Changes in abundance of heterotrophic and coliform bacteria resident in stored water bodies in relation to incoming bacterial loads following rain events.

Microbial properties of harvested rainwater were assessed at two study sites at Newcastle on the east coast of Australia. The investigation monitored daily counts of heterotrophic bacteria (HPC), total coliforms and E. coli during a mid-winter month (July). Immediately after a major rainfall event, increases in bacterial loads were observed at both sites, followed by gradual reductions in numbers to prior baseline levels within 7 days. Baseline HPC levels ranged from 500-1000 cfu/mL for the sites evaluated, and the loads following rain peaked at 3590-6690 cfu/mL. Baseline levels of total coliforms ranged from 0-100 cfu/100 mL and peaked at 480-1200 cfu/100 mL following rain. At Site 1, there was no evidence of E. coli loading associated with the rain events assessed, and Site 2 had no detectable E.coli colonies at baseline, with a peak load of 17 cfu/100 mL following rain which again diminished to baseline levels. It was concluded that rainfall events contributed to the bacterial load in rainwater storage systems, but processes within the rainwater storage ensured these incoming loads were not sustained.

Extensive bacterial diversity indicates the potential operation of a dynamic micro-ecology within domestic rainwater storage systems.

The concept that domestic rainwater storage tanks may host sustainable microbial ecosystems has not previously been addressed. The bacterial diversity, cultivated from more than 80 samples from 22 tanks at various locations across eastern Australia, is presented here as prima facie evidence for the potential operation of a functional micro-ecology within rainwater storage systems. Cultivated isolates were found to comprise members of four major bacterial divisions; Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes, including more than 200 species from 80 different genera. The pattern of abundance distribution was typical of that observed in most natural communities, comprising a small number of abundant taxa and a multitude of rare taxa, while the specific composition resembled that previously described in a number of natural aquatic systems. Although Proteobacteria from alpha, beta and gamma sub-classes were dominant, a set of core taxa comprising representative genera from all four phyla could be identified. Coliform and other species specifically associated with faecal material comprised <15% of the species identified, and represented <1.5% of total average abundance. The composition of the cultivated populations and scope of diversity present, suggested that rainwater tanks may support functional ecosystems comprising complex communities of environmental bacteria, which may have beneficial implications for the quality of harvested rainwater.

Evaluating potential applications of faecal sterols in distinguishing sources of faecal contamination from mixed faecal samples.

Faecal samples from humans, herbivores, carnivores and birds as well as samples from septic tanks and effluents from a sewage treatment plant (STP) were extracted and analyzed by gas chromatography-mass spectrometry for faecal sterols including coprostanol, epicoprostanol, cholestanol, cholesterol, stigmasterol, campesterol, 24-ethylcoprostanol and beta-sitosterol. Coprostanol was observed in the highest concentrations from the human derived samples, but it was also present in substantial quantities in a range of herbivores. There was no unique marker of human faecal contamination. Multivariate analyses revealed that the faecal sterol profiles were significantly different between the four groups of animals (Wilks' lambda=0.007, P<0.002), and coprostanol and 24-ethylcoprostanol were the major discriminant factors. However, when faecal samples were mixed, the confounding of faecal sterol levels prevented accurate identification of contributing species. Conversely, faecal sterol ratios were highly efficient at identifying which mixtures contained human contribution, but could not appropriately determine percentage contributions of sources.

Comparisons of water quality parameters from diverse catchments during dry periods and following rain events.

In this study, 12 catchments sites located along the north coast of New South Wales in Australia were grouped into the four categories of septic, cattle, sewage treatment plant (STP) and forested sites via cluster analysis based on their land use patterns. Water samples from all these sites were collected between October 2004 and June 2006 at a regular monthly interval and within 48 h of rain events. The samples were analyzed for bacterial counts including faecal coliform and total coliform; faecal sterols including coprostanol, epicoprostanol, cholesterol, cholestanol, 24-ethylcoprostanol, campesterol, stigmasterol and beta-sitosterol; and the elements including Na, Rb, Sr, Ag, Cd, Sn, Cs, Ba, Hg, Tl, Pb, Bi, U, Mg, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, K, As, Se, P and Mo. Over the course of the sampling period, the STP site had the highest average coprostanol level of 1693+/-567 ng/L which was significantly higher (p<0.05) than the septic sites (190+/-71 ng/L), the cattle sites (163+/-94 ng/L) and forested sites (14+/-4 ng/L). As expected, the forested sites had significantly lower average level of faecal coliforms (373+/-87 cfu/100 mL) compared with the STP (1395+/-574 cfu/100 mL), septic (1243+/-494 cfu/100 mL) and cattle sites (535+/-112 cfu/100 mL). The concentrations of coprostanol were not correlated with the numbers of faecal coliform bacteria when the entire data set was evaluated. The forested sites generally had the lowest average levels of elemental compositions, with significantly lower levels noted for Na, U, Mg, V, Cu, Sr, K, As, P and Mo, whereas Fe was the only element notably higher in the forested sites. Temporal and rain events analyses of the data set revealed that elevated levels of both coprostanol and faecal coliforms were not exclusive to rain events. The average coprostanol levels in rain event samples at each site were not significantly different compared with the corresponding dry event samples. Conversely, faecal coliform numbers increased by 2-4 times in rain events samples from septic, cattle and forested sites, but did not alter in the STP site. Multivariate analyses identified coprostanol and Sr as major contributing factors for the discrimination of septic, cattle, STP and forested sites for both rain and dry events samples. It was clear that each land use type of catchment could be characterized by biochemical, bacteriological and elemental parameters.

Bacterial source tracking from diverse land use catchments by sterol ratios.

Water samples from sites potentially impacted by septic tanks, cattle, sewage treatment plant (STP) and natural forests were collected at regular monthly intervals and within 48 h of rainfall events between October 2004 and June 2006. All samples (n=296) were analysed for faecal coliforms and faecal sterols including coprostanol, epicoprostanol, cholestanol, cholesterol and 24-ethylcoprostanol. Faecal sterol ratios were used to assign human and/or herbivore contamination sources and to estimate their percentage relative contributions in water samples. The catchments had significantly different profiles of designated contamination origins (p<0.05), which were consistent with land use patterns. The STP impacted site had the highest incidence of human contamination assignations and the highest mean levels of coprostanol, whilst the forested site had the highest incidence of uncontaminated samples and the lowest mean concentration of coprostanol. Coprostanol concentrations were not always correlated with faecal coliform counts.