High Free Ammonia Concentrations Research Articles

Enhancing blackwater methane production by enriching hydrogenotrophic methanogens through hydrogen supplementation

Source diverted blackwater (i.e., toilet wastewater with or without food waste residuals) represents an ideal waste stream for bioenergy recovery through anaerobic digestion. However, due to the high free ammonia concentration contained in blackwater collected from water conserving toilets (e.g., vacuum toilets), CH4 recovery can be significantly reduced because methanogenesis is inhibited. Our current study evaluated the feasibility of enriching hydrogenotrophic methanogens (known to be more resistant than acetoclastic methanogens toward free ammonia inhibition) in anaerobic sludge with H2 supplementation for blackwater treatment. Our results demonstrated that hydrogenotrophic methanogens can be enriched by supplementing anaerobic sludge with H2 and CO2 in a volume ratio of 4:1 for 3 months. The dominance of hydrogenotrophic methanogens after 3-month enrichment was confirmed with quantitative PCR studies. It was observed that the hydrogenotrophic methanogen dominant anaerobic sludge significantly alleviated blackwater free ammonia inhibition, and led to enhanced biochemical methane production (BMP) (e.g., from 30% to 53%). Our current study offers a new treatment option for anaerobic treatment of wastewater with high free ammonia content.

Application of a synthetic zeolite as a storage medium in SBRs to achieve the stable partial nitrification of ammonium

Free ammonia (FA) is the growth substrate for ammonia-oxidizing bacteria (AOB), but a high concentration of FA could also inhibit AOB activities.

Heterotrophic Nitrifiers Dominate Reactors Treating Incineration Leachate with High Free Ammonia Concentrations

Anaerobically treated leachate from municipal solid waste incineration plants contains extremely high free ammonia (FA) concentrations, which can hinder short-cut nitrification. pH adjustments made...

Nitrogen removal by simultaneous partial nitrification, anammox and denitrification (SNAD) in a structured-bed reactor treating animal feed processing wastewater: Inhibitory effects and bacterial community

The aim of this study was to investigate the post-treatment of UASB effluent by treating animal feed production wastewater using simultaneous partial nitrification, anammox and denitrification (SNAD) in a structured-bed reactor subjected to low aeration and recirculation. The average nitrogen loads applied were 0.307, 0.249 and 0.149 kgN m−3d−1, correlated to COD/N ratios of 0.28, 0.41 and 0.26 (Phases 1, 2 and 3, respectively). The best mean values for removal efficiencies of total-N and COD were obtained in Phase 1 with 48 ± 24% and 63 ± 20%, respectively, reaching a maximum total-N removal efficiency of 79%. The anammox process was the main pathway of nitrogen removal, as pointed out in the nitrogen mass balance. High free ammonia (FA) concentrations in Phases 2 and 3, associated to the limitation of oxygen supply, caused inhibition of nitrite oxidizing bacteria (NOB) activity, leading to NO2 accumulation, having an impact on the denitrifying activity. At the end of the operational period, sequencing analysis detected sequences related to heterotrophic denitrifiers (22.5%), anammox bacteria, Candidatus Anammoximicrobium (2%) and ammonia oxidizing bacteria (AOB) belonging to Nitrosomonadales and Nitrosomonas (0.6%). These results demonstrated that nitritation, denitrification and anammox were likely in the processes involved in nitrogen removal in this reactor.

Effect of CaO2 addition on anaerobic digestion of waste activated sludge at different temperatures and the promotion of valuable carbon source production under ambient condition

The effect of calcium peroxide (CaO2) addition on anaerobic digestion (AD) of waste activated sludge (WAS) at different temperatures (20 °C, 35 °C, and 55 °C) were investigated. The results show that CaO2 addition had significant positive effect on short-chain fatty acids (SCFAs) production under ambient and mesophilic conditions. Polysaccharides and proteins embedded in extracellular polymeric substances (EPS) were effectively released from inner fraction to outer fraction, and non-biodegradable humic-like substances were decreased while easily biodegradable tryptophan-like proteins increased. These effects were most remarkable under ambient conditions. However, CaO2 addition was unfavorable to thermophilic AD because of high free ammonia concentrations and the accumulation of humic-like substances. Temperature showed a stronger effect than CaO2 on microbial community structure, but CaO2 addition was more effective than temperature in enhancing hydrolytic and acidifying microorganisms. Predictive functional profiling indicated that microbial hydrolysis, metabolism and acidification were promoted by CaO2 under ambient conditions.

Effects of Environmental Factors on the Synergy of Functional Bacteria in Completely Autotrophic Granular Sludge

To obtain experimental evidences for optimizing a completely autotrophic nitrogen removal process based on granules, the effects of dissolved oxygen (DO) concentration, temperature (t), initial ammonium (NH4+-N) concentration, and solution pH conditions on the synergy between the aerobic and anaerobic ammonium-oxidizing bacteria (AOB and AMX) were investigated using a single factor batch experiment, while the analysis of the microbial community structure within them was conducted using MiSeq high-throughput pyrosequencing. Results revealed that AOB (genus Nitrosomonas) and AMX (genus Candidatus Kuenenia) dominated in the granules, representing relative abundances of 32.9% and 9.8%, respectively. For the granules, the highest specific nitrogen removal rate of q(TN)=(17.7±1.0) mg·(g·h)-1 was obtained at a DO concentration of 2 mg·L-1, while the initial NH4+-N concentration was set at 100 mg·L-1. And a lower DO level resulted in partial nitritation became the rate-limiting step of process, otherwise, it would be the ANAMMOX reaction instead. According to the free energy of the reactions, the activity of AMX was more sensitive to low temperature than that of AOB. When the reaction temperature was lower than 30℃, nitrite accumulation could be observed in bulk liquid, with the significant decrease of q(TN) for the granules. Under the same oxygen supply conditions, an initial NH4+-N concentration lower than 100 mg·L-1 could inhibit the activity of AMX partly. However, with an initial NH4+-N concentration over 150 mg·L-1, either oxygen-limiting or high free ammonia concentration could lead to the dramatic decrease of q(TN). In addition, the effective synergy of the two types of ammonium oxidizers in granules was always achieved at solution pH in the range of 7.0-8.5.

Rapid start-up of a nitritation granular reactor using activated sludge as inoculum at the influent organics/ammonium mass ratio of 2/1

Partial oxidation of ammonium to nitrite is a pre- and crucial step to achieve shortcut biological nitrogen removal from ammonium-rich wastewater. In the present study, a nitritation granular reactor using activated sludge as inoculum was started up in a sequencing batch reactor (SBR) at a fixed influent C/N ratio of 2:1. Variations in the reactor performance, functional bacteria activities, sludge morphology and bacterial community structure were investigated. Results showed the formation of compact granules was achieved in 55 days, and a stable nitrite accumulation rate of 0.68 kg N·m−3·d−1 was maintained in the following period. With a rapid growth of granular size, the total nitrogen removal by simultaneous nitritation/denitritation was progressively increased to 50%. In sludge granulation, the significant enrichment of r-strategist ammonium oxidizing bacteria (Nitrosomonas) was identified. Additionally, both high free ammonia concentration and extra nitrite competition by heterotrophic denitrifiers were critical to suppress nitrite oxidizing bacteria effectively.

Effect of Free Ammonia on Ammonia Escape During an Ammonia Oxidation Process

This study used short process nitrification sludge to investigate the effect of high free ammonia (FA) on ammonia escape during ammonia oxidation. A sequencing batch reactor (SBR) was used to study the ammonia escape process under different FA concentrations. The results indicate that FA in water is usually combined with water molecules at lower FA concentrations to produce stable NH3·H2O with almost no ammonia escape. With high FA concentration (FA>687.1 mg·L-1), ammonia nitrogen is not oxidized to oxidized nitrogen (NO2--N concentration is <0.1 mg·L-1 at the end of aeration), but the total nitrogen (TN) reached 269.7 mg·L-1. Therefore, when there is a high FA concentration, NH4+-N will evaporate in the form of FA, resulting in ammonia escape. In addition, when 226.6 ≤ FA ≤ 711.8 mg·L-1, the free ammonia escape rate (FEV) grows rapidly with the increase in FA concentration.

ADAPTATION OF THE ACTIVATED SLUDGE TO THE DIGESTATE LIQUORS DURING THE NITRIFICATION AND DENITRIFICATION PROCESSES

The activated sludge process of the digestate liquors after chemical separation was conducted using a 10 L lab-scale sequencing batch reactor (SBR) and a 0.50 m3 pilotscale SBR independently (with pH control). Due to the relatively high concentration of free ammonia (FA), clear inhibitory effects of the digestate liquors on the nitrifying bacteria were observed. The adaptation of the activated sludge to the toxicity was evaluated with the trends of ammonia uptake rate (AUR) and nitrate utilization rate (NUR). The lab-scale AUR values decreased from 5.3 to 2.6 g N/(kg VSS·h) over time after the addition of digestate liquors (5–10% of the reactor working volume), indicating an apparent FA inhibition on the nitrification process in the FA concentration range of 0.3–0.5 mg N/L. The pilot-scale AUR values increased from 1.8 to 3.6 g N/ (kg VSS·h) in the first two weeks and then decreased to 2.4 g N/(kg VSS·h), showing a lag of the inhibition on the nitrifying bacteria at the FA concentration ≈ 0.15 mg N/L. The lab-scale NURs increased from 2.6 to 10.4 g N/(kg VSS·h) over time, and the pilot-scale NURs increased from 1.0 to 4.0 g N/(kg VSS·h) in a similar pattern. The clear dependence of both the lab- and pilot-scale NURs on time indicated the adaptation of the heterotrophic biomass to the digestate liquors. Ethanol – used instead of fusel oil – was found to be a more efficient external carbon source for better adaptation of the activated sludge under unfavorable conditions.

Enrichment and Physiological Characterization of a Cold-Adapted Nitrite-Oxidizing Nitrotoga sp. from an Eelgrass Sediment.

Nitrite-oxidizing bacteria (NOB) are responsible for the second step of nitrification in natural and engineered ecosystems. The recently discovered genus Nitrotoga belongs to the Betaproteobacteria and potentially has high environmental importance. Although environmental clones affiliated with Nitrotoga are widely distributed, the limited number of cultivated Nitrotoga spp. results in a poor understanding of their ecophysiological features. In this study, we successfully enriched the nonmarine cold-adapted Nitrotoga sp. strain AM1 from coastal sand in an eelgrass zone and investigated its physiological characteristics. Multistep-enrichment approaches led to an increase in the abundance of AM1 to approximately 80% of the total bacterial population. AM1 was the only detectable NOB in the bacterial community. The 16S rRNA gene sequence of AM1 was 99.6% identical to that of "Candidatus Nitrotoga arctica," which was enriched from permafrost-affected soil. The highest nitrogen oxidation rate of AM1 was observed at 16°C. The half-saturation constant (Km ) and the generation time were determined to be 25 μM NO2- and 54 h, respectively. The nitrite oxidation rate of AM1 was stimulated at concentrations of <30 mM NH4Cl but completely inhibited at 50 mM NH4Cl. AM1 can grow well under specific environmental conditions, such as low temperature and in the presence of a relatively high concentration of free ammonia. These results help improve our comprehension of the functional importance of NitrotogaIMPORTANCE Nitrite-oxidizing bacteria (NOB) are key players in the second step of nitrification, which is an important process of the nitrogen cycle. Recent studies have suggested that the organisms of the novel NOB genus Nitrotoga were widely distributed and played a functional role in natural and engineered ecosystems. However, only a few Nitrotoga enrichments have been obtained, and little is known about their ecology and physiology. In this study, we successfully enriched a Nitrotoga sp. from sand in a shallow coastal marine ecosystem and undertook a physiological characterization. The laboratory experiments showed that the Nitrotoga enrichment culture could adapt not only to low temperature but also to relatively high concentrations of free ammonia. The determination of as-yet-unknown unique characteristics of Nitrotoga contributes to the improvement of our insights into the microbiology of nitrification.

Erratum to: Effects of rapid temperature rising on nitrogen removal and microbial community variation of anoxic/aerobic process for ABS resin wastewater treatment.

ABS resin wastewater is a high-temperature nitrogenous organic wastewater. It can be successfully treated with anoxic/aerobic (A/O) process. In this study, the effect of temperature on nitrogen removal and microbial community after quick temperature rise (QTR) was investigated. It was indicated that QTR from 25 to 30 °C facilitated the microbial growth and achieved a similar effluent quality as that at 25 °C. QTR from 25 to 35 °C or 40 °C resulted in higher effluent concentration of chemical oxygen demand (COD), biochemical oxygen demand (BOD5), total nitrogen (TN), and total phosphorus (TP). Illumina MiSeq pyrosequencing analysis illustrated that the richness and diversity of the bacterial community was decreased as the temperature was increased. The percentage of many functional groups was changed significantly. QTR from 25 to 40 °C also resulted in the inhibition of ammonia oxidation rate and high concentration of free ammonia, which then inhibited the growth of NOB (Nitrospira), and thus resulted in nitrite accumulation. The high temperature above 35 °C promoted the growth of a denitrifying bacterial genus, Denitratisoma, which might increase N2O production during the denitrification process.

Effect of high free ammonia concentration on microalgal growth and substrate uptake

Effect of high free ammonia concentration on microalgal growth and substrate uptake

Effects of rapid temperature rising on nitrogen removal and microbial community variation of anoxic/aerobic process for ABS resin wastewater treatment.

ABS resin wastewater is a high-temperature nitrogenous organic wastewater. It can be successfully treated with anoxic/aerobic (A/O) process. In this study, the effect of temperature on nitrogen removal and microbial community after quick temperature rise (QTR) was investigated. It was indicated that QTR from 25 to 30°C facilitated the microbial growth and achieved a similar effluent quality as that at 25°C. QTR from 25 to 35°C or 40°C resulted in higher effluent concentration of chemical oxygen demand (COD), biochemical oxygen demand (BOD5), total nitrogen (TN), and total phosphorus (TP). Illumina MiSeq pyrosequencing analysis illustrated that the richness and diversity of the bacterial community was decreased as the temperature was increased. The percentage of many functional groups was changed significantly. QTR from 25 to 40°C also resulted in the inhibition of ammonia oxidation rate and high concentration of free ammonia, which then inhibited the growth of NOB (Nitrospira), and thus resulted in nitrite accumulation. The high temperature above 35°C promoted the growth of a denitrifying bacterial genus, Denitratisoma, which might increase N2O production during the denitrification process.

Recovery of ammonia from anaerobically digested manure using gas-permeable membranes

Nitrogen (N) can be recovered from different types of wastewaters. Among these wastewaters, anaerobically digested swine manure (digestate) has the highest N content in ammonia form (NH3). It is desirable to reduce N in digestate effluents to safely incorporate them in arable soil in N vulnerable zones (NVZ) and to mitigate NH3 emissions during N land application. Additional benefit is to minimize inhibition of the anaerobic process by removing NH3 during the anaerobic digestion process. This work aimed to apply the gas-permeable membrane technology to evaluate ammonia (NH3) recovery from high-ammonia digested swine manure. Anaerobically digested swine manure with NH4+ content of 4,293 mg N L−1 was reduced by 91 % (to 381 mg N L−1) during the 32-day experiment. Although the results showed a total N recovery efficiency of 71 %, it is possible to increase this recovery efficiency to > 90 % by adjusting the area of the membrane system to match the high free ammonia concentration (FA) in digested swine manure. Moreover, final digestate pH and alkalinity were kept around 8.1 and 8,923 mgCaCO3 L−1, which are convenient for the anaerobic process or incorporation in arable soil when the process is finished.

Cultivation and characteristics of partial nitrification granular sludge in a sequencing batch reactor inoculated with heterotrophic granules.

The aim of this study was to develop a simple operation strategy for the cultivation of partial nitrification granules (PNGs) treating an autotrophic medium. For this strategy, aerobic granular sludge adapted to high concentration organics removal was seeded in a sequencing batch reactor (SBR) with a height/diameter ratio of 3.8, and the ratio of organics to the ammonia nitrogen-loading rate (C/N ratio) in the influent was employed as the main control parameter to start up the partial nitrification process. After 86days of operation, the nitrite accumulation rate reached 1.44kg/(m3day) in the SBR, and the removal efficiency of ammonia nitrogen (NH4+-N) was over 95%. The PNGs showed a dense and compact structure, with an excellent settling ability, a typical extracellular polymeric substance (EPS) composition, and a high ammonia oxidation activity. The high-throughput pyrosequencing results indicated that the microbial community structure in the granules was significantly influenced by the C/N ratio, and ammonia-oxidizing bacteria (AOB), including the r-strategist Nitrosomonas and k-strategist Nitrosospira genre, which accounted for approximately 40% of the total biomass at the end of operation. The effective suppression of nitrite-oxidizing bacteria (NOB) growth was attributed to oxygen competition on the granular surface among functional bacteria, as well as the high free ammonia or free nitrous acid concentrations during the aeration period.

Effects of free ammonia on volatile fatty acid accumulation and process performance in the anaerobic digestion of two typical bio-wastes

The effect of free ammonia on volatile fatty acid (VFA) accumulation and process instability was studied using a lab-scale anaerobic digester fed by two typical bio-wastes: fruit and vegetable waste (FVW) and food waste (FW) at 35°C with an organic loading rate (OLR) of 3.0kg VS/(m3·day). The inhibitory effects of free ammonia on methanogenesis were observed due to the low C/N ratio of each substrate (15.6 and 17.2, respectively). A high concentration of free ammonia inhibited methanogenesis resulting in the accumulation of VFAs and a low methane yield. In the inhibited state, acetate accumulated more quickly than propionate and was the main type of accumulated VFA. The co-accumulation of ammonia and VFAs led to an “inhibited steady state” and the ammonia was the main inhibitory substance that triggered the process perturbation. By statistical significance test and VFA fluctuation ratio analysis, the free ammonia inhibition threshold was identified as 45mg/L. Moreover, propionate, iso-butyrate and valerate were determined to be the three most sensitive VFA parameters that were subject to ammonia inhibition.

Improve biogas production from low-organic-content sludge through high-solids anaerobic co-digestion with food waste

Anaerobic co-digestion of sewage sludge and food waste was tested at two different total solid (TS) concentrations. In the low-solids group with TS 4.8%, the biogas production increased linearly as the ratio of food waste in substrate increased from 0 to 100%, but no synergetic effect was found between the two substrates. Moreover, the additive food waste resulted in the accumulation of volatile fatty acids and decelerated biogas production. Thus, the blend ratio of food waste should be lower than 50%. While in the high-solids group with TS 14%, the weak alkaline environment with pH 7.5–8.5 avoided excessive acidification but high concentration of free ammonia was a potential risk. However, good synergetic effect was found between the two substrates because the added food waste improved mass transfer in sludge cake. Thus, 50% was recommended as the optimum ratio of food waste in substrate because of the best synergetic effect.

Startup strategy for partial nitritation treatment of anaerobically digested effluent of swine wastewater in a sand filter

This research investigated the startup of the partial nitritation (PN) process in a sand filter at ambient room temperature (5.5–27°C) to produce an effluent suitable for the subsequent anaerobic ammonium oxidation (anammox) process. A naturally ventilated sand filter was used to treat anaerobically digested effluent of swine wastewater. Startup strategy of gradual step increases in the influent ammonium nitrogen concentration and pH successfully achieved the PN process. The principal cause for nitrite accumulation was high free ammonia concentration (up to about 20mgL−1). An effluent with a nitrite-to-ammonium ratio of 1.02 was obtained, with the optimal influent ammonium nitrogen concentration of 600.0mgL−1 and pH of 8.09. Overall, the PN process can be maintained in a sand filter without forced aeration, and coupling with the subsequent anammox process to achieve completely unpowered autotrophic nitrogen removal.

Effect of ammonia on the photosynthetic activity of Arthrospira and Chlorella: A study on chlorophyll fluorescence and electron transport

Although ammoniacal nitrogen is the preferred nitrogen source for microalgae/cyanobacteria, at elevated concentrations and high pH values it may negatively impact photosynthesis and growth. Chlorophyll (Chl) fluorescence analysis is a useful tool to monitor the influence of various stress conditions on the photosynthetic activity of plants or microalgae/cyanobacteria. In this study, we investigated the effect of ammoniacal nitrogen on Chl fluorescence in microalgae/cyanobacteria. Chl fluorescence analysis revealed that the parameters related to flux ratios and specific energy fluxes of photochemistry were gradually inhibited as the free ammonia (FA) concentration increased. Photosynthetic electron transport activity was measured using artificial electron acceptors, donors or inhibitors. These analyses suggest that ammonia has multiple impacts on the photosynthetic apparatus; photosystems I (PSI) and II (PSII), the electron transport chain, the oxygen-evolution complex (OEC) as well the dark respiration were gradually inhibited by increasing FA concentration. At high FA concentration, the PSI/PSII activity increased, suggesting that PSI was more tolerant to FA than PSII. Non-photochemical quenching (NPQ) decreased to zero at elevated FA concentrations. The Chl fluorescence data obtained in the presence of DCMU (diuron) suggest that the decrease of NPQ under ammonia inhibition/toxicity is due to the increase of PSI/PSII activity. The rapid response of Chl fluorescence transients to increases in FA may allow one to use pulse amplitude modulation (PAM) fluorescence as a tool to monitor ammonia inhibition/toxicity in cultures of microalgae/cyanobacteria.

Nitrogen Removal from Anaerobically Digested Swine Waste Centrate Using a Laboratory-Scale Chabazite-Sequencing Batch Reactor

Abstract Anaerobic digestion (AD) is widely used for the treatment of livestock waste because the biogas produced can be utilized as energy source. Further treatment of AD effluents is often required for total nitrogen (TN) removal, which is typically carried out using biological nitrogen removal processes. However, the high free ammonia (FA) concentrations present in AD effluents can inhibit nitrification processes. This research tested a sequencing batch reactor (SBR) amended with particulate chabazite (chabazite-SBR) for removal of TN from anaerobically digested swine waste centrate. In the chabazite-SBR, zeolitic material with a high ion exchange (IX) capacity for NH4+ is used to ease nitrification inhibition. Once nitrification is complete, an external organic electron donor is supplied during an anoxic stage to promote denitrification. An overall TN removal efficiency of 84% was achieved in a laboratory-scale chabazite-SBR, with specific nitrification and denitrification rates of 0.43 and 1.49 mg-N/...