Expression of the nirS, hzsA, and hdh Genes in Response to Nitrite Shock and Recovery in Candidatus Kuenenia stuttgartiensis

In this study, Candidatus Kuenenia stuttgartiensis were subjected to distinct nitrite shocks (66 (control), 200, 300, 400, and 500 mg N/L), and the responses of mRNA levels of cytochrome cd₁ nitrite/nitric oxide oxidoreductase (nirS), hydrazine synthase (hzsA), and hydrazine dehydrogenase (hdh) were assessed. Changes in the hydrazine dehydrogenase (HDH) protein level were monitored. At 200 mg NO₂^--N/L, the normalized specific anaerobic ammonium-oxidizing activity (nSAA) slightly increased relative to the control despite a significant decrease in nirS, hzsA, and hdh mRNA levels. When nitrite increased to 300 and 400 mg N/L, increased nirS, hzsA, and hdh mRNA levels were observed, but the nSAA decreased, relative to the 200 mg NO₂^--N/L exposure. HDH protein detection revealed that Candidatus Kuenenia stuttgartiensis attempted to yield high enzyme levels by stimulating mRNA synthesis to resist the nitrite-induced stress. On 500 mg NO₂^--N/L shock, the nirS, hzsA, and hdh mRNA levels decreased, alongside decreased nSAA and HDH levels. Although the mRNA levels did not always coincide with activities, our findings advance understanding of the mechanisms that anammox bacteria use to cope with nitrite inhibition at the transcriptional and translational levels, which will improve the diagnostic accuracy of bioreactor failures when nitrite accumulation occurs.