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Relationship between Measured Indicator Microbes, Pathogens, and Human Health Effect Levels in a Non-point Source Subtropical Marine Recreational Beach A. M. Abdelzaher [email protected] H.M. Solo-Gabriele [email protected] 305-284-2908 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. MATERIALS & METHODS ABSTRACT 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. Epidemiology Correlation coefficients (p<0.05) between indicator microbes, pathogens, and environmental factors 3 Health Data + Environmental Data 15 Sampling Days: December 2007 to June 2008 2.5 DryHealth sand Interview 70 Organism Correlated with: R p 60 Enterococci (MF) E. coli C. perfringens 0.57 0.57 0.02 0.03 50 Enterococci (CS) Bacteroidales (human UCD) 0.68 0.005 solar radiation Fecal coliform -0.56 0.68 0.03 0.005 C. perfringens solar radiation turbidity 0.70 0.53 -0.58 0.004 0.04 0.02 40 1.5 30 E. coli Water Temperature 1 20 Non-Bathers Randomized 0.5 10 0 CFU/100 ml Water sampling Bathers GEU/100 ml 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. CFU/500 ml CFU/100 ml % excess illness pH, temperature, salinity, turbidity solar radiation, tidal height, rain, wind speed, camara images Investigator Collected GEU/100 ml PFU/100 ml Analysis of pathogens: protozoa and viruses Bather Collected TSC/100 ml per 500 ml 10 L samples collected periodically over 3.5 hr sampling duration. Environmental Measurements #organisms /100L per 1 L Composite Samples (investigator collected) Determine association (if any) between health effects and water quality (indicator microbes, pathogens, and environmental factors). % m Environ. Measures STUDY SITE 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. 0 <2 <2 3 <2 55 68 1.6 1.1 1.6 391 64 3 neg neg pos <0.3 <0.3 pos 11 0.0% 10.3% 0.5% 0% 0.41 15 <2 104 <2 52 81 <0.6 <0.6 <0.5 2090 51 <1 pos neg neg <0.3 1 pos 16 9.7% 3.0% 0.0% 0% 0.50 4 31 27 7 61 233 <0.5 <0.5 <0.5 1640 59 5 neg neg neg <0.3 1.7 pos 24 4.8% 4.8% 5.1% 0% 0.60 15 3 10 <2 18 512 <0.5 <0.5 <0.5 532 15 <1 pos neg neg <0.3 <0.3 neg 11 0.1% -2.3% -4.7% 0% 0.05 rising rising rising ebbing (W/m2) Hobie Cat Beach Rainfall (0-6 hours) Rainfall ( 6-24 hrs) Rainfall (24-48 hrs) Turbidity (NTU) 2 Total sample volume = 225 L Indicators and pathogen association with rain and turbidity NTU Study Design: Randomized exposure assessment study Determine daily cumulative health effects (Gastrointestinal illness as well as skin, eye, ear and respiratory infection) for bathers. Identify possible implications on beach regulation practices. RESULTS & DISCUSSION cm of rain 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. Presented by: H.M. Solo-Gabriele 241 230 325 449 13 7 20 1295 13 1647 <0.5 <0.5 <0.5 4738 43 <1 pos pos neg <0.3 <0.3 pos 4 2.2% 4.2% 0.0% 21% 0.47 99 83 196 1653 69 483 <0.5 <0.5 <0.6 120 25 <1 neg pos neg <0.3 <0.3 pos <2 2.4% 8.6% 0.0% 84% 0.45 109 27 61 402 100 89 <0.5 2.1 <0.6 463 28 <1 neg neg neg <0.3 <0.3 neg 8 5.8% 8.3% 5.9% 29% 0.57 <2 3 66 13 23 124 <0.5 <0.5 <0.5 <1 25 <1 pos pos neg <0.3 <0.3 neg <2 0.2% 7.7% 7.3% 0% 0.15 3 <2 <2 4 13 46 <0.5 <0.5 <0.5 314 6 <1 neg pos neg <0.3 <0.3 pos 11 0.5% 2.8% 2.2% 0% 0.27 29 3 5 5 124 2361 0.9 2.3 2.2 12900 273 <1 neg neg neg <0.3 <0.3 pos <2 4.2% 15.6% 2.0% 0% 0.03 50 23 9 34 43 974 <0.7 <0.7 0.5 1102 407 <1 neg neg neg <0.3 <0.3 neg <2 1.9% 11.5% 2.0% 77% 0.53 13 25 127 24 15 472 1 <0.5 <0.6 119 4 <1 neg neg pos <0.3 0.3 pos 19 -0.1% 5.1% 0.0% 0% 0.33 <2 <2 7 6 11 449 <0.5 1.5 <0.5 <1 <1 <1 neg neg pos <0.3 0.3 pos 14 4.1% 5.9% -2.0% 0% 0.28 14 30 159 159 48 285 <0.5 <0.5 0.5 516 37 <1 neg neg neg <0.3 <0.3 pos 2 0.0% 0.0% 0.0% 0% 0.19 <2 Enterococci (MF) 17 E.coli 147 Fecal coliform 16 C. perfringens 68 Enterococci(CS) 637 Enterococci (qPCR) 0.5 Cryptosporidium oocysts 1 Giardia cysts 0.53 Enterovirus 538 Bacteroidales Dog-Specific 28 Bacteroidales UCD <1 Bacteroidales HF8 neg B. thetaiotaomicron neg Polyomavirus neg Enterococci esp gene <0.3 F+ coliphage <0.3 F- coliphage pos V.vulnificus 13 S. aureus -4.9% GI Illness -1.1% Skin Illness 0.0% AFRI 0% Post-HT Sampling 0.45 Tidal Height rising ebbing rising rising rising rising ebbing ebbing rising ebbing rising Tidal Stage 383 260 190 352 312 366 378 393 380 113 236 Solar Radiation SUMMARY 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. RECOMMENDATIONS Virginia Key within Miami-Dade County, Florida, USA. Rainfall, tide, and solar radiation impact microbe levels Pets are allowed Beach admission is free Indicators and pathogens likely de-coupled after release in the environment Subtropical beach No known pt source of pollution Frequently exceeds enterococci standard (104/100 ml) for single samples. 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 Microbial Analysis of Water Enterococci EPA recommended marine bacterial indicator Analyzed by MF, chromogenic substrate, and qPCR. Fecal coliform, E. coli, and C. perfringens Indicator Microbes 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 . Coliphage (F+ and F-) Human Polyomavirus (HPyVs) & ESP gene for enterococci Bacteriodes (B. thetaiotaomicron , human UCD, human HF8, dog TSC), Microbial Source Tracking S. aureus & Vibrio vulnificus Composite Sampling Area Daily Sampling Transect Study Beach Composite Sampling Area Daily Sampling Transect Cryptosporidium & Giardia enterovirus, norovirus , reovirus & hepatitus A ACKNOWLEDGEMENTS Pathogens