Anammox Activity Research Articles

Resistance of anammox granular sludge to copper nanoparticles and oxytetracycline and restoration of performance

Nanoparticles and antibiotics, the two most frequently detected emerging pollutants from different wastewater sources, are eventually discharged into wastewater treatment plants. In this study, the widely used materials CuNPs and oxytetracycline (OTC) were selected as target pollutants to investigate their joint effects on anaerobic ammonium oxidation (anammox). The results indicated that the environmental concentration slightly inhibited the performance of the reactors, while the performance rapidly deteriorated within a week under high-level combined shocks (5.0 mg L−1 CuNPs and 2.0 mg L−1 OTC). After the second shock (2.5 mg L−1 CuNPs and 2.0 mg L−1 OTC), the resistance of anammox bacteria was enhanced, with an elevated relative abundance of Candidatus Kuenenia and absolute abundance of hzsA, nirS, and hdh. Moreover, the extracellular polymeric substance (EPS) content and specific anammox activity (SAA) showed corresponding changes. Improved sludge resistance was observed with increasing CuNP and OTC doses, which accelerated the recovery of performance.

A novel strategy for anammox consortia preservation: Transformation into anoxic sulfide oxidation consortia

The preservation of anaerobic ammonia oxidation (anammox) consortia is crucial for the rapid start-up and the process stability of the anammox based bioreactor. This work proposed and evaluated the feasibility of an anammox consortia preservation strategy, in which the anammox sludge was transformed into intermediate anoxic sulfide oxidation (ASO) functional microorganisms. Initially, the ASO process was successfully started up by inoculating anammox sludge and the overall sulfide and nitrate removal rates stabilized at 57.5 ± 0.22 and 10.0 ± 0.18 kg m−3 day−1, respectively. Then, the bioreactor function was reversely transformed into anammox, whose nitrogen removal rate reached 1.68 kg m−3 day−1. Granule characteristics analysis revealed that both biomass and extracellular polymeric substance content returned to their original states after the reverse start-up. Although the population of Candidatus_Kuenenia was greatly declined during ASO process, its richness was successfully recovered after the reverse start-up of the anammox process. The inferred metagenomes analysis demonstrated that the shifts in functional microorganisms were related to variation in the main metabolic pathways. The specific activities of anammox and ASO both are regarded as key indicators for the successful start-up of bioreactor. This work revealed a novel technique for the preservation of anammox consortia and might be a potential strategy for overcoming the drawback of long start-up time.

A novel anammox reactor with a nitrogen gas circulation: performance, granule size, activity, and microbial community.

Anammox process was regarded to be one of the vital links to achieve energy-saving or energy-producing wastewater treatment plant. In the study, an anammox reactor with the nitrogen gas circulation was constructed to culture anammox granules, and the performance, granule size distribution, and microbial community were investigated. Dissolved oxygen loading is found to be an important factor for the start-up of the anammox process, and the nitrogen removal rate of 2.12kgNm-3day-1 was achieved under the average nitrogen loading rate of 2.6kgNm-3day-1. The activity test showed that the highest specific anammox activity of 345.9mgNgVSS-1day-1 was achieved for granules with size of 0.5-1.0mm. The Illumina high-throughput sequencing analysis revealed the consistent variation of Candidatus Brocadia and Denitratisoma abundance in granues of all sizes, suggesting possible synergistic mechanism between heterotrophic bacteria Denitratisoma and anammox bacteria Ca. Brocadia. Furthermore, the results indicated the reactor with the nitrogen gas circulation is an efficient strategy to start-up anammox.

The importance of integrated fixed film activated sludge reactor and intermittent aeration in nitritation-anammox systems: Understanding reactor optimization for lagoon supernatant treatment

A single stage nitritation-anammox process was studied in a continuous flow reactor fed with raw lagoon municipal-sludge supernatant containing 835 ± 45 mg N L−1 of NH4+-N in the feed. Reactor operation was divided into five periods for a total of 438 days. Application of intermittent aeration with a 105 min aeration phase followed by a 15 min non-aeration phase reduced the aerobic nitritation rate by 13% but enhanced the anammox activity by 1.9 times. Under the dual effect of IFAS and intermittent aeration, the nitrogen removal rate (NRR) was improved from 0.12 ± 0.012 kg N (m3 d)−1 to 0.28 ± 0.03 kg N (m3 d)−1, representing an increased removal of NH4+ from 56 ± 4.3% to 92.2 ± 2.1%. The evolution of bacterial activity and communities showed that anammox activity significantly improved with the addition of the non-aeration phase, and the AOB population was largely improved with an increase of suspended biomass concentration in the reactor. Microbial community composition through 16S rRNA gene sequencing analysis showed that Nitrosomonas was the predominant AOB in the flocs phase, while Candidatus Brocadia was the dominant anammox bacteria in the biofilm phase.

Impact of gas-water ratios on N2O emissions in biological aerated filters and analysis of N2O emissions pathways

Biological aerated filter (BAF) is a widely applied biofilm process for wastewater treatment. However, characteristics of nitrous oxide (N2O) production in BAF are rarely reported. In this study, two tandem BAFs treating domestic wastewater were built up, and different gas-water ratios were controlled to explore N2O production pathway. Results showed that N2O production increased with increasing gas-water ratio in both BAFs; higher gas-water ratio promoted more N2O releasing from hydroxylamine oxidation process. To improve nitrogen removal performance and reduce N2O emission, the optimal gas-water ratios for BAF1 and BAF2 were 5:1 and 1.5:1, respectively. Most of N2O was produced from ammonia oxidizing bacteria (AOB) denitrification and hydroxylamine oxidation in BAF1, and heterotrophic denitrification contributed to relieve N2O emission. In BAF2, N2O was emitted from AOB denitrification and hydroxylamine oxidation by 87.8% and 12.2%, respectively. Heterotrophic denitrification is a N2O sink in BAF, causing BAF1 produced less N2O than BAF2 with the same gas-water ratio. Enhancing heterotrophic denitrification and anaerobic ammonium oxidation (Anammox) activity could reduce the release of N2O in BAFs.

Role and application of quorum sensing in anaerobic ammonium oxidation (anammox) process: A review

Anaerobic ammonium oxidation (anammox) is a promising biological nitrogen removal process, due to its advantages of high efficiency and low cost. However, problems remain in the application, such as long startup period and susceptibility to environmental variations. Quorum sensing (QS), as a means of bacterial communication, attracts more attentions in regulating aggregation behavior and microbial density. Many physiological characteristics of anammox bacteria have been confirmed to be associated with QS, including specific anammox activity, growth rate and the production of extracellular polymeric substances (EPS), which directly affect the performance of anammox process. Therefore, a comprehensive understanding of the QS in anammox process is prerequisite. This work systematically reviewed the role and application of QS in the anammox process with the focus on mechanism. Additionally, current challenges and research needs were proposed.

A novel strategy for accelerating the recovery of a Fe(II)-inhibited anammox reactor by intermittent addition of betaine: Performance, kinetics and statistical analysis

In this manuscript, Fe(II) inhibition of anammox and its recovery were investigated, and the performance, kinetics and statistical features were comprehensively studied simultaneously. Anammox was suppressed and completely inhibited by the addition of 109.29 and 378.57 mg/L Fe(II), respectively, via uncompetitive inhibition. Nitrite inhibition of anammox was best fitted by the Edwards model and Aiba model. EDTA-2Na wash (0.5, 1.0, 1.5, and 2.0 mM) had a limited effect on anammox recovery, while the addition of 2.0 mM betaine accelerated anammox recovery. Prolonged betaine addition caused an unintended reduction of anammox activity, though it self-recovered after the withdrawal of betaine. The modified Boltzmann model most accurately simulated the processes of anammox recovery using the EDTA-2Na wash, betaine regulation and self-recovery, and the modified Stover-Kincannon model was able to assess the results of anammox recovery. The one-sample t-test was successfully applied to determine the effects of these three recovery strategies on inhibited anammox, which were short-term disinhibition or long-term recovery effects. The above-mentioned results demonstrate that an intermittent addition of betaine, which is a better alternative to frequently-used but poorly-degradable EDTA, may be a useful and environmentally friendly recovery strategy for Fe(II)-inhibited anammox reactor.

Optimizing of operation strategies of the single-stage partial nitrification-anammox process

To achieve efficient and stable nitrogen removal and low N2O emission in the single-stage partial nitrification (PN)-anaerobic ammonium oxidation (anammox) process, a sequence batch reactor (SBR) was operated for 223 days, the optimal operation strategies were investigated. Aeration rate control was applied to achieve the synergy between anammox and partial nitrification. The variation of specific anammox activity (SAA), specific ammonia oxidizing bacteria activity (SAOA), and specific nitrite oxidizing bacteria activity (SNOA) revealed that the anammox process was a rate-limiting step under low-strength ammonium loading condition. With the long-term acclimation, the anammox process showed more tolerance to substrate variation, in contrast, the inhibitory threshold concentration of partial nitrification to ammonium was decreased. The evolution of specific activities and substrate tolerance abilities of the functional microbes verified the changes in the nitrogen removal performance and proved that the aeration control strategy was efficient for operating a low nitrogen loading PN-anammox reactor. The nitrous oxide (N2O) isotopic composition analysis revealed that 73% of N2O was produced via the nitrifier denitrification pathway. N2O production could be mitigated by increasing the SAA in the PN-anammox process. The reveal of the synergy effect between functional microbes and the N2O pathway provide a guidance for operating a PN-anammox system for treating low nitrogen loading wastewater.

Molecular and stable isotopic evidence for anaerobic ammonium-oxidizing bacteria in tidal flow constructed wetlands

The process of anaerobic ammonium oxidation (anammox) plays a significant part in global nitrogen cycling, yet its depth-specific distribution and importance in tidal flow constructed wetlands (TF CWs) remained unclear. Currently, the abundance, biodiversity, vertical distribution, rates and potential role of the anammox process in two TF CWs of the Yangtze Estuary, China, were explored. The results indicated that anammox bacteria were widely distributed in the TF CWs, and Candidatus Brocadia was the dominant group among the anammox genera. The widespread presence of the anammox bacteria was observed at different water depths of the soil cores, and the abundance of total bacterial 16S rRNA and hzsB genes was 4.47–20.08 × 109 and 0.81–5.91 × 105 copies per gram of dry soil (copies g−1 ds), respectively. The homogenized soils were incubated with stable nitrogen isotope to confirm the rates of anammox in the TF CWs, and the anammox activities were 2.38–6.72 nmol of N2 per gram of dry soil per day (nmol N2 g−1 dry soil d−1), with contributions of 2.7–7.5% nitrogen gas production and anammox that was more active in the deeper layer of the two TF CWs.

Vacuum lyophilization preservation and rejuvenation performance of anammox bacteria

The storage of anaerobic ammonia oxidizing bacteria (anammox) plays an important role in the application of anammox. Glycerol, sodium alginate and DMSO were used as the cryoprotectant, and vacuum lyophilization was used to prepare the anammox bacteria powder. Simultaneously, the control experiment was set up with the same protectant and preservation time. Bacteria powders were preserved using vacuum lyophilization and preserved at 4°C for 60 days. During the 54 days of rejuvenation, the reactors that were inoculated with bacteria powder preserved by different methods showed significant difference. The results show that the anammox bacteria powder with 3wt% DMSO as the cryoprotectant and without the substrate solution presented the best rejuvenation effect. The average specific anammox activity was 115.84mg-N·(g VSS·d)-1 with an activity recovery rate of 89%, and its stoichiometric ratio (Rs and Rp) was 1.33 and 0.21, which were very close to the theoretical values. The vacuum lyophilization method for the long-term preservation of anammox bacteria was effective.

Evaluation of Sludge Digester Effluent as Feeding Solution for the Enrichment of Anammox Bacteria

AbstractThe aim of this study is to evaluate the feasibility of sludge digester effluent as feeding solution to enrich anaerobic ammonium oxidation (anammox) bacteria. The performance of the two parallel pilot scale‐upflow packed bed anammox reactors (UPBAn1 and UPBAn2) are examined in terms of the enrichment of anammox bacteria. The control experiment is set up conducting synthetic wastewater as feeding solution in the UPBAn1 reactor whereas, the sludge digester effluent is fed to the nitritation reactor and then the partially nitrated digester effluent to the UPBAn2 reactor. Anammox activities are evaluated by mass balances based on ammonium (NH4+), nitrite (NO2−), and nitrate (NO3−) analysis and NRR. Microbial community of anammox bacteria is analyzed using real‐time polymerase chain reaction (PCR). The results demonstrate that UPBAn 1 and UPBAn2 reactors are successfully enriched on days 64 and 40 with NRRs of 19.54 and 19.43 g N m−3 per day, respectively. This study reveals that both synthetic wastewater and digester effluent are suitable for the enrichment of anammox bacteria; however, digester effluent as feeding solution for enrichment of anammox bacteria based on the ease of process control and process stability is more advisable.

Deciphering the microbial community and functional genes response of anammox sludge to sulfide stress

Sulfide has attracted increasing attention due to its odor nuisance, toxicity and corrosion. Although variations in the nitrogen removal performance of anammox under sulfide stress have been reported previously, understanding the microorganisms at the molecular level is of greater significance. This study first deciphered the microbial community and functional gene response of anammox sludge to sulfide stress. Results showed that 20 mg L−1 sulfide could reduce specific anammox activity by 61.7%. The protein-like substances within extracellular polymeric substances were quenched at the end of the experiment. Moreover, the relative abundance of Candidatus Kuenenia significantly decreased from 28.7% to 6.4% while Thiobacillus increased from 0 to 7.2% due to sulfide stress. Furthermore, the abundances of functional genes (hzsA, hdh, nirK and nirS) significantly decreased when the sulfide concentration reached 20 mg L−1. These findings provide a further theoretical basis for the anammox process for nitrogen removal from wastewater containing sulfide.

Enhance the treatment of low strength wastewater at low temperature with the coexistence system of AnAOB and heterotrophic bacteria: Performance and bacterial community

The application of anammox process in mainstream wastewater treatment process is still facing challenges especially at the low temperature. To resolve this problem, the coexistence system of anaerobic ammonia-oxidizing bacteria (AnAOB) and heterotrophic bacteria (HB) was built in this study. The nitrogen removal efficiency mainly maintained at above 90% during the process of temperature reducing from 35 °C to 10 °C. The nitrogen removal rate were 0.30 g N·L−1·d−1 at both 25 and 15 °C and 0.10 g N·L−1·d−1 at 10 °C, respectively. Analysis of 16S rRNA genus sequencing revealed that as the temperature reduced to 10 °C, the Denutrotisoma genera presented a downward trend but Comamonadaceae genera showed an upward trend. At 10 °C, the contrast of anammox activities between granular and flocculent sludge in the system revealed that although the abundance of anammox genera was much lower in flocculent sludge than that in granular sludge, the anammox activities showed no significant discrepancy. And the abundance of Comamonadaceae and Chloroflexales genera were much higher in flocculent sludge than those in granular sludge, presenting their key roles to anammox activity at low temperature. The Circos diagram and Cluster of orthologous Group of protein functional predication showed that the functional abundance related to interaction among microbial communities were higher in flocculent sludge but those related to self-growth was higher in granular sludge. This result indicated the significance of the interactions based on the microbial diversity in the application of annamox process at low temperature.

What's the variation in anammox reactor performance after single and joint temperature based shocks?

A laboratory-scale up-flow anaerobic sludge blanket reactor was used to investigate the effects of temperature-based (single and combined with loading) shocks on the performance of anaerobic ammonium oxidation (anammox) reactor. The reactor was tolerant to 15 °C and 25 °C shocks; however, temperature shock of 55 °C led to severe accumulation of effluent NO2−-N (>100 mg L−1), which induced substrate inhibition. Although the shock experiments achieved a maximum effluent NO2−-N concentration of 205.4 mg L−1 after a hydraulic retention time-substrate concentration shock at 25 °C, the inhibition was still reversible. During the experiments, the content of extracellular polymeric substances (EPS) increased significantly after each shock to protect the anammox bacteria, and the value decreased correspondingly at the end of the recovery phase. The specific anammox activity showed the opposite tendency compared with that of the EPS. The performance of anammox reactor under a series of short-term temperature shocks was investigated, and the results can provide new ideas for future research.

Adaptation and characterization of thermophilic anammox in bioreactors

Anammox, the oxidation of ammonium with nitrite, is a key microbial process in the nitrogen cycle. Under mesophilic conditions (below 40 °C), it is widely implemented to remove nitrogen from wastewaters lacking organic carbon. Despite evidence of the presence of anammox bacteria in high-temperature environments, reports on the cultivation of thermophilic anammox bacteria are limited to a short-term experiment of 2 weeks. This study showcases the adaptation of a mesophilic inoculum to thermophilic conditions, and its characterization. First, an attached growth technology was chosen to obtain the process. In an anoxic fixed-bed biofilm bioreactor (FBBR), a slow linear temperature increase from 38 to over 48 °C (0.05–0.07 °C d−1) was imposed to the community over 220 days, after which the reactor was operated at 48 °C for over 200 days. Maximum total nitrogen removal rates reached up to 0.62 g N L−1 d−1. Given this promising performance, a suspended growth system was tested. The obtained enrichment culture served as inoculum for membrane bioreactors (MBR) operated at 50 °C, reaching a maximum total nitrogen removal rate of 1.7 g N L−1 d−1 after 35 days. The biomass in the MBR had a maximum specific anammox activity of 1.1 ± 0.1 g NH4+-N g−1 VSS d−1, and the growth rate was estimated at 0.075–0.19 d−1. The thermophilic cultures displayed nitrogen stoichiometry ratios typical for mesophilic anammox: 0.93–1.42 g NO2--Nremoved g−1 NH4+-Nremoved and 0.16–0.35 g NO3--Nproduced g−1 NH4+-Nremoved. Amplicon and Sanger sequencing of the 16S rRNA genes revealed a disappearance of the original “Ca. Brocadia” and “Ca. Jettenia” taxa, yielding Planctomycetes members with only 94–95% similarity to “Ca. Brocadia anammoxidans” and “Ca. B. caroliniensis”, accounting for 45% of the bacterial FBBR community. The long-term operation of thermophilic anammox reactors and snapshot views on the nitrogen stoichiometry, kinetics and microbial community open up the development path of thermophilic partial nitritation/anammox. A first economic assessment highlighted that treatment of sludge reject water from thermophilic anaerobic digestion of sewage sludge may become attractive.

Effect of COD on mainstream anammox: Evaluation of process performance, granule morphology and nitrous oxide production

The effect of COD addition into an anammox reactor was assessed during short and long term exposure. The short term exposure was assessed via batch tests and lasted 48 h. Results indicated the presence of a very active denitrifying community able to consume COD using nitrate and nitrite as electron acceptors. However, the presence of COD did not result in an increase of the ammonium concentration at the end of the tests indicating that anammox activity was not suppressed by the addition of COD. Different COD concentrations (125, 225 and 175 mg COD/L) were also added in the reactor during 3 periods within its operation (period II, III and V respectively). Long term COD addition (up to 102 d of continuous addition during period II and III) caused a decrease of the anammox activity and a shift on the microbial community, with a decrease on the anammox fraction. However, the anammox process was never lost and it fully recovered as soon as COD addition stopped. Finally, dissolved N2O was monitored under periods with and without COD addition, showing higher concentrations during transient periods from COD addition to no addition. The results of this paper provide evidence of how a long term COD exposure into an anammox reactor affect the overall nitrogen removal process, the granular structure of the anammox biomass, its microbial composition and for the first time, its N2O emissions.

Cell Density-dependent Anammox Activity of Candidatus Brocadia sinica Regulated by N-acyl Homoserine Lactone-mediated Quorum Sensing.

The activity of anaerobic ammonia-oxidizing (anammox) bacteria is considered to depend on cell density; however, this has not yet been confirmed due to the fastidious nature of anammox bacteria (e.g., slow growth, oxygen sensitivity, and rigid aggregate formation). In the present study, the cell density-dependent occurrence of anammox activity (14-15N2 gas production rate) was investigated using planktonic enrichment cultures of Candidatus Brocadia sinica. This activity was detectable when the density of cells was higher than 107‍ ‍cells‍ ‍mL–1 and became stronger with increases in cell density. At the cell densities, the transcription of the BROSI_A1042 and BROSI_A3652 genes, which are potentially involved in the biosynthesis and reception of N-acyl homoserine lactone (AHL), was detectable in Brocadia sinica cells. The presence of AHL molecules in the MBR culture of B. sinica was confirmed by an AHL reporter assay and gas chromatography mass spectrometry analysis. The exogenous addition of the MBR culture extract and AHL molecules (a cocktail of C6, C8, C10, and C12-homoserine lactones) increased the specific 14-15N2 production rate of B. sinica. These results suggest that the specific anammox activity of B. sinica is regulated by AHL-mediated quorum sensing.

Effect of Fe(II) on simultaneous marine anammox and Feammox treating nitrogen-laden saline wastewater under low temperature: Enhanced performance and kinetics

A sequencing batch reactor (SBR) was used to study enhanced nitrogen removal of marine anammox bacteria (MAB) treating nitrogen-laden saline wastewater with Fe(II) addition. The reactor was operated at 15 °C with influent pH of 7.5 ± 0.1. At 4 mg/L Fe(II), ammonia removal rate (ARR) and nitrite removal rate (NRR) increased to 0.85 and 0.90 kg/(m3·d), respectively. Specific anammox activity (SAA) sharply increased to 0.76 kg/(kg·d). Both marine anammox and marine Feammox reaction occurred simultaneously. ΔNO2−-N/ΔNH4+-N was lower than theoretical ratio due to marine Feammox. When Fe(II) was >25 mg/L, the maximal substrate conversion rate increased from 2.97 to 3.47 kg/(m3·d) within 1 h. High Fe(II) concentration (> 25 mg/L) had no negative effect on marine anammox but could inhibit marine Feammox process. The optimal Fe(II) dose was 25 mg/L for marine Feammox. The remodified Logistic model was appropriate to analyze nitrogen removal independent of Fe(II) concentration, and the modified Boltzmann model could only be used to analyze nitrogen removal at high Fe(II) concentration. At 10 mg/L Fe(II), the fitted λ value declined from 0.52 h to 0.34 h. The lag time of MAB was shorten greatly.

Performance of Anammox Processes for Wastewater Treatment: A Critical Review on Effects of Operational Conditions and Environmental Stresses

The anaerobic ammonium oxidation (anammox) process is well-known as a low-energy consuming and eco-friendly technology for treating nitrogen-rich wastewater. Although the anammox reaction was widely investigated in terms of its application in many wastewater treatment processes, practical anammox application at the pilot and industrial scales is limited because nitrogen removal efficiency and anammox activity are dependent on many operational factors such as temperature, pH, dissolved oxygen concentration, nitrogen loading, and organic matter content. In practical application, anammox bacteria are possibly vulnerable to non-essential compounds such as sulfides, toxic metal elements, alcohols, phenols, and antibiotics that are potential inhibitors owing to the complexity of the wastewater stream. This review systematically summarizes up-to-date studies on the effect of various operational factors on nitrogen removal performance along with reactor type, mode of operation (batch or continuous), and cultured anammox bacterial species. The effect of potential anammox inhibition factors such as high nitrite concentration, high salinity, sulfides, toxic metal elements, and toxic organic compounds is listed with a thorough interpretation of the synergistic and antagonistic toxicity of these inhibitors. Finally, the strategy for optimization of anammox processes for wastewater treatment is suggested, and the importance of future studies on anammox applications is indicated.

Toxicity of azoles towards the anaerobic ammonium oxidation (anammox) process

AbstractBACKGROUNDAzoles are an important class of compounds that are widely used as corrosion inhibitors in aircraft de‐icing agents, cooling towers, semiconductor manufacturing and household dishwashing detergents. They also are important moieties in pharmaceutical drugs and fungicides. Azoles are widespread emerging contaminants occurring frequently in water bodies. Azole compounds can potentially cause inhibition towards key biological processes in natural ecosystems and wastewater treatment processes. Of particular concern is the inhibition of azoles to the nitrification process (aerobic oxidation of ammonium). This study investigated the acute toxicity of azole compounds towards the anaerobic ammonia oxidation (anammox) process, which is an important environmental biotechnology gaining traction for nutrient‐nitrogen removal during wastewater treatment. In this study, using batch bioassay techniques, the anammox toxicity of eight commonly occurring azole compounds was evaluated.RESULTSThe results show that 1H‐benzotriazole and 5‐methyl‐1H‐benzotriazole had the highest inhibitory effect on the anammox process, causing 50% decrease in anammox activity (IC50) at concentrations of 19.6 and 17.8 mg L−1, respectively. 1H‐imidazole caused less severe toxicity with an IC50 of 79.4 mg L−1. The other azole compounds were either nontoxic (1H‐pyrazole, 1H‐1,2,4‐triazole and 1‐methyl‐pyrazole) or at best mildly toxic (1H‐benzotriazole‐5‐carboxylic acid and 3,5‐dimethyl‐1H‐pyrazole) towards the anammox bacteria at the concentrations tested.CONCLUSIONSThis study showed that most azole compounds tested displayed mild to low or no toxicity towards the anammox bacteria. The anammox bacteria were found to be far less sensitive to azoles compared to nitrifying bacteria. © 2019 Society of Chemical Industry