First comparison of conventional activated sludge versus root-zone treatment for SARS-CoV-2 RNA removal from wastewaters: statistical and temporal significance

In the initial pandemic phase, effluents from wastewater treatment facilities were reported mostly free from Severe Acute Respiratory Coronavirus 2 (SARS-CoV-2) RNA, and thus conventional wastewater treatments were generally considered effective. However, there is a lack of first-hand data on i) comparative efficacy of various treatment processes for SARS-CoV-2 RNA removal; and ii) temporal variations in the removal efficacy of a given treatment process in the backdrop of active COVID-19 cases. This work provides a comparative account of the removal efficacy of conventional activated sludge (CAS) and root zone treatments (RZT) based on weekly wastewater surveillance data, consisting of forty-four samples, during a two-month period. The average genome concentration was higher in the inlets of CAS-based wastewater treatment plant in the Sargasan ward (1.25 x 103 copies/ L), than that of RZT plant (7.07 x 102 copies/ L) in an academic institution campus of Gandhinagar, Gujarat, India. ORF 1ab and S genes appeared to be more sensitive to treatment i.e., significantly reduced (p 0.05). CAS treatment exhibited better RNA removal efficacy (p=0.014) than RZT (p=0.032). Multivariate analyses suggested that the effective genome concentration should be calculated based on the presence/absence of multiple genes. The present study stresses that treated effluents are not always free from SARS-CoV-2 RNA, and the removal efficacy of a given WWTPs is prone to exhibit temporal variability owing to variations in active COVID-19 cases in the vicinity and genetic material accumulation over time. Disinfection seems less effective than the adsorption and coagulation processes for SARS-CoV-2 removal. Results stress the need for further research on mechanistic insight on SARS-CoV-2 removal through various treatment processes taking solid-liquid partitioning into account. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=97 SRC="FIGDIR/small/21256898v1_ufig1.gif" ALT="Figure 1"> View larger version (45K): org.highwire.dtl.DTLVardef@1cc204aorg.highwire.dtl.DTLVardef@5c17b7org.highwire.dtl.DTLVardef@192e23org.highwire.dtl.DTLVardef@124168d_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIWastewater treatments may not completely remove the SARS-CoV-2 RNA. C_LIO_LIThe activated sludge process exhibited better RNA removal efficacy than root-zone treatment. C_LIO_LIORF 1ab and S genes appeared more sensitive to treatment than N genes. C_LIO_LITemporal variability is observed in the removal efficacy of wastewater treatment plants. C_LI