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S. aureus & Vibrio vulnificus Cryptosporidium & Giardia
enterovirus, norovirus , reovirus & hepatitus A
National Center for Environmental Health (NCEH), Centers for Disease Control and Prevention (CDC), Florida Dept of Health (FL DOH), Florida Dept of Environmental Protection (FL DEP), Environmental Protection Agency (EPA) Internship Program, National Science Foundation (NSF) & National Institute of Environmental Health Sciences (NIEHS) Oceans and Human Health Center at the University of Miami Rosenstiel School (NSF 0CE0432368/0911373; NIEHS 1 P50 ES12736), NSF REU in OHH, NSF SGER (NSF SGER 0743987) in OHH. We would also like to thank IDEXX and the numerous students which assisted in this study for their support .
Relationship between Measured Indicator Microbes, Pathogens, and Human Health Effect Levels in a Non-point Source Subtropical Marine Recreational Beach
Amir Abdelzaher, Mary Wright, Cristina Ortega, A. Rasem Hasan, Helena Solo-Gabriele , Tomoyoku Shibata, Jonathan Kish, Kelly Withum, Guoqing He, Samir Elmir, J. Alfredo Bonilla, Tonya D. Bonilla, Carol Palmer, Troy Scott, George Lukasik, Valerie J. Harwood, Shannon McQuaig, Chris Sinigalliano, Maribeth Gidley, David Wanless,
Kelly Goodwin, Lisa Plano, Cristy A. Garza, Xiaofang Zhu, John D. Wang, Jill Stewart, Helen Yampara-Iquise, Charles Carson, Jerold Dickerson, Jay Fleisher, Lora Fleming.
A. M. [email protected]
H.M. [email protected]
305-284-2908
Presented by:H.M. Solo-Gabriele
OBJECTIVES
Determine beach water quality in terms of indicator microbe and pathogen levels. Determine if associations (if any) exist between indicator microbes, pathogens,
and environmental factors. Determine daily cumulative health effects (Gastrointestinal illness as well as skin,
eye, ear and respiratory infection) for bathers. Determine association (if any) between health effects and water quality (indicator
microbes, pathogens, and environmental factors). Identify possible implications on beach regulation practices.
MATERIALS & METHODS RESULTS & DISCUSSION
SUMMARY
Studies which evaluate the relationship between indicator microbes, pathogens, and human health at recreational beaches, especially at non-point source beaches, are rare, although such studies are necessary for establishing criteria which would protect public health while minimizing economic burdens. The objectives of this study were to evaluate water quality (indicator microbes, pathogens, and environmental factors) and daily cumulative health effects (gastrointestinal, skin, and respiratory illnesses) for bathers at a non-point source subtropical marine recreational beach in order to better understand the inter-relationships between these factors and hence improve monitoring and pollution prevention techniques. Daily composite samples were collected during the BEACHES epidemiology study conducted in Miami (FL) at a non-point source subtropical marine beach. These samples were analyzed for several pathogens, microbial source tracking markers, indicator microbes, and environmental parameters. Analysis demonstrated that rainfall and tide were the more influential factors in determining the presence of both indicator microbes and pathogens, while direct relationships between indicators and pathogens were not identified. Rainfall and F+ coliphage should be further assessed to confirm their possible relationship with skin and GI illness, respectively. The results of this research demonstrated the complexity of beach systems characterized by non-point sources, and how more novel and comprehensive approaches are needed to assess beach water quality for the purpose of protecting bather health.
Indicators and pathogen association with rain and turbidity
Dry sand
Epidemiology
Enterococci EPA recommended marine bacterial indicator Analyzed by MF, chromogenic substrate, and qPCR.
Fecal coliform, E. coli, and C. perfringens Coliphage (F+ and F-)
Microbial Analysis of Water
Indicator Microbes
Microbial Source
Tracking
Pathogens
STUDY SITE
Virginia Key within Miami-Dade County, Florida, USA.
Pets are allowed Beach admission is free Subtropical beach No known pt source of pollution Frequently exceeds enterococci standard (104/100
ml) for single samples.
Hobie CatBeach
Low tide knee depth40m
10m
Composite Sampling Area
Daily Sampling Transect
Study Beach
Inter-tidal zone
High Tide Line
Low Tide Line
Low tide knee depth
High tide knee depth
40m
10m
Composite Sampling Area
Daily Sampling Transect
Study Design: Randomized exposure assessment study Health Data + Environmental Data 15 Sampling Days: December 2007 to June 2008
Non-Bathers
Health Interview
Bathers
Randomized
Composite Samples (bather collected) 30-60 bathers per day collected 5 L each over approximately 3.5 hr sample
duration. 1 L from each 5L collected per bather pooled together for a total of 30-60 L
sample per day. Analysis of pathogens, indicator microbes, & MST.
Composite Samples (investigator collected) 10 L samples collected periodically over 3.5 hr sampling duration. Total sample volume = 225 L Analysis of pathogens: protozoa and viruses
Environmental Measurements pH, temperature, salinity, turbidity solar radiation, tidal height, rain, wind speed, camara images
Water sampling
ABSTRACT
Organism Correlated with: R p
Enterococci (MF)E. coli 0.57 0.02
C. perfringens 0.57 0.03
Enterococci (CS)Bacteroidales (human UCD) 0.68 0.005
solar radiation -0.56 0.03
E. coliFecal coliform 0.68 0.005
C. perfringens 0.70 0.004
Water Temperaturesolar radiation 0.53 0.04
turbidity -0.58 0.02
Correlation coefficients (p<0.05) between indicator microbes, pathogens, and environmental factors
ACKNOWLEDGEMENTS
Human Polyomavirus (HPyVs) & ESP gene for enterococci
Bacteriodes (B. thetaiotaomicron , human UCD, human HF8, dog TSC),
Bather Collected Investigator Collected Environ. Measures
0
10
20
30
40
50
60
70
0
0.5
1
1.5
2
2.5
3
NT
U
cm o
f ra
in
Rainfall (0-6 hours)
Rainfall ( 6-24 hrs)
Rainfall (24-48 hrs)
Turbidity (NTU)
<2 15 4 15 13 99 109 <2 3 29 50 13 <2 14 <2 Enterococci (MF)<2 <2 31 3 7 83 27 3 <2 3 23 25 <2 30 17 E.coli
3 104 27 10 20 196 61 66 <2 5 9 127 7 159 147 Fecal coliform <2 <2 7 <2 1295 1653 402 13 4 5 34 24 6 159 16 C. perfringens55 52 61 18 13 69 100 23 13 124 43 15 11 48 68 Enterococci(CS)
GEU/100 ml 68 81 233 512 1647 483 89 124 46 2361 974 472 449 285 637 Enterococci (qPCR)1.6 <0.6 <0.5 <0.5 <0.5 <0.5 <0.5 <0.5 <0.5 0.9 <0.7 1 <0.5 <0.5 0.5 Cryptosporidium oocysts1.1 <0.6 <0.5 <0.5 <0.5 <0.5 2.1 <0.5 <0.5 2.3 <0.7 <0.5 1.5 <0.5 1 Giardia cysts1.6 <0.5 <0.5 <0.5 <0.5 <0.6 <0.6 <0.5 <0.5 2.2 0.5 <0.6 <0.5 0.5 0.53 Enterovirus
TSC/100 ml 391 2090 1640 532 4738 120 463 <1 314 12900 1102 119 <1 516 538 Bacteroidales Dog-Specific 64 51 59 15 43 25 28 25 6 273 407 4 <1 37 28 Bacteroidales UCD3 <1 5 <1 <1 <1 <1 <1 <1 <1 <1 <1 <1 <1 <1 Bacteroidales HF8
neg pos neg pos pos neg neg pos neg neg neg neg neg neg neg B. thetaiotaomicron neg neg neg neg pos pos neg pos pos neg neg neg neg neg neg Polyomavirus
per 500 ml pos neg neg neg neg neg neg neg neg neg neg pos pos neg neg Enterococci esp gene<0.3 <0.3 <0.3 <0.3 <0.3 <0.3 <0.3 <0.3 <0.3 <0.3 <0.3 <0.3 <0.3 <0.3 <0.3 F+ coliphage<0.3 1 1.7 <0.3 <0.3 <0.3 <0.3 <0.3 <0.3 <0.3 <0.3 0.3 0.3 <0.3 <0.3 F- coliphage
CFU/500 ml pos pos pos neg pos pos neg neg pos pos neg pos pos pos pos V.vulnificusCFU/100 ml 11 16 24 11 4 <2 8 <2 11 <2 <2 19 14 2 13 S. aureus
0.0% 9.7% 4.8% 0.1% 2.2% 2.4% 5.8% 0.2% 0.5% 4.2% 1.9% -0.1% 4.1% 0.0% -4.9% GI Illness10.3% 3.0% 4.8% -2.3% 4.2% 8.6% 8.3% 7.7% 2.8% 15.6% 11.5% 5.1% 5.9% 0.0% -1.1% Skin Illness
0.5% 0.0% 5.1% -4.7% 0.0% 0.0% 5.9% 7.3% 2.2% 2.0% 2.0% 0.0% -2.0% 0.0% 0.0% AFRI% 0% 0% 0% 0% 21% 84% 29% 0% 0% 0% 77% 0% 0% 0% 0% Post-HT Sampling m 0.41 0.50 0.60 0.05 0.47 0.45 0.57 0.15 0.27 0.03 0.53 0.33 0.28 0.19 0.45 Tidal Height
rising rising rising ebbing rising ebbing rising rising rising rising ebbing ebbing rising ebbing rising Tidal Stage
(W/m2) 241 230 325 449 383 260 190 352 312 366 378 393 380 113 236 Solar Radiation
% excess illness
per 1 L
CFU/100 ml
#organisms/100L
GEU/100 ml
PFU/100 ml
LEFT: Beach “snapshots” including microbial indicators, source tracking markers, pathogen levels and excess illness percentages between bathers and non-bathers rates, and their potential association with environmental parameters. Data highlighted to indicate possible influence of rainfall. Data circled or with a rectangle to indicate possible relationships between factors (see results). Excess illness percentage rates of 0% represent sampling days where there was neither bather nor non-bathers illness/infection while negative values represent days in which the number of cases in non-bathers exceeded the number in bathers.
Note: Parameters that did not meet normalcy requirements (such as rainfall, post-HT sampling %, pathogens, coliphage, and all of the source tracking markers except Bacteriodales UCD), mostly due to being frequently below detection limits, were not evaluated statistically.
RECOMMENDATIONS Rainfall, tide, and solar radiation impact microbe levels Indicators and pathogens likely de-coupled after release in the
environment Further EPI studies to include 24-hr prior rainfall and coliphage
(F+ & F-) Complexity of non-point source beach system calls for a
different approach Focus not on correlating single organisms with health or
organisms with each other
The need for more sampling coupled with measures of human health outcomes
Obvious source prevention runoff, bather shedding, animal inputs
“Comprehensive Tool Box with Approval Process Approach” use multiple lines of evidence instead of relying on only one
measure of a fecal indicator
