Microbial ecology of nitrifying simulated premises plumbing

Abstract

Because of the Stage 2 Disinfectants and Disinfection Rule limiting then concentration of disinfection by products in drinking water, the use of chloramine as an alternative to chlorine has been increasing. However, the ammonia introduced by chloramination can lead to nitrification which results in the production of nitrite and nitrate, leading to regulatory violations. Nitrification in reactors with copper and polyvinyl chloride (PVC) surfaces was established by indigenous organisms from Bozeman tap water and has been stably maintained for more than 6 years. Statistical analyses of polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) profiles determined that the active bacterial populations were different in the two systems. The assemblage of the organisms was also different from the starting population (BAC influent) suggesting both material and ammonia/carbon source affect the population. No known ammonia oxidizing bacteria were detected suggesting the role of different group for ammonia oxidation. Fluorescence in situ hybridization (FISH) detected archaea in the biofilm from both reactors. Archaeal 16S rRNA gene sequences were found to be phylogenetically affiliated with known archaeal ammonia oxidizers. Two archaeal amoA sequences were amplified from the system as determined by DGGE. We propose to provisionally classify a detected archaeon as Candidatus Nitrosotenuis bozemanii, based on its affinity to Nitrosotenuis uzonensis (Hatzenpichler et al., in preparation). Bacterial abundances were comparable in the two systems but archaeal abundances were higher in the PVC reactor suggesting material effect on the overall microbial population composition and density. Enrichment in modified synthetic Crenarchaeota medium yielded a culture of archaea and bacteria that consistently oxidizes ammonia to nitrate. Attempts to isolate the archaeal component using antibiotics failed, suggesting the disruption of a possible beneficial relationship between the archaea and bacteria. Genes involved in the transformation of nitrogen within the system were also investigated and hao distantly related to that of ammonia oxidizing bacteria was detected but its potential role remains unknown. This study provides evidence of archaea associated with biofilms in drinking water and while further analysis is needed to definitively elucidate their role, results of this study prompts the reevaluation of the current concept of nitrification in drinking water.