Coliforms and other microbial indicators occurrence in water and biofilm in full-scale distribution systems.

Biofilm and microbial water quality were studied in four middle size full-scale distribution systems (DS) in France serving 5,000-30,000 inhabitants (maximum residence time 23-160h) through three sampling campaigns over 1 year. Three of these DSs were chosen because of a quite high occurrence of bacterial indicators (i.e. total coliforms), the last DS was considered as a reference. Biofilm was studied on cast iron coupons incubated for more than 1 month in devices continuously fed with water from the DS in conditions imitating those met in DS. The devices were located at different points (4-6) along each DS. The abundance of bacteria in biofilm was estimated by heterotrophic plate counts (HPC) after detachment of the biofilm from the support by sonication. Microbiological water quality was estimated in parallel; analysis of total coliforms, E. coli, enterococci and anaerobic sulphide-reducing bacteria spores (ASRB spores) was carried out in biofilm and water. Over the period of the study, 171 water samples and 57 biofilm samples were collected. Over these 171 waters, 19 (11%) were positive for at least one of the measured indicators while two biofilm samples were positive (3.5%). Significant differences were observed in the levels of contamination between the DSs. High residence time in the DS, low disinfectant residual and high temperature increased the risk of indicator occurrence in the water phase. Due to the low number of biofilm samples positive for bacterial indicators, the data collected in the present study did not allow observation of a direct association between biofilm and water contaminations, even if the occurrence of indicators in water appeared on DSs with the highest density of biofilm (HPC).

Influence of phosphate and disinfection on the composition of biofilms produced from drinking water, as measured by fluorescence in situ hybridization.

Biofilms were grown in annular reactors supplied with drinking water enriched with 235 microg C/L. Changes in the biofilms with ageing, disinfection, and phosphate treatment were monitored using fluorescence in situ hybridization. EUB338, BET42a, GAM42a, and ALF1b probes were used to target most bacteria and the alpha (alpha), beta (beta), and gamma (gamma) subclasses of Proteobacteria, respectively. The stability of biofilm composition was checked after the onset of colonization between T = 42 days and T = 113 days. From 56.0% to 75.9% of the cells detected through total direct counts with DAPI (4'-6-diamidino-2-phenylindole) were also detected with the EUB338 probe, which targets the 16S rRNA of most bacteria. Among these cells, 16.9%-24.7% were targeted with the BET42a probe, 1.8%-18.3% with the ALF1b probe, and <2.5% with the GAM42a probe. Phosphate treatment induced a significant enhancement to the proportion of gamma-Proteobacteria (detected with the GAM42a probe), a group that contains many health-related bacteria. Disinfection with monochloramine for 1 month or chlorine for 3 days induced a reduction in the percentage of DAPI-stained cells that hybridized with the EUB338 probe (as expressed by percentages of EUB338 counts/DAPI) and with any of the ALF1b, BET42a, and GAM42a probes. The percentage of cells detected by any of the three probes (ALF1b+BET42a+GAM42a) tended to decrease, and reached in total less than 30% of the EUB338-hybridized cells. Disinfection with chlorine for 7 days induced a reverse shift; an increase in the percentage of EUB338 counts targeted by any of these three probes was noted, which reached up to 87%. However, it should be noted that the global bacterial densities (heterotrophic plate counts and total direct counts) tended to decrease over the duration of the experiment. Therefore, those bacteria that could be considered to resist 7 days of chlorination constituted a small part of the initial biofilm community, up to the point at which the other bacterial groups were destroyed by chlorination. The results suggest that there were variations in the kinetics of inactivation by disinfectant, depending on the bacterial populations involved.

Effect of adding phosphate to drinking water on bacterial growth in slightly and highly corroded pipes.

The effect of phosphate addition in drinking water was tested under static conditions as batch tests and under dynamic conditions using continuously fed reactors. Phosphate supplements in batch tests from 0.1 to 2 mg P-PO4 L(-1) did not show any relationship between bacterial growth and phosphate concentration. Dynamic tests in slightly corroded reactor (stainless steel) treated at 1 mg P-PO4 L(-1) showed only a moderate improvement in the growth of microorganisms. On the contrary, phosphate treatment applied to the highly corroded reactor (unlined cast iron) led to an immediate, drastic drop in iron oxide release and bacterial production. Phosphate uptake by the reactor wall was less than 14% with the stainless-steel reactor and 70-90% with the corroded cast iron reactor. Moreover, about 5% of the phosphate associated to corroded iron pipe walls was released for 20 days after the end of treatment.