Removal of Escherichia coli, Enterococcus fecalis, coliphage MS2, poliovirus, and hepatitis A virus from oysters (Crassostrea virginica) and hard shell clams (Mercinaria mercinaria) by depuration.

Filter-feeding bivalve mollusks (shellfish) can bioaccumulate pathogenic microorganisms in up to 1000-fold higher levels than overlying waters, and therefore disease risks are associated with consuming raw or partially cooked shellfish. Many of these shellfish-borne diseases are due to enteric bacteria and viruses associated with fecal contamination. To control shellfish-borne diseases, guidelines for shellfish harvest waters and shellfish meat have been devised, which include cleansing of contaminated shellfish by depuration in controlled systems, heat pasteurization, or relay to clean waters. This study examines the depuration of oysters (Crassostrea virginica) and hard shell clams (Mercinaria mercinaria) in a flow-through depuration system under variable temperature (12 °C, 18 °C, and 25 °C), salinity (8 ppt, 18 ppt, and 28 ppt), turbidity (<1NTU, 10NTU, and 20NTU), pH (pH 7 and pH 8), and algae conditions (0 cells/mL and 50,000 cells/mL), with constant dissolved oxygen (5-7 mg/L). Oysters and hard shell clams were artificially contaminated with enteric microorganisms: Escherichia coli, Enterococcus faecalis, coliphage MS2, Poliovirus type-1 and Hepatitis A virus HM-175 (HAV), then depurated in 5-day trials with daily sampling. In oysters, optimizing environmental parameters of water temperature improved E. coli, MS2, poliovirus and HAV depuration, and optimized salinity improved E. coli, E. faecalis, and MS2 depuration rates. In hard shell clams, salinity improved E. coli and E. faecalis depuration rates. Adjusting turbidity, pH or algae did not improve microorganism depuration in either oysters or hard shell clams, with the exception of turbidity on E. faecalis in hard shell clams. Microorganism depuration rates in oysters from greatest to least were: MS2>E. coli>E. faecalis>poliovirus>HAV, and in clams depuration rates from greatest to least were: E. coli>E. faecalis>HAV>MS2>poliovirus. Because E. coli and E. faecalis were removed at faster rates than HAV and poliovirus, these fecal bacteria appear to be poor process indicators of the virological quality of depurated oysters and hard shell clams.

Water quality indicators and the risk of illness at beaches with nonpoint sources of fecal contamination.

BACKGROUND: Indicator bacteria are a good predictor of illness at marine beaches that have point sources of pollution with human fecal content. Few studies have addressed the utility of indicator bacteria where nonpoint sources are the dominant fecal input. Extrapolating current water-quality thresholds to such locations is uncertain. METHODS: In a cohort of 8797 beachgoers at Mission Bay, California, we measured baseline health at the time of exposure and 2 weeks later. Water samples were analyzed for bacterial indicators (enterococcus, fecal coliforms, total coliforms) using both traditional and nontraditional methods, ie, chromogenic substrate or quantitative polymerase chain reaction. A novel bacterial indicator (Bacteroides) and viruses (coliphage, adenovirus, norovirus) also were measured. Associations of 14 health outcomes with both water exposure and water quality indicators were assessed. RESULTS: Diarrhea and skin rash incidence were the only symptoms that were increased in swimmers compared with nonswimmers. The incidence of illness was not associated with any of the indicators that traditionally are used to monitor beaches. Among nontraditional water quality indicators, associations with illness were observed only for male-specific coliphage, although a low number of participants were exposed to water at times when coliphage was detected. CONCLUSIONS: Traditional fecal indicators currently used to monitor these beaches were not associated with health risks. These results suggest a need for alternative indicators of water quality where nonpoint sources are dominant fecal contributors.

Use of viral pathogens and indicators to differentiate between human and non-human fecal contamination in a microbial source tracking comparison study.

Assays for the detection and typing of adenoviruses, enteroviruses and F+ specific coliphages were performed on samples created as part of a national microbial source tracking methods comparison study. The samples were created blind to the researchers, and were inoculated with a variety of types of fecal contamination source (human, sewage, dog, seagull and cow) and mixtures of sources. Viral tracer and pathogen assays demonstrated a general ability to discriminate human from non-human fecal contamination. For example, samples inoculated with sewage were correctly identified as containing human fecal contamination because they contained human adenovirus or human enterovirus. In samples containing fecal material from individual humans, human pathogen analysis yielded negative results probably because the stool samples were taken from healthy individuals. False positive rates for the virus-based methods (0-8%) were among the lowest observed during the methods comparison study. It is suggested that virus-based source tracking methods are useful for identification of sewage contamination, and that these methods may also be useful as an indication of the public health risk associated with viral pathogens. Overall, virus-based source tracking methods are an important approach to include in the microbial source tracking 'toolbox'.