Environmental Science and Pollution Research

, Volume 25, Issue 11, pp 10977–10987 | Cite as

Molecular detection and genotypic characterization of enteric adenoviruses in a hospital wastewater

  • Chourouk Ibrahim
  • Abdennaceur Hassen
  • Pierre Pothier
  • Selma Mejri
  • Salah Hammami
Research Article
  • 67 Downloads

Abstract

Hospital wastewater (HWW) represents a major source of the diffusion of many antibiotics and some toxic pathogenic microorganisms in the aquatic environment. Sanitation services play a critical role in controlling transmission of numerous waterborne pathogens, especially enteric human adenoviruses (HAdVs) that can cause acute gastroenteritis. This study intended to evaluate the human adenoviruses (HAdVs) detection rates, to determine the genotype of these viruses and to assess the efficiency of HAdVs removal in hospital pilot wastewater treatment plant (PWWTP) in Tunis City, Tunisia. Therefore, hospital wastewater samples (n = 102) were collected during the study year from the two biological wastewater treatment techniques: natural oxidizing ponds and the rotating biological disks or biodisks. Nested polymerase chain reaction (Nested PCR) was used to evaluate the HAdVs detection rates. The genotype of HAdVs positive samples was achieved by the sequencing of the PCR products. HAdVs were detected in 64% (65/102) of positive wastewater samples. A substantial increase in the frequencies of HAdVs was observed at the exit of the two wastewater treatment techniques studied. The typing of HAdVs species F showed the occurrence of only HAdVs type 41. This data acquired for the first time in Tunisia showed high persistence and survival of HAdVs in the two biological wastewater treatment techniques experienced, and mainly highlighted the poor virological quality of the treated wastewater intended for recycling, agriculture reuse, and discharges into the natural receiving environments. Consequently, tertiary wastewater treatment appeared necessary in this case to decrease the load of enteric viruses flowing in the water environment.

Keywords

Enteric human adenoviruses Hospital wastewater Wastewater treatment plant Nested-PCR Waterborne diseases Environmental virology 

Notes

Acknowledgements

Our sincere gratitude and thank are addressed to the Tunisian Ministry of Higher Education and Scientific Research and to the National Reference Centre of Enteric Viruses (CNR) in France for their technical and financial support.

The HAdVs sequences found in the present study from wastewater samples have been submitted to the International Centre for Biotechnology Information and received the following accession numbers: KT369590—KT369610.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. AFNOR (1992) Mise en œuvre des dispositifs d’assainissement autonome. Normalisation Française, DTU 64.1, FranceGoogle Scholar
  2. Adefisoye MA, Nwodo UU, Green E, Okoh AI (2016) Quantitative PCR detection and characterization of human adenovirus, rotavirus and hepatitis a virus in discharged effluents of two wastewater treatment facilities in the Eastern Cape, South Africa. Food Environ Virol 8(4):262–274.  https://doi.org/10.1007/s12560-016-9246-4 CrossRefGoogle Scholar
  3. Akusjärvi G, Aleström P, Pettersson M, Lager M, Jörnvall H, Pettersson U (1984) The gene for the adenovirus 2 hexon polypeptides. J Biol Chem 259(22):13976–13979Google Scholar
  4. Allard A, Albinsson B, Wadell G (2001) Rapid typing of human adenoviruses by a general PCR combined with restriction endonuclease analysis. J Clin Microbiol 39(2):498–505CrossRefGoogle Scholar
  5. Amdiouni H, Faouzi A, Fariat N, Hassar M, Soukri A, Nourlil J (2012) Detection and molecular identification of human adenoviruses and enteroviruses in wastewater from Morocco. Lett Appl Microbiol 54(4):359–366.  https://doi.org/10.1111/j.1472-765X.2012.03220.x CrossRefGoogle Scholar
  6. Aw TG, Gin KY (2010) Environmental surveillance and molecular characterization of human enteric viruses in tropical urban wastewater. J Appl Microbiol 109(2):716–730.  https://doi.org/10.1111/j.1365-2672.2010.04701.x Google Scholar
  7. Carducci A, Battistini R, Rovini E, Verani M (2009) Viral removal by wastewater treatment: monitoring of indicators and pathogens. Food Environ Virol 1(2):85–91.  https://doi.org/10.1007/s12560-009-9013-x CrossRefGoogle Scholar
  8. Carter MJ (2005) Enterically infecting viruses: pathogenicity, transmission and significance for food and waterborne infection. J Appl Microbiol 98(6):1354–1380.  https://doi.org/10.1111/j.1365-2672.2005.02635.x CrossRefGoogle Scholar
  9. Dey RS, Ghosh S, Chawla-Sarkar M, Panchalingam S, Nataro JP, Sur D, Manna B, Ramamurthy T (2011) Circulation of a novel pattern of infections by enteric adenovirus serotypes 41 among children below 5 years of age in Kolkata, India. J Clin Microbiol 49(2):500–505.  https://doi.org/10.1128/JCM.01834-10 CrossRefGoogle Scholar
  10. Enriquez CE, Hurst CJ, Gerba CP (1995) Survival of the enteric adenoviruses 40 and 41 in tap, sea and waste water. Water Res 29(11):2548–2553.  https://doi.org/10.1016/0043-1354(95)00070-2 CrossRefGoogle Scholar
  11. Environmental Protection Agency (1992) Standards for the disposal of sewage sludge. Fed Regist 503:9387–9404Google Scholar
  12. Felsenstein J (1993) PHYLIP (phylogeny inference package) version 3.5.c. Department of Genetics, University of Washington: http://www.Caos.kun.nl/cammsa/PHYLIP
  13. Fernandez-Cassi X, Silvera C, Cervero-Aragó S, Rusiñol M, Latif-Eugeni F, Bruguera-Casamada C, Civit S, Araujo RM, Figueras MJ, Girones R, Bofill-Mas S (2016) Evaluation of the microbiological quality of reclaimed water produced from a lagoon system. Environ Sci Pollut Res Int 23(16):16 816–16 833CrossRefGoogle Scholar
  14. Filho EP, da Costa Faria NR, Fialho AM, de Assis RS, Almeida MM, Rocha M, Galvão M, dos Santos FB, Barreto ML, Leite JP (2007) The adenoviruses associated with acute gastroenteritis in hospitalized and community children up to 5 years old in Rio de Janeiro and Salvador, Brazil. J Med Microbiol 56(Pt 3):313–319.  https://doi.org/10.1099/jmm.0.46685-0 CrossRefGoogle Scholar
  15. Fodha I, Chouikha A, Dewar J, Trabelsi A, Boujaafar N, Steele AD (2007) Prevalence of adenovirus antigens in children presenting with acute diarrhea. Med Trop 67(3):256–258Google Scholar
  16. Fong TT, Phanikumar MS, Xagoraraki I, Rose JB (2010) Quantitative detection of human adenoviruses in wastewater and combined sewer overflows influencing a Michigan river. Appl Environ Microbiol 76(3):715–723.  https://doi.org/10.1128/AEM.01316-09 CrossRefGoogle Scholar
  17. Fumian TM, Vieira CB, Leite JP, Miagostovich MP (2013) Assessment of the burden of virus agents in an urban sewage treatment plant in Rio de Janeiro, Brazil. J Water Health 11(1):110–119.  https://doi.org/10.2166/wh.2012.123 CrossRefGoogle Scholar
  18. Grøndahl-Rosado RC, Yarovitsyna E, Trettenes E, Myrmel M, Robertson LJ (2014) A one-year study on the concentrations of norovirus and enteric adenoviruses in wastewater and a surface drinking water source in Norway. Food Environ Virol 6(4):232–245CrossRefGoogle Scholar
  19. Hassen A, Jedidi N, Kallali H, Ferchichi M, Ghrabi A, Chebbi F, Saidi N, Shayeb H, Ennabli M (1994) Élimination des indicateurs et des métaux lourds au cours de traitement des eaux usées domestiques dans une station pilote semi-industrielle. Sci Techn de l’eau 27(4):34–41Google Scholar
  20. He XQ, Cheng L, Zhang DY, Xie XM, Wang DH, Wang Z (2011) One-year monthly survey of rotavirus, astrovirus and norovirus in three sewage treatment plants (STPs) in Beijing, China and associated health risk assessment. Water Sci Technol 64(6):1202–1210.  https://doi.org/10.2166/wst.2011.080 CrossRefGoogle Scholar
  21. Hewitt J, Greening GE, Leonard M, Lewis GD (2013) Evaluation of human adenovirus and human polyomavirus as indicators of human sewage contamination in the aquatic environment. Water Res 47(17):6750–6761.  https://doi.org/10.1016/j.watres.2013.09.001 CrossRefGoogle Scholar
  22. Iaconelli M, Valdazo-González B, Equestre M, Ciccaglione AR, Marcantonio C, Della Libera S, La Rosa G (2017) Molecular characterization of human adenoviruses in urban wastewater using the next generation and sanger sequencing. Water Res 121:240–247.  https://doi.org/10.1016/j.watres.2017.05.039 CrossRefGoogle Scholar
  23. Ibrahim C, Hammami S, Mejri S, Mehri I, Pothier P, Hassen A (2017a) Detection of Aichi virus genotype B in two lines of wastewater treatment processes. Microb Pathog 109:109–512CrossRefGoogle Scholar
  24. Ibrahim C, Mehri I, Hammami S, Mejri S, Hassen A, Pothier P (2017b) Removal of human astroviruses from hospital wastewater by two biological treatment methods: natural oxidizing lagoons and rotating biodisks. Desalin Water Treat 89:287–296CrossRefGoogle Scholar
  25. Ibrahim C, Hammami S, Pothier P, Hassen A (2016) Quantification and genotyping of rotavirus a within two wastewater treatment processes. Clean Soil Air Water 44(4):393–401.  https://doi.org/10.1002/clen.201400588 CrossRefGoogle Scholar
  26. Ibrahim C, Chérif N, Hammami S, Pothier P, Hassen A (2015) Quantification and molecular characterization of norovirus after two wastewater treatment procedures. Water Air Soil Pollut 226:187–193CrossRefGoogle Scholar
  27. Jothikumar N, Cromeans TL, Hill VR, Lu X, Sobsey MD, Erdman DD (2005) Quantitative real-time PCR assays for detection of human adenoviruses and identification of serotypes 40 and 41. Appl Environ Microbiol 71(6):3131–3136.  https://doi.org/10.1128/AEM.71.6.3131-3136.2005 CrossRefGoogle Scholar
  28. Lim MCY, Wang YF, Huang SW, Yang JY, Wang JR (2015) High incidence of mammalian orthoreovirus identified by environmental surveillance in Taiwan. PLoS One 10(11):e0142745.  https://doi.org/10.1371/journal.pone.0142745 CrossRefGoogle Scholar
  29. Lin J, Ganesh A (2013) Water quality indicators: bacteria, coliphages, enteric viruses. Int J Environ Health Res 23(6):484–506.  https://doi.org/10.1080/09603123.2013.769201 CrossRefGoogle Scholar
  30. Lipp EK, Kurz R, Vincent R, Rodriguez-Palacios C, Farrah SR, Rose JB (2001) The effects of seasonal variability and weather on microbial fecal pollution and enteric pathogens in a subtropical estuary. Estuar Coasts 24(2):266–276.  https://doi.org/10.2307/1352950 CrossRefGoogle Scholar
  31. Kaas L, Gourinat AC, Urbès F, Langlet J (2016) A 1-year study on the detection of human enteric viruses in New Caledonia. Food Environ Virol 8(1):46–56.  https://doi.org/10.1007/s12560-015-9224-2 CrossRefGoogle Scholar
  32. Khoshdel A, Parvin N, Doosti A, Famouri F (2015) Prevalence of nosocomial diarrhea due to adenoviruses 40 and 41 in a pediatric ward in Iran. J Clin Diagn Res 9(12):SC15–SC17.  https://doi.org/10.7860/JCDR/2015/15353.6976 Google Scholar
  33. Kokkinos P, Mandilara G, Nikolaidou A, Velegraki A, Theodoratos P, Kampa D, Blougoura A, Christopoulou A, Smeti E, Kamizoulis G, Vantarakis A, Mavridou A (2015) Performance of three small-scale wastewater treatment plants. A challenge for possible reuse. Environ Sci Pollut Res Int 22(22):17744–17752CrossRefGoogle Scholar
  34. Kimura M (1980) A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16(2):111–120.  https://doi.org/10.1007/BF01731581 CrossRefGoogle Scholar
  35. Kumar S, Tamura K, Nei M (1994) MEGA: molecular evolutionary genetics analyzes software for microcomputers. Comput Appl Biosci 10:189–191Google Scholar
  36. Kuo HW, Chen LZ, Shih MH (2015) High prevalence of type 41 and high sequence diversity of partial hexon genes of human adenoviruses in municipal raw sewage and activated sludge. J Appl Microbiol 119(4):1181–1195.  https://doi.org/10.1111/jam.12907 CrossRefGoogle Scholar
  37. La Rosa G, Sanseverino I, Della Libera S, Iaconelli M, Ferrero VEV, Barra Caracciolo A, Lettieri T (2017) The impact of anthropogenic pressure on the virological quality of water from the Tiber river, Italy. Lett Appl Microbiol 65(4):298–305.  https://doi.org/10.1111/lam.12774 CrossRefGoogle Scholar
  38. La Rosa G, Pourshaban M, Iaconelli M, Muscillo M (2010) Quantitative real-time PCR of enteric viruses in influent and effluent samples from wastewater treatment plants in Italy. Ann Ist Super Sanita 46(3):266–273.  https://doi.org/10.4415/ANN_10_03_07 Google Scholar
  39. Le TH, Ng C, Chen H, Yi XZ, Koh TH, Barkham TM, Zhou Z, Gin KY (2016) Occurrences and characterization of antibiotic resistant bacteria and genetic determinants of hospital wastewater in a tropical country. Antimicrob Agents Chemother 60(12):7449–7456.  https://doi.org/10.1128/AAC.01556-16 Google Scholar
  40. Lewis GD, Melcaft TG (1988) Polyethylene glycol precipitation for recovery of pathogenic viruses, including hepatitis A virus and human rotavirus, from oyster, water and sediment samples. Appl Environ Microbiol 54(8):1983–1988Google Scholar
  41. Liu L, Qian Y, Zhang Y, Deng J, Jia L, Dong H (2014) The adenoviruses associated with acute diarrhea in children in Beijing, China. PLoS One 9(2):e88791.  https://doi.org/10.1371/journal.pone.0088791 CrossRefGoogle Scholar
  42. Lin J, Singh A (2015) Detection of human enteric viruses in Umgeni River, Durban, South Africa. J Water Health 13(4):1098–1112CrossRefGoogle Scholar
  43. Miagostovish MP, Ferreira FF, Guimarares FR, Fumian TM, Diniz-Mendes L, Luz SL, Silva LA, Leite JP (2008) Molecular detection and characterization of gastroenteritis viruses occurring naturally in the stream waters of Manaus, Central Amazonia, Brazil. Appl Environ Microbiol 74(2):375–382.  https://doi.org/10.1128/AEM.00944-07 CrossRefGoogle Scholar
  44. Myrmel M, Lange H, Rimstad E (2015) A 1-year quantitative survey of noro-, adeno-, human boca-, and hepatitis E viruses in raw and secondarily treated sewage from two plants in Norway. Food Environ Virol 7(3):213–223CrossRefGoogle Scholar
  45. Osuolale O, Okoh A (2015) Incidence of human adenoviruses and hepatitis a virus in the final effluent of selected wastewater treatment plants in Eastern Cape Province, South Africa. Virol J 12(1):98.  https://doi.org/10.1186/s12985-015-0327-z CrossRefGoogle Scholar
  46. Page RDM (1996) TREVIEW: an application to display phylogenetic trees on personal computers. Comput Appl Biosci 12(4):357–358Google Scholar
  47. Prado T, Silva DM, Guilayn WC, Rose TL, Gaspar AM, Miagostovich MP (2011) Quantification and molecular characterization of enteric viruses detected in effluents from two hospital wastewater treatment plants. Water Res 45(3):1287–1297.  https://doi.org/10.1016/j.watres.2010.10.012 CrossRefGoogle Scholar
  48. Rames E, Roiko A, Stratton H, Macdonald J (2016) Technical aspects of using human adenovirus as a viral water quality indicator. Water Res 96:308–326.  https://doi.org/10.1016/j.watres.2016.03.042 CrossRefGoogle Scholar
  49. Reis TA, Assis AS, do Valle DA, Barletta VH, de Carvalho IP, Rose TL, Portes SA, Leite JP, da Rosa e Silva ML (2016) The role of human adenoviruses type 41 in acute diarrheal disease in Minas Gerais after rotavirus vaccination. Braz J Microbiol 47(1):243–250.  https://doi.org/10.1016/j.bjm.2015.11.011 CrossRefGoogle Scholar
  50. Rodier (1978) L’analyse de l’eau. Eaux naturelles, eaux résiduaires, eaux de mer. 6th Édn, Dunod, Paris, ISBN: 2,100,496,360, 1383Google Scholar
  51. Sdiri-Loulizi K, Gharbi-Khelifi H, de Rougemont A, Hassine M, Chouchane S, Sakly N, Pothier P, Guédiche MN, Aouni M, Ambert-Balay K (2009) Molecular epidemiology of human astrovirus and adenovirus serotypes 40/41 strains related to acute diarrhea in Tunisian children. J Med Virol 81(11):1895–1902.  https://doi.org/10.1002/jmv.21586 CrossRefGoogle Scholar
  52. Sdiri-Loulizi K, Hassine M, Aouni Z, Gharbi-Khelifi H, Chouchane S, Sakly N, Guédiche MN, Pothier P, Aouni M, Ambert-Balay K (2010) Detection and molecular characterization of enteric viruses in environmental samples in Monastir, Tunisia between January 2003 and April 2007. J Appl Microbiol 109(3):1093–1104.  https://doi.org/10.1111/j.1365-2672.2010.04772.x CrossRefGoogle Scholar
  53. Seto D, Chodosh J, Brister JR, Jones MS, Members of the Adenovirus Research Community (2011) Using the whole-genome sequence to characterize and name human adenoviruses. J Virol 85(11):5701–5702CrossRefGoogle Scholar
  54. Sibley SD, Goldberg TL, Pedersen JA (2011) Detection of known and novel adenoviruses in cattle wastes via broad-spectrum primers. Appl Environ Microbiol 77(14):5001–5008.  https://doi.org/10.1128/AEM.00625-11 CrossRefGoogle Scholar
  55. Soares CC, Volotão EM, Albuquerque MC, Nozawa CM, Linhares RE, Volokhov D, Chizhikov V, Lu X, Erdman D, Santos N (2004) Genotyping of enteric adenoviruses by using single-stranded conformation polymorphism analysis and hetero duplex mobility assay. J Clin Microbiol 42(4):1723–1726.  https://doi.org/10.1128/JCM.42.4.1723-1726.2004 CrossRefGoogle Scholar
  56. Souza FS, Féris LA (2016) Hospital and municipal wastewater: identification of relevant pharmaceutical compounds. Water Environ Res 88(9):871–877.  https://doi.org/10.2175/106143016X14609975747603 CrossRefGoogle Scholar
  57. Uhrbrand K, Schultz AC, Koivisto AJ, Nielsen U, Madsen AM (2017) Assessment of airborne bacteria and noroviruses in air emission from a new highly advanced hospital wastewater treatment plant. Water Res 112:110–119.  https://doi.org/10.1016/j.watres.2017.01.046 CrossRefGoogle Scholar
  58. Vecchia AD, Fleck JD, Kluge M, Comerlato J, Bergamaschi B, Luz RB, Arantes TS, Silva JV, Thewes MR, Spilki FR (2012) Assessment of enteric viruses in a sewage treatment plant located in Porto Alegre, southern Brazil. Braz J Biol 72(4):839–946.  https://doi.org/10.1590/S1519-69842012000500009 CrossRefGoogle Scholar
  59. Vergara GG, Rose JB, Gin KY (2016) Risk assessment of noroviruses and human adenoviruses in recreational surface waters. Water Res 103:276–282.  https://doi.org/10.1016/j.watres.2016.07.048 CrossRefGoogle Scholar
  60. Wieczorek M, Krzysztoszek A, Witek A (2015) Species-specific identification of human adenoviruses in sewage. Pol J Microbiol 64(1):23–28Google Scholar
  61. WHO (2005) Water recreation and disease. Plausibility of associated infections: acute effects, sequelae and mortality. IWA Publishing, LondonGoogle Scholar
  62. Zhang QL, Wang HB, Wang YL, Zhou JS, Cai J, He RR (2016) Genotypes of adenoviruses in infants and young children with diarrhea. Zhongguo Dang Dai Er Ke Za Zhi 18(8):718–720Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Chourouk Ibrahim
    • 1
    • 2
  • Abdennaceur Hassen
    • 2
  • Pierre Pothier
    • 3
  • Selma Mejri
    • 4
  • Salah Hammami
    • 5
  1. 1.Faculty of Mathematical, Physical and Natural Sciences of TunisUniversity of Tunis El ManarTunisTunisia
  2. 2.Laboratory of Treatment and Wastewater ValorizationCentre of Research and Water Technologies (CERTE)Techno Park of Borj-CédriaTunisia
  3. 3.Laboratory of VirologyNational Reference Centre for Enteric Viruses, University Hospital of DijonDijonFrance
  4. 4.Laboratory of VirologyUniversity of Tunis El Manar, IRESA, Veterinary Research Institute of TunisiaLa Rabta, TunisTunisia
  5. 5.Laboratory of MicrobiologyUniversity of Manouba, IRESA, National School of Veterinary Medicine at Sidi ThabetTunisTunisia

Personalised recommendations