One-year survey of opportunistic premise plumbing pathogens and free-living amoebae in the tap-water of one northern city of China

In this study, qPCR was used to quantify opportunistic premise plumbing pathogens(OPPPs) and free-living amoebae in 11 tap water samples collected over four seasons from a city in northern China. Results demonstrated that the average numbers of gene copies of Legionella spp. and Mycobacterium spp. were significantly higher than those of Aeromonas spp.(p < 0.05). Legionella spp. and Mycobacterium spp. were 100%(44/44) positively detected while P. aeruginosa and Aeromonas spp. were 79.54%(35/44) and 77.27%(34/44) positively detected. Legionella pneumophila was only detected in 4 samples(4/44), demonstrating its occasional occurrence. No Mycobacterium avium or Naegleria fowleri was detected in any of the samples. The average gene copy numbers of target OPPPs were the highest in summer,suggesting seasonal prevalence of OPPPs. Average gene copy numbers of OPPPs in the taps of low-use-frequency were higher than in taps of high-use-frequency, but the difference was not significant for some OPPPs(p > 0.05). Moderate negative correlations between the chlorine concentration and the gene copy numbers of OPPPs were observed by Spearman analysis(rsranged from -0.311 to -0.710, p < 0.05). However, no significant correlations existed between OPPPs and AOC, BDOC, or turbidity. Moderate positive correlations were observed between the target microorganisms, especially for Acanthamoeba spp., through Spearman analysis(p < 0.05). Based on our studies, it is proposed that disinfectant concentration, season, taps with different-use frequency, OPPP species, and potential microbial correlations should be considered for control of OPPPs in tap water.

Effects of O3/Cl2 disinfection on corrosion and opportunistic pathogens growth in drinking water distribution systems

The effects of O3/Cl2 disinfection on corrosion and the growth of opportunistic pathogens in drinking water distribution systems were studied using annular reactors （ARs）. The corrosion process and most probable number （MPN） analysis indicated that the higher content of iron-oxidizing bacteria and iron-reducing bacteria in biofilms of the AR treated with O3/Cl2 induced higher Fe304 formation in corrosion scales. These corrosion scales became more stable than the ones that formed in the AR treated with Cl2 alone. O3/Cl2 disinfection inhibited corrosion and iron release efficiently by changing the content of corrosion-related bacteria. Moreover, ozone disinfection inactivated or damaged the opportunistic pathogens due to its strong oxidizing properties. The damaged bacteria resulting from initial ozone treatment were inactivated by the subsequent chlorine disinfection. Compared with the AR treated with Cl2 alone, the opportunistic pathogens M. auium and L. pneumophila were not detectable in effluents of the AR treated with O3/Cl2, and decreased to （4.60 ± 0.14） and （3.09 ± 0.12） loglo （gene copies/g corrosion scales） in biofilms, respectively. The amoeba counts were also lower in the AR treated with O3/Cl2. Therefore, O3/Cl2 disinfection can effectively control opportunistic pathogens in effluents and biofilms of an AR used as a model for a drinking water distribution system.

Characterization of bacterial community and iron corrosion in drinking water distribution systems with O3-biological activated carbon treatment

Bacterial community structure and iron corrosion were investigated for simulated drinking water distribution systems（DWDSs） composed of annular reactors incorporating three different treatments： ozone, biologically activated carbon and chlorination（O3-BAC-Cl2）;ozone and chlorination（O3-Cl2）; or chlorination alone（Cl2）. The lowest corrosion rate and iron release, along with more Fe3O4 formation, occurred in DWDSs with O3-BAC-Cl2 compared to those without a BAC filter. It was verified that O3-BAC influenced the bacterial community greatly to promote the relative advantage of nitrate-reducing bacteria（NRB）in DWDSs. Moreover, the advantaged NRB induced active Fe（III） reduction coupled to Fe（II） oxidation, enhancing Fe3O4 formation and inhibiting corrosion. In addition, O3-BAC pretreatment could reduce high-molecular-weight fractions of dissolved organic carbon effectively to promote iron particle aggregation and inhibit further iron release. Our findings indicated that the O3-BAC treatment, besides removing organic pollutants in water, was also a good approach for controlling cast iron corrosion and iron release in DWDSs.

O_3-BAC-Cl_2: A multi-barrier process controlling the regrowth of opportunistic waterborne pathogens in drinking water distribution systems

Simulated drinking water distribution system(DWDS) treated with O_3-BAC-Cl_2(ozonebiological activated carbon-chlorine) was constructed to study its effects on the regrowth of five typical opportunistic pathogens(OPs). It was found that O_3-BAC-Cl_2 could significantly reduce the regrowth of target OPs in the effluents of DWDS compared with Cl_2 and O_3-Cl_2 with the same residual chlorine levels. However, the effect of O_3-BAC-Cl_2 on the average numbers of target OPs gene markers in the biofilms of DWDS was not apparent, suggesting that OPs in the biofilms of DWDS were tolerant to the upstream disinfection process. The quantification of target OPs in the BAC-filter column demonstrated that OPs decreased with the increase of depth, which was likely due to the organic nutrient gradient and microbial competition inside the BAC-filter. Increase in the ozone dose could further reduce the OPs at the bottom of the BAC-filter. Spearman correlation analysis demonstrated that some significant correlations existed between target microorganisms, suggesting potential microbial ecological relationships. Overall, our results demonstrated that the BAC-filter may act as a "battlefield"suppressing the OPs through microbial competition. O_3-BAC-Cl_2 could be an effective multibarrier process to suppress the proliferation of OPs in the bulk water of DWDS. However, OPs protected by the biofilms of DWDS should receive further attention because OPs may be detached and released from the biofilms.