Combining partial nitrification, granular activated carbon (GAC) filtration, and distillation is a well-studied approach to convert urine into a fertilizer. To evaluate the environmental sustainability of a technology, the operational carbon footprint and therefore nitrous oxide (N₂O) emissions should be known, but N₂O emissions from urine nitrification have not been assessed yet. Therefore, N₂O emissions of a decentralized urine nitrification reactor were monitored for 1 month. During nitrification, 0.4-1.2% of the total nitrogen load was emitted as N₂O-N with an average N₂O emission factor (EF_N2O) of 0.7%. Additional N₂O was produced during anoxic storage between nitrification and GAC filtration with an estimated EF_N2O of 0.8%, resulting in an EF_N2O of 1.5% for the treatment chain. N₂O emissions during nitrification can be mitigated by 60% by avoiding low dissolved oxygen or anoxic conditions and nitrite concentrations above 5 mg-N L^-1. Minimizing the hydraulic retention time between nitrification and GAC filtration can reduce N₂O formation during intermediate storage by 100%. Overall, the N₂O emissions accounted for 45% of the operational carbon footprint of 14 kg-CO_2,equiv kg-N^-1 for urine fertilizer production. Using electricity from renewable sources and applying the proposed N₂O mitigation strategies could potentially lower the carbon footprint by 85%.

Acid-tolerant ammonia-oxidizing bacteria (AOB) can open the door to new applications, such as partial nitritation at low pH. However, they can also be problematic because chemical nitrite oxidation occurs at low pH, leading to the release of harmful nitrogen oxide gases. In this publication, the role of acid-tolerant AOB in urine treatment was explored. On the one hand, the technical feasibility of ammonia oxidation under acidic conditions for source-separated urine with total nitrogen concentrations up to 3.5 g-N L^-1 was investigated. On the other hand, the abundance and growth of acid-tolerant AOB at more neutral pH was explored. Under acidic conditions (pH of 5), ammonia oxidation rates of 500 mg-N L^-1 d^-1 and 10 g-N g-VSS^-1 d^-1 were observed, despite high concentrations of 15 mg-N L^-1 of the AOB-inhibiting compound nitrous acid and low concentration of 0.04 mg-N L^-1 of the substrate ammonia. However, ammonia oxidation under acidic conditions was very sensitive to process disturbances. Even short periods of less than 12 h without oxygen or without influent resulted in a complete cessation of ammonia oxidation with a recovery time of up to two months, which is a problem for low maintenance applications such as decentralized treatment. Furthermore, undesirable nitrogen losses of about 10% were observed. Under acidic conditions, a novel AOB strain was enriched with a relative abundance of up to 80%, for which the name “Candidatus (Ca.) Nitrosacidococcus urinae” is proposed. While Nitrosacidococcus members were present only to a small extent (0.004%) in urine nitrification reactors operated at pH values between 5.8 and 7, acid-tolerant AOB were always enriched during long periods without influent, resulting in an uncontrolled drop in pH to as low as 2.5. Long-term experiments at different pH values showed that the activity of “Ca. Nitrosacidococcus urinae” decreased strongly at a pH of 7, where they were also outcompeted by the acid-sensitive AOB Nitrosomonas halophila. The experiment results showed that the decreased activity of “Ca. Nitrosacidococcus urinae” correlated with the limited availability of dissolved iron at neutral pH.

Urine nitrification is pH-sensitive due to limited alkalinity and high residual ammonium concentrations. This study aimed to investigate how the pH affects nitrogen conversion and the microbial community of urine nitrification with a pH-based feeding strategy. First, kinetic parameters for NH₃, HNO₂, and NO₂^- limitation and inhibition were determined for nitrifiers from a urine nitrification reactor. The turning point for ammonia-oxidizing bacteria (AOB), i.e., the substrate concentration at which a further increase would lead to a decrease in activity due to inhibitory effects, was at an NH₃ concentration of 12 mg-N L^-1, which was reached only at pH values above 7. The total nitrite turning point for nitrite-oxidizing bacteria (NOB) was pH-dependent, e.g., 18 mg-N L^-1 at pH 6.3. Second, four years of data from two 120 L reactors were analyzed, showing that stable nitrification with low nitrite was most likely between pH 5.8 and 6.7. And third, six 12 L urine nitrification reactors were operated at total nitrogen concentrations of 1300 and 3600 mg-N L^-1 and pH values between 2.5 and 8.5. At pH 6, the AOB Nitrosomonas europaea was found, and the NOB belonged to the genus Nitrobacter. At pH 7, nitrite accumulated, and Nitrosomonas halophila was the dominant AOB. NOB were inhibited by HNO₂ accumulation. At pH 8.5, the AOB Nitrosomonas stercoris became dominant, and NH₃ inhibited NOB. Without influent, the pH dropped to 2.5 due to the growth of the acid-tolerant AOB “Candidatus Nitrosacidococcus urinae”. In conclusion, pH is a decisive process control parameter for urine nitrification by influencing the selection and kinetics of nitrifiers.

Nitrification is well-suited for urine stabilization. No base dosage is required if the pH is controlled within an appropriate operating range by urine feeding, producing an ammonium-nitrate fertilizer. However, the process is highly dependent on the selected pH set-points and is susceptible to process failures such as nitrite accumulation or the growth of acid-tolerant ammonia-oxidizing bacteria. To address the need for a robust and reliable process in decentralized applications, two different strategies were tested: operating a two-position pH controller (inflow on/off) with a narrow pH control band at 6.20/6.25 (∆pH = 0.05, narrow-pH) vs. a wider pH control band at 6.00/6.50 (∆pH = 0.50, wide-pH). These variations in pH also cause variations in the chemical speciation of ammonia and nitrite and, as shown, the microbial production of nitrite. It was hypothesized that the higher fluctuations would result in greater microbial diversity and, thus, a more robust process. The diversity of nitrifiers was higher in the wide-pH reactor, while the diversity of the entire microbiome was similar in both systems. However, the wide-pH reactor was more susceptible to tested process disturbances caused by increasing pH or temperature, decreasing dissolved oxygen, or an influent stop. In addition, with an emission factor of 0.47%, the nitrous oxide (N₂O) emissions from the wide-pH reactor were twice as high as the N₂O emissions from the narrow-pH reactor, most likely due to the nitrite fluctuations. Based on these results, a narrow control band is recommended for pH control in urine nitrification.

Separation of urine at the source allows for the efficient recovery of nitrogen, phosphorus and other nutrients and reduces the load of these nutrients on wastewater treatment plants. Before urine can be used as a fertilizer, a treatment for nitrogen stabilization is required. Nitrification is a well-suited process for urine stabilization and is also the targeted approach in bioregenerative life support systems for Space such as in the MELiSSA project. However, due to the limited alkalinity and the high residual ammonium concentration, urine nitrification without alkalinity addition is very pH sensitive, which can compromise stable urine nitrification. Usually, the pH is controlled with the influent. Nevertheless, process failures can occur, namely nitrite accumulation, complete cessation of nitrification, and the growth of acid-tolerant ammonia-oxidizing bacteria (AOB), which can further lower the pH during periods of limited urine and lead to the emission of harmful nitrogen oxide gases. [...]

Die Urinseparierung ermöglicht eine effiziente Rückgewinnung von Stickstoff, Phosphor und anderen Nährstoffen und führt zur Entlastung der Kläranlagen. Bevor Urin als Düngemittel verwendet werden kann, ist jedoch eine Stickstoffbehandlung erforderlich. Dafür bietet sich die Nitrifikation an, welche auch im Rahmen der bioregenerative Lebenserhaltungssysteme für Weltraumanwendungen wie das MELiSSA Projekt untersucht werden. Aufgrund der begrenzten Alkalinität und der hohen Ammoniumkonzentration ist die Nitrifikation von Urin ohne Erhöhung der Alkalinität jedoch sehr pH-empfindlich, was die Stabilität der Urinnitrifikation gefährdet. Normalerweise wird der pH-Wert mit dem Zufluss geregelt. Dennoch kann es zu Prozessstörungen kommen, wie zur Akkumulation von Nitrit, zum vollständigen Erlegen der Nitrifikation und zum Wachstum säuretoleranter Ammoniakoxidierenden Bakterien (AOB), die den pH-Wert bei geringem Urinzuflus weiter senken und zur Emission schädlicher Stickoxidgase führen können. [...]