Detection and Characterization of Hepatitis E Virus Genotype 3 in Wastewater and Urban Surface Waters in Germany

Abstract

In highly populated areas, environmental surveillance of wastewater and surface waters is a key factor to control the circulation of viruses and risks for public health. Hepatitis E virus (HEV) genotype 3 is considered as an emerging pathogen in industrialized countries. Therefore, this study was carried out to determine the prevalence of HEV in environmental waters in urban and suburban regions in Germany. HEV was monitored in water samples using quantitative RT-PCR (RT-qPCR) and nested RT-PCR without or with virus concentration via polyethylene glycol precipitation or ultracentrifugation. By RT-qPCR, 84-100% of influent samples of wastewater treatment plants were positive for HEV RNA. Genotypes HEV-3c and 3f were identified in wastewater, with HEV-3c being the most prevalent genotype. These data correlate with subtypes identified earlier in patients from the same area. Comparison of wastewater influent and effluent samples revealed a reduction of HEV RNA of about 1 log 10 during passage through wastewater treatment plants. In addition, combined sewer overflows (CSOs) after heavy rainfalls were shown to release HEV RNA into surface waters. About 75% of urban river samples taken during these CSO events were positive for HEV RNA by RT-qPCR. In contrast, under normal weather conditions, only around 30% of river samples and 15% of samples from a bathing water located at an urban river were positive for HEV. Median concentrations of HEV RNA of all tested samples at this bathing water were below the limit of detection.

First molecular characterization of the hepatitis E virus in humans in Cameroon: Confirmation of the HEV outbreak in Touboro, North‐Cameroon

“…HEV genotype 3 has been detected in wastewater in several studies. 20 All these results show that due to the absence of routine screening of HEV in the health system, HEV infection remains globally unknown in the human population and when this infection becomes symptomatic, the diagnosis is directed toward other diseases as was the case with these patients suspected to having yellow fever virus in 2013. 9 The limit of our study is that the samples used for the amplification had not arrived at the laboratory under the optimal temperature conditions required, which would have influenced the results obtained with a possible degradation of RNA.…”

Occurrence of various viruses and recent evidence of SARS-CoV-2 in wastewater systems

“… Ultracentrifugation followed PEG Total of 184 (111 influent and 83 effluent) wastewater samples analysed, 93/111 (84%) influents and 26/83 (31%) effluents. Median 3× 10 3 (influent) and 1× 10 3 (effluent) ( Beyer et al, 2020 ) Mexico Poliovirus Viral nucleic acid obtained from concentrated wastewater samples. Poliovirus serotype detected by qRT-PCR.…”

“…HEV presents an incubation period ranging from 2 to 9 weeks and is clinically indistinguishable from hepatitis A. This virus has been detected in wastewater in France ( Miura et al, 2016 , Courault et al, 2017 , Kaas et al, 2019 ), China ( Li et al, 2017 ), Colombia ( Baez et al, 2017 ), Portugal ( Matos et al, 2018 ), Italy ( Di Profio et al, 2019 ), Germany ( Beyer et al, 2020 ), and some other industrialized countries ( Fenaux et al, 2019 ).…”

Occurrence of Antibiotics, Antibiotic Resistance Genes and Viral Genomes in Wastewater Effluents and Their Treatment by a Pilot Scale Nanofiltration Unit

“…Similar results were obtained in a Spanish study [ 35 ], which detected HEV genomes only in the influent samples of four WWTPs, with average concentrations of 10 3 genome copies·L −1 . Additionally, a study from de-Beyer et al revealed the presence of the HEV genomes in the effluent samples of several German WWTPs, with an average concentration of 10 3 genome copies·L −1 [ 36 ].…”