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

Analysis of Water and Sediment Samples taken from Hurricane Katrina Flooded Homes

for the Presence of EnterococciSarah Haight

Department of Biological Sciences, York College of Pennsylvania

REVIEW OF LITERATURE

Hurricane Katrina

•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 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).

•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

•Gram positive cocci (Weaver et. al 2005).

•Present in fecal matter

•Gallert et. al were able to isolate 84 enterococci colonies from raw sewage samples (2005)

•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.

•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

INTRODUCTION

•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).

•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).

•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.

EXPECTED RESULTS

•Enterococci will be present in both sediment and water samples

•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 CITEDHurricane 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.

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 Solo-Gabriele, 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

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.

Houses selected based on government “Failed

to Clean” list

Sediment samples collected from the

kitchen and the living room.

Water samples collected from

refrigerator, swimming pool (if present), and one other location.

Analyzed using cultures and

biochemical tests

Water analyzed using Standard Methods for the identification and

quantification of enterococci

Identification confirmed using qPCR

Identification confirmed using qPCR

RESEARCH DESIGN

Sediment located in a home in Violet, LA

Bath tub filled with water left from Katrina

Uncleaned home with refrigerator

Undrained swimming pool