Influence of detachment on substrate removal and microbial ecology in a heterotrophic/autotrophic biofilm

Competition between heterotrophic bacteria oxidizing organic substrate and autotrophic nitrifying bacteria in a biofilm was evaluated. The biofilm was grown in a tubular reactor under different shear and organic substrate loading conditions. The reactor was initially operated without organic substrate in the influent until stable ammonia oxidation rates of 2.1 g N/(m2 d) were achieved. A rapid increase of fluid shear in the tubular reactor on day 156 resulted in biofilm sloughing, reducing the biofilm thickness from 330 to 190 μm. This sloughing event did not have a significant effect on ammonia oxidation rates. The addition of acetate to the influent of the reactor resulted in decreased ammonia oxidation rates (1.8 g N/(m2 d)) for low influent acetate concentrations (17 mg COD/L) and the breakdown of nitrification at high influent acetate concentrations (55 mg COD/L). Rapidly increasing fluid shear triggered biofilm sloughing in some cases—but maintaining constant shear did not prevent sloughing events from occurring. With the addition of acetate to the influent of the reactor, the biofilm thickness increased up to 1350 μm and individual sloughing events removed up to 50% of the biofilm. Biofilm sloughing had no significant influence on organic substrate removal or ammonia oxidation. During 325 days of reactor operation, ammonia was oxidized only to nitrite; no nitrate production was observed. This lack of nitrite oxidation was confirmed by fluorescent in situ hybridization (FISH) analysis, which detected betaproteobacterial ammonia oxidizers but not nitrite oxidizers. Mathematical modeling correctly predicted breakdown of nitrification at high influent acetate concentrations. Model predictions deviated systematically from experimental results, however, for the case of low influent acetate concentrations.