Laboratory microcosm experiments were used to assess the effects of environmental parameters on the persistence of the Bacteroides 16S rRNA genes derived from equine fecal samples in stream water to investigate the utility of Bacteroides spp. as fecal indicator organisms. Quantitative real-time polymerase chain reaction (qPCR) was used to measure gene concentrations over time with treatments designed to compare filtered vs. unfiltered stream water, fecal aggregate size, initial fecal concentrations, and water temperatures. Comparison of Bacteroides16S rRNA genes/mL in microcosms constructed with unfiltered stream water and filtered stream water indicated that stream water filtration to remove indigenous microorganisms followed by temperature had the largest effects on gene persistence. First-order exponential decay functions were fitted to the data from each microcosm constructed using unfiltered stream water, and the decay constants (k) ranged from 0.0071 h(-1) in the microcosms incubated at 5 degrees C to 0.0336 h(-1) in a set of microcosms incubated at 25 degrees C. Analysis of k calculated from the 10 experimental treatments indicated that k is more highly correlated to temperature than initial Bacteroides 16S rRNA gene starting concentrations. The equation resulting from graphing k (as the dependent variable) vs. temperature (as the independent variable) best fit a peak, Gaussian, 3 parameter function with a maximum decay at 30 degrees C, a r(2) of 0.83 and all parameters were significant (P < 0.0015). Thus this data suggest that factors that reduce biological activity, such as physical removal of stream microorganisms by filtration and low temperature, result in slower Bacteroides 16S rRNA gene decay.