Noroviruses (NoVs) are important human pathogens associated with foodborne and waterborne gastroenteritis. These viruses are genetically highly heterogeneous, with more than forty genotypes within three genogroups (GI, GII, and GIV) identified in humans. However, the vast majority of human infections are associated with variants of a unique genotype, GII.4. Aside from these NoV strains of epidemiological relevance, NoV strains of genogroup GIV (Alphatron-like) are reported in a sporadic fashion and their overall prevalence in the community is unknown and this likely reflects the lack of specific diagnostic tools. We analyzed raw sewages collected from 32 wastewater treatment plants distributed throughout Italy (307 samples) and stool specimens collected from hospitalized patients with clinical signs of diarrhea of unknown etiology (285 samples). By using specific qualitative and quantitative RT-PCR assays, 21.8 % of the sewage samples and 3.2 % of the stool specimens tested positive for GIV NoVs. The number of genome copies in fecal samples ranged from 5.08 × 104 to 1.73× 106/g of feces. Sequence analysis showed limited genetic variability in human GIV viruses. The presence of GIV NoV both in sewage and in clinical samples confirms that not only GI and GII NoVs but also GIV strains are circulating in humans. Monitoring of GIV NoV is recommended in order to understand the dynamics of circulation in human populations, environmental contamination, and potential health risks.
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This study was financed by the joint Italian–American Project “Assessing the Impact of GIV Norovirus on Human Health: a Molecular Epidemiological Investigation on Environmental and Clinical Samples as a Basis for the Design of Novel Diagnostic Tools for an Emerging Pathogen,” and partially by the Grant “Calicivirus nei carnivori e nell’uomo: caratterizzazione molecolare, epidemiologia, implicazioni zoonosiche”—PRIN 2008. We thank Professor Herbert W. Virgin, Washington University (St. Louis, Missouri, United States) for providing the murine NoV stain used as sample process control. We gratefully acknowledge for wastewater sample collection: 1–2. E. Lorenzi, L. Meucci, M. Deceglie (SMAT Spa, Torino), E. Garrou, M. Morello, G. Mantovani (ARPA Piemonte, Torino); 3–4. G. Manassero (Arpa Valle d’Aosta), A. Martello (Corpo Forestale Valdostano); 5. W. Bodini, C. Amadasi (Vettabbia Spa, Milano); 6. L. Boscolo (Amiacque Spa, Milano); 7. M. Tomasoni, D. Monteverdi (A2A Spa, Brescia); 8–9. M. Poli, M. Dekas (Eco Center Spa AG, Bolzano), W. Strobl, E. Scarperi (APPA, Bolzano); 10–11. L. Bruni, G. Gatti, L.Tomasi, G. Cimadon (APPA,Trento); 12–13. P. Parati, E. Dell’Andrea, G. Gambillara (Arpa Veneto, Venezia); 14–15. S. Gaiter, L. Sola (ARPA Liguria, Genova); 16. F.Cornia, M.A. Corvaglia, P. Albertelli (ARPA Emilia Romagna, Bologna); 17. A. Gambaccioni, M. Razzolini (Publiacqua S.p.A. Firenze); 18. E. Renna, G. Saltalamacchia, M. Lucarini (ARPA Umbria, Perugia); 19–20. C. Mengarelli, Trimboli (ARPA Marche, Ancona); 21–24. C. Gala, R. Tomassini (Arpalazio, Roma), G. Ranalletta (ACEA ATO2 S.p.A., Roma); 25. E. Rufolo, R. Martino (ARPA Campania, Sezione provinciale di Napoli); 26–27. G. Assennato, G. Blonda, (Regione Puglia, Direzione Scientifica), V. Perrino, M. Mariani (ARPA Puglia, Bari); 28. R. Vita, R. Masotti, R. Martoccia (ARPA Basilicata, Potenza); 29. F. Pedulla, G. Belmusto (ARPA Calabria, Reggio Calabria); 30. L. Librici G. Abbate (ARPA Sicilia, Palermo); 31–32. M. G. Mulas, G. Campus (Regione Sardegna), A.M. Mereu, M. Secci (ARPA Sardegna, Cagliari).
Conflict of interest
There is no conflict of interest for all the authors. All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the 1975 Helsinki Declaration, as revised in 2008. Informed consent was obtained from all patients included in the study.
M. Muscillo and M. Fratini have contributed equally to this study.
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Muscillo, M., Fratini, M., Graffeo, R. et al. GIV Noroviruses in Wastewaters and in Stool Specimens from Hospitalized Patients. Food Environ Virol 5, 194–202 (2013). https://doi.org/10.1007/s12560-013-9121-5
- Norovirus genogroup IV
- Nested PCR
- Real-time PCR
- Stool samples