DEVELOPMENT OF A PCR-BASED ASSAY FOR THE DETECTION OF CHIKEN FECES

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Water quality and safety are major public health issues. The water in our homes and recreational areas is expected to be safe to drink or swim in and to be relatively free of pollutants including fecal pollutants. Many human diseases such as cholera, typhoid, salmonellosis, and dysentery can be transmitted through fecal contamination of water. The goal of this research project is to develop a PCR-based assay for chicken fecal contamination using fecal anaerobic bacteria from the order Bacteroidales. Previous studies have shown that Bacteroidales strains present in the digestive tracts of different species, or in a group of species sharing a common characteristic (e.g. ruminants), are genetically distinct. Assays based on differences in 16S rRNA gene sequences have already been developed for the identification of human and ruminant fecal contamination, but an assay to distinguish chicken fecal contamination from other types of fecal pollution is not yet available. Therefore, the focus of this project is to identify a genetic marker specific for chicken feces and develop a PCR-based assay for chicken fecal contamination. The assay will be tested for specificity using fecal DNA samples from other animals. The assay will be further tested by analysis of water samples containing known fecal contaminants and by testing water samples taken from local watersheds. Finally, the assay will be tested for its ability to quantitate different levels of fecal contamination and for limits of sensitivity (upper and lower). The long-term objective of this research is to develop and implement rapid, specific, sensitive, and inexpensive molecular methods for the determination of the source or sources of fecal contamination in water samples.