case studies that have been reported from India, across the nation, including, i.e., Uttarakhand and Rajasthan from Northern India (Arora et al, 2020 , 2021 ), Hyderabad (Hemalatha et al, 2021 ; Kopperi et al, 2021 ), and Chennai from Southern India (Chakraborty et al, 2021 ); and Gujarat (Kumar et al, 2020 , 2021a , b , c ) and Maharashtra from Central India (Sharma et al, 2021 ); most of which have successfully demonstrated the usefulness of WBE but on a limited scale. The awareness of WBE has been increasing during the COVID-19 pandemic; however, WBE is still not an established practice in low- and middle-income countries (LMICs) such as India.…”

Monitoring of SARS-CoV-2 Variants by Wastewater-Based Surveillance as a Sustainable and Pragmatic Approach—A Case Study of Jaipur (India)

“…The samples for RNA isolation were produced with minor changes to the methodology previously reported [25,26]. The wastewater samples were exposed to UV light for 30 min to sterilize the surface before being thoroughly mixed.…”