Inactivation kinetics of nitrite-oxidizing bacteria by free nitrous acid

Recent studies have shown that free nitrous acid (FNA, i.e., HNO₂) is biocidal to many microorganisms, promoting the development of FNA-based technology in biological wastewater treatment. Suppression of nitrite-oxidizing bacteria (NOB) is a critical step for autotrophic nitrogen removal via anammox. In this study, the biocidal effect of FNA on NOB was determined by developing a model methodology combined with NOB incubation. Sixteen groups of FNA exposure tests were conducted at five different FNA concentrations from 0 to 4 mg HNO₂-N/L, obtained from three pH values (5.0, 5.5 and 6.0) with nitrite ranged from 21 to 1680 mg NO₂^--N/L, with one as a control. Nitrate production curves were tracked during incubations of the FNA-exposed sludge, and then used to estimate active NOB concentrations by the kinetic model-based fitting. The results showed that with 24-hour exposure to FNA at a level of over 1 mg HNO₂-N/L, the active NOB decreased around two orders of magnitude compared with that in the primordial sludge. The Weibull model can well describe the biocidal effect, which would be useful for the optimization of FNA conditions. The maximum NOB growth rate was increased after FNA exposure. This result suggests that long-term implementation of FNA-based technology can select fast-growing NOB in activated sludge, causing a 'NOB adaptation' issue.