Nitrogen removal characteristics of heterotrophic nitrification-aerobic denitrification by Alcaligenes faecalis C16

Alcaligenes faecalis C16 was found to have the ability to heterotrophically nitrify and aerobically denitrify. In order to further understand its nitrogen removal ability and mechanism, the growth and ammonium removal response were investigated at different C/N ratios and ammonium concentrations in the medium with citrate and acetate as carbon source separately. Furthermore, experiments of nitrogen sources, production of nitrogen gas and enzyme assay were conducted. Results show that the bacterium converts NH+4-N and produces NH2 OH during the growing phase and nitrite accumulation is its distinct metabolic feature. A. faecalis C16 is able to tolerate not only high ammonium concentration but also high C/N ratio, and the ammonium tolerance is associated with carbon source and C/N ratio. The nitrogen balance under different conditions shows that approximately28%–45% of the initial ammonium is assimilated into the cells, 44%–60% is denitrified and several percent is converted to nitrification products. A. faecalis C16 cannot utilize hydroxylamine, nitrite or nitrate as the sole nitrogen source for growth. However, nitrate can be used when ammonium is simultaneously present in the medium. A possible pathway for nitrogen removal by C16 is suggested. The preliminary enzyme assay provides more evidence for this nitrogen removal pathway.

Study on Characteristics in the Removal Process of Ammonia Nitrogen and Nitrate Nitrogen by an Isolated Heterotrophic Nitrification-Aerobic Denitrification Strain Rhodococcus Sp.

Removal of ammonia nitrogen and nitrate nitrogen by an heterotrophic nitrification-aerobic denitrification strain is an economical and effective method. In this article, a kind of heterotrophic nitrification-aerobic denitrification strain which has aerobic denitrification and heterotrophic nitrification ability was selected, and then was identified as rhodococcus sp. by 16S rRNA sequencing analysis and morphological observation. After that, carbon source utilization and nitrification- denitrification activity of this strain in different C/N, initial nitrogen concentration were studied. In addition, the assimilation and denitrification activities of ammonia and nitrate were also researched under the condition of nitrate and ammonia coexisted in the solution. The results show that the strain can grow in sodium acetate, glucose, sodium succinate and sodium citrate solutions, and it can not survive in sodium oxalate, sucrose and soluble starch solutions. Initial concentration and C/N were important for nitrogen removal rate. This strain can completely remove nitrate/ammonia when nitrate/ammonia concentration was lower than 15 mg l-1/80 mg l-1. the C/N of 10 and of 12 were the optimum C/N ratio in the nitrate and ammonia removal process respectively. pH value rose up sharply in the denitrification process and it increased relatively slowly in the nitrification process, which shows that pH is one of the most important factor inhibiting the denitrification removal process. Nitrite concentration was much higher in denitrification process than in nitrification process. In addition, this strain gave priority to utilizing ammonia as nitrogen source when ammonia and nitrate coexisted in the solution.

Quorum sensing systems regulate heterotrophic nitrification-aerobic denitrification by changing the activity of nitrogen-cycling enzymes

Heterotrophic nitrification-aerobic denitrification(HNAD)is essential in diverse nitrogen-transforming processes.How HNAD is modulated by quorum sensing(QS)systems is still ambiguous.The QS system in Pseudomonas aeruginosa manipulates colony behavior.Here,we described the influence of the Pseudomonas quinolone signal(PQS)and N-acyl-L-homoserine lactone(AHL)on HNAD.The HNAD of P.aeruginosa was inhibited by the oversecretion of PQS.AHL-or PQS-deficient P.aeruginosa mutants had a higher ability for nitrogen removal.QS inhibited heterotrophic nitrification mainly via controlling the activity of nitrite oxidoreductase(NXR)and the depressed aerobic denitrification by regulating the catalytic abilities of nitric oxide reductase(NOR),nitrite reductase(NIR),and nitrate reductase(NAR).The addition of citrate as the sole carbon source increased the nitrogen removal efficiency compared with other carbon sources.Nitrite,as the sole nitrogen source,could be used entirely with only the moderate concentration of PQS contained.AHL and PQS controlled both nitrification and denitrification,suggesting that QS plays an important role in nitrogen cycle under aerobic conditions.

A heterotrophic nitrification-aerobic denitrification bacterium Halomonas venusta TJPU05 suitable for nitrogen removal from high-salinity wastewater

A novel salt-tolerant heterotrophic nitrification and aerobic denitrification(HN-AD)bacterium was isolated and identified as Halomonas venusta TJPU05(H.venusta TJPU05).The nitrogen removal performance of H.venusta TJPU05 in simulated water(SW)with sole or mixed nitrogen sources and in actual wastewater(AW)with high concentration of salt and nitrogen was investigated.The results showed that 86.12%of NH_(4)^(+)-N,95.68%of NO_(3)^(-)-N,100%of NO_(2)^(-)-N and 84.57%of total nitrogen(TN)could be removed from SW with sole nitrogen sources within 24 h at the utmost.H.venusta TJPU05 could maximally remove 84.06%of NH_(4)^(+)-N,92.33%of NO_(3)^(-)-N,92.9%of NO_(2)^(-)-N and 77.73%of TN from SW with mixed nitrogen source when the salinity was above 8%.The application of H.venusta TJPU05 in treating AW with high salt and high ammonia nitrogen led to removal efficiencies of 50.96%,47.28%and 43.19%for NH_(4)^(+)-N,NO_(3)^(-)-N and TN respectively without any optimization.Furthermore,the activities of nitrogen removal–related enzymes of the strain were also investigated.The successful detection of high level activities of ammonia oxygenase(AMO),hydroxylamine oxidase(HAO),nitrate reductase(NAR)and nitrite reductase(NIR)enzymes under high salinity condition further proved the HN-AD and salt-tolerance capacity of H.venusta TJPU05.These results demonstrated that the H.venusta TJPU05 has great potential in treating high-salinity nitrogenous wastewater.

Simultaneous heterotrophic nitrification and aerobic denitrification at high initial phenol concentration by isolated bacterium Diaphorobacter sp. PD-7

A strain capable of phenol degradation, hetemtrophic nitrification and aerobic denitrification was isolated from activated sludge of coking-plant wastewater ponds under aerobic condition. Based on its morphology, physiology, biochemical analysis and phylogenetic characteristics, the isolate was identified as Diaphorobacter sp. PD-7. Biodegradation tests of phenol showed that the maximum phenol degradation occurred at the late phase of exponential growth stages, with 1400 mg·L^-1 phenol completely degraded within 85 h. Diaphorobacter sp. PD-7 accumulated a vast quantity of phenol hydroxylase in this physiological phase, ensuring that the cells quickly utilize phenol as a sole carbon and energy source. The kinetic behavior ofDiaphorobacter sp. PD-7 in batch cultures was investigated over a wide range of initial phenol concentrations （0-1400mg·L^-1） by using the Haldane model, which adequately describes the dynamic behavior of phenol biodegradation by strain Diaphombacter sp. PD-7. At initial phenol concentration of 1400mg· L^-l, batch experiments （0.25 L flask） of nitrogen removal under aerobic condition gave almost entirely removal of 120.69mg· L^- 1 ammonium nitrogen within 75 h, while nitrate nitrogen removal reached 91% within 65 h. Moreover, hydroxylamine oxidase, periplasmic nitrate reductase and nitrite reductase were successfully expressed in the isolate.

Heterotrophic ammonium removal characteristics of an aerobic heterotrophic nitrifying-denitrifying bacterium, Providencia rettgeri YL

Bacterium Providencia rettgeri YL was found to exhibit an unusual ability to heterotrophically nitrify and aerobically denitrify various concentrations of ammonium （NH4^＋-N）. In order to further understand its removal ability, several experiments were conducted to identify the growth and ammonium removal response at different carbon to nitrogen （C/N） mass ratios, shaking speeds, temperatures, ammonium concentrations and to qualitatively verify the production of nitrogen gas using gas chromatography techniques. Results showed that under optimum conditions （C/N 10, 30℃, 120 r/min）, YL can significantly remove low and high concentrations of ammonium within 12 to 48 h of growth, respectively. The nitrification products hydroxylamine （NHzOH）, nitrite （NO2^-） and nitrate （NO3^-） as well as the denitrification product, nitrogen gas （N2）, were detected under completely aerobic conditions.

Isolation and characterization of heterotrophic nitrifying bacteria in MBR

The study presented the method for isolating the heterotrophic nitrifiers and the characterization of heterotrophic nitrification. Continuous tests via a membrane bioreactor (MBR) were operated under the controlled conditions to proliferate the nitrifiers. Heterotrophic nitrifying bacteria were isolated from the system in which the efficiency of total nitrogen(TN) removal was up to 80%. Since no autotrophic ammonium and nitrite oxidizers could be detected by fluorescence in situ hybridization(FISH), oxidized-N production was unlikely to be catalyzed by autotrophic nitrifiers during the heterotrophic nitrifiers' isolation in this study. The batch test results indicate that the isolated heterotrophic bacteria were able to nitrify. After 3 weeks incubation, the efficiencies of the COD removal by the three isolated bacterial strains B1, B2, and B3 were 52 6%, 71 7%, and 77 7%, respectively. The efficiencies of the TN removal by B1, B2, and B3 were 35 6%, 61 2% and 68 7%, respectively.

Isolation and Characterization of a New Heterotrophic Nitrifying Bacillus sp. Strain

Objective To characterize the heterotrophic nitrifying bacteria. Methods The bacteria were isolated from membrane bioreactor for treating synthetic wastewater using the method newly introduced in this study. Fluorescence in situ hybridization （FISH） was used to validate the nonexistence of autotrophic ammonia oxidizers and nitrite oxidizers. Batch tests were carried out to investigate the capability of heterotrophic nitrification by the pure culture. Phylogenetic analysis of the pure culture was performed. Results A heterotrophic nitrifier, named Bacillus sp. LY, was newly isolated from the membrane bioreactor system in which the efficiency of TN removal was up to 80%. After 24-day, incubation, the removal efficiency of COD by Bacillus sp. LY was 71.7 %. The ammonium nitrogen removal rate after assimilation nearly ceased by Bacillus sp. LY was 74.7% The phylogenetic tree of Bacillus sp. LY and the neighbouring nitrifiers were given. Conclusions The batch test results indicate that Bacillus sp. LY can utilize the organic carbon as the source of assimilation when it grows on glucose and ammonium chloride medium accompanying the formation of oxidized-nitrogen. It also can denitrify nitrate while nitrifying. Bacillus sp. LY may become a new bacterial resource for heterotrophic nitrification and play a bioremediation role in nutrient removal.

Simultaneous nitrification and denitrification conducted by Halomonas venusta MA-ZP17-13 under low temperature conditions

Nitrification is a key step in the global nitrogen cycle.Compared with autotrophic nitrification,heterotrophic nitrification remains poorly understood.In this study,Halomonas venusta MA-ZP17-13,isolated from seawater in shrimp aquaculture (Penaeus vannamei),could simultaneously undertake nitrification and denitrification.With the initial ammonium concentration at 100 mg/L,the maximum ammonium-nitrogen removal rate reached98.7%under the optimal conditions including C/N concentration ratio at 5.95,p H at 8.93,and Na Cl at 2.33%.The corresponding average removal rate was 1.37 mg/(L·h)(according to nitrogen) in 3 d at 11.2℃.By whole genome sequencing and analysis,nitrification-and denitrification-related genes were identified,including ammonia monooxygenase,nitrate reductase,nitrite reductase,nitric oxide dioxygenase and nitric oxide synthase;while no gene encoding hydroxylamine oxidase was identified,it implied the existence of a novel nitrification pathway from hydroxylamine to nitrate.These results indicate heterotrophic bacterium H.venusta MA-ZP17-13 can undertake simultaneous nitrification and denitrification at low temperature and has potential for NH_(4)^(+)-N/NH_(3)-N removal in marine aquaculture systems.

Stimulation impact of electric currents on heterotrophic denitrifying microbial viability and denitrification performance in high concentration nitrate-contaminated wastewater

Electric current stimulation has been shown to have a positive influence on heterotrophic denitrifying microbial viability and has the potential to improve wastewater denitrification performance. This study investigated the effects of varying current densities on microbial activity and NO_3^- removal efficiency under heterotrophic conditions.NO_3^-removal rate was highest at an applied current density of 400 mA/m^2. However, the optimum removal efficiency of total inorganic nitrogen(TIN; 99%) was achieved when the current density was fixed at 200 m A/m^2. Accumulation of NH_4^+-N and NO_2^--N byproducts were also minimized at this current density. The activity of heterotrophic denitrifying microorganisms was much higher at both 200 and 400 mA/m^2. Moreover, the average adenosine-5′-triphosphate(ATP)content(an indicator of cell metabolism) at a current density of 1600 mA/m^2 was lower than that under no current, indicating heterotrophic denitrifying microbial activity can be inhibited at high current densities. Hence, direct electrical stimulation on the activity of heterotrophic denitrifying microorganisms in the developed system should be lower than 1600 mA/m^2. This study improves the understanding of electric current influence on heterotrophic denitrifying microorganisms and promotes the intelligent application of direct electrical stimulation on wastewater treatment processes.

固定化异养硝化-好氧反硝化菌在污水生物强化中的研究进展

异养硝化-好氧反硝化(Heterotrophic nitrification-aerobic denitrification,HN-AD)菌可以在有机碳存在的好氧条件下实现同时硝化和反硝化,广泛应用于各类污水处理过程中。综述了HN-AD菌株的脱氮特性和代谢途径,总结了其在污水处理中的应用和研究现状,比较了不同载体材料的优缺点,重点讨论了固定HN-AD菌株提高反应器处理效果和稳定性的作用机理。最后,展望了固定化HN-AD菌株在污水处理中面临的挑战和未来的研究方向。

一株脱氮毕赤酵母Y 4的分离鉴定及其氨氮降解性能

当前水体氨氮污染严重,环境治理迫在眉睫,筛选高效安全的脱氮菌株可为废水脱氮提供更多选择。本研究从沼气发酵污泥中分离筛选能降解氨氮的菌株,使用18S rDNA序列分析,对筛选出的优良菌株进行种类鉴定;同时采用单因素试验和正交试验,探究菌株最佳脱氮条件[培养温度、初始pH值、盐浓度、碳氮比(C/N)]及其硝化、反硝化能力。结果表明,分离筛选得到1株具有较强脱氮能力的野生型毕赤酵母(Pichia sp.)菌株Y 4。在试验优化后,得到菌株Y 4的最适脱氮条件:培养温度25℃,初始pH值8.0、盐浓度(氯化钠浓度)30 g/L、C/N为30。在培养24 h后菌株Y 4对亚硝酸氮(NO-2 N)的降解率为80.50%,在培养48 h后菌株Y 4对氨氮(NH+4 N)的降解率达99.92%,且在24 h内菌株Y 4的异养硝化和好氧反硝化的速率分别达到6.25和6.71 mg/(L·h),表明毕赤酵母菌株Y 4对废水氮污染处理以及生物修复具有一定的应用价值与潜力。

一株低温异养硝化-好氧反硝化细菌的分离鉴定与脱氮特性

微生物脱氮技术是污水氮素净化的有效方法之一,但低温条件下微生物活性和代谢能力降低,导致脱氮效果变差。为提升低温生活污水的氮素净化效果,从黑龙江龙凤湿地(46°53′N,125°18′E)中分离筛选到1株低温异养硝化-好氧反硝化细菌J1-2,16S rRNA基因序列分析结果显示为不动杆菌属(Acinetobacter)。该菌株在8℃低温条件下能有效去除不同形态氮源,对单一氨氮、硝酸盐氮和亚硝酸盐氮的60 h去除率分别为93.88%、81.46%、96.31%,对这3种氮源的利用顺序为氨氮、硝酸盐氮、亚硝酸盐氮。该菌株在8℃下去除氨氮的最佳条件如下:pH为7、转速为150 r/min、碳源为柠檬酸钠、碳氮质量比为20,该条件应用于实际生活污水处理,总氮(TN)、氨氮和硝酸盐氮去除率分别为55.79%、86.47%、65.40%,具有良好的应用前景。

好氧反硝化细菌Burkholderia sp.ZH8的脱氮特性与生物强化作用

针对传统生物脱氮处理含氮废水工艺流程复杂、脱氮效率低的问题,从实验室运行稳定的生物滤池筛选得到一株好氧反硝化菌株ZH8,该菌株具有较强的异养硝化和好氧反硝化能力.通过形态学、生理生化特征及16S rDNA序列分析,鉴定菌株ZH8为伯克氏菌(Burkholderia sp.).单因素试验表明,好氧反硝化菌株ZH8的最佳培养条件为:碳源为丁二酸钠,C/N为15,温度为30℃,转速为160r/min和pH值为7~8.此外,菌株ZH8不仅能以NH_(4)^(+)-N、NO_(3)^(-)-N或NO_(2)^(-)-N分别为单一氮源进行生长,而且在混合氮源条件下也可实现同步硝化反硝化作用.通过全基因组分析,菌株ZH8的异养硝化脱氮为谷氨酸脱氢酶途径的铵同化过程,好氧反硝化途径为:NO_(3)^(-)-N→NO_(2)^(-)-N→NO→N_(2)O→N_(2),碳代谢途径包括三羧酸循环、乙醛酸循环、糖酵解和戊糖磷酸.在采用纯菌ZH8为菌种培养的生物膜反应器中,反应运行至8~15d即可完成挂膜,系统可维持高效稳定的脱氮性能,NH_(4)^(+)-N、NO_(3)^(-)-N及TN的去除率均可达到90%以上.该菌株为生物处理领域提供了新型菌种资源,能够有效处理含氮废水,具有实际应用价值.

HN-AD菌群特性及其在水产养殖废水处理中的应用

我国水产养殖业正朝着高密度、集约化的方向发展,而养殖水体中NH_(4)^(+)-N、NO_(2)^(-)-N、NO_(3)^(-)-N的积累问题严重影响了水资源循环效率及水产品品质。异养硝化-好氧反硝化(HN-AD)菌群可以在较高溶解氧条件下去除养殖水体中的氮素,是净化养殖水质的关键核心。简述了具有HN-AD功能的微生物分类现状,阐述了具有HN-AD功能的微生物群落特征,分析了HN-AD菌在耐盐、降解农药和去除抗生素等方面的拓展效能,同时探究了基于环境因子的HN-AD过程调控效率,最后预测HN-AD技术在未来的发展方向。在可持续发展与“碳达峰”双重目标的驱动下,HN-AD技术正朝着更高效、可持续性和低碳排放的方向发展,助力水产养殖业不断提高水循环利用率和养殖密度。

异养/硫自养集成短程反硝化工艺影响因素研究

为保证厌氧氨氧化(Anammox)过程的亚硝态氮(NO2--N)稳定来源,提升自养脱氮系统的运行效能。本研究采用上流式厌氧污泥床(UASB)反应器,构建异养/硫自养集成短程反硝化系统,探究碳源浓度、pH、水力停留时间(HRT)、温度对系统的影响。结果表明,碳源浓度与系统短程反硝化效能成反比。碳源浓度为200 mg/L时NO2--N积累率(NAR)最高,此时NO_(2)^(-)-N平均积累率及其速率分别为76.99%和1.63 mg/(L·h),异养反硝化功能菌属Thauera丰度由3.26%升至19.73%。系统短程反硝化效能随pH的升高先升后降,pH为8.0±0.5时NAR最高,此时NO2--N平均积累率及其速率分别为79.46%和1.90 mg/(L·h);HRT与系统短程反硝化效能成反比。当HRT=9.7左右时NAR最高,此时NO2--N平均积累率及其速率分别为77.35%和1.68 mg/(L·h)。温度与系统短程反硝化效能成正比,与Thauera、Terrimonas等反硝化功能菌属丰度成反比。当温度在35℃左右时NAR最高,此时NO_(2)^(-)-N平均积累率及其速率分别为71.21%和1.68 mg/(L·h)。集成短程反硝化为自养脱氮工艺提供NO2--N的过程具有NAR高且运行稳定性强等优点,更适用于自养脱氮工艺稳定高效运行,为该领域提供更多思考。

低温对异养硝化菌Acinetobacter sp.生长代谢及群体感应影响研究

为进一步探究低温条件下HN-AD菌氮素转化机理及其群体感应系统对低温响应机制,采用GC-MS测定HN-AD过程中释放的信号分子,结果表明菌株JQ1004至少分泌包括C_(6)-,C_(8)-,和C_(10)-HSL在内3种高丝氨酸内酯(AHLs),其浓度变化均随细菌浓度增加不断累积,呈现显著密度依赖性.当温度由30℃降低至10℃时,C_(6)-和C_(8)-HSL呈不断下降趋势,而C_(10)-HSL变化为先上升后下降,并在20℃达到最大值10.89nmol/L,表明C_(10)-HSL在菌株适应低温环境中发挥重要作用.低温添加10nmol/L C_(10)-HSL可显著缩短细菌迟滞期,提高脱氮速率.20℃和10℃时添加C_(10)-HSL使amoA表达量分别提高了35%和22%,napA表达量分别增加了14%和9.4%.C_(10)-HSL通过提高luxR调控的受体蛋白活性来提高菌株群感能力.低温添加C_(10)-HSL可促进胞外蛋白分泌,降低多糖含量,提高生物膜活性,进而强化其对低温恶劣环境适应能力.

异养硝化-好氧反硝化细菌MF1的生长及脱氮特性研究

从某对虾养殖池塘采集的底泥中筛出一株菌株MF1,经16S rDNA鉴定为Klebsiella michiganensis(密歇根克雷伯氏菌)。各菌株生长特性试验表明:氨氮培养基中菌株MF1的最适碳源为丙酮酸钠。在温度30℃、pH 7、转速100 r/min及C/N比例为10的条件下,菌株MF1的脱氮效率达到最大值。

耐高温异养硝化-好氧反硝化菌的分离鉴定及其脱氮特性研究

经过16S rDNA以及各项菌株生理生化特性研究试验,寻找一种耐热性较好的异养硝化-好氧反硝化菌(HN-AD)菌株,并将菌株接种至模拟污水环境的硝酸盐氮培养基中,研究其在不充足碳源及充足碳源的情况下菌株的脱氮效率及硝酸盐氮去除率。结果表明,在温泉中筛选出一种可使污水的pH更适合自身反硝化作用的HN-AD菌株,经鉴定所筛选出的HN-AD菌为斯氏假单胞菌。该菌株在模拟污水环境下反硝化最适pH为8.0,在充足碳源的情况下,其硝酸盐氮去除率能到达93%以上。菌株不仅拥有着较为高效的脱氮效率以及较高的硝酸盐氮去除率,而且拥有着自我调节环境pH的能力,能在充足碳源的情况下调节至最佳氮降解的pH。

异养硝化-好氧反硝化细菌的研究进展及在水产养殖中的应用

异养硝化-好氧反硝化细菌可以同时实现硝化和反硝化,具有高效的脱氮性能,是目前废水生物脱氮处理的研究热点之一。文章从异养硝化-好氧反硝化细菌的研究现状、作用机制、脱氮影响因素、在水产养殖中的应用等4个方面进行了综述,可为异养硝化-好氧反硝化细菌的研究提供理论依据,为异养硝化-好氧反硝化细菌在水产养殖中的应用提供新的思路。