Transcript Haight - York College of Pennsylvania

Analysis of Water and Sediment Samples taken from
Hurricane Katrina Flooded Homes for the Presence
of Enterococci
Sarah Haight
Department of Biological Sciences, York College of Pennsylvania
Sediment located in a home in Violet, LA
INTRODUCTION
REVIEW OF LITERATURE
•On August 29, 2005, the eye of Hurricane Katrina
passed over St. Bernard Parish, LA. The Parish,
consisting of mostly residential neighborhoods, was
flooded with anywhere from 2 to 29 feet of water
when the levees, designed to protect the below-sea
level area, breached (Hurricane Katrina).
Hurricane Katrina
•After the flooding, major concerns were raised
regarding the possibility that the flood water would
act as a reservoir for potentially harmful bacteria,
especially those found in human fecal matter.
Testing of the water revealed elevated levels of
pathogenic bacteria, including enterococci, in the
New Orleans area (Schwab et al. 2007).
•Water samples from near the Superdome showed
total coliform levels of 8,083,000 CFU/mL, with
1,716,700 CFU/mL of presumptive Aeromonas and
Vibrio (Presley et. al 2006).
•Enterococci levels were examined in the majority of
studies, which could be because of the bacterium’s
growing resistance to normal antibiotic treatments
(Gallert et. al 2005).
•Despite the clean-up process in the city of New
Orleans, St. Bernard Parish has not returned to its
pre-Katrina state, with many houses still containing
sediment and water deposited during the flooding.
This sediment and water could pose a threat to
those attempting to rehabilitate the area.
•However, little to no research has been done on the
Parish.
•August 30, 2005: New Orleans covered in
approximately 34 billion liters of water (Sinigalliano
et. al 2007)
•September 24, 2005: Rita re-floods the area
•Water samples from near Charity Hospital showed
total coliform levels of 259,000 CFU/mL, with 3,700
presumptive Aeromonas and Vibrio
•Lake Pontchartrain enterococci levels were found to
be at 1,400 CFU/100mL of water after the flood
water from the city was pumped into the lake
(Sinigalliano et. al 2007).
•Sediment samples from along the banks found to
contain fecal indicator organisms in levels ranging
from 0.7 to 970 MPN/g.
•Residential sediment samples were found to contain
enterococci levels of 270-980 MPN/g (Sinigalliano et.
al 2007).
•Enterococci levels increased with the amount of
organic matter present.
Enterococci
Bath tub filled with water left from Katrina
Uncleaned home with refrigerator
OBJECTIVES
•To determine if water and sediment
samples collected from St. Bernard
Parish, LA homes contain enterococci
•To determine the location where
enterococci is most prevalent in the
house
•If a colony was resistant to one drug, it was
normally resistant to multiple drugs (normally one
to three in total).
•A major health concern is vancomycin-resistant
enterococci (VRE) (Trick et. al 2004).
•VRE normally have a vanA or vanB genotype, both
of which can be tested for using PCR
•The highest levels of enterococci in water will
be located in the refrigerator samples, with
expected levels around 300-400 CFU/g
•The highest levels of enterococci in sediment
samples will be in the kitchen samples, with
expected numbers over 350 MPN/g
LITERATURE CITED
Houses selected based on
government “Failed to
Clean” list
Hurricane Katrina: The Essential Timeline. Available from:
http://news.nationalgeographic.com/news/2005/09/0914_050914_katrina_timeline
.html. Accessed: 2007 November 3.
Gallert, C., Fund, K., and Winter, J. 2005. Antibiotic resistance of bacteria in raw
and
biologically treated sewage and in groundwater below leaking sewers. Applied
Microbiology and Biotechnology [serial online] 69:106-112. Available from:
EbscoHost.
Sediment samples
collected from the kitchen
and the living room.
Analyzed using cultures
and biochemical tests
Water samples collected
from refrigerator,
swimming pool (if
present), and one other
location.
Water analyzed using
Standard Methods for the
identification and
quantification of
enterococci
•Present in fecal matter
•The enterococci isolated were found to be highly
resistant to penicillin and ampicillin, and resistant
to vancomycin, triple sulfa (sulfadicin,
sulfamethacin, sulfamericin), and
trimethoprim/sulfamethoxazole.
•Enterococci will be present in both sediment
and water samples
RESEARCH DESIGN
•Gram positive cocci (Weaver et. al 2005).
•Gallert et. al were able to isolate 84 enterococci
colonies from raw sewage samples (2005)
EXPECTED RESULTS
Presley, S.M., Rainwater, T.R., Austin, G.P., Platt, S.G., Zak, J.C., Cobb, G.P.,
Marsland,
E.J., Tian, K., Zhang, B., Anderson, T.A., Cox, S.B., Abel, M.T., Leftwich, B.B.,
Huddleston, J.R., Jeter, R.M., and Kendall, R.J. 2006. Assessment of pathogens and
toxicants in New Orleans, LA following Hurricane Katrina. Environmental Science
and Technology [serial online] 40:468-474.
Schwab, K.J., Gibson, K.E., Williams, D.L., Kulbicki, K.M., Lo, C.P., Mihalic, J.N.,
Breysse, P.N., Gurriero, F.C. and Geyh, A.S. 2007. Microbial and chemical
assessment of regions within New Orleans, LA impacted by Hurricane Katrina.
Environmental Science Technology. 41:2401-2406.
Sinigalliano, C.D., Gidley, M.L., Shibata, T., Whitman, D., Dixon, T.H., Laws, E., Hou,
A., Bachoon, D., Brand, L., Amaral-Zettier, L., Gast, R.J., Steward, G.F., Nigro, O.D.,
Fujioka, R., Betancourt, W.Q., Vithanage, G., Mathews, J., Fleming, L.E., and SoloGabriele, H.M. 2007. Impacts of Hurricanes Katrina and Rita on the microbial
landscape of the New Orleans area. Proceedings of the National Academy of
Sciences [serial online] 104(21):9029-9034. Available from: www.pnas.org.
Trick, W.E., Paule, S.M., Cunningham, S., Cordell, R.L., Lankford, M., Stosor, V.,
Solomon, S.L., and Peterson, L.R. 2004. Detection of vancomycin-resistant
enterococci before and after antimicrobial therapy: use of conventional culture and
polymerase chain reaction. Clinical Infectious Disease [serial online] 38:780-786.
Available from: EbscoHost.
Weaver, R.W., Entry, J.A., and Graves, A. 2005. Numbers of fecal streptococci and
Identification confirmed
using qPCR
Identification confirmed
using qPCR
Escherichia coli in fresh and dry cattle, horse, and sheep manure. Canadian Journal
of Microbiology [serial online] 51:847-851. Available from: EbscoHost.
Acknowledgements
I would like to thank the residents of St. Bernard Parish who were so hospitable to
the volunteers after having lost so much. Thanks also goes to Dr. Mathur.
Undrained swimming pool