The inhibitory effects of nitrite (NO2-)/free nitrous acid (HNO2-FNA) on the metabolism of Nitrobacter were investigated using a method allowing the decoupling of the growth and energy generation processes. A lab-scale sequencing batch reactor was operated forthe enrichment of a Nitrobacter culture. Fluorescent in situ hybridization (FISH) analysis showed that 73% of the bacterial population was Nitrobacter. Batch tests were carried out to assess the oxygen and nitrite consumption rates of the enriched culture at low and high nitrite levels, in the presence or absence of inorganic carbon. It was observed that in the absence of CO2, the Nitrobacter culture was able to oxidize nitrite at a rate that is 76% of that in the presence of CO2, with an oxygen consumption rate that is 85% of that measured in the presence of CO2. This enabled the impacts of nitrite/FNA on the catabolic and anabolic processes of Nitrobacter to be assessed separately. FNA rather than nitrite was likely the actual inhibitor to the Nitrobacter metabolism. It was revealed that FNA of up to 0.05 mg HNO2-N x L(-1) (3.4 microM), which was the highest FNA concentration used in this study, did not have any inhibitory effect on the catabolic processes of Nitrobacter. However, FNA initiated its inhibition to the anabolic processes of Nitrobacterat approximately 0.011 mg HNO2-N x L(-1) (0.8 microM), and completely stopped biomass synthesis at a concentration of approximately 0.023 mg HNO2-N x L(-1) (1.6 microM). The inhibitory effect could be described by an empirical inhibitory model proposed in this paper, but the underlying mechanisms remain to be revealed.
In laboratory experiments, source-separated urine was stabilised with nitrification and denitrified via nitritation and anaerobic ammonium oxidation. The highest total ammonia concentration in the influent was 7,300 gN/m3, the maximum pH 9.2. In a moving bed biofilm reactor (MBBR) with Kaldnes biofilm carriers, we stabilised urine as a 1:1 ammonium nitrate solution. The maximum nitrification rate was 380 gN/m3/d corresponding to 1.7 gN/m2(biofilm)/d. Nitrite ammonium solutions were produced in a continuous flow stirred tank reactor (CSTR) with 4.8 days sludge retention time (SRT) at 30 degrees C and in a sequencing batch reactor (SBR) with more than 30 days SRT. Nitrate build-up was negligible in both reactors. Nitritation rates were 780 gN/m3/d in the CSTR and 280 gN/m3/d in the SBR, respectively. However, shortening the cycles would increase nitritation in the SBR. High concentrations of nitrous acid, salts, and presumably hydroxylamine suppressed nitrite oxidation in the nitritation reactors. In all three nitrification reactors, maximally 50% of the influent total ammonia was oxidised without pH control. None of the common inhibition or limitation approaches could explain why ammonia oxidation always stopped at pH values around 6. In a batch experiment, we showed that source-separated urine can be denitrified autotrophically by anammox bacteria.
In wastewater treatment plants with anaerobic sludge digestion, 15-20% of the nitrogen load is recirculated to the main stream with the return liquors from dewatering. Separate treatment of this ammonium-rich digester supernatant significantly reduces the nitrogen load of the activated sludge system. Two biological applications are considered for nitrogen elimination: (i) classical autotrophic nitrification/heterotrophic denitrification and (ii) partial nitritation/autotrophic anaerobic ammonium oxidation (anammox). With both applications 85-90% nitrogen removal can be achieved, but there are considerable differences in terms of sustainability and costs. The final gaseous products for heterotrophic denitrification are generally not measured and are assumed to be nitrogen gas (N2). However, significant nitrous oxide (N2O) production can occur at elevated nitrite concentrations in the reactor. Denitrification via nitrite instead of nitrate has been promoted in recent years in order to reduce the oxygen and the organic carbon requirements. Obviously this "achievement" turns out to be rather disadvantageous from an overall environmental point of view. On the other hand no unfavorable intermediates are emitted during anaerobic ammonium oxidation. A cost estimate for both applications demonstrates that partial nitritation/anammox is also more economical than classical nitrification/denitrification. Therefore autotrophic nitrogen elimination should be used in future to treat ammonium-rich sludge liquors.
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