Detection of Enteroviruses and SARS-CoV-2 in Tunisian Wastewater

Monitoring the circulation of enteric viruses in environmental wastewater is a valuable tool for preventing the emergence of waterborne and food-borne diseases in humans. The detection of viruses was performed in five Tunisian wastewater treatment plants, three located in the Grand Tunis City (WWTP 1, WWTP 2, WWTP 3) and two in the Sahel of Tunisia (WWTP 4, WWTP 4), known as very developed and crowded zones, to assess the effectiveness of three biological wastewater treatment procedures namely natural oxidizing lagoons, rotating biodisks procedure, activated sludge procedure, and one tertiary sewage treatment using UV-C₂₅₄ reactor for this enteric viruses' removal. Thus, 242 sewage samples were collected between June 2019 and May 2020 from different lines of wastewater treatment procedures implemented in the five wastewater treatment plants investigated. SARS-CoV-2 was analyzed using real-time multiplex reverse-transcription polymerase chain reaction (multiplex real-time RT-PCR) and enteroviruses using reverse-transcription polymerase chain reaction (RT-PCR). The enteroviruses detection showed 93% and 73% respective high frequencies only in the two WWTPs of the Grand Tunis (WWTP 1 and WWTP 2). SARS-CoV-2 was detected in 58% of the all wastewater samples collected from the five studied WWTPs with a respective dominance of N gene (47%), S gene (42%), RdRp gene (42%) and at last E gene (20%). These enteroviruses and SARS-CoV-2 detection were revealed in all steps of the wastewater treatment procedures, so poor virological quality is found at the exit of each biological and tertiary step of treatment investigated. For the first time in Tunisia, these results highlighted the enterovirus and SARS-CoV-2 detection with high rates, and the ineffectiveness of the biological and UV-C254 treatment implemented to remove these viruses. The preliminary results of SARS-CoV-2 circulation in Tunisian wastewater confirmed the wide positivity rate underlined by other works worldwide and allowed showing a move towards integrating wastewater as a way for this virus to spread in different areas and environments. So, this last result about SARS-CoV-2 circulation allowed us to caution about the strong probability of diffusion of this hazardous virus through water and sewage; despite its enveloped character and nature, as a labile and sensitive virus in these environments. Thus, establishing a national surveillance strategy is needed to improve the sanitary quality of treated wastewater and prevent public health problems related to these viruses in treated wastewater.