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Volume 13
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10.3390/microorganisms13102319
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Article

Rotavirus Quantification and Genotyping in Wastewater: A Molecular Surveillance Study in Italy (2024–2025)

by
Giusy Bonanno Ferraro
1,*,
Carolina Veneri
1,
Agata Franco
1,
David Brandtner
2,
Daniele Congiu
1,
Pamela Mancini
1,
Marcello Iaconelli
1,
Elisabetta Suffredini
3 and
Giuseppina La Rosa
1
1
National Center for Water Safety (CeNSiA), Istituto Superiore di Sanità, 00161 Rome, Italy
2
Department of Infectious Disease, Istituto Superiore di Sanità, 00161 Rome, Italy
3
Department of Food Safety, Nutrition and Veterinary Public Health, Istituto Superiore di Sanità, 00161 Rome, Italy
*
Author to whom correspondence should be addressed.
Microorganisms 2025, 13(10), 2319; https://doi.org/10.3390/microorganisms13102319
Submission received: 20 August 2025 / Revised: 29 September 2025 / Accepted: 3 October 2025 / Published: 7 October 2025
(This article belongs to the Special Issue Surveillance of Health-Relevant Pathogens Employing Wastewater)
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Abstract

Rotavirus remains a leading cause of acute gastroenteritis worldwide, particularly in young children, despite widespread vaccination efforts. This study aims to evaluate rotavirus circulation at the population level through wastewater-based epidemiology (WBE), offering a non-invasive, complementary approach to clinical surveillance. Between 2024 and 2025, a total of 172 composite 24 h samples were collected from eight urban wastewater treatment plants across Northern, Central, and Southern Italy. Viral RNA was concentrated by PEG precipitation and quantified using digital RT-PCR, while genotypes were determined via nested PCR targeting VP7 and VP4 genes. Rotavirus RNA was detected in 143 out of 172 samples (83.1%), with viral loads ranging between 4.2 × 102 to 7.3 × 105 genome copies per liter (g.c./L). Genotyping revealed G3 as the predominant VP7 type, followed by G1, G2, G6, and G9. All VP4-positive samples were classified as P8. This investigation expands current knowledge of rotavirus epidemiology in Italy using molecular surveillance of urban wastewater. By combining digital RT-PCR and genotyping, it offers a robust framework for integrating WBE into rotavirus monitoring programs, especially in settings where clinical surveillance is limited.
Keywords: rotavirus; wastewater-based epidemiology (WBE); quantification; genotyping; urban wastewater; digital RT-PCR rotavirus; wastewater-based epidemiology (WBE); quantification; genotyping; urban wastewater; digital RT-PCR

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MDPI and ACS Style

Bonanno Ferraro, G.; Veneri, C.; Franco, A.; Brandtner, D.; Congiu, D.; Mancini, P.; Iaconelli, M.; Suffredini, E.; La Rosa, G. Rotavirus Quantification and Genotyping in Wastewater: A Molecular Surveillance Study in Italy (2024–2025). Microorganisms 2025, 13, 2319. https://doi.org/10.3390/microorganisms13102319

AMA Style

Bonanno Ferraro G, Veneri C, Franco A, Brandtner D, Congiu D, Mancini P, Iaconelli M, Suffredini E, La Rosa G. Rotavirus Quantification and Genotyping in Wastewater: A Molecular Surveillance Study in Italy (2024–2025). Microorganisms. 2025; 13(10):2319. https://doi.org/10.3390/microorganisms13102319

Chicago/Turabian Style

Bonanno Ferraro, Giusy, Carolina Veneri, Agata Franco, David Brandtner, Daniele Congiu, Pamela Mancini, Marcello Iaconelli, Elisabetta Suffredini, and Giuseppina La Rosa. 2025. "Rotavirus Quantification and Genotyping in Wastewater: A Molecular Surveillance Study in Italy (2024–2025)" Microorganisms 13, no. 10: 2319. https://doi.org/10.3390/microorganisms13102319

APA Style

Bonanno Ferraro, G., Veneri, C., Franco, A., Brandtner, D., Congiu, D., Mancini, P., Iaconelli, M., Suffredini, E., & La Rosa, G. (2025). Rotavirus Quantification and Genotyping in Wastewater: A Molecular Surveillance Study in Italy (2024–2025). Microorganisms, 13(10), 2319. https://doi.org/10.3390/microorganisms13102319

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