Ammonia nitrogen affects bacterial virulence and conditional pathogenic bacterial growth by regulating biofilm...

The control of conditional pathogenic bacteria and inhibition of their virulence factors (VFs) in drinking water distribution systems (DWDSs) is vital for drinking water safety. This study adopted two groups of DWDSs to investigate how ammonia nitrogen affects bacterial VFs and conditional pathogenic bacterial growth in biofilms. Our results indicated that Acidimicrobium (95,916.62 ± 119.24 TPM), Limnohabitans (30,338.81 ± 139.14 TPM), and Sediminibacterium (10,658.01 ± 48.94 TPM) were predominant in the biofilm bacterial community of DWDSs with NH₃-N addition. Under these conditions, the abundances of various bacterial metabolites, such as L-glutamate (1.45-fold), 2-oxoglutarate (1.24-fold), pyruvate (2.10-fold), and adenosine monophosphate (AMP, 5.29-fold), were significantly upregulated, which suggested the upregulation of amino acid, carbohydrate, nucleotide, lipid, pyrimidine and purine metabolism. These metabolic pathways accelerated extracellular polymeric substance (EPS) secretion. The protein concentration in EPS also increased to 187.59 ± 0.58 μg/cm². The increased EPS secretion promoted the amide I CO group of the EPS protein to interact with the surface of the DWDSs, thus enhancing the ability of bacteria (especially conditional pathogenic bacteria) to adhere to the pipe surface to form biofilms. Due to EPS protection, the abundance of the adherence subtype of VFs and the plate counts of Pseudomonas aeruginosa increased to 5912.8 ± 21.89 TPM and 655.78 ± 27.10 CFU/cm², respectively. Therefore, NH₃-N in DWDSs increased bacterial VFs levels and promoted the growth of some conditional pathogenic bacteria by regulating biofilm microbial metabolic pathways and EPS secretion, ultimately impacting the interaction between EPS and the pipe surface.