The microbial population and physicochemical process parameters of a sequencing batch reactor for nitrogen removal from urine were monitored over a 1.5-year period. Microbial community fingerprinting (automated ribosomal intergenic spacer analysis), 16S rRNA gene sequencing, and quantitative PCR on nitrogen cycle functional groups were used to characterize the microbial population. The reactor combined nitrification (ammonium oxidation)/anammox with organoheterotrophic denitrification. The nitrogen elimination rate initially increased by 400%, followed by an extended period of performance degradation. This phase was characterized by accumulation of nitrite and nitrous oxide, reduced anammox activity, and a different but stable microbial community. Outwashing of anammox bacteria or their inhibition by oxygen or nitrite was insufficient to explain reactor behavior. Multiple lines of evidence, e.g., regime-shift analysis of chemical and physical parameters and cluster and ordination analysis of the microbial community, indicated that the system had experienced a rapid transition to a new stable state that led to the observed inferior process rates. The events in the reactor can thus be interpreted to be an ecological regime shift. Constrained ordination indicated that the pH set point controlling cycle duration, temperature, airflow rate, and the release of nitric and nitrous oxides controlled the primarily heterotrophic microbial community. We show that by combining chemical and physical measurements, microbial community analysis and ecological theory allowed extraction of useful information about the causes and dynamics of the observed process instability.
The combination of nitritation and autotrophic denitrification (anammox) in a single sequencing batch reactor (SBR) is an energy efficient process for nitrogen removal from high-strength ammonia wastewaters. So far, the process has been successfully applied to digester supernatant. However, the process could also be suitable to treat source-separated urine, which has very high ammonium and organic substrate concentrations (up to 8,200 gN/m3 and 10,000 gCOD/m3). In this study, reactor performance was tested for digester supernatant and diluted source-separated urine. Ammonium concentrations in both solutions were similar (between 611 and 642 gN/m3), thus reactor performance could be directly compared. Differences were mainly due to higher activity of heterotrophic bacteria in urine. Nitrogen removal was slightly higher for source-separated urine, because heterotrophic bacteria denitrified the nitrate that was produced by anammox bacteria. In spite of higher heterotrophic growth with source-separated urine, calculated sludge concentrations at steady state were higher with digester supernatant due to accumulation of inert particulate organic matter from the influent. Although the sludge concentrations are less problematic for source-separated urine, process instabilities are more likely, because lower pH values are reached and heterotrophic denitrification can cause sudden increases of nitrite concentrations and/or nitric oxide. Both compounds inhibit aerobic ammonium oxidizing bacteria, heterotrophic bacteria and, most importantly, anammox bacteria. Nitrite and nitric oxide production by heterotrophic denitrification must be better understood to optimize nitritation/anammox for source-separated urine.
Electrodes for nitric and nitrous oxide have been on the market for some time, but have not yet been tested for an application in wastewater treatment processes. Both sensors were therefore assessed with respect to their (non)linear response, temperature dependence and potential cross sensitivity to dissolved compounds, which are present and highly dynamic in nitrogen conversion processes (nitric oxide, nitrous oxide, nitrogen dioxide, ammonia, hydrazine, hydroxylamine, nitrous acid, oxygen, and carbon dioxide). Off-gas measurements were employed to differentiate between cross sensitivity to interfering components and chemical nitric oxide or nitrous oxide production. Significant cross sensitivities were detected for both sensors: by the nitrous oxide sensor to nitric oxide and by the nitric oxide sensor to ammonia, hydrazine, hydroxylamine and nitrous acid. These interferences could, however, be removed by correction functions. Temperature fluctuations in the range of ±1 °C lead to artifacts of ±3.5% for the nitric oxide and ±3.9% for the nitrous oxide sensor and can be corrected with exponential equations. The results from this study help to significantly shorten and optimize the determination of the correction functions and are therefore relevant for all users of nitric and nitrous oxide electrodes.
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