Transcript Vibrio - University of New Hampshire

Monitoring Vibrio Bacteria in Great Bay Estuary
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J. Lemaire , S. Jones , S. Hale M. Hartwick
Northeast Center for Vibrio Disease and Ecology, University of New Hampshire, 56 College Road, Durham, NH
Conclusions
Materials & Methods
Introduction
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
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 1. Vibrio Cholerae
(http://www.flickr.com/photos/equi
noxgraphics/4494306873/)
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.
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
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.
Figure 6. Heart infusion broth
PCR and Gel Electrophoresis
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
Objectives
•Monitor Vibrio in The Great Bay through bi-weekly and bi- monthly
surveillance of oyster, sediment, plankton and water from 3 locations
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.
Results
•Microbial and genetic verification of presence
of Vibrio species
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/
Positive Vibrio Isolates
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•Identify patterns in environmental
factors contributing to concnetrations
of Vibrio occurrence
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Isolates
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.
V.p.
40
V.v.
V.c.
30
•Sustain safe shellfish beds and
safe beaches
Qualitatively the amount of isolates seem to be
increasing throughout the 2014 summer season,
with V. parahaemolyticus being consistently
more prevalent.
•Protect public health
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April
May
June
July
Month
•Collect remote sensing data for
the development of statisitcal models
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/)
Acknowledgements
This research was supported with funding from the
National Science Foundation’s grant to NH EPSCoR (IIA1330641).
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.