Model-based process analysis of partial nitrification efficiency under dynamic nitrogen loading

In this study, the ammonia removal efficiency for high ammonia-containing wastewaters was evaluated via partial nitrification. A nitrifier biocommunity was first enriched in a fill-and-draw batch reactor with a specific ammonium oxidation rate of 0.1 mg NH₄ ⁻-N/mg VSS.h. Partial nitrification was established in a chemostat at a hydraulic retention time (HRT) of 1.15 days, which was equal to the sludge retention time (SRT). The results showed that the critical HRT (SRT) was 1.0 day for the system. A maximum specific ammonium oxidation rate was achieved as 0.280 mg NH₄ ⁻-N/mg VSS.h, which is 2.8-fold higher than that obtained in the fill-and-draw reactor, indicating that more adaptive and highly active ammonium oxidizers were enriched in the chemostat. Dynamic modeling of partial nitrification showed that the maximum growth rate for ammonium oxidizers was found to be 1.22 day⁻¹. Modeling studies also validated the recovery period as 10 days.     

短程硝化启动运行中功能菌群变化研究

【目的】短程硝化-厌氧氨氧化是可实现的最短生物脱氮工艺,短程硝化是实现该工艺的重要环节和必要条件。【方法】采用序批式反应器(SBR)来实现短程硝化过程的启动和稳定运行,并对该过程中的相关功能菌群变化进行检测分析。【结果】通过控制低DO浓度(<1 mg/L)和逐步提高氨氮进水负荷,可抑制氨氧化细菌(NOB)菌群增殖并促进亚硝酸氧化菌(AOB)菌群规模显著扩大,实现短程硝化过程的启动和稳定运行。在氨氮进水负荷为0.055 kg/(m3.d)时,平均氨氮去除容积负荷和污泥负荷可达到0.043kg/(m3.d)和0.16 kg/(kg.d),平均亚硝酸盐积累率可达到83.4%。在短程硝化启动和稳定运行过程中,NOB菌群密度从2.0×105CFU/mL降至1.5×104CFU/mL,相对丰度从5.51%降至2.14%;AOB菌群密度从4.5×104CFU/mL增加至1.5×107CFU/mL,相对丰度从0.18%增加至7.25%。【结论】AOB菌群规模的扩大是实现短程硝化和氨氮去除能力提高的主要原因,同时较高的进水氨氮浓度和负荷也会造成亚硝化活性的抑制。     

Distinctive microbial ecology and biokinetics of autotrophic ammonia and nitrite oxidation in a partial nitrification bioreactor

Biological nitrogen removal (BNR) based on partial nitrification and denitrification via nitrite is a cost-effective alternate to conventional nitrification and denitrification (via nitrate). The goal of this study was to investigate the microbial ecology, biokinetics, and stability of partial nitrification. Stable long-term partial nitrification resulting in 82.1 +/- 17.2% ammonia oxidation, primarily to nitrite (77.3 +/- 19.5% of the ammonia oxidized) was achieved in a lab-scale bioreactor by operation at a pH, dissolved oxygen and solids retention time of 7.5 +/- 0.1, 1.54 +/- 0.87 mg O(2)/L, and 3.0 days, respectively. Bioreactor ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) populations were most closely related to Nitrosomonas europaea and Nitrobacter spp., respectively. The AOB population fraction varied in the range 61 +/- 45% and was much higher than the NOB fraction, 0.71 +/- 1.1%. Using direct measures of bacterial concentrations in conjunction with independent activity measures and mass balances, the maximum specific growth rate (micro(max)), specific decay (b) and observed biomass yield coefficients (Y(obs)) for AOB were 1.08 +/- 1.03 day(-1), 0.32 +/- 0.34 day(-1), and 0.15 +/- 0.06 mg biomass COD/mg N oxidized, respectively. Corresponding micro(max), b, and Y(obs) values for NOB were 2.6 +/- 2.05 day(-1), 1.7 +/- 1.9 day(-1), and 0.04 +/- 0.02 mg biomass COD/mg N oxidized, respectively. The results of this study demonstrate that the highly selective partial nitrification operating conditions enriched for a narrow diversity of rapidly growing AOB and NOB populations unlike conventional BNR reactors, which host a broader diversity of nitrifying bacteria. Further, direct measures of microbial abundance enabled not only elucidation of mixed community microbial ecology but also estimation of key engineering parameters describing bioreactor systems supporting these communities.     

The ammonium nitrogen oxidation process in horizontal subsurface flow constructed wetlands

The ammonium nitrogen oxidation process (ANOP) is the first and most important step for nitrogen removal in constructed wetlands (CWs). The process was investigated by observing the products generated from the ANOP in on-site aerobic systems with selective inhibition of nitrite-oxidizing bacteria (NOB) through appropriate regulation of the pH, temperature and dissolved oxygen concentrations. The effects of season, plant type and density on ANOP were also studied to determine the optimal conditions for the ANOP. Nitrite accumulation was found in the aerobic experiments and greater ammonia-oxidizing bacteria than NOB numbers, showing that partial nitrification to nitrite was occurring in the studied CWs. The nitrogen removal rate was positively linearly correlated with the nitrite accumulation rate, and so the more NH₃-N removed by ANOP, the greater the resulting nitrogen removal. Season and plant density had a significant effect on the ANOP. However, there were no significant differences between the units planted with common reed and cattail.     

PVA-alginate immobilized cells for anaerobic ammonium oxidation (anammox) process

The feasibility of an anaerobic ammonium oxidation (anammox) process combined with a cell-immobilization technique for autotrophic nitrogen removal was investigated. Anammox biomass was cultivated from local activated sludge and achieved significant anammox activity in 6 months. The development of a mature anammox biomass was confirmed by fluorescence in situ hybridization (FISH) analysis and off-line activity measurements. The abundance fraction of the anammox bacteria determined by FISH analysis was estimated by software. The anaerobic ammonia oxidizers occupied almost half of the total cells. Additionally, the anammox biomass was granulated as spherical gel beads of 3-4 mm in diameter by using a cell-immobilization technique. The nitrogen removal activity was proved to be successfully retained in the beads, with about 80% of nitrogenous compounds (NH(4) (+), NO(2) (- )and total nitrogen) removed after 48 h. These results offer a promising technique for the preservation of anammox microorganisms, the protection of them against the unfavorable surroundings, and the prevention of biomass washout towards the implementation of sustainable nitrogen elimination biotechnology. This is the first report on the immobilization of anammox biomass as gel beads.     

Identification of anammox bacteria in a full-scale deammonification plant making use of anaerobic ammonia oxidation

The existence of anaerobic ammonia-oxidizing (anammox) bacteria was postulated in the late 1970s. Approximately 20 years later, these lithotrophic members of the nitrogen cycle were identified as deep-branching members of the planctomycetes. Recently, full-scale implementation of biological deammonification was successfully achieved in the DEMON reactor at the wastewater treatment plant in Strass, Austria. The sludge of this reactor contains red granules and brownish flocs that can be physically separated. The two fractions yielded different banding patterns in denaturing gradient gel electrophoresis of PCR products obtained with primer sets targeting the 16S rRNA genes of planctomycetes. Comparative analysis of partial sequences of almost full-length 16S rRNA gene clones obtained from the granules and flocs confirms the differences in the community composition of the two fractions. The sequences retrieved from the red granules were 93% similar to those of Candidatus Brocadia anammoxidans, a bacterium known to catalyze the anaerobic ammonia oxidation.     

海洋氮循环中细菌的厌氧氨氧化

细菌厌氧氨氧化过程是在一类特殊细菌的厌氧氨氧化体内完成的以氨作为电子供体硝酸盐作为电子受体的一种新型脱氮反应.厌氧氨氧化菌的发现,改变人们对传统氮的生物地球化学循环的认识:反硝化细菌并不是大气中氮气产生的唯一生物类群.而且越来越多的证据表明,细菌厌氧氨氧化与全球的氮物质循环密切相关,估计海洋细菌的厌氧氨氧化过程占到全球海洋氮气产生的一半左右.由于氮与碳的循环密切相关,因此可以推测,细菌的厌氧氨氧化会影响大气中的二氧化碳浓度,从而对全球气候变化产生重要影响.另外,由于细菌厌氧氨氧化菌实现了氨氮的短程转化,缩短了氮素的转化过程,因此为开发更节约能源、更符合可持续发展要求的废水脱氮新技术提供了生物学基础.     

Improved simultaneous nitrification and denitrification in a single reactor by using two different immobilization carriers with specific oxygen transfer characteristics

To maximize nitrogen utilization rates during nitrification and denitrification in a simultaneous reaction for direct nitrogen removal from ammonia–nitrogen in a single reactor, two different carriers were applied that immobilized nitrifiers and denitrifiers separately. With the optimized DO concentration and mixing ratio of immobilization carriers, ammonium–nitrogen was successfully removed as designed until the middle phase of treatment where nitrogen removal rate was higher than 83% of the theoretical value, although an imbalance between nitrification and denitrification occurred at a later phase of treatment where residual nitrate–nitrogen concentration was less than 2 mg/l. The new approach using two different carriers to immobilize nitrifiers and denitrifiers separately was proved useful for controlling both nitrification and denitrification rates, enabling the utilization of maximum treatment ability of both nitrifiers and denitrifiers in a single reactor for direct nitrogen removal from ammonium–nitrogen.     

Comparison of quasisteady-state performance of the DEAMOX process under intermittent and continuous feeding and different nitrogen loading rates

The recently developed denitrifying ammonium oxidation (DEAMOX) process combines the anammox reaction with autotrophic denitrifying conditions using sulfide as an electron donor for the production of nitrite from nitrate within an anaerobic biofilm. This paper compares a quasisteady-state performance of this process for treatment of baker's yeast wastewater under intermittent and continuous feeding and increasing nitrogen loading rate (NLR) from 300 till 858 mg N/L/d. The average total nitrogen removal slightly decreased on increasing the NLR: from 86 to 79% (intermittent feeding) and from 87 to 84% (continuous feeding). The better performance under continuous feeding was due to a more complete nitrate removal in the former case whereas the ammonia removal was similar for both feeding regimes under the comparable NLR. A possible explanation can be that, during continuous feeding (simultaneous supply of nitrate and sulfide), there were less mass transfer limitations for sulfide oxidizing denitrifiers presumably located in the outer layer of sludge aggregates. On the contrary, the ammonia oxidisers presumably located inside the aggregates apparently suffered from nitrite mass transfer limitations under both the feedings. The paper further describes some characteristics of the DEAMOX sludge.     

异养硝化细菌的生物脱氮

正氮类污染物是造成水体污染和富营养化的主要原因,水体中氮的去除对于清洁水体有着重要的意义~([1-2])。因此,高效与便捷地去除污水中的氮是污水处理中的关键问题,而生物脱氮又被认为是目前废水脱氮中最经济有效的方法之一~([3])。最初研究者们认为只有自养硝化细菌才能进行高效的硝化作用,近年来研究表明,异养硝化微生物同样在生物硝化过程中起着不可忽视的作用~([4])。异养硝化微生物相比传统的自养微生物在废水生物脱氮中占有明显优势,具有良好的应用前景。     

The development of simultaneous partial nitrification, ANAMMOX and denitrification (SNAD) process in a single reactor for nitrogen removal

The simultaneous partial nitrification, ANAMMOX and denitrification (SNAD) process was validated to potentially remove ammonium and COD from wastewater in a single, oxygen-limited, non-woven rotating biological contactor (NRBC) reactor. An ammonium conversion efficiency of 79%, TN removal efficiency of 70% and COD removal efficiency of 94% were obtained with the nitrogen and COD loading rate of 0.69 kgN/m(3)d and 0.34 kg/m(3)d, respectively. Scanning electron microscopy (SEM) observation and fluorescence in situ hybridizations (FISH) analysis revealed the existence of the dominant groups of bacteria. As a result, the aerobic ammonia-oxidizing bacteria (AOB), with a spot of aerobic heterotrophic bacteria were mainly distributed in the aerobic outer part of the biofilm. However, ANAMMOX bacteria with denitrifying bacteria were present and active in the anaerobic inner part of the SNAD biofilm. These bacteria were found to exist in a dynamic equilibrium to achieve simultaneous nitrogen and COD removal in NRBC system.     

Application, eco-physiology and biodiversity of anaerobic ammonium-oxidizing bacteria

The demand for new and sustainable systems for nitrogen removal has increased dramatically in the last decade. It is clear that the conventional systems cannot deal with the increasing nitrogen loads in a cost effective way. As an alternative, the implementation of the anammox (anaerobic ammonium oxidation) process in the treatment of wastewater with high ammonium concentrations has been started. The compact anammox reactors can sustain high nitrogen loads without any problems. The highest observed anammox capacity is 8.9 kg N removed m^-3 reactor day^-1. The first 75 m³ anammox reactor is operating in Rotterdam, the Netherlands, combined with the partial nitrification process Single reaction system for High Ammonium Removal Over Nitrite (SHARON). Partial nitrification and anammox can also be combined in one reactor systems like Completely Autotrophic Nitrogen removal Over Nitrite (CANON) or Oxygen Limited Ammonium removal via Nitrification Denitrification (OLAND) where aerobic ammonium-oxidizing bacteria (AOB) and anammox bacteria cooperate under oxygen-limitation. These systems remove about 1.5 kg N m^-3 reactor day^-1. In addition to ammonium, urea can also be converted in the CANON system after a two-week adaptation period. The ecophysiological properties of the anammox bacteria make them very well suited to convert ammonium and nitrite. The K_s values for ammonium and nitrite are below 5 M. However, nitrite above 10 mM is detrimental for the anammox process, and oxygen reversibly inhibits the process at concentrations as low as 1 M. Acetate and propionate can be used by the anammox bacteria to convert nitrite and nitrate, whereas methanol and ethanol severely inhibit the anammox reaction. The enzyme hydroxylamine/hydrazine oxidoreductase (HAO), one of the key enzymes, is located in the anammoxosome, which is a membrane bound organelle. The membranes of the anammox bacteria contain unique ladderane lipids and hopanoids. The bacteria responsible for the anammox reaction are related to the Planctomycetes. The first anammox bacteria were isolated via Percoll centrifugation and characterized as Candidatus Brocadia anammoxidans. Survey of different wastewater treatment plants using anammox specific 16S rRNA gene primers and anammox specific oligonucleotide probes has revealed the presence of at least three other anammox bacteria, which have been tentatively named Candidatus Kuenenia stuttgartiensis, Candidatus Scalindua wagneri and Candidatus Scalindua brodae. A close relative of the latter, Candidatus Scalindua sorokinii was found to be responsible for about 50% of the nitrogen conversion in the anoxic zone of the Black Sea, making the anammox bacteria an important player in the oceanic nitrogen cycle.     

Activated sludge under free ammonia treatment using gel immobilization technology for long-term partial nitrification with different initial biomass

To achieve stable partial nitrification, activated sludge from a wastewater treatment plant using free ammonia (FA) inhibition was immobilized in a polyvinyl alcohol carrier. After FA treatment at 16.44 mg L⁻¹ for 1 day, due to the increased growth rate gap between ammonium-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB), AOB enrichment and NOB inhibition were achieved within 12 days, with AOB and NOB accounting for 65.61 and 0.05%, respectively. Subsequently, with dissolved oxygen concentrations of 4−5 mg L⁻¹, pH of 7.6–7.8 and temperature of 25 ± 1 °C, the immobilized carrier made of activated sludge achieved more than 90% and more than 86% of nitrite accumulation rate at the influent ammonia concentration of 90−110 mg L⁻¹ and 35−50 mg L⁻¹, respectively. After 50 days operation, the NOB content was 0.10%, indicating the immobilized carrier provided favorable conditions for maintaining the low NOB content. Furthermore, due to the low NOB content in the inoculum and the oxygen-limited environment formed by the increase in the AOB numbers in the carrier, immobilized carrier with different initial biomass (1, 2.5 and 5%) can achieve stable partial nitrification.     

Stability and microbial community structure of a partial nitrifying fixed-film bioreactor in long run

A partial nitrification system was investigated for 471 days under DO varying concentrations for assessing its stability and population dynamics. Within 130 days of operation at feed DO concentration of 1.0 ± 0.1 mg/L, more than 85% of nitrite was accumulated. Efficiency deteriorated when the feed DO concentration was increased to 4.2 ± 0.3 mg/L. Nitrite accumulation could not be re-established on decreasing feed DO to 1.0 ± 0.1 mg/L. Even at DO concentration of <0.05 mg/L, nitrate production was observed; a condition termed as anoxic nitrification. NOB was detected in the biomass even under this condition by Fluorescence in-situ hybridization (FISH) analysis. Through 16S rRNA gene sequencing a major fraction of unknown bacterial sequences closely resembling haloalkalophilic bacteria of marine origin were detected. The study indicated that these bacterial species might play a role in anoxic nitrification and that NOB could survive extreme low DO condition.     

An investigation of a process for partial nitrification and autotrophic denitrification combined desulfurization in a single biofilm reactor

In this study, a vertical submerged biofilm reactor was applied to investigate autotrophic partial nitrification/denitrification and simultaneous sulfide removal by using synthetic wastewater. The appropriate influent ratios of ammonia and sulfide needed to achieve partial autotrophic nitrification and denitrification were evaluated with influent ammonium nitrogen ranging from 54.6 to 129.8 mg L^?1 and sulfide concentrations ranging from 52.7 to 412.4 mg S L^?1. The results demonstrated that the working parameter was more stable when the sulfur/nitrogen ratio was set at 3:2, which yielded the maximum sulfur conversion. Batch experiments with different phosphate concentrations proved that a suitable phosphate buffer solution to control pH values could improve synchronous desulfurization denitrification process performance.     

Effects of temperature on simultaneous nitrification and denitrification via nitrite in a sequencing batch biofilm reactor

A laboratory scale experiment was described in this paper to enhance biological nitrogen removal by simultaneous nitrification and denitrification (SND) via nitrite with a sequencing batch biofilm reactor (SBBR). Under conditions of total nitrogen (TN) about 30 mg/L and pH ranged 7.15–7.62, synthetic wastewater was cyclically operated within the reactor for 110 days. Optimal operation conditions were established to obtain consistently high TN removal rate and nitrite accumulation ratio, which included an optimal temperature of 31 °C and an aeration time of 5 h under the air flow of 50 L/h. Stable nitrite accumulation could be realized under different temperatures and the nitrite accumulation ratio increased with an increase of temperature from 15 to 35 °C. The highest TN removal rate (91.9%) was at 31 °C with DO ranged 3–4 mg/L. Process control could be achieved by observing changes in DO and pH to judge the end-point of oxidation of ammonia and SND.     

Biological nitrogen removal with nitrification and denitrification via nitrite pathway

Presently, the wastewater treatment practices can be significantly improved through the introduction of new microbial treatment technologies. To meet increasingly stringent discharge standards, new applications and control strategies for the sustainable removal of ammonium from wastewater have to be implemented. Partial nitrification to nitrite was reported to be technically feasible and economically favorable, especially when wastewater with high ammonium concentrations or low C/N ratios is treated. For successful implementation of the technology, the critical point is how to maintain partial nitrification of ammonium to nitrite. Partial nitrification can be obtained by selectively inhibiting nitrite oxidizing bacteria through appropriate regulation of the system’s DO concentration, microbial SRT, pH, temperature, substrate concentration and load, operational and aeration pattern, and inhibitor. The review addressed the microbiology, its consequences for their application, the current status regarding application, and the future developments.