Bacterial strains were isolated from beach water samples using the initial Environmental Protection Agency method for enumeration and analyzed by pulsed-field gel electrophoresis (PFGE). procedurerecommended by the U.S. Environmental Protection Agency (EPA)is based on epidemiological studies that demonstrate a direct relationship between the density of organisms in water and the occurrence of swimming-associated gastroenteritis (6, 21, 29). Although is the EPA-recommended indicator of fecal pollution, fecal coliforms (FC) continue to be widely used for monitoring of recreational waters, according to data reported by the EPA Beaches Environmental Assessment, Closure, and Health Program (http://www.epa.gov/ost/beaches/)is considered a more specific indicator of fecal pollution than FC, as FC 172152-19-1 manufacture have been found in ambient waters in the absence of apparent fecal pollution and may establish viable populations when high levels of carbohydrates are available as a nutrient source (28). Byamukama et al. attributed the discriminatory power of as an indicator organism to its weak ability to replicate 172152-19-1 manufacture in the natural environment (4). 172152-19-1 manufacture However, recent findings suggest that may occur in ambient waters in the absence of apparent fecal pollution, pointing to prolonged survival or replication of in subtropical or tropical environments (5, 13, 16, 23, 24). Solo-Gabriele et al. (25) reported multiplication in riverbank soil during drying and wetting cycles in laboratory experiments that simulate tidal activity. Conflicting conclusions regarding the ability of to replicate outside its host may arise from factors specific to each data set, such as climate and differing isolation methods for and FC among studies (4). If it occurs, however, bacterial replication in such systems would lead to elevated counts beyond what actually is introduced from fecal contamination events (25). A complication in assessing the environmental viability of is the difficulty in differentiating replication from simple accumulation of cells in waters subject to regular contamination. The aim of this work was to determine if replication in the environment contributed to high levels of this indicator organism at a beach site that historically has had poor water quality. The main area of study, South Shore Beach in Milwaukee, Wis., is subject to contamination by combined sewer overflows (CSOs) and urban and agricultural runoff from the Milwaukee River Basin that empties into the Milwaukee Harbor via three major tributaries. A large population of ring-billed gulls at the site, at times reaching up to 300 individuals, may also adversely affect water quality (18). In addition, the beach area is enclosed on three sides with a break wall that reduces dilution and mixing effects. THE TOWN of Milwaukee Wellness Department reported amounts more than EPA limitations for recreational drinking water (<235 100 ml?1) on 32 times in 1999 and 42 times in 2000 through the going swimming season (data supplied by the town of Milwaukee Wellness Division). In 1999, ARF6 raised amounts didn’t coincide with known elements such as for example rainfall or CSOs often, and many out-of-limit times happened during popular in fact, dry climate. In 2000, there have been high matters pursuing CSO and rainfall occasions, but matters were detected at high amounts when 172152-19-1 manufacture rainfall occurred without CSO event equally. June Drinking water examples had been gathered on five consecutive times during each one of the weeks, July, and August in 2000 (Desk ?(Desk1).1). Examples were also used 24 h after two CSO occasions (3 July and 13 Sept 2000). Samples had been acquired 172152-19-1 manufacture 2 m from shoreline at a depth of 45 cm using 20 sterile 50-ml polypropylene centrifuge pipes for each test to make sure that replication inside the container wouldn’t normally affect results. Examples were put into the dark on snow.