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Factors Influencing the Persistence of Fecal Bacteroides in Stream Water

^a Dep. of Earth and Planetary Sciences, The Univ. of Tennessee Center for Environ. Biotechnology, The Univ. of Tennessee
^b Center for Environ. Biotechnology, The Univ. of Tennessee
^c Civil and Environ. Eng., The Univ. of Tennessee

View larger version (17K): [in this window] [in a new window]   Fig. 1. Master standard curve generated from 31 individual standard curves resulting in a linear equation y = –0.31x + 11.49, where y = log (Bacteroides 16S rRNA genes) and x = CT.

View larger version (31K): [in this window] [in a new window]   Fig. 2. Comparison of fecal particle size and the persistence of Bacteroides 16S rRNA genes from three separate slurries: coarse, medium, and fine. (A) visual comparison of the finest and coarsest particle size intervals evaluated in the particle size analysis, (B) size interval distribution for three aggregate size intervals (coarse, medium, and fine) as reflected in percent of total weight for each filtrate.

View larger version (17K): [in this window] [in a new window]   Fig. 3. Comparison of Bacteroides 16S rRNA genes/mL for fine, medium, and coarse fecal slurries in unfiltered water (experimental), in filtered water (filtered control) and stream water without fecal slurry (background control). All microcosms were incubated at 25°C. Error bars in the plots represent the standard deviations between triplicate microcosms for the microcosms containing fecal slurries and the standard deviations in triplicate PCR reactions for the filtered stream water control and the stream water only control. a* = samples from unfiltered stream water microcosms with significantly different Bacteroides 16S rRNA genes/ml concentrations than the corresponding stream water background controls. b* = samples from unfiltered stream water microcosms with significantly different Bacteroides 16S rRNA genes/mL concentrations than the corresponding filtered stream water control.

View larger version (17K): [in this window] [in a new window]   Fig. 4. Comparison of Bacteroides 16S rRNA genes/mL with initial starting fecal concentrations in microcosms of 10, 100, and 1000 mg/L in unfiltered water (experimental), filtered water (filtered control) and in stream water without fecal slurry (background control). All microcosms were incubated at 25°C. Error bars in the plots represent the standard deviations between triplicate microcosms for the unfiltered stream water microcosms containing fecal slurries and the standard deviations in triplicate PCR reactions for the filtered stream water microcosm containing fecal slurries and stream water only microcosm. a* = samples from unfiltered stream water microcosms with significantly different Bacteroides 16S rRNA genes/mL concentrations than the corresponding stream water background controls. b* = samples from unfiltered stream water microcosms with significantly different Bacteroides 16S rRNA genes/mL concentrations than the corresponding filtered stream water controls.

View larger version (39K): [in this window] [in a new window]   Fig. 5. Bacteroides 16S rRNA genes/mL in microcosms incubated with variable temperature ranging from 5 to 35°C. For each temperature three microcosms were used: (1) A fecal slurry of 100 mg/L in unfiltered stream water (experimental) (2) A fecal slurry of 100 mg/L in filtered stream water (filtered control), and (3) the unfiltered stream water without fecal slurry (background control). Error bars in the plots represent the standard deviations between triplicate microcosms for the microcosms containing fecal slurries and the standard deviations in triplicate PCR reactions for the stream water only microcosm. a* = samples from unfiltered stream water microcosms with significantly different Bacteroides 16S rRNA genes/mL concentrations than the corresponding stream water background controls. b* = samples from unfiltered stream water microcosms with significantly different Bacteroides 16S rRNA genes/mL concentrations than the corresponding filtered stream water controls.

View larger version (11K): [in this window] [in a new window]   Fig. 6. Relationship between decay rate (k) of the log concentration Bacteroides 16S rRNA genes/mL with temperature in microcosms in unfiltered stream water using a Peak, Gaussian, 3 Parameter equation (y = a x exp{–0.5 x [(x – x0)/b]2}) and a linear equation (y = y0 + a x x). The error bars represent the standard deviation of k for all experimental microcosms. For the peak Gaussian equation: Parameter a = 0.0304 ± 0.0032 (P = <0.001), parameter b = 15.2113 ± 3.0242 (P = 0.0015) and X0 = 29.8817 ± 3.4623 (P < 0.001). For the linear equation Y0 = 0.0057 ± 0.0032 (P = 0.2132) and a = 0.0009 ± 0.0002 (P = 0.0016).