Biofilm Responses to Toxic Shocks in Closed Pipes: Using Nitrous Oxide Emissions as an Early Warning of Toxicity Ahead of a Wastewater Treatment Works

Wastewater treatment works can receive toxic substances that can kill microorganisms responsible for waste degradation. Implementation of toxicity monitors in-sewer, as part of an early warning system to help prevent toxic substances entering treatment works, is, however, very rare. This work presents results from a pilot-scale study using an in-sewer early warning system based on detection of nitrous oxide (N₂O) gas emitted by nitrifying bacteria naturally present in sewer biofilm. Nitrous oxide has potential to be an indicator of nitrification inhibition as it is typically emitted when nitrifiers are under stress. The biofilm was allowed to develop over 14 days under fixed wastewater flow and level. Presence of nitrifying bacteria was verified on day 13 followed by a 90 min toxic shock on day 14 by four different known nitrification inhibitors. Pre-shock nitrification rates averaged 0.78 mg-NH₄⁺-N mg-VS⁻¹ d⁻¹and were significantly reduced post shock to <0.2 mg-NH₄⁺-N mg-VS⁻¹ d⁻¹. Nitrous oxide emissions were found to vary with influent wastewater quality, suggesting a more complex data processing algorithm is needed instead of a simple threshold emission value. The extent of nitrification inhibition differed from the recorded response for suspended growth biomass with allylthiourea resulting in a 77 and 81 % nitrification inhibition for literature suspended growth EC₅₀and EC₇₅concentrations, respectively. Results from this study suggest nitrifying biofilms in closed pipes can be used as part of an early warning system but will likely require amplification of the response to be of practical use. Further research is required to better understand the biofilm response and calibrate the early warning system for differentiating its unique baseline from true toxicity events.