Advertisement

Estuaries and Coasts

, Volume 37, Issue 2, pp 421–435 | Cite as

Quantification of Vibrio vulnificus in an Estuarine Environment: a Multi-Year Analysis Using QPCR

  • Jennifer J. WetzEmail author
  • A. Denene Blackwood
  • J. Stephen Fries
  • Zachary F. Williams
  • Rachel T. Noble
Article

Abstract

Using a refined quantitative polymerase chain reaction assay, we compared surface and bottom water concentrations of Vibrio vulnificus with total Vibrio spp. concentrations, fecal indicator bacteria (Escherichia coli and Enterococcus sp.), and environmental parameters (salinity, temperature, chlorophyll a, and turbidity) over 4 years at a mesohaline site in the Neuse River Estuary in North Carolina. V. vulnificus was not detected when water temperature was below 20 °C and 93 % of positive samples were from salinities 10–20 psu. V. vulnificus was detected in 50 % of summer samples with peak concentrations in summer bottom waters. Significant positive correlations for V. vulnificus and temperature, salinity, and bacterial group abundance were found. Significant negative correlations with chlorophyll a were also observed. Interannual comparisons indicated a significant decrease in mean V. vulnificus since 2005, corresponding with increasing salinity due to severe drought conditions in 2007 and 2008. Total Vibrio spp. abundance also decreased in 2007 but returned to the previously observed abundance by 2008. Although a significant positive relationship between total Vibrio spp. and V. vulnificus was documented, interannual comparisons indicate that total Vibrio spp. densities may not be indicative of V. vulnificus in all environmental conditions and that long-term drought conditions may alter community composition.

Keywords

Vibrio vulnificus Estuary Drought Total Vibrio 

Notes

Acknowledgments

The authors would like to thank the members of the Paerl and Noble Laboratories at UNC’s Institute of Marine Science for ModMon sample collection and processing related to this project. Support for this work was provided by the National Science Foundation and the National Institutes of Health through the Ecology of Infectious Disease Program grants OCE-0327056 and 082193 and an associated REU supplement grant to Z.W.

References

  1. Baker-Austin, C., J.A. Trinanes, N.G.H. Taylor, R. Hartnell, A. Siitonen, and J. Martinez-Urtaza. 2012. Emerging Vibrio risk at high latitudes in response to ocean warming. Nature Climate Change 3: 73–77.CrossRefGoogle Scholar
  2. Blackwell, K.D., and J.D. Oliver. 2008. The ecology of Vibrio vulnificus, Vibrio cholerae, and Vibrio parahaemolyticus in North Carolina estuaries. Journal of Microbiology 46: 146–153.CrossRefGoogle Scholar
  3. Campbell, M.S., and A.C. Wright. 2003. Real-time PCR analysis of Vibrio vulnificus from oysters. Applied and Environmental Microbiology 69: 7137–7144.CrossRefGoogle Scholar
  4. Cazorla, C., A. Guigon, M. Noel, M. Quilici, and F. Lacassin. 2011. Fatal Vibrio vulnificus infection associated with eating raw oysters, New Caledonia. Emerging Infectious Diseases 17: 136–137.CrossRefGoogle Scholar
  5. Centers for Disease Control and Prevention (CDC). 2005. Vibrio illnesses after Hurricane Katrina-multiple states, August–September 2005. Morbidity Mortality Weekly Report 54: 928–931.Google Scholar
  6. Centers for Disease Control and Prevention (CDC). 2007. Preliminary FoodNet data on the incidence of infection with pathogens transmitted commonly through food—10 states, United States. Morbidity Mortality Weekly Report 56: 336–339.Google Scholar
  7. Chase, E., and V.J. Harwood. 2011. Comparison of the effects of environmental parameters on growth rates of Vibrio vulnificus biotypes I, II, and II by culture and quantitative PCR analysis. Applied and Environmental Microbiology 77: 4200–4207.CrossRefGoogle Scholar
  8. Christensen JH et al. (2007) Regional climate projections. In: S Solomon, D Qin, M Manning, Z Chen, M Marquis, KB Averyt, M Tignor, and HL Miller (eds.). Climate change 2007: the physical basis. Contribution of Working Group I to the Fourth Assessment Report of the Intergovernmental Panel on Climate Change. Cambridge University Press: Cambridge, UK. pp. 847–940Google Scholar
  9. Colwell, R.R. 1996. Global climate and infectious diseases: the cholera paradigm. Science 274: 2025–2031.CrossRefGoogle Scholar
  10. Converse, R.R., A.D. Blackwood, M. Kirs, J.F. Griffith, and R.T. Noble. 2009. Rapid QPCR-based assay for fecal Bacteroides spp. as a tool for assessing fecal contamination in recreational waters. Water Research 43: 4828–4837.CrossRefGoogle Scholar
  11. Dechet, A.M., P.A. Yu, N. Koram, and J. Painter. 2008. Nonfoodborne Vibrio infections: an important cause of morbidity and mortality in the United States, 1997–2006. Clinical Infectious Disease 46: 970–976.CrossRefGoogle Scholar
  12. Eiler, A., C. Gonzalez-Rey, S. Allen, and S. Bertilsson. 2007. Growth response of Vibrio cholerae and other Vibrio spp. to cyanobacterial dissolved organic matter and temperature in brackish water. FEMS Microbiology Ecology 60: 411–416.CrossRefGoogle Scholar
  13. Fries, J.S., G.W. Characklis, and R.T. Noble. 2006. Attachment of fecal indicator bacteria to particles in the Neuse River Estuary. Journal of Environmental Engineering 132: 1338–1345.CrossRefGoogle Scholar
  14. Fries, J.S., R.T. Noble, H.W. Paerl, and G.W. Characklis. 2007. Particle suspensions and their regions of effect in the Neuse River Estuary: implications for water quality monitoring. Estuarine, Coastal and Shelf Science 30: 359–364.Google Scholar
  15. Froelich, B., T. Williams, R.T. Noble, and J.D. Oliver. 2012. Apparent loss of Vibrio vulnificus in North Carolina oysters coincides with drought-induced increase in salinity. Applied and Environmental Microbiology 78: 3885–3889.CrossRefGoogle Scholar
  16. Froelich, B., J. Bowen, J. R. Gonzalez, A. Snedeker, and R. T. Noble. 2013. Mechanistic and statistical models of total Vibrio abundance in the Neuse River Estuary. Water Research doi: 10.1016/j.watres.2013.06.050.
  17. Fukushima, H., Y. Tsunomori, and R. Seki. 2003. Duplex real-time SYBR green PCR assays for detection of 17 species of food- or waterborne pathogens in stools. Journal of Clinical Microbiology 41: 5134–5514.CrossRefGoogle Scholar
  18. Hall, N.S., R.W. Litaker, E. Fensin, J.E. Adolf, H.A. Bowers, A.R. Place, and H.W. Paerl. 2008. Environmental factors contributing to the development and demise of a toxic dinoflagellate (Karlodinium veneficum) bloom in a shallow, eutrophic, lagoonal estuary. Estuarine, Coastal and Shelf Science 31: 402–418.Google Scholar
  19. Hoi, L., J.L. Larsen, I. Dalsgaard, and A. Dalsgaard. 1998. Occurrence of Vibrio vulnificus biotypes in Danish marine environments. Applied and Environmental Microbiology 64: 7–13.Google Scholar
  20. Hsieh, J.L., J.S. Fries, and R.T. Noble. 2008. Dynamics and predictive modeling of Vibrio spp. in the Neuse River Estuary, North Carolina, USA. Environmental Microbiology 10: 57–64.Google Scholar
  21. Hurley, M.A., and M.E. Roscoe. 1983. Automated statistical analysis of microbial enumeration by dilution series. Journal of Applied Bacteriology 55: 159–164.CrossRefGoogle Scholar
  22. Jacobs, J.M., M. Rhodes, C.W. Brown, R.R. Hood, A.K. Leight, W. Long, and R.J. Wood. 2010. Predicting the distribution of Vibrio vulnificus in Chesapeake Bay. NOAA NOS NCCOS Technical Memorandum 112: 1–12.Google Scholar
  23. Kaspar, C.W., and M.L. Tamplin. 1993. Effects of salinity and temperature on the survival of Vibrio vulnificus in seawater and shellfish. Applied and Environmental Microbiology 59: 2425–2429.Google Scholar
  24. Kaysner, C.A., C. Abeyta Jr., M.M. Wekell, A. DePaola Jr., R.F. Stott, and J.M. Leitch. 1987. Virulent strains of Vibrio vulnificus isolated from estuaries of the United States West coast. Applied and Environmental Microbiology 53: 1349–1351.Google Scholar
  25. Lin, M., D.A. Payne, and J.R. Schwarz. 2003. Intraspecific diversity of Vibrio vulnificus in Galveston Bay water and oysters as determined by randomly amplified polymorphic DNA PCR. Applied and Environmental Microbiology 69: 3170–3175.CrossRefGoogle Scholar
  26. Lipp, E.K., C. Rodriguez-Palacios, and J.B. Rose. 2001. Occurrence and distribution of the human pathogen Vibrio vulnificus in a subtropical Gulf of Mexico estuary. Hydrobiologia 460: 165–173.CrossRefGoogle Scholar
  27. Macian, C.M., C.R. Arias, R. Aznar, E. Garay, and M.J. Pujalte. 2000. Identification of Vibrio spp. (other than V. vulnificus) recovered on CPC agar from marine natural samples. International Microbiology 3: 51–53.Google Scholar
  28. Meehl, G.A., T.F. Stocker, W.D. Collins, P. Friedlingstein, A.T. Gaye, J.M. Gregory, et al. 2007. Global climate projections. In: Solomon, S., D. Qin, M. Manning, Z. Chen, M. Marquis, K.B. Averyt, M. Tignor, and H.L. Miller (eds.). Climate change 2007: the physical basis. Contribution of Working Group I to the Fourth Assessment Report of the Intergovernmental Panel on Climate Change. Cambridge University Press: Cambridge, UK pp. 747–845Google Scholar
  29. Morris Jr., J.G., A.C. Wright, D.M. Roberts, P.K. Wood, L.M. Simpson, and J.D. Oliver. 1987a. Identification of environmental Vibrio vulnificus isolates with a DNA probe for the cytotoxin-hemolysin gene. Applied and Environmental Microbiology 53: 193–195.Google Scholar
  30. Morris Jr., J.G., A.C. Wright, L.M. Simpson, P.K. Wood, P.E. Johnson, and J.D. Oliver. 1987b. Virulence of Vibrio vulnificus: association with utilization of transferrin-bound iron, and lack of correlation with levels of cytotoxin or protease production. FEMS Microbiology Letters 40: 55–59.CrossRefGoogle Scholar
  31. Motes, M.L., A. DePaola, D.W. Cook, J.E. Veazey, J.C. Hunsucker, W.E. Garthright, R.J. Blodgett, and S.J. Chirtel. 1998. Influence of water temperature and salinity on Vibrio vulnificus in Northern Gulf and Atlantic coast oysters (Crassostrea virginica). Applied and Environmental Microbiology 64: 1459–1465.Google Scholar
  32. Mourin˜o-Pe’rez, R.R., A.Z. Worden, and F. Azam. 2003. Growth of Vibrio cholerae 01 in red tide waters off California. Applied and Environmental Microbiology 69: 6923–6931.CrossRefGoogle Scholar
  33. Newton, A., M. Kendall, D.J. Vugia, O.L. Henao, and B.E. Mahon. 2012. Increasing rates of Vibriosis in the United States, 1996–2010: review of surveillance data from 2 systems. Clinical Infectious Diseases 54(Suppl. 5): S391–395.CrossRefGoogle Scholar
  34. Nigro, O.D., A.X. Hou, G. Vithanage, R.S. Fujioka, and G.F. Steward. 2011. Temporal and spatial variability in culturable pathogenic Vibrio spp. in Lake Pontchartrain, Louisiana, following Hurricanes Katrina and Rita. Applied and Environmental Microbiology 77: 5384–5393.CrossRefGoogle Scholar
  35. Noble, R.T., and J.A. Fuhrman. 1998. Use of SYBR green I for rapid epifluorescence counts of marine viruses and bacteria. Aquatic Microbial Ecology 14: 113–118.CrossRefGoogle Scholar
  36. North Carolina Drought Management Advisory Council. 2009. Annual activities report. North Carolina: North Carolina Division of Water Resources, Department of Environmental and Natural Resources. 14 pp.Google Scholar
  37. Oliver, J.D. 2005. Vibrio vulnificus. In Oceans and health: pathogens in the marine environment, ed. S. Belkin and R.R. Colwell, 253–276. New York, USA: Springer.CrossRefGoogle Scholar
  38. Oliver, J.D., and R. Bockian. 1995. In vivo resuscitation, and virulence towards mice, of viable but nonculturable cells of Vibrio vulnificus. Applied and Environmental Microbiology 61: 2620–2623.Google Scholar
  39. O’Neill, K.R., S.H. Jones, and D.J. Grimes. 1992. Seasonal incidence of Vibrio vulnificus in the Great Bay estuary of New Hampshire and Maine. Applied and Environmental Microbiology 58: 3257–3262.Google Scholar
  40. Paerl, H.W., L.M. Valdes, A.R. Joyner, B.L. Peierls, M.F. Piehler, S.R. Riggs, R.R. Christian, L.A. Eby, L.B. Crowder, J.S. Ramus, E.J. Clesceri, C.P. Buzzelli, and R.A.J. Luettich. 2006. Ecological response to hurricane events in the Pamlico Sound System, North Carolina, and implications for assessment and management in a regime of increased frequency. Estuarine, Coastal and Shelf Science 29: 1033–1045.Google Scholar
  41. Paerl, H.W., K.L. Rossignol, N.S. Hall, B.L. Peierls, and M.S. Wetz. 2010. Phytoplankton community indicators of short and long-term ecological change in the anthropogenically and climatically impacted Neuse River Estuary, North Carolina, USA. Estuarine, Coastal and Shelf Science 33: 485–497.Google Scholar
  42. Panicker, G., and A.K. Bej. 2005. Real-time PCR detection of Vibrio vulnificus in oysters: comparison of oligonucleotide primers and probes targeting vvhA. Applied and Environmental Microbiology 7: 5702–5709.CrossRefGoogle Scholar
  43. Panicker, G., M.L. Myers, and A.K. Bej. 2004. Rapid detection of Vibrio vulnificus in shellfish and Gulf of Mexico water by real-time PCR. Applied and Environmental Microbiology 70: 498–507.CrossRefGoogle Scholar
  44. Paz, S., N. Bisharat, E. Paz, O. Kidar, and D. Cohen. 2006. Climate change and the emergence of Vibrio vulnificus disease in Israel. Environmental Research 103: 390–396.CrossRefGoogle Scholar
  45. Peierls, B.L., R.R. Christian, and H.W. Paerl. 2003. Water quality and phytoplankton as indicators of hurricane impacts on a large estuarine ecosystem. Estuaries 26: 1329–1343.CrossRefGoogle Scholar
  46. Pfeffer, C.S., M.F. Hite, and J.D. Oliver. 2003. Ecology of Vibrio vulnificus in estuarine waters of eastern North Carolina. Applied and Environmental Microbiology 69: 3526–3531.CrossRefGoogle Scholar
  47. Randa, M.A., M.F. Polz, and E. Lim. 2004. Effects of temperature and salinity on Vibrio vulnificus population dynamics as assessed by quantitative PCR. Applied and Environmental Microbiology 70: 5469–5476.CrossRefGoogle Scholar
  48. Rozen, S., and H.J. Skaletsky. 2000. Primer3 on the WWW for general users and for biologist programmers. In Bioinformatics methods and protocols: methods in molecular biology, ed. S. Krawetz and S. Misener, 365–386. Totowa, NJ: Humana Press. Source code available at http://primer3.sourceforge.net/.Google Scholar
  49. Tamplin, M., G.E. Robdrick, N.J. Blake, and T. Cuba. 1982. Isolation and characterization of Vibrio vulnificus from two Florida estuaries. Applied and Environmental Microbiology 44: 1466–1470.Google Scholar
  50. Tantillo, G.M., M. Fontanarosa, A. Di Pinto, and M. Musti. 2004. Updated perspectives on emerging vibrios associated with human infections. Letters in Applied Microbiology 39: 117–126.CrossRefGoogle Scholar
  51. Thompson, J.R., M.A. Randa, L.A. Marcelino, A. Tomita-Mitchell, E. Lim, and M.F. Polz. 2004. Diversity and dynamics of a North Atlantic coastal Vibrio community. Applied and Environmental Microbiology 70: 4103–4110.CrossRefGoogle Scholar
  52. Valdes-Weaver, L.M., M.F. Piehler, J.L. Pinckney, K.E. Howe, K. Rossignol, and H.W. Paerl. 2006. Long-term temporal and spatial trends in phytoplankton biomass and class-level taxonomic composition in the hydrologically variable Neuse–Pamlico estuarine continuum, North Carolina, U.S.A. Limnology and Oceanography 51: 1410–1420.CrossRefGoogle Scholar
  53. Vezzulli, L., R.R. Colwell, and C. Pruzzo. 2012. Ocean warming and spread of pathogenic Vibrios in the aquatic environment. Microbial Ecology. doi: 10.1007/s00248-012-0163-2.Google Scholar
  54. Weis, K.E.R.M., R. Hutchinson Hammond, and C.G.M. Blackmore. 2011. Vibrio illness in Florida, 1998–2007. Epidemiology and Infection 139: 591–598.CrossRefGoogle Scholar
  55. Wetz, J.J., A.D. Blackwood, J.S. Fries, Z.F. Williams, and R.T. Noble. 2008. Trends in total Vibrio spp. and Vibrio vulnificus concentrations in the eutrophic Neuse River Estuary, North Carolina, during storm events. Aquatic Microbial Ecology 53: 141–149.Google Scholar
  56. Wetz, M.S., E.A. Hutchinson, R.S. Lunetta, H.W. Paerl, and J.C. Taylor. 2011. Severe droughts reduce estuarine primary productivity with cascasding effects on higher trophic levels. Limnology and Oceanography 56: 627–638.CrossRefGoogle Scholar
  57. Wetz, M.S. and D. W. Yaskowitz (2013) An ‘extreme’ future for estuaries? Effects of extreme climatic events on estuarine water quality and ecology. Marine Pollution Bulletin 69: 7–18.Google Scholar
  58. World Health Organization, Food and Agriculture Organization of the United Nations. 2005. Risk assessment of Vibrio vulnificus in raw oysters. Interpretive summary and technical report. Geneva, Switzerland: WHO.Google Scholar
  59. Wright, A.C., R.T. Hill, J.A. Johnson, M.C. Roghman, R.R. Colwell, and J.G. Morris Jr. 1996. Distribution of Vibrio vulnificus in the Chesapeake Bay. Applied and Environmental Microbiology 62: 717–724.Google Scholar
  60. Wright, A.C., and J.G. Morris Jr. 1991. The extracellular cytolysin of Vibrio vulnificus: inactivation and relationship to virulence in mice. Infection and Immunity 59: 192–197.Google Scholar

Copyright information

© Coastal and Estuarine Research Federation 2013

Authors and Affiliations

  • Jennifer J. Wetz
    • 1
    • 2
    Email author
  • A. Denene Blackwood
    • 1
  • J. Stephen Fries
    • 1
    • 3
  • Zachary F. Williams
    • 4
  • Rachel T. Noble
    • 1
  1. 1.Institute of Marine ScienceUniversity of North Carolina-Chapel HillMorehead CityUSA
  2. 2.Harte Research Institute for Gulf of Mexico StudiesTexas A&M University-Corpus ChristiTexasUSA
  3. 3.Science, Engineering, and Policy Analysis Group, Computer Sciences CorporationAlexandriaUSA
  4. 4.University of North Carolina at Chapel Hill, School of MedicineChapel HillUSA

Personalised recommendations