Detection of enteric viruses in wastewater of Monastir region by RTPCR method

##plugins.themes.academic_pro.article.main##

Mouna Hassine
khira Sdiri
Samira Riabi
Abir Beji
Zaidoun Aouni
Mahjoub Aouni

Abstract

Background: Wastewater frequently has been contaminated by enteric viruses, which can cause gastroenteritis, hepatitis, and other viral diseases. Since current water treatments do not ensure their complete removal and they become contaminants of the water environment.
Aim: In order to monitor and assess the prevalence and the seasonal incidence of enteric viruses contaminating wastewater, a total of 93 samples were collected from the stations of Sayada-Lamta-Bouhjar and El Frina.
Methods: The purpose of this study is to establish a rapid, sensitive and specific RT-PCR method using highly conserved primers to detect enteric viruses (caliciviruses, enteroviruses and hepatitis A viruses) circulating in the area of Monastir, during the period from October 2005 to July 2006.
Results: Our study shows that the caliciviruses were detected in 67.56% of the wastewater samples and present an essentially winter distribution with a specific peak in spring. The enteroviruses were detected in 35.13% of the samples tested and present a regular circulation during the year with a predominance in the summer and early fall. Finally, the presence of hepatitis A viruses was noted especially in winter in 10.81% of the wastewater samples.
Conclusion: We notice that current wastewater treatment methods are sometimes not efficient to eliminate enteric viruses.

Keywords:

Caliciviruses, Enteroviruses, Hepatitis A virus, Wastewater, RT, PCR

##plugins.themes.academic_pro.article.details##

References

  1. Bosch A, Lucena F, Diez JM, Gajardo R, Blasi M, and Jofre J. Waterborne viruses associated with hepatitis outbreak. Res. Technol. Manag. 1991; 3: 80-83.
  2. Metcalf T, Melnick J, and Estes M. Environmental virology: from detection of virus in sewage and water by isolation to identification by molecular biology- a trip of over 50 years. Annu. Rev. Microbiol. 1995; 49: 461-487.
  3. Brugha R, Vipond I, and Evans M. A community outbreak of foodborne small round-structured virus gastroenteritis caused by a contamination water supply. Epidemiol. Infect. 1999; 122: 145-154.
  4. Gerba CP, and Goyal SM. Methods in environmental virology. Microbiology Series. Ed. Marcel Dekker, Inc. New York and Basel. 1982; 7: 151-160.
  5. Chomczynski P, and Sacchi N. Single step method of isolation of RNA by guanidium thiocyanate-phenol-chlorophorm extraction. Anal. Biochem. 1987; 162: 156-159.
  6. Vinjé J, and Koopmans MP. Molecular detection and epidemiology of small round structured viruses in outbreaks of gastroenteritis in the Netherlands. J. Inf. Dis. 1996; 174: 610-615.
  7. Kämmerer U, Kunkel B, and Kun K. Nested PCR for specific detection and rapid identification of human picornaviruses. J. Clin. Microbiol. 1994; 32: 285-291.
  8. Traoré O, Arnal C, Mignotte B, Maul A, Laveran H, Billaudel S, and Schwartzbrod L. Reverse transcriptase PCR detection of Astrovirus, Hepatitis A virus and Poliovirus in experimentally contaminated mussels: comparison of several extraction and concentration methods. Appl. Environ. Microbiol. 1998; 64: 3118-3122.
  9. Puig M, Jofre J, Lucena F, Allard A, Wadell G, and Girones R. Detection of adenoviruses and enteroviruses in polluted waters by nested PCR amplification. Appl. Environ. Microbiol. 1994; 60: 2963-2970.
  10. Abbaszadegen M, Stewart P, and Le Chevalier M. A strategy for detection of viruses in groundwater by PCR. Appl. Environ. Microbiol. 1999; 65: 444-449.
  11. Schvoerer E, Ventura M, Dubos O, Cazaux G, Serceau R, Gournier N, Dubois V, Caminade P, Fleury H, and Lafon ME. Qualitative and quantitative molecular detection of enteroviruses in water from bathing areas and from a sewage treatment plant. Res. Microbiol. 2001; 152: 179-186.
  12. Tsai YL, Tran B, and Palmer CJ. Analysis of viral RNA persistence in seawater by reverse transcriptase-PCR. Appl. Environ. Microbiol. 1995; 61: 363-366.
  13. Limsawat S, and Ohgaki S. Fate of liberated viral RNA in wastewater determined by PCR. Appl. Environ. Microbiol. 1997; 63: 2932-2933.
  14. Pina S, Puig M, Lucena F, Jofre J, and Girones R. Viral pollution in the environment and in shellfish: Human adenovirus detection by PCR as an index of human viruses. Appl. Environ. Microbiol. 1998; 9: 3376-3382.
  15. Koopmans M, Von Bonsdorff CH, Vinjé J, De Medici D, and Monroe S. Food-borne viruses. FEMS. Microbiol. Rev. 2002; 26: 187-205.
  16. Sdiri K, Khelifi H, Belguith K, and Aouni M. Comparaison de la culture cellulaire et de la RT-PCR pour la détection des entérovirus dans les eaux usées et les coquillages en Tunisie. Pathol. Biol. 2006; 54: 280-284.
  17. Belguith K, Bouslama L, Nsaibia S, Sdiri K, Ben Othman H, and Aouni M. Comportement des entérovirus au cours des traitements des eaux usées. Microbiol. Hyg. Alim. 2006; 52: 27-33.
  18. Nairn C, and Clements GB. A study of enterovirus isolation in Glasgow from 1977 to1997. J. Med. Virol. 1999: 58: 304-312.
  19. Belguith K, Hassen A, Gharbi J, and Aouni M. Dominance de la circulation des echovirus dans les eaux usées de la région de Monastir. Microbiol. Hyg. Alim. 2004; 47: 49-53.
  20. Melnick JL. Enteroviruses. In: Evans SE. Viral infections of humans. Epidemiology and control. 3rd Edition. New York: Plenum Publishing corporation. 1991: 191-263.
  21. Lopman B, Vennema H, Kohli E, Pothier P, Sanchez A, Negredo A, Buesa J, Schreier E, Reacher M, and Brown D. Increase in viral gastroenteritis outbreaks in Europe and epidemic spread of new norovirus variant. The Lancet 2004; 363: 682-688.
  22. Bosch A, Pinto R, Blanch A, and Jofre J. Detection of human rotavirus in sewage through two concentration procedures. Wat. Res. 1988; 22: 343-48.
  23. Green D, and Lewis G. Comparative detection of enteric viruses in wastewaters, sediments and oysters by reverse transcription-PCR and cell culture. Wat. Res. 1999; 33: 1195-1200.
  24. Chung H, Jaykus L, and Sobsey M. Detection of human enteric viruses in oysters by in vivo and in vitro amplification of nucleic acids. Appl. Environ. Microbiol 1996; 62: 3772-3778.