Human intestinal enteroids platform to assess the infectivity of gastroenteritis viruses in wastewater

Noelia Carmona-Vicente; Annamaria Pandiscia; Cristina Santiso-Bellón; Alba Perez-Cataluña; Jesús Rodríguez-Díaz; Veronica P Costantini; Javier Buesa; Jan Vinjé; Gloria Sánchez; Walter Randazzo

doi:10.1016/j.watres.2024.121481

Human intestinal enteroids platform to assess the infectivity of gastroenteritis viruses in wastewater

Water Res. 2024 May 15:255:121481. doi: 10.1016/j.watres.2024.121481. Epub 2024 Mar 18.

Affiliations

¹ Department of Microbiology and Ecology, University of Valencia, Valencia, Spain.
² Department of Preservation and Food Safety Technologies, IATA-CSIC, Valencia, Spain; Department of Veterinary Medicine, University of Bari, Valenzano, Italy.
³ Department of Preservation and Food Safety Technologies, IATA-CSIC, Valencia, Spain.
⁴ Department of Microbiology and Ecology, University of Valencia, Valencia, Spain; INCLIVA Health Research Institute, Valencia, Spain.
⁵ National Calicivirus Laboratory, Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
⁶ Department of Preservation and Food Safety Technologies, IATA-CSIC, Valencia, Spain. Electronic address: wrandazzo@iata.csic.es.

PMID: 38520776
DOI: 10.1016/j.watres.2024.121481

Abstract

Fecal-orally transmitted gastroenteritis viruses, particularly human noroviruses (HuNoVs), are a public health concern. Viral transmission risk through contaminated water results underexplored as they have remained largely unculturable until recently and the robust measuring of gastroenteritis viruses infectivity in a single cell line is challenging. This study primarily aimed to test the feasibility of the human intestinal enteroids (HIE) model to demonstrate the infectivity of multiple gastroenteritis viruses in wastewater. Initially, key factors affecting viral replication in HIE model were assessed, and results demonstrated that the reagent-assisted disruption of 3D HIE represents an efficient alternative to syringe pass-through, and the filtering of HuNoV stool suspensions could be avoided. Moreover, comparable replication yields of clinical strains of HuNoV genogroup I (GI), HuNoV GII, rotavirus (RV), astrovirus (HAstV), and adenoviruses (HAdV) were obtained in single and multiple co-infections. Then, the optimized HIE model was used to demonstrate the infectivity of multiple naturally occurring gastroenteritis viruses from wastewater. Thus, a total of 28 wastewater samples were subjected to (RT)-qPCR for each virus, with subsequent testing on HIE. Among these, 16 samples (57 %) showed replication of HuNoVs (n = 3), RV (n = 5), HAstV (n = 8), and/or HAdV (n = 5). Three samples showed HuNoV replication, and sequences assigned to HuNoV GI.3[P13] and HuNoV GII.4[P16] genotypes. Concurrent replication of multiple gastroenteritis viruses occurred in 4 wastewater samples. By comparing wastewater concentrate and HIE supernatant sequences, diverse HAstV and HAdV genotypes were identified in 4 samples. In summary, we successfully employed HIE to demonstrate the presence of multiple infectious human gastroenteritis viruses, including HuNoV, in naturally contaminated wastewater samples.

Keywords: Cell culture; Enteric virus; Environmental contamination; Human norovirus; Viral infectivity; Virus isolation.