Citation:

STANG, D., K. P. BRENNER, AND M. R. RODGERS. A CRITICAL EVALUATION OF A FLOW CYTOMETER USED FOR DETECTING ENTEROCOCCI IN RECREATIONAL WATERS. JOURNAL OF WATER AND HEALTH. IWA Publishing, London, Uk, 5(2):295-305, (2007).

Impact/Purpose:

The objectives of this research are: (1) to evaluate rapid state-of-the-art measuement methods of pathogens that may indicate the presence of fecal pollution in recreational waters (beaches); (2) to obtain, jointly with a sister laboratory (NHEERL), a new set of water quality data and related health effects data at a variety of beaches across the U.S., in both marine and non-marine waters; (3) to analyze the research data set to evaluate the utility of the tested measurement methods, the new EMPACT monitoring protocol, and the health effects data / questionnaire, in order to establish a relationship between measured pathogens and observed health effects; and (4) to communicate the results to the Office of Water in support of their efforts to develop new state and/or federal guidelines and limits for water quality indicators of fecal contamination, so that beach managers and public health officials can alert the public about the potential health hazards before exposure to unsafe water can occur.

Description:

The current U. S. Environmental Protection Agency-approved method for enterococci (Method 1600) in recreational water is a membrane filter (MF) method that takes 24 hours to obtain results. If the recreational water is not in compliance with the standard, the risk of exposure to enteric pathogens may occur before the water is identified as hazardous. Because flow cytometry combined with specific fluorescent antibodies has the potential to be used as a rapid detection method for microorganisms, this technology was evaluated as a rapid, same-day method to detect enterococci in bathing beach waters. The flow cytometer chosen for this study was a laser microbial detection system designed to detect labeled antibodies. A comparison of MF counts with flow cytometry counts of enterococci in phosphate buffer and sterile-filtered recreational water showed good agreement between the two methods. However, when flow cytometry was used, the counts were several orders of magnitude higher than the MF counts with no correlation to Enterococcus spike concentrations. The unspiked sample controls frequently had higher counts than the samples spiked with enterococci. Particles within the spiked water samples were probably counted as target cells by the flow cytometer because of autofluorescence or non-specific adsorption of antibody and carryover to subsequent samples. For these reasons, this technology may not be suitable for enterococci detection in recreational waters. Improvements in research and instrument design that will eliminate high background and carryover may make this a viable technology in the future.

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