1.  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. Abdelzaher a.zaher@miami.edu H.M. Solo-Gabriele hmsolo@miami.edu305-284-2908 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 Cat Beach Low tide knee depth 40m 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 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 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