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Recent studies have shown that free nitrous acid (FNA) is biocidal to a broad range of microorganisms. Microorganisms residing in anaerobic sewer biofilms were found to be inactivated after a short (6-24 h) exposure to FNA. In this study, we investigate the inactivation kinetics of anaerobic sewer biofilms grown in real wastewater. Microbial viability of biofilms was determined using LIVE/DEAD staining. A two-fraction kinetic model was developed to simulate the inactivation of mixed culture in biofilms. The kinetic parameters were estimated by using Bayesian statistics. Model simulation found that a fraction (85 %) of the biofilm community was highly sensitive to FNA with a high inactivation rate, and a fraction (15 %) was tolerant to FNA and persisted after FNA treatment. This different susceptibility to FNA treatment was likely due to the diverse microbial community and biofilm protection. The fact that nearly 85 % microbes were inactivated confirmed that FNA is a strong biocide to mixed-culture biofilms. It was found that the inactivation rate constant was not affected by pH levels. The kinetic model was successfully used to optimize FNA dosage for sulfide control in sewer biofilms. Also, results suggest that a high FNA concentration is preferred than long exposure time to reduce the total chemical consumption. [ABSTRACT FROM AUTHOR]

Free nitrous acid (FNA) has been identified to be a ubiquitous inhibitor of a wide range of microorganisms, including bacteria involved in wastewater treatment. The FNA-induced inhibition on the anoxic (nitrite as electron acceptor) metabolism of denitrifying poly-phosphate accumulating organisms (DPAOs) was investigated using sludge from a sequencing batch reactor performing carbon, nitrogen, and phosphorus removal from synthetic wastewater. We found that FNA had a much stronger inhibitory effect on phosphorus (P) uptake and glycogen production than on poly-β-hydroxyalkanoate degradation and nitrite reduction. The intracellular adenosine triphosphate levels decreased sharply during the FNA incubation, and the decreasing rates were positively correlated with increasing FNA concentrations. The electron transport activity of DPAOs when exposed to FNA displayed a similar trend. Further, at FNA concentrations above 0.044 mg HNO2-N/L, the anaerobic metabolism of DPAOs was initiated despite of the presence of nitrite, as evidenced by the release of phosphorus and the consumption of glycogen. DPAO metabolism did not recover completely from FNA inhibition in the subsequent FNA-free environment. The recovery rate depended on the concentration of FNA applied in the previous anoxic period. These results suggest that the inhibitory effects are diverse and may be attributable to different mechanisms operating simultaneously. [ABSTRACT FROM AUTHOR]

Free nitrous acid (FNA i.e. HNO 2 ) was revealed to be effective in enhancing biodegradability of secondary sludge. Also, nitrite-oxidizing bacteria were found to be more susceptible to FNA than ammonium-oxidizing bacteria. Based on these findings, a novel FNA-based sludge treatment technology is proposed to enhance energy recovery from wastewater/sludge. Energy analysis indicated that the FNA-based technology would make wastewater treatment become an energy generating process (yielding energy at 4 kWh/PE/y; kWh/PE/y: kilowatt hours per population equivalent per year), rather than being a large energy consumer that it is today (consuming energy at 24 kWh/PE/y). Importantly, FNA required for the sludge treatment could be produced as a by-product of wastewater treatment. This proposed FNA-based technology is economically and environmentally attractive, and can be easily implemented in any wastewater treatment plants. It only involves the installation of a simple sludge mixing tank. This article presents the concept of the FNA-based technology. [ABSTRACT FROM AUTHOR]

• Stable NOB suppression is maintained in MABR treating low-strength wastewater for > 200 days. • Acidic pH of 5.0–5.2 and in situ FNA of 1 mg N/L jointly result in NOB suppression. • An unprecedentedly high AOB activity at around 2.4 kg N/(m3 d) is achieved at a short HRT of a mere 30 min. • Acid-tolerant AOB in MABR-biofilms show different apparent kinetics and responses to environmental factors. Shortcut nitrogen removal holds significant economic appeal for mainstream wastewater treatment. Nevertheless, it is too difficult to achieve the stable suppression of nitrite-oxidizing bacteria (NOB), and simultaneously maintain the activity of ammonia-oxidizing bacteria (AOB). This study proposes to overcome this challenge by employing the novel acid-tolerant AOB, namely " Candidatus Nitrosoglobus", in a membrane-aerated biofilm reactor (MABR). Superior partial nitritation was demonstrated in low-strength wastewater from two aspects. First, the long-term operation (256 days) under the acidic pH range of 5.0 to 5.2 showed the successful NOB washout by the in situ free nitrous acid (FNA) of approximately 1 mg N/L. This was evidenced by the stable nitrite accumulation ratio (NAR) close to 100 % and the disappearance of NOB shown by 16S rRNA gene amplicon sequencing and fluorescence in situ hybridization. Second, oxygen was sufficiently supplied in the MABR, leading to an unprecedentedly high ammonia oxidation rate (AOR) at 2.4 ± 0.1 kg N/(m3 d) at a short hydraulic retention time (HRT) of a mere 30 min. Due to the counter diffusion of substrates, the present acidic MABR displayed a significantly higher apparent oxygen affinity (0.36 ± 0.03 mg O 2 /L), a marginally lower apparent ammonia affinity (14.9 ± 1.9 mg N/L), and a heightened sensitivity to FNA and pH variations, compared with counterparts determined by flocculant acid-tolerant AOB. Beyond supporting the potential application of shortcut nitrogen removal in mainstream wastewater, this study also offers the attractive prospect of intensifying wastewater treatment by markedly reducing the HRT of the aerobic unit. [Display omitted] [ABSTRACT FROM AUTHOR]

Abstract: In full-scale wastewater treatment systems, phosphorus removal typically occurs together with nitrogen removal. Nitrite, an intermediate of both the nitrification and denitrification processes, can accumulate in the reactor. The inhibitory effect of nitrite/free nitrous acid (FNA) on the aerobic metabolism of poly-phosphate accumulating organisms (PAOs) is investigated. A culture highly enriched (90 ± 5%) in Candidatus “Accummulibacter phosphatis”, a well-known PAO, was used to perform a series of batch experiments at various nitrite and pH levels. FNA was found to inhibit all key aerobic metabolic processes performed by PAOs, namely PHA oxidation, phosphate uptake, glycogen replenishment and growth. The inhibitory effect on the anabolic processes (growth, phosphate uptake and glycogen production) was much stronger than that on the catabolic processes (PHA oxidation). 50% inhibition on all anabolic processes occurred at FNA concentrations of approximately 0.5 × 10−3 mg HNO2–N/L (equivalent to 2.0 mg NO2 −–N/L at pH 7.0), while full inhibition occurred at FNA concentrations of approximately 6.0 × 10−3 mg HNO2–N. These concentrations could be found in full-scale wastewater treatment systems that achieve nitrogen removal via the nitrite pathway. In comparison, PHA oxidation remained at 40–50% of the highest rate at FNA concentrations in the range 2.0 × 10−3–10.0 × 10−3 mg HNO2–N/L. Interestingly, PAOs were able to reduce nitrite under aerobic conditions (DO ≈ 3 mg/L), with the rate increasing substantially with the FNA concentration. The inhibition on phosphate uptake was found to be reversible. [ABSTRACT FROM AUTHOR]

Sidestream sludge treatment approaches have been developed in recent years to achieve mainstream nitrite shunt or partial nitritation, where NOB are selectively inactivated by biocidal factors such as free nitrous acid (FNA) or free ammonium (FA) in a sidestream reactor. The existence of NOB in raw wastewater has been increasingly realized and could pose critical challenge to stable NOB suppressions in those systems. This study, for the first time, evaluated the impact of influent NOB on the NOB suppressions in a mainstream nitrite shunt system achieved through sidestream sludge treatment. An over 500-day sequential batch reactor operation with six experimental phases rigorously demonstrated the negative effects of influent NOB on mainstream NOB control. Continuously seeding of NOB contained in influent stimulated NOB community shifts, leading to different extents of ineffective NOB suppression. The role of primary wastewater treatment in NOB removal from raw wastewater was also investigated. Results suggest primary settling and High Rate Activated Sludge system could remove a large part of NOB contained in raw wastewater. Primary treatment for raw wastewater is necessary for ensuring stable mainstream NOB suppressions. Image 1 • Influent NOB challenge NOB suppression by stimulating community shift and resistance. • Primary settling substantially reduces NOB in raw wastewater. • High-rate activated sludge treatment substantially reduces NOB in raw wastewater. • This study expanded the current knowledge of mainstream NOB suppression. [ABSTRACT FROM AUTHOR]

Abstract Inline sludge treatment using free nitrous acid (FNA) was recently shown to be effective in establishing the nitrite pathway in a biological nitrogen removal system. However, the effects of FNA treatment conditions on the nitrite pathway performance remained to be investigated. In this study, three different FNA treatment frequencies (daily sludge treatment ratios of 0.22, 0.31 and 0.38, respectively), two FNA concentrations (1.35 mgN/L and 4.23 mgN/L, respectively) and two influent feeding regimes (one- and two-step feeding) were investigated in four laboratory-scale sequencing batch reactors. The nitrite accumulation ratio was positively correlated to the FNA treatment frequency. However, when a high treatment frequency was used e.g., daily sludge treatment ratio of 0.38, a significant reduction in ammonia oxidizing bacteria (AOB) activity occurred, leading to poor ammonium oxidation. AOB were able to acclimatise to FNA concentrations up to of 4.23 mgN/L, whereas nitrite oxidizing bacteria (NOB) were limited by an FNA concentration of 1.35 mgN/L over the duration of the study (up to 120 days). This difference in sensitivity to FNA could be used to further enhance nitrite accumulation, with 90% accumulation achieved at an FNA concentration of 4.23 mgN/L and a daily sludge treatment ratio of 0.31 in this study. However, this high level of nitrite accumulation led to increased N 2 O emission, with emission factors of up to 3.9% observed. The N 2 O emission was mitigated (reduced to 1.3%) by applying two-step feeding resulting in a nitrite accumulation ratio of 45.1%. Economic analysis showed that choosing the optimal FNA treatment conditions depends on a combination of the wastewater characteristics, the nitrogen discharge standards, and the operational costs. This study provides important information for the optimisation and practical application of FNA-based sludge treatment technology for achieving the mainstream stable nitrite pathway. Graphical abstract Unlabelled Image Highlights • Study result and analysis guide free nitrous acid treatment condition determination • Stable mainstream nitrite pathway reached with nitrite accumulation ratio over 90%. • Distinct nitrifying bacteria acclimation behaviours to high treatment concentration • Treatment frequency negatively related to in-reactor nitrifying bacteria activities. • N 2 O emission mainly from nitrifier denitrification, can be reduced by multi-feeding. [ABSTRACT FROM AUTHOR]

Abstract: Sludge treatment and disposal is one of the major challenges for biological wastewater treatment plants and can represent up to 60% of their total operating costs. This study presents a novel strategy based on free nitrous acid (FNA or HNO2) treatment to achieve sludge reduction. Two sequencing batch reactors treating synthetic domestic wastewater were used, with one serving as an experimental reactor and the other as a control. In the experimental system, 50% of the excess sludge was exposed to FNA at 2.0 mg N/L for 24–42 h in an FNA treatment unit and then returned to the parent reactor. The sludge wastage flow from the experimental reactor was adjusted to obtain a similar mixed liquor suspended solids concentration to that in the control reactor, which was operated at a sludge retention time of 15 days. It was found that sludge production in the experimental system was 28% lower in comparison to the control. The addition of the FNA-treated sludge did not negatively affect the treatment performance and sludge properties of the experimental reactor, namely the organic carbon and nitrogen removal, nitrous oxide (N2O) emission and sludge settleability. Endogenous denitrification batch tests indicate that FNA-treated sludge is available as a carbon source for denitrification. [Copyright &y& Elsevier]

• An acidic aerobic digestion system treating anaerobically-digested sludge was established. • The acidic condition (4.8 ± 0.2) was self-sustained by Ca. Nitrosoglobus. • The total solids of anaerobically-digested sludge was markedly reduced by 22.6 ± 5.5%. • The stabilization level of anaerobically-digested sludge was improved to class A. Anaerobic digestion is a commonly used process for the reduction and stabilization of wasted activated sludge generated in wastewater treatment plants. However, anaerobically-digested (AD) sludge is still a problematic waste stream due to its large volume and often poor quality. In this study, two aerobic digesters were set up to treat anaerobically-digested sludge, with one digester operated in self-generated acidic condition as the experimental reactor, and one at neutral pH as the control reactor. The acidic condition in the experimental reactor was driven by an inoculated special ammonia-oxidizing bacterium, ' Candidatus Nitrosoglobus', which can tolerate low pH. As a result of ammonium oxidation by Ca. Nitrosoglobus, the pH decreased to 4.8 ± 0.2 and nitrite accumulated to and stayed at 200.0 ± 17.2 mg N L−1, from which free nitrous acid (FNA) at 8.5 ± 1.8 mg HNO 2 N L−1 formed in-situ. As a combined effect of low pH and high concentration of FNA, the experimental reactor reduced the total solids (TS), volatile solids (VS) and non-volatile solids (NVS) in the AD sludge by 25.2 ± 7.0%, 29.8 ± 4.3%, and 22.6 ± 5.5%, respectively. In contrast, the control reactor without Ca. Nitrosoglobus inoculation (operated at a near-neutral pH of 6.8 ± 0.3 and no FNA formation) only reduced VS in the AD sludge by 10.4 ± 4.3%, along with negligible NVS reduction. Additionally, the acidic aerobic digestion in the experimental reactor significantly stabilized AD sludge, decreasing the specific oxygen uptake rate (SOUR) to 0.5 ± 0.1 mg O 2 g−1VS h−1 and the most probable number (MPN) of Faecal Coliforms to 2.4 ± 0.1 log(MPN g−1TS), both of which meet USEPA standards for Class A biosolids. In comparison, the control reactor produced biosolids at Class B level only, with an SOUR of 1.8 ± 0.2 mg O 2 g−1VS h−1 and a Faecal Coliforms MPN of 3.6 ± 0.1 log(MPN g−1TS). By reducing the volume and improving the quality of the AD sludge, the acidic aerobic digestion of AD sludge enabled by Ca. Nitrosoglobus has the potential to significantly save the sludge disposal costs in wastewater treatment. [Display omitted] [ABSTRACT FROM AUTHOR]

Free nitrous acid (FNA), the protonated form of nitrite, has historically been an unwanted substance in wastewater systems due to its inhibition on a wide range of microorganisms. However, in recent years, advanced understanding of FNA inhibitory and biocidal effects on microorganisms has led to the development of a series of FNA-based applications that improve wastewater management practices. FNA has been used in sewer systems to control sewer corrosion and odor; in wastewater treatment to achieve carbon and energy efficient nitrogen removal; in sludge management to improve the sludge reduction and energy recovery; in membrane systems to address membrane fouling; and in wastewater algae systems to facilitate algae harvesting. This paper aims to comprehensively and critically review the current status of FNA-based applications in improving wastewater management. The underlying mechanisms of FNA inhibitory and biocidal effects are also reviewed and discussed. Knowledge gaps and current limitations of the FNA-based applications are identified; and perspectives on the development of FNA-based applications are discussed. We conclude that the FNA-based technologies have great potential for enhancing the performance of wastewater systems; however, further development and demonstration at larger scales are still required for their wider applications. Image 1 • Underlying mechanisms of FNA inhibitory and biocidal effects are reviewed. • FNA is used to control sewer corrosion and odor, and membrane fouling. • FNA improves wastewater treatment, sludge management and algae harvesting. • Knowledge gaps and current limitations of the FNA-based applications are identified. • Perspectives on the development of FNA-based applications are discussed. [ABSTRACT FROM AUTHOR]