Effect of GAC pre-treatment and disinfectant on microbial community structure and opportunistic pathogen occurrence

Opportunistic pathogens in potable water systems are an emerging health concern; however, the factors influencing their proliferation are poorly understood. Here we investigated the effects of prior granular activated carbon (GAC) biofiltration [GAC-filtered water, unfiltered water, and a blend (30% GAC filtered and 70% unfiltered water)] and disinfectant type (chlorine, chloramine) on opportunistic pathogen occurrence using five annular reactors (ARs) to simulate water distribution systems, particularly premise plumbing. GAC pre-treatment effectively reduced total organic carbon (TOC), resulting in three levels of influent TOC investigated. Quantitative polymerase chain reaction (q-PCR) provided molecular evidence of natural colonization of Legionella spp., Mycobacterium spp., Acanthamoeba spp., Hartmannella vermiformis and Mycobacterium avium on AR coupons. Cultivable mycobacteria and amoeba, including pathogenic species, were also found in bulk water and biofilm samples. While q-PCR tends to overestimate live cells, it provided a quantitative comparison of target organisms colonizing the AR biofilms in terms of gene copy numbers. In most cases, total bacteria and opportunistic pathogens were higher in the three undisinfected ARs, but the levels were not proportional to the level of GAC pre-treatment/TOC. Chlorine was more effective for controlling mycobacteria and Acanthamoeba, whereas chloramine was more effective for controlling Legionella. Both chlorine and chloramine effectively inhibited M. avium and H. vermiformis colonization. Pyrosequencing of 16S rRNA genes in coupon biofilms revealed a significant effect of GAC pre-treatment and disinfectant type on the microbial community structure. Overall, this study provides insights into the potential of different disinfectants and GAC biofilters at the treatment plant and in buildings to control downstream opportunistic pathogens and broader drinking water microbial communities. [Display omitted] •Colonization of drinking water biofilms by Legionella spp., Mycobacterium spp., and Acanthamoeba spp. was investigated.•Effects of upstream bio-granular activated carbon pre-treatment and low doses of chorine versus chloramine were compared.•Microbial community was characterized by quantitative polymerase chain reaction and pyrosequencing.•Differences in opportunistic pathogens and microbial community structure were observed based on influent water treatment.•No water treatment method was effective for controll