A PCR Assay To Discriminate Human and Ruminant Feces on the Basis of Host Differences in Bacteroides-Prevotella Genes Encoding 16S rRNA

ABSTRACT Our purpose was to develop a rapid, inexpensive method of diagnosing the source of fecal pollution in water. In previous research, we identified Bacteroides-Prevotella ribosomal DNA (rDNA) PCR markers based on analysis. These markers length heterogeneity PCR and terminal restriction fragment length polymorphism distinguish cow from human feces. Here, we recovered 16S rDNA clones from natural waters that were close phylogenetic relatives of the markers. From the sequence data, we designed specific PCR primers that discriminate human and ruminant sources of fecal contamination.

[1]  B A Wiggins,et al.  Discriminant analysis of antibiotic resistance patterns in fecal streptococci, a method to differentiate human and animal sources of fecal pollution in natural waters , 1996, Applied and environmental microbiology.

[2]  Gerwin C. Raangs,et al.  Variations of Bacterial Populations in Human Feces Measured by Fluorescent In Situ Hybridization with Group-Specific 16S rRNA-Targeted Oligonucleotide Probes , 1998, Applied and Environmental Microbiology.

[3]  Hans H. Cheng,et al.  Characterization of microbial diversity by determining terminal restriction fragment length polymorphisms of genes encoding 16S rRNA , 1997, Applied and environmental microbiology.

[4]  J. T. Staley,et al.  Survival and detection of Bacteroides spp., prospective indicator bacteria , 1985, Applied and environmental microbiology.

[5]  R. B. Reneau,et al.  Determining Sources of Fecal Pollution in a Rural Virginia Watershed with Antibiotic Resistance Patterns in Fecal Streptococci , 1999, Applied and Environmental Microbiology.

[6]  C. Kreader,et al.  Design and evaluation of Bacteroides DNA probes for the specific detection of human fecal pollution , 1995, Applied and environmental microbiology.

[7]  D. Stickler,et al.  An assessment of Bacteroides fragilis group organisms as indicators of human faecal pollution. , 1985, Journal of Applied Bacteriology.

[8]  Awwa,et al.  Standard Methods for the examination of water and wastewater , 1999 .

[9]  N. Saitou,et al.  The neighbor-joining method: a new method for reconstructing phylogenetic trees. , 1987, Molecular biology and evolution.

[10]  Katharine G. Field,et al.  Identification of Nonpoint Sources of Fecal Pollution in Coastal Waters by Using Host-Specific 16S Ribosomal DNA Genetic Markers from Fecal Anaerobes , 2000, Applied and Environmental Microbiology.

[11]  S. Giovannoni,et al.  Kinetic Bias in Estimates of Coastal Picoplankton Community Structure Obtained by Measurements of Small-Subunit rRNA Gene PCR Amplicon Length Heterogeneity , 1998, Applied and Environmental Microbiology.

[12]  Ross A. Overbeek,et al.  The ribosomal database project , 1992, Nucleic Acids Res..

[13]  R. A. Conway,et al.  Use of Antibiotic Resistance Analysis To Identify Nonpoint Sources of Fecal Pollution , 1999, Applied and Environmental Microbiology.

[14]  W. Moore,et al.  Human fecal flora: variation in bacterial composition within individuals and a possible effect of emotional stress , 1976, Applied and environmental microbiology.