Tracking SARS-CoV-2 and its variants in wastewater in Tunisia

Author:

Othman Ines1ORCID,Bisseux Maxime23,Helmi Amna4,Hamdi Rawand1,Nahdi Imen5,Slama Ichrak1,Mastouri Maha16,Bailly Jean Luc2,Aouni Mahjoub1

Affiliation:

1. a Faculty of Pharmacy, University of Monastir, LR99-ES27, Monastir, Tunisia

2. b CHU Clermont-Ferrand, 3IHP, Virology Laboratory, National Reference Centre for Enteroviruses and Parechoviruses-Associated Laboratory, F-63003 Clermont-Ferrand, France

3. c Université Clermont Auvergne, CNRS 6023-LMGE, EPIE, F-63001 Clermont-Ferrand, France

4. d Directorate of Milieu Hygiene and Environmental Protection at the Health Ministry, Tunis, Tunisia

5. e African Biotechnology Society - ABS Advanced, Tunis, Tunisia

6. f Laboratory of Microbiology, Fattouma Bourguiba University Hospital, Monastir, Tunisia

Abstract

ABSTRACT Wastewater-based genomic surveillance can improve community prevalence estimates and identify emerging variants of pathogens. Wastewater influents and treated effluents from six wastewater treatment plants (WWTPs) in Tunisia were analyzed between December 2021 and July 2022. Wastewater samples were analyzed with reverse transcription solid digital PCR (RT-sdPCR) and whole-genome sequencing to determine the amount of SARS-CoV-2 RNA and assign SARS-CoV-2 lineages. The virus variants detected in wastewater samples were compared with COVID-19 prevalence data. The quantitative results in wastewater influents revealed that viral RNA concentrations at the treatment plants corroborate with locally reported clinical cases and show an increase before the increment of clinically diagnosed new COVID-19 cases between April and July 2022. Delta and Omicron variants were identified in the Tunisian wastewater. Interestingly, the presence of variant BA.5 was detected in samples prior to its inclusion as a variant of concern (VOC) by the Tunisian National Health Authorities. SARS-CoV-2 was detected in wastewater effluents, indicating that the wastewater treatment techniques used in the majority of Tunisian WWTPs are inefficient in removing the virus traces. This study reports the first identification of SARS-CoV-2 VOCs in Tunisian wastewater samples.

Funder

Swiss Agency for Development and Cooperation

United Nations Development Programme in Tunisia

Publisher

IWA Publishing

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