J. Lemaire1, S. Jones1, S. Hale1 M. Hartwick1 Monitoring Vibrio Bacteria in Great Bay Estuary J. Lemaire1, S. Jones1, S. Hale1 M. Hartwick1 Northeast Center for Vibrio Disease and Ecology, University of New Hampshire, 56 College Road, Durham, NH Conclusions Introduction Materials & Methods Collection Vibrio species are present in The Great Bay, Durham, NH. All three species which are monitored, V. parahaemolyticus, V. vulnificus, and V. cholerae, were detected V. Parahaemolyticus seems to be the most prevalent of the three species of Vibrio in Great Bay (Figure 4) . Qualitatively, the number of isolates collected increased as the summer progressed. Environmental parameters including salinity, dissolved oxygen, and pH, are measured throughout sampling but more data analysis needs to be completed before any conclusive evidence of trends can be made. Data is continuously being established and will be compared to previous years data. This enables the ability to prepare statistical analysis to determine patterns in environmental factors during Vibrio season peaks. Once environmental modeling methods are developed with this data, they will be used to to forewarn vibrio outbreaks before illness occurs will be made. This could enable beach closures and shellfish bed closures before bacteria and diseases have the chance cause infection and disease. Culture Vibrios are motile bacteria found worldwide in marine, fresh and brackish water environments. Species within the Vibrio genus can be human pathogens. Pathogenic V. cholerae causes the disease cholera. There have been 7 great cholera pandemics recorded since 1817. Vibrio parahaemolyticus, and Vibrio vulnificus can cause food borne illnesses through the consumption of raw or undercooked shellfish. V. Parahaemolyticus, V. vulnificus, and V. cholerae disease originate from environmental exposure. Vibrio presence and concentration is monitored here in the Great Bay Estuary in water, sediment, oyster, and plankton in Great Bay, NH. Physical data, including: Temperature, Salinity, pH, Chlorophyll, Dissolved Oxygen, Rainfall and optical measurements, are collected. Microbiological techniques aid in the quantification of Vibrio in The Great Bay. This monitoring program is aimed at better understanding Vibrio ecology and improving methods to protect public health. Figure 3. Water sampling at Adams Point, on Great Bay. Figure 4a. ChromAgar differential/selective media. b. Potential V.p. colonies will grow purple. Potential V.v./V.c will grow blue Figure 6. Heart infusion broth PCR and Gel Electrophoresis Figure 1. Vibrio Cholerae (http://www.flickr.com/photos/equinoxgraphics/4494306873/) Lane 1: 100 bp ladder Lane 2: Negative control Lane 3: Positive control All other lanes: Environmental isolate Collection date 6/27/14 All lanes with DNA bands size 400-500 bp are positive for the tlh gene in V. Parahaemolyticus. References Figure 7. Electrophoresis 1.2 % Agarose Gel. Centers for Disease Control and Prevention. (2014).  Vibrios. Retrieved Dziejman, M., Balon, E., Boyd, D., Fraser, C. M., Heidelberg, J. F., & Mekalanos, J. J. (2002). Comparative genomic analysis of Vibrio cholerae: genes that correlate with cholera endemic and pandemic disease. Proceedings of the National Academy of Sciences, 99(3), 1556-1561. Ellis, C. N., Schuster, B. M., Striplin, M. J., Jones, S. H., Whistler, C. A., & Cooper, V. S. (2012). Influence of seasonality on the genetic diversity of Vibrio parahaemolyticus in New Hampshire shellfish waters as determined by multilocus sequence analysis. Applied and environmental microbiology, 78(10), 3778-3782. Finkelstein RA. Cholera, Vibrio cholerae O1 and O139, and Other Pathogenic Vibrios. In: Baron S, editor. Medical Microbiology. 4th edition. Galveston (TX): University of Texas Medical Branch at Galveston; 1996. Chapter 24. Available from: http://www.ncbi.nlm.nih.gov/books/NBK8407/ from http://www.cdc.gov/vibrio/ Objectives Monitor Vibrio in The Great Bay through bi-weekly and bi- monthly surveillance of oyster, sediment, plankton and water from 3 locations Microbial and genetic verification of presence of Vibrio species Identify patterns in environmental factors contributing to concnetrations of Vibrio occurrence Sustain safe shellfish beds and safe beaches Protect public health Collect remote sensing data for the development of statisitcal models Results The positive V. parahaemolyticus, V. vulnificus, and V. cholerae isolates collected during surveillance and bi-weekly sampling are shown here grouped into the amount of positive isolates per month. Qualitatively the amount of isolates seem to be increasing throughout the 2014 summer season, with V. parahaemolyticus being consistently more prevalent. Acknowledgements This research was supported with funding from the National Science Foundation’s grant to NH EPSCoR (IIA-1330641). Thank you to Dr. Stephen Jones, Dr. Stephen Hale and Meg Hartwick for their mentorship as well as Emily Shultz, Elizabeth Deyett and Stephanie Rodriguez for research assistance. Figure 4. Confirmed Vibrio species counts per month of 2014 summer season. Figure 2. NH State and Map of Great Bay. (http://ciceet.unh.edu/great_bay/)