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Frequent Detection and Genetic Diversity of Human Bocavirus in Urban Sewage Samples

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Abstract

The prevalence and genetic diversity of human bocaviruses (HBoVs) in sewage water samples are largely unknown. In this study, 134 raw sewage samples from 25 wastewater treatment plants (WTPs) in Italy were analyzed by nested PCR and sequencing using species-specific primer pairs and broad-range primer pairs targeting the capsid proteins VP1/VP2. A large number of samples (106, 79.1 %) were positive for HBoV. Out of these, 49 were classified as HBoV species 2, and 27 as species 3. For the remaining 30 samples, sequencing results showed mixed electropherograms. By cloning PCR amplicons and sequencing, we confirmed the copresence of species 2 and 3 in 29 samples and species 2 and 4 in only one sample. A real-time PCR assay was also performed, using a newly designed TaqMan assay, for quantification of HBoVs in sewage water samples. Viral load quantification ranged from 5.51E+03 to 1.84E+05 GC/L (mean value 4.70E+04 GC/L) for bocavirus 2 and from 1.89E+03 to 1.02E+05 GC/L (mean value 2.27E+04 GC/L) for bocavirus 3. The wide distribution of HBoV in sewages suggests that this virus is common in the population, and the most prevalent are the species 2 and 3. HBoV-4 was also found, representing the first detection of this species in Italy. Although there is no indication of waterborne transmission for HBoV, the significant presence in sewage waters suggests that HBoV may spread to other water environments, and therefore, a potential role of water in the HBoV transmission should not be neglected.

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Acknowledgments

We thank Professor Herbert W. Virgin, Washington University (St. Louis, Missouri, United States) for providing the murine norovirus strain used as sample process control.

We thank the assistance of the following members of the Italian Wastewater Network for wastewater sample collection: E. Lorenzi, M. De Ceglia (SMAT Spa, Castiglione T.se, Torino), L. Meucci, D. Giacosa, A. Poncino (Centro Ricerche SMAT, Torino); N. Mesiano (SMAT, Collegno); W. Bodini, C. Amadasi (Vettabbia Spa, Milano); F. Montanelli, L. Boscolo, P. Viola, F. Pizza (Amiacque Spa, Milano); M. Tomasoni, D. Monteverdi, L. Massafra (A2A Spa, Brescia); Werner Strobl, M. Poli; E. Scarperi, (APPA, Bolzano), M. Poli, M. Dekas (Eco Center Spa AG, Bolzano); L. Bruni, G. Gatti, L. Tomasi, G. Cimadon (Provincia Autonoma di Trento, Trento); I. Pellegrini, A. Ventura, C. Zorzetto (ARPA FVG, Trieste); P. Parati, E. Dell’Andrea, G. Gambillara (Arpa Veneto, Venezia); C. Galas, R. Tomassini (Arpalazio, Roma), G. Ranalletta (ACEA ATO2 S.p.A., Roma); V. Perrino, M. Mariani (ARPA Puglia, Bari); G. Manassero (Arpa Valle D’Aosta, Aosta); A. Martello (Corpo Forestale Aldostano); S. Gaiter, L. Sola (ARPA Genova); M.A. Corvaglia, P. Albertelli (ARPA Emilia Romagna, Bologna); C. Mengarelli, A. Trimboli (ARPA Marche, Ancona); A. Gambaccioni, M. Razzolini (Publiacqua S.p.A. Firenze); E. Renna, G. Saltalamacchia, M. Lucarini (ARPA Umbria, Perugia); A. Renzi, D. Rosoni, C. Spatola Mayo (ARTA Abruzzo, Pescara); A. Lucci, V. Colecchia, C. Iorio (ARPA Molise, Campobasso); E. Rufolo, R. Martino (ARPA Campania, Napoli); R. Vita, R. Masotti, R. Martoccia (ARPA Basilicata, Potenza); F. Pedulla, G. Belmusto (ARPA Calabria, Reggio Calabria); L. Librici, G. Abbate (ARPA Sicilia, Palermo); A.M. Mereu, M. Secci (ARPA Sardegna, Cagliari); R. Bucci, A. Schito, T. Carpinelli (ARPA Puglia, Lecce).

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Authors and Affiliations

  1. Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy

    M. Iaconelli, S. Della Libera & Giuseppina La Rosa

  2. Department Experimental Medicine and Surgery, University of Rome “Tor Vergata”, Rome, Italy

    M. Divizia

  3. Department of Infectious Parasitic Immune-Mediated Diseases, Istituto Superiore di Sanità, Rome, Italy

    P. Di Bonito

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  1. M. Iaconelli
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  2. M. Divizia
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  3. S. Della Libera
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Correspondence to Giuseppina La Rosa.

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Iaconelli, M., Divizia, M., Della Libera, S. et al. Frequent Detection and Genetic Diversity of Human Bocavirus in Urban Sewage Samples. Food Environ Virol 8, 289–295 (2016). https://doi.org/10.1007/s12560-016-9251-7

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