Skip to main content

Advertisement

SpringerLink
Log in

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

  • Published:
Estuaries and Coasts Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • 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.

    Article  Google Scholar 

  • 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.

    Article  Google Scholar 

  • Campbell, M.S., and A.C. Wright. 2003. Real-time PCR analysis of Vibrio vulnificus from oysters. Applied and Environmental Microbiology 69: 7137–7144.

    Article  CAS  Google Scholar 

  • 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.

    Article  Google Scholar 

  • 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 

  • 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 

  • 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.

    Article  CAS  Google Scholar 

  • 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. 847–940

  • Colwell, R.R. 1996. Global climate and infectious diseases: the cholera paradigm. Science 274: 2025–2031.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • 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 

  • Hurley, M.A., and M.E. Roscoe. 1983. Automated statistical analysis of microbial enumeration by dilution series. Journal of Applied Bacteriology 55: 159–164.

    Article  Google Scholar 

  • 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 

  • 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.

    CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • 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. 747–845

  • 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.

    CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • 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.

    Article  Google Scholar 

  • 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.

    Article  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  Google Scholar 

  • 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 

  • 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.

    Chapter  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • 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 

  • 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 

  • 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.

    CAS  Google Scholar 

  • 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.

    Article  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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 

  • 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.

    CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  Google Scholar 

  • 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 

  • 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.

    Article  CAS  Google Scholar 

  • 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 

  • 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.

    Article  CAS  Google Scholar 

  • 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 

  • 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 

  • 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.

    CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

Download references

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.

Author information

Authors and Affiliations

  1. Institute of Marine Science, University of North Carolina-Chapel Hill, 3431 Arendell Street, Morehead City, NC, 28557, USA

    Jennifer J. Wetz, A. Denene Blackwood, J. Stephen Fries & Rachel T. Noble

  2. Harte Research Institute for Gulf of Mexico Studies, Texas A&M University-Corpus Christi, Corpus Christi, Texas, 78412, USA

    Jennifer J. Wetz

  3. Science, Engineering, and Policy Analysis Group, Computer Sciences Corporation, 6101 Stevenson Avenue, Alexandria, VA, 22304, USA

    J. Stephen Fries

  4. University of North Carolina at Chapel Hill, School of Medicine, Chapel Hill, NC, 27599, USA

    Zachary F. Williams

Authors
  1. Jennifer J. Wetz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. Denene Blackwood
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. Stephen Fries
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zachary F. Williams
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Rachel T. Noble
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jennifer J. Wetz.

Additional information

Communicated by Bongkeun Song

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wetz, J.J., Blackwood, A.D., Fries, J.S. et al. Quantification of Vibrio vulnificus in an Estuarine Environment: a Multi-Year Analysis Using QPCR. Estuaries and Coasts 37, 421–435 (2014). https://doi.org/10.1007/s12237-013-9682-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12237-013-9682-4

Keywords

Search

Navigation