Corroding copper and steel exposed to intermittently flowing tap water promote biofilm formation and growth of Legionella pneumophila.

The information about the impact of copper pipes on the growth of Legionella pneumophila in premise plumbing is controversial. For this reason, pipe segments of copper, stainless steel (SS), mild steel (MS), polyethylene, chlorinated polyvinylchloride (CPVC) and glass (controls) were exposed to intermittently flowing (20 min stagnation time) nonchlorinated tap water of 37 °C or 16 °C (ambient temperature) during six months to study the impact of metals on biofilm formation and growth of L. pneumophila. Biofilm concentrations (BfC, measured as ATP) on copper were 3 (at 37 °C) to 6 (at 16 °C) times higher than on SS. The maximum colony counts of L. pneumophila on the materials tested at 37 °C showed a quadratic relationship with the associated BfCs, with highest values on copper and MS. The average Cu concentration on the glass control of copper (glass-copper) was more than two log units lower than the Fe concentration on glass-MS, suggesting that copper released less corrosion by-products than MS. The release of corrosion by-products with attached biomass from MS most likely enhanced biofilm formation on glass-MS. Cloning and 16S RNA gene sequence analysis of the predominating biofilm bacteria revealed that an uncultured Xanthobacteraceae bacterium and Reyranella accounted for 75% of the bacterial community on copper at 37 °C. The nitrite-oxidizing Nitrospira moscoviensis, which can also utilize hydrogen (H) and formate, accounted for >50% of the bacterial abundance in the biofilms on MS and glass-MS at 37 °C. The predominating presence of the strictly anaerobic non-fermentative Fe(III)-reducing Geobacter and the Fe(II)-oxidizing Gallionella on MS exposed to tap water of 16 °C indicated anoxic niches and the availability of H, low molecular weight carboxylic acids (LMWCAs) and Fe(II) at the MS surface. LMWCAs likely also promoted bacterial growth on copper, but the release mechanisms from natural organic matter at the surface of corroding metals are unclear. The effects of water stagnation time and flow dynamics on biofilm formation on copper requires further investigation.