厌氧氨氧化甲烷化反硝化耦合的机理及动力学研究
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摘要
“可持续发展”和“节能减排”是当今废水处理技术研究与开发的指导思想和重要原则。论文通过实验室EGSB和BAF废水处理反应器及其耦合的连续运行试验、序批式反应动力学试验和分子生物学测试与分析,配合进行理论论证和模型研究,对厌氧氨氧化甲烷化反硝化的动力学规律、耦合机理及影响因素,O2和或微量NO_2下氨氧化菌的氨代谢特性及其动力学,EGSB与限制氧曝气BAF耦合处理废水的技术特点,进行了深入和系统研究;以便为研究和开发集好氧氨氧化、厌氧氨氧化、甲烷化和反硝化为一体的废水生物处理新技术,在高容积负荷速率下同时去除COD和氨组分的前提下节约能源和有机碳源,提供科学依据,积累重要技术资料。论文得到如下主要研究结果:
①以好氧活性污泥和厌氧活性污泥接种于2个膨胀颗粒污泥床(EGSB)反应器中,进水流量为10 mL/min,回流量为180 mL/min,进水COD浓度在180 mg/L,有机负荷率(OLR)和污泥负荷率(SLR)分别为2.51 kgCOD/(m~3.d)和0.19 kgCOD/(kgMLSS.d),出水COD维持在约40 mg/L,COD去除率达80%以上。控制温度在32～35℃,pH在6.8～7.2,反应器内氧化还原电位在-340 mV以下,水力停留时间(HRT)4.2 h,上升流速4.86 m/h以及加入80 mg/L絮凝剂(硫酸铝钾),进而缩短了启动时间,促进了颗粒污泥的形成。分别经过60 d和120 d运行,反应器启动成功。试验结果表明,上升流速、絮凝剂和污泥类型对颗粒污泥的形成有明显影响;接种好氧活性污泥在低浓度COD下,合理控制负荷率仍能成功启动EGSB反应器。
②完成启动的EGSB反应器控制温度在32℃～35℃、pH值在7.3～8.3、氧化还原电位在-40 mV～-150 mV,水力停留时间(HRT)和上升流速与启动期相同,进水中添加亚硝酸盐和氨盐组分,经过240 d运行,逐步耦合厌氧氨氧化甲烷化反硝化。EGSB1#和EGSB2#进水有机容积负荷速率(OLR)分别为4.8 kgCOD/(m~3.d)和2.88 kgCOD/(m~3.d),污泥负荷速率(SLR)分别为0.55 kgCOD/(kgMLSS.d)和0.33 kgCOD/(kgMLSS.d),COD去除率分别达到85%和76%,氨氮去除率分别达到35%和20%,亚硝态氮去除率都达到99.9%,总氮去除率分别达到67%和59%。结果表明,pH、温度、溶解氧、氧化还原电位、亚硝酸盐和有机物COD对EGSB反应器中厌氧氨氧化甲烷化反硝化的耦合和颗粒污泥的特性有影响。
③用间歇式试验同时测定颗粒污泥的氨氮和亚硝态氮的消耗量,求得污泥厌氧氨氧化活性为9.84×10~(-4) mgNH+4-N/(mgMLSS.h)或者12.9×10~(-4) mgNO_2~--N/(mgMLSS.h);通过16S-rRNA的分子生物学基因序列测试,发现了一种新的厌氧氨氧化菌(anaerobic ammonium-oxidizing Planctomycete cquenviron-1)。
④在间歇式试验反应器中接种颗粒污泥,对厌氧氨氧化、甲烷化和反硝化动力学特性进行分别研究。求得厌氧氨氧化动力学参数:最大氨氮反应速率为6.65×10~(-3)(mg NH+4-N/(mgMLSS.h)),氨氮半饱和常数为87.1 mg/L,氨氮抑制常数为1123 mg/L;亚硝态氮半饱和常数为15.39 mg/L,亚硝态氮抑制常数为159.5 mg/L;甲烷化动力学参数:最大比基质降解速率为0.158(mgCOD/(mgVSS.h)),半饱和常数为464.27 mg/L,甲烷的产率系数为0.254 ml/mg;短程反硝化动力学参数:最大反硝化速率为0.0069 mgNO_2~--N/(mgMLSS.h),亚硝酸盐氮半饱和常数为2.26 mg/L,有机物半饱和常数为4.72 mg/L。三者耦合时厌氧氨氧化速率、甲烷化速率和反硝化速率比单独研究时均减小。
⑤采用间歇式试验方法,分别研究微量NO_2对厌氧氨氧化、甲烷化和反硝化的影响。微量NO_2对厌氧氨氧化具有强化作用,保持厌氧氨氧化动力学参数值不变,基于Haldane模型建立了厌氧氨氧化的NO_2强化函数,估计了强化函数中的最大强化系数( 48.79)、NO_2半饱和常数(2570 mg/m~3)、NO_2抑制常数(3.9 mg/m~3)和基础速率系数(0.0188)。微量NO_2对甲烷化和反硝化的影响可用反竞争性抑制动力学方程进行描述;甲烷化的最大比乙酸盐去除速率为0.149 mgCOD/(mgVSS·h),乙酸盐半饱和常数为396 mgCOD/L,NO_2抑制系数为231 mg/m~3;反硝化的亚硝酸盐氮最大去除速率0.006865/h,亚硝酸盐氮半饱和常数0.1 mg/L,NO_2抑制系数为1530 mg/m~3。试验中大部分的NOX出现损失。
⑥采用间歇式试验方法,研究微量NO_2对厌氧氨氧化甲烷化反硝化三者耦合的影响。保持厌氧氨氧化动力学参数值不变,基于Haldane模型建立了厌氧氨氧化的NO_2强化函数,估计了强化函数中的最大强化系数(30.55)、NO_2半饱和常数(1960 mg/m~3)、NO_2抑制常数(8.2 mg/m~3)和基础速率系数(0.0314)。微量NO_2对三者耦合时甲烷化和反硝化的影响可用反竞争性抑制动力学方程进行描述;甲烷化的最大比乙酸盐去除速率为0.15 mgCOD/(mgVSS·h),乙酸盐半饱和常数为395 mgCOD/L,NO_2抑制系数为623 mg/m~3;反硝化的亚硝酸盐氮最大去除速率0.00685/h,亚硝酸盐氮半饱和常数0.214 mg/L,NO_2抑制系数为22400 mg/m~3。试验中大部分的NOX气体物质出现损失。三者耦合时甲烷化和反硝化受到NO_2的抑制作用明显低于独立试验;当NO_2小于50 ppm时,三者耦合时厌氧氨氧化速率比独立实验时小;在当NO_2大于50 ppm时,三者耦合时厌氧氨氧化速率与独立实验时大。
⑦采用SBR反应器,pH值控制在7.8～8.2,DO控制在0.8 mg/L～1.0 mg/L,温度控制在30±2℃。初始NH_4~+-N浓度为50 mg/L,待稳定后逐渐增加进水氨氮浓度至250 mg/L,每次增加幅度50 mg/L,MPN计数结果显示,经120 d富集,好氧氨氧化菌的浓度提高了300倍。间歇式试验结果表明,pH值、DO浓度和温度对好氧氨氧化菌的富集有显著影响。在富集过程中,控制pH值、DO浓度和温度是关键因素,游离氨和HNO_2进行适当控制,以保证抑制亚硝酸盐氧化菌而不抑制好氧氨氧化菌。
⑧采用批式呼吸法测得好氧氨氧化菌产率系数为0.2119 mg COD/mg NH4+-NOD(或者0.7268 mg COD/mg NH4+-N)氨氧化菌最大氨氮降解速率为0.1 mg NOD/(mg COD.h)(或者0.0292 mg N/(mg COD.h))。在间歇式试验中加入24μM NaN3抑制NO_2~--N氧化,建立氨氧化反应动力学方程,得到氨氮半饱和系数为18.38 mg NOD/L(或者5.36 mg NH4+-N/L),DO半饱和系数为0.494 mg/L。
⑨采用批式试验,在无分子氧条件下,确定了好氧氨氧化菌的NO_2型氨氧化动力学方程,得到了最大氨氧化速率( q NO_2,max=0.144 mgNOD/(mgCOD·h))、二氧化氮半饱和常数(K NO_2=0.821μmol/L )和二氧化氮抑制性常数( K I =1.721μmol/L)。在微量NO_2气体中添加2%O2氧气后,氨氧化速率明显提高,最大氨氧化速率发生在2%O2和50 ppm NO_2的条件下,达到0.198 mgNOD/(mgCOD·h)。在21%O2和微量NO_2条件下,氨氧化速率继续大幅度提高;在21%O2和100 ppmNO_2时氨氧化速率达到0.477 mgNOD/(mgCOD·h),比无NO_2空气曝气条件下氨氧化速率高3倍。提出了NO_2表观强化氨氧化函数的概念,建立了在O2和微量NO_2混合气体下的氨氧化动力学方程,利用2%O2和微量NO_2条件下的实验结果验证了动力学方程,讨论了NO_2强化氨氧化的机理。
⑩进行充填陶瓷滤料的曝气生物滤池去除COD和氨氧化试验。进水氨氮为50mg/L左右、COD为100 mg/L左右和回流比为200%时,经过20多天的运行,出水氨氮小于0.1mg/L、COD小于30mg/L、亚硝态氮为45.55 mg/L和硝态氮为4.18 mg/L;过大和过小的回流比对BAF的运行性能都是不利的。
⑾将EGSB与BAF耦合,实现了好氧氨氧化、厌氧氨氧化、甲烷化和反硝化的集成。当进水流量为13 mL/min,进水氨氮浓度为50 mg/L、COD浓度为500 mg/L,外回流比为200%时,系统的总氮去除率达到80.6%,出水COD浓度保持在40 mg/L以下,出水氨氮1 mg/L、亚硝态氮4.3 mg/L和硝态氮2.56 mg/L。系统的COD去除负荷速率:1.7122 (kg/m~3.d),氮去除负荷速率为:0.1363(kg/m~3.d);氨氮去除负荷速率为:0.1632 (kg/m~3.d);节约曝气量0.2405 (kg O2/m~3.d)和碳源0.1775 (kg COD /m~3.d),回收甲烷554.94(L/m~3.d)。
“Sustainable development”and“energy saving and emission reduction”are the guiding thought and important principle for the study and development of wastewater treatment technique, nowadays. The following contents were studyed by integrating of EGSB reactor technique and BAF reactor technique, batch experiment, molecule biology determination, theoretic argumentation and the research of model: the kinetic characteristics, mechanism and influencing factors of anaerobic ammonium oxidation, methanogenesis and denitrification; ammonia oxidation characteristics and kinetics of ammonia oxidizer mixed culture under the conditions of O2 and trace NO_2 mixed gasses; the characteristics of integration with EGSB reactor technique and BAF reactor technique for wastewater treatment. It provided important and scientific data conveniently for developing this technique with integration of the aerobic ammonium oxidation, anaerobic ammonium oxidation, methanogensis and denitrification, which could be of capability to save energy and organic substances requirements under the conditions of simultaneous removal of COD and ammonium at high loadings. The results were given as follows:
①Granular sludge was formed in two EGSB reactors inoculated with aerobic activated sludge and anaerobic activated sludge after the start-up operation of 120d and 60d,respectively. The effluent COD concentration about 40 mg/L and COD removal efficiency up to 80% were obtained with the influent flux 10 mL/min, inner recycle flux 180 mL/min, COD concentration 180 mg/L, the organic loading rate up to 1.728 kgCOD/(m~3.d) and the sludge loading rate 0.19 kgCOD/(kgMLSS.d). The temperature was controlled between 32℃and 35℃, pH at 6.8～7.2, redox potential below -340mV, HRT 4.2h, the up-velocity 4.86m/h and the flocculant (aluminium potassium sulfate) 80mg/L. The formation of the granular sludge was accelerated. Results demonstrated that up-velocity,flocculant and the type of sludge affected the formation of granular sludge.The start-up of the reactor which was inoculated with aerobic sludge was succeeded if the loading rate was controlled rationally in low strength COD.
②Anaerobic ammonium oxidation, methanogensis and denitrification were integrated after 240 d with adding nitrite and ammonium in EGSB reactor. The pH was controlled between 7.5 and 8.3, redox potential at -40 mV～-150 mV and temperature at 32℃～35℃in the reactor;HRT and the up-velocity were controlled as same as start-up. The organic loading rate (OLR) was 4.8 kgCOD/(m~3.d) and 2.88 kgCOD/(m~3.d), the sludge loading rate (SLR) was 0.55 kgCOD/(kgMLSS.d) and 0.33 kgCOD/(kgMLSS.d), COD removal efficiency was 85% and 76%, the ammonium nitrogen removal efficiency was 35% and 20%,the nitrite nitrogen was both 99.9% and the total nitrogen was 67% and 59% in EGSB1# and EGSB2#, respectively. Results demonstrated that pH, temperature, DO, redox potential, nitrite and COD affected the integration of anaerobic ammonium oxidation bacteria, Methanogenesis bacteria and denitrification bacteria and the characteristic of granular sludge in EGSB reactor.
③The anaerobic ammonium oxidation activity about 9.84×10~(-4) mg NH_4~+-N/(mgMLSS.h) or 12.9×10~(-4) mgNO_~2-N/(mgMLSS.h) was obtained by measuring the simultaneous consumption of ammonium and nitrite under anoxic conditions. A new anammox bacterial species, denominated as anaerobic ammonium-oxidizing Planctomycete cquenviron-1, was discovered by 16S-rRNA based molecule biology determination.
④The kinetic characteristics of anaerobic ammonium oxidation,methanogenesis and denitrification inoculated with the granular sludge from EGSB reactor, in which anaerobic ammonium oxidation, methanogenesis and denitrification were integrated, were investigated by batch experiment,respectively. The kinetic parameters of anaerobic ammonium oxidation were determined, where the maximum anaerobic ammonium oxidation rate was 6.65×10~(-3) mg NH+4-N/(mgMLSS.h), the half saturate coefficient of ammonium nitrogen and nitrite nitrogen were 87.1 mg/L and 15.39 mg/L,respectively;inhibition coefficient of ammonium nitrogen and nitrite nitrogen were 1123 mg/L and 159.5 mg/L,respectively.The kinetic parameters of methanogenesis were determined, where the maximum specific degradation rate, half saturation constant and yield coefficient were 0.158 mgCOD/(mgVSS·h), 464 mgCOD/L and 0.254 mL CH4/mgCOD respectively. The kinetic parameters of denitrification were determined, where the maximum denitrification rate, half saturation constant of NO-2-N and COD were 0.0069 mg NO_2~--N/(mgMLSS.h), 2.26 mg NO_2~--N/L and 4.72 mgCOD/L respectively.The rates of anaerobic ammonium oxidation, methanogenesis and denitrification in coupling experiments of them all decreased.
⑤The effect of trace NO_2 on anaerobic ammonium oxidation, methanogenesis and denitrification was investigated by batch experiment. Trace NO_2 apparently enhanced the anaerobic ammonium oxidation, the kinetic parameters of anaerobic ammonium oxidation were kept invariable, the function for NO_2 apparently to enhance ammonium oxidation was suggested using the Haldane based model. The parameters were estimated, where the half saturate coefficient of NO_2 was 2570 mg/m~3, inhibition coefficient of NO_2 was 3.9 mg/m~3, the maximum enhanced rate coefficient was 48.79, the basic rate coefficient was 0.0188.The methanogenesis and denitrification kinetics could be described using uncompetitive inhibition model. The maximum NaAc removal rate, NaAc half saturation constant and NO_2 inhibition coefficient in methanogenesis were 0.149 mgCOD/mgVSS·h, 396 mgCOD/L and 231 mg NO_2/ m~3,respectively. The maximum nitrite removal rate, half saturation constant of NO_2~--N and NO_2 inhibition coefficient in denitrification were 0.006865/h, 0.1 mg NO_2~--N/L and 1530 mg NO_2/m~3,respectively. The most of NOX in the experiment was lost.
⑥The effect of trace NO_2 on integrating anaerobic ammonium oxidation, methanogenesis and denitrification was investigated by batch experiment. The kinetic parameters of anaerobic ammonium oxidation were kept invariable, the function for NO_2 apparently to enhance ammonium oxidation was suggested using the Haldane based model. The parameters were estimated, where the half saturate coefficient of NO_2 was 1960 mg/m~3, inhibition coefficient of NO_2 was 8.2 mg/m~3, the maximum enhanced rate coefficient was 30.55, the basic rate coefficient was 0.0314 . The methanogenesis and denitrification kinetics the integration system could be described using uncompetitive inhibition model. The maximum NaAc removal rate, NaAc half saturation constant and NO_2 inhibition coefficient in methanogenesis were 0.15 mgCOD/mgVSS·h, 395 mgCOD/L and 623 mg NO_2/m~3 ,respectively. The maximum nitrite removal rate, half saturation constant of NO_2~--N and NO_2 inhibition coefficient in denitrification were 0.00685 /h, 0.214 mg NO_2~--N /L and 22400 mg NO_2/m~3,respectively. Most of the NOX in the experiment was lost.The rates of methanogenesis and denitrification in the coupling experiments of them were decreased. When NO_2 was lower than 50 ppm, the rate of anaerobic ammonium oxidation in the coupling experiments of them was smaller than it in the separate experiment, but larger when NO_2 was higher than 50 ppm.
⑦The mixed ammonium oxidizer culture was enriched from normal activated sludge in SBR reactor, in which the pH was controlled between 7.8 and 8.2 ,DO between 0.8 and 1.0 mg/L and temperature at 30±2℃. The experimentation was carried out through synthetic wastewater,in which the initial NH_4~+-N was 50 mg/L and was increased up to 250 mg/L gradually by the step of 50 mg/L.The MPN test was run every 15 days to characterize the enrichment of ammonium oxidizer.The concentration of the ammonium oxidizer culture increased by 300 times after the cultivation for 120 days. The results of batch experiment indicated that pH, DO and temperature had significant influence on the enrichment of aerobic ammonia oxidation bacteria. During the enrichment cultivation, pH, DO and temperature were the key factors, FA and nitrous acid could be properly controlled to inhibit the growth of nitrite oxidation bacteria, but not the ammonia oxidation bacteria.
⑧The biomass yield coefficient of 0.2119 mg COD produced/mg NH4+-NOD (or 0.7268 mg COD/mg NH_4~+-N)and the maximum specific ammonium nitrogen consumption rate of 0.1 mg NOD/(mg COD.h)(or 0.0292 mg N/(mg COD.h)) for autotrophic aerobic ammonia oxidation bacteria were obtained from batch respirometry. The kinetics of aerobic ammonia oxidation was established by adding 24μM NaN3 to the sequence batch reactor. The kinetic parameters were determined, where the half saturate coefficient of NH4+-N was 18.38 mg NOD/L (or 5.36 mg NH_4~+-N/L) and the half saturate coefficient of DO was 0.494 mg/L.
⑨The kinetics of the NO_2-dependent ammonia oxidation was developed for ammonia oxidizer mixed culture when there was no molecular oxygen in the batch tests. The kinetic parameters were determined, where the half saturate coefficient of NO_2 was 0.821μmol/L, inhibition coefficient of NO_2 concentration was 1.721μmol/L, the maximum ammonium oxidation rate was 0.144 mgNOD/(mgCOD·h). After adding 2%O_2 to trace NO_2, the ammonium oxidation rates increased obviously. The maximum ammonium oxidation rate, 0.198 mgNOD/(mgCOD·h) occurred under the condition of the mixed gasses containing 2%O_2 and 50 ppm NO_2. Under the condition of mixed gasses containing 21%O_2 and trace NO_2, the ammonium oxidation rates further increased greatly. The maximum ammonium oxidation rate, 0.477 mgNOD/(mgCOD·h) occurred under the condition of 21%O_2 and 100 ppm NO_2 in the mixed gas, which is 3 times higher than the general aerobic ammonium oxidation rate. The function for NO_2 apparently to enhance ammonium oxidation was suggested. The kinetic models of ammonium oxidation under the conditions of O_2 and trace NO_2 mixed gasses was developed. The model was validated by the results of ammonium oxidation experiments under the conditions of the mixed gasses containing 2%O_2 and trace NO_2. The mechanism for NO_2 to enhance ammonium oxidation under the conditions of O_2 and trace NO_2 mixed gasses was discussed.
⑩COD removal and aerobic ammonium oxidation were performed in BAF reactor filled with expanded clay.After the start-up operation of 20d, the effluent COD concentration below 30 mg/L ,ammonium nitrogen below 0.1 mg/L ,nitrite nitrogen 45.55 mg/L and nitrate nitrogen 4.18 mg/L were obtained with the influent ammonium nitrogen concentration about 50 mg/L, COD concentration about 100 mg/L and inner recycle ratio 200% in BAF reactor. Bigger or smaller inner recycle ratio was unsuitable for running of BAF reactor.
⑾The aerobic ammonium oxidation,anaerobic ammonium oxidation, methanogensis and denitrification were coupled in the integration of EGSB reactor technique and BAF reactor technique. The removal efficiency of total nitrogen was 80.6% and COD concentration was below 40 mg/L, ammonium nitrogen below 1 mg/L, nitrite nitrogen 4.3 mg/L, nitrate nitrogen 2.56 mg/L in the effluent under the performance conditions of influent flux 13 mL/min,ammonium nitrogen concentration 50 mg/L , COD concentration 500 mg/L and outer recycle ratio 200%. Because of application for EGSB-BAF system, the removal rate of total COD, total nitrogen and total ammonium nitrogen were 1.7122 (kg/m~3.d), 0.1363 (kg/m~3.d) and 0.1632 (kg/m~3.d) ,respectively; it can saved 0.2405 (kg O_2/m~3.d) and 0.1775 (kg COD /m~3.d), received 554.94(L CH4 /m~3.d).
①以好氧活性污泥和厌氧活性污泥接种于2个膨胀颗粒污泥床(EGSB)反应器中,进水流量为10 mL/min,回流量为180 mL/min,进水COD浓度在180 mg/L,有机负荷率(OLR)和污泥负荷率(SLR)分别为2.51 kgCOD/(m~3.d)和0.19 kgCOD/(kgMLSS.d),出水COD维持在约40 mg/L,COD去除率达80%以上。控制温度在32～35℃,pH在6.8～7.2,反应器内氧化还原电位在-340 mV以下,水力停留时间(HRT)4.2 h,上升流速4.86 m/h以及加入80 mg/L絮凝剂(硫酸铝钾),进而缩短了启动时间,促进了颗粒污泥的形成。分别经过60 d和120 d运行,反应器启动成功。试验结果表明,上升流速、絮凝剂和污泥类型对颗粒污泥的形成有明显影响;接种好氧活性污泥在低浓度COD下,合理控制负荷率仍能成功启动EGSB反应器。
②完成启动的EGSB反应器控制温度在32℃～35℃、pH值在7.3～8.3、氧化还原电位在-40 mV～-150 mV,水力停留时间(HRT)和上升流速与启动期相同,进水中添加亚硝酸盐和氨盐组分,经过240 d运行,逐步耦合厌氧氨氧化甲烷化反硝化。EGSB1#和EGSB2#进水有机容积负荷速率(OLR)分别为4.8 kgCOD/(m~3.d)和2.88 kgCOD/(m~3.d),污泥负荷速率(SLR)分别为0.55 kgCOD/(kgMLSS.d)和0.33 kgCOD/(kgMLSS.d),COD去除率分别达到85%和76%,氨氮去除率分别达到35%和20%,亚硝态氮去除率都达到99.9%,总氮去除率分别达到67%和59%。结果表明,pH、温度、溶解氧、氧化还原电位、亚硝酸盐和有机物COD对EGSB反应器中厌氧氨氧化甲烷化反硝化的耦合和颗粒污泥的特性有影响。
③用间歇式试验同时测定颗粒污泥的氨氮和亚硝态氮的消耗量,求得污泥厌氧氨氧化活性为9.84×10~(-4) mgNH+4-N/(mgMLSS.h)或者12.9×10~(-4) mgNO_2~--N/(mgMLSS.h);通过16S-rRNA的分子生物学基因序列测试,发现了一种新的厌氧氨氧化菌(anaerobic ammonium-oxidizing Planctomycete cquenviron-1)。
④在间歇式试验反应器中接种颗粒污泥,对厌氧氨氧化、甲烷化和反硝化动力学特性进行分别研究。求得厌氧氨氧化动力学参数:最大氨氮反应速率为6.65×10~(-3)(mg NH+4-N/(mgMLSS.h)),氨氮半饱和常数为87.1 mg/L,氨氮抑制常数为1123 mg/L;亚硝态氮半饱和常数为15.39 mg/L,亚硝态氮抑制常数为159.5 mg/L;甲烷化动力学参数:最大比基质降解速率为0.158(mgCOD/(mgVSS.h)),半饱和常数为464.27 mg/L,甲烷的产率系数为0.254 ml/mg;短程反硝化动力学参数:最大反硝化速率为0.0069 mgNO_2~--N/(mgMLSS.h),亚硝酸盐氮半饱和常数为2.26 mg/L,有机物半饱和常数为4.72 mg/L。三者耦合时厌氧氨氧化速率、甲烷化速率和反硝化速率比单独研究时均减小。
⑤采用间歇式试验方法,分别研究微量NO_2对厌氧氨氧化、甲烷化和反硝化的影响。微量NO_2对厌氧氨氧化具有强化作用,保持厌氧氨氧化动力学参数值不变,基于Haldane模型建立了厌氧氨氧化的NO_2强化函数,估计了强化函数中的最大强化系数( 48.79)、NO_2半饱和常数(2570 mg/m~3)、NO_2抑制常数(3.9 mg/m~3)和基础速率系数(0.0188)。微量NO_2对甲烷化和反硝化的影响可用反竞争性抑制动力学方程进行描述;甲烷化的最大比乙酸盐去除速率为0.149 mgCOD/(mgVSS·h),乙酸盐半饱和常数为396 mgCOD/L,NO_2抑制系数为231 mg/m~3;反硝化的亚硝酸盐氮最大去除速率0.006865/h,亚硝酸盐氮半饱和常数0.1 mg/L,NO_2抑制系数为1530 mg/m~3。试验中大部分的NOX出现损失。
⑥采用间歇式试验方法,研究微量NO_2对厌氧氨氧化甲烷化反硝化三者耦合的影响。保持厌氧氨氧化动力学参数值不变,基于Haldane模型建立了厌氧氨氧化的NO_2强化函数,估计了强化函数中的最大强化系数(30.55)、NO_2半饱和常数(1960 mg/m~3)、NO_2抑制常数(8.2 mg/m~3)和基础速率系数(0.0314)。微量NO_2对三者耦合时甲烷化和反硝化的影响可用反竞争性抑制动力学方程进行描述;甲烷化的最大比乙酸盐去除速率为0.15 mgCOD/(mgVSS·h),乙酸盐半饱和常数为395 mgCOD/L,NO_2抑制系数为623 mg/m~3;反硝化的亚硝酸盐氮最大去除速率0.00685/h,亚硝酸盐氮半饱和常数0.214 mg/L,NO_2抑制系数为22400 mg/m~3。试验中大部分的NOX气体物质出现损失。三者耦合时甲烷化和反硝化受到NO_2的抑制作用明显低于独立试验;当NO_2小于50 ppm时,三者耦合时厌氧氨氧化速率比独立实验时小;在当NO_2大于50 ppm时,三者耦合时厌氧氨氧化速率与独立实验时大。
⑦采用SBR反应器,pH值控制在7.8～8.2,DO控制在0.8 mg/L～1.0 mg/L,温度控制在30±2℃。初始NH_4~+-N浓度为50 mg/L,待稳定后逐渐增加进水氨氮浓度至250 mg/L,每次增加幅度50 mg/L,MPN计数结果显示,经120 d富集,好氧氨氧化菌的浓度提高了300倍。间歇式试验结果表明,pH值、DO浓度和温度对好氧氨氧化菌的富集有显著影响。在富集过程中,控制pH值、DO浓度和温度是关键因素,游离氨和HNO_2进行适当控制,以保证抑制亚硝酸盐氧化菌而不抑制好氧氨氧化菌。
⑧采用批式呼吸法测得好氧氨氧化菌产率系数为0.2119 mg COD/mg NH4+-NOD(或者0.7268 mg COD/mg NH4+-N)氨氧化菌最大氨氮降解速率为0.1 mg NOD/(mg COD.h)(或者0.0292 mg N/(mg COD.h))。在间歇式试验中加入24μM NaN3抑制NO_2~--N氧化,建立氨氧化反应动力学方程,得到氨氮半饱和系数为18.38 mg NOD/L(或者5.36 mg NH4+-N/L),DO半饱和系数为0.494 mg/L。
⑨采用批式试验,在无分子氧条件下,确定了好氧氨氧化菌的NO_2型氨氧化动力学方程,得到了最大氨氧化速率( q NO_2,max=0.144 mgNOD/(mgCOD·h))、二氧化氮半饱和常数(K NO_2=0.821μmol/L )和二氧化氮抑制性常数( K I =1.721μmol/L)。在微量NO_2气体中添加2%O2氧气后,氨氧化速率明显提高,最大氨氧化速率发生在2%O2和50 ppm NO_2的条件下,达到0.198 mgNOD/(mgCOD·h)。在21%O2和微量NO_2条件下,氨氧化速率继续大幅度提高;在21%O2和100 ppmNO_2时氨氧化速率达到0.477 mgNOD/(mgCOD·h),比无NO_2空气曝气条件下氨氧化速率高3倍。提出了NO_2表观强化氨氧化函数的概念,建立了在O2和微量NO_2混合气体下的氨氧化动力学方程,利用2%O2和微量NO_2条件下的实验结果验证了动力学方程,讨论了NO_2强化氨氧化的机理。
⑩进行充填陶瓷滤料的曝气生物滤池去除COD和氨氧化试验。进水氨氮为50mg/L左右、COD为100 mg/L左右和回流比为200%时,经过20多天的运行,出水氨氮小于0.1mg/L、COD小于30mg/L、亚硝态氮为45.55 mg/L和硝态氮为4.18 mg/L;过大和过小的回流比对BAF的运行性能都是不利的。
⑾将EGSB与BAF耦合,实现了好氧氨氧化、厌氧氨氧化、甲烷化和反硝化的集成。当进水流量为13 mL/min,进水氨氮浓度为50 mg/L、COD浓度为500 mg/L,外回流比为200%时,系统的总氮去除率达到80.6%,出水COD浓度保持在40 mg/L以下,出水氨氮1 mg/L、亚硝态氮4.3 mg/L和硝态氮2.56 mg/L。系统的COD去除负荷速率:1.7122 (kg/m~3.d),氮去除负荷速率为:0.1363(kg/m~3.d);氨氮去除负荷速率为:0.1632 (kg/m~3.d);节约曝气量0.2405 (kg O2/m~3.d)和碳源0.1775 (kg COD /m~3.d),回收甲烷554.94(L/m~3.d)。
“Sustainable development”and“energy saving and emission reduction”are the guiding thought and important principle for the study and development of wastewater treatment technique, nowadays. The following contents were studyed by integrating of EGSB reactor technique and BAF reactor technique, batch experiment, molecule biology determination, theoretic argumentation and the research of model: the kinetic characteristics, mechanism and influencing factors of anaerobic ammonium oxidation, methanogenesis and denitrification; ammonia oxidation characteristics and kinetics of ammonia oxidizer mixed culture under the conditions of O2 and trace NO_2 mixed gasses; the characteristics of integration with EGSB reactor technique and BAF reactor technique for wastewater treatment. It provided important and scientific data conveniently for developing this technique with integration of the aerobic ammonium oxidation, anaerobic ammonium oxidation, methanogensis and denitrification, which could be of capability to save energy and organic substances requirements under the conditions of simultaneous removal of COD and ammonium at high loadings. The results were given as follows:
①Granular sludge was formed in two EGSB reactors inoculated with aerobic activated sludge and anaerobic activated sludge after the start-up operation of 120d and 60d,respectively. The effluent COD concentration about 40 mg/L and COD removal efficiency up to 80% were obtained with the influent flux 10 mL/min, inner recycle flux 180 mL/min, COD concentration 180 mg/L, the organic loading rate up to 1.728 kgCOD/(m~3.d) and the sludge loading rate 0.19 kgCOD/(kgMLSS.d). The temperature was controlled between 32℃and 35℃, pH at 6.8～7.2, redox potential below -340mV, HRT 4.2h, the up-velocity 4.86m/h and the flocculant (aluminium potassium sulfate) 80mg/L. The formation of the granular sludge was accelerated. Results demonstrated that up-velocity,flocculant and the type of sludge affected the formation of granular sludge.The start-up of the reactor which was inoculated with aerobic sludge was succeeded if the loading rate was controlled rationally in low strength COD.
②Anaerobic ammonium oxidation, methanogensis and denitrification were integrated after 240 d with adding nitrite and ammonium in EGSB reactor. The pH was controlled between 7.5 and 8.3, redox potential at -40 mV～-150 mV and temperature at 32℃～35℃in the reactor;HRT and the up-velocity were controlled as same as start-up. The organic loading rate (OLR) was 4.8 kgCOD/(m~3.d) and 2.88 kgCOD/(m~3.d), the sludge loading rate (SLR) was 0.55 kgCOD/(kgMLSS.d) and 0.33 kgCOD/(kgMLSS.d), COD removal efficiency was 85% and 76%, the ammonium nitrogen removal efficiency was 35% and 20%,the nitrite nitrogen was both 99.9% and the total nitrogen was 67% and 59% in EGSB1# and EGSB2#, respectively. Results demonstrated that pH, temperature, DO, redox potential, nitrite and COD affected the integration of anaerobic ammonium oxidation bacteria, Methanogenesis bacteria and denitrification bacteria and the characteristic of granular sludge in EGSB reactor.
③The anaerobic ammonium oxidation activity about 9.84×10~(-4) mg NH_4~+-N/(mgMLSS.h) or 12.9×10~(-4) mgNO_~2-N/(mgMLSS.h) was obtained by measuring the simultaneous consumption of ammonium and nitrite under anoxic conditions. A new anammox bacterial species, denominated as anaerobic ammonium-oxidizing Planctomycete cquenviron-1, was discovered by 16S-rRNA based molecule biology determination.
④The kinetic characteristics of anaerobic ammonium oxidation,methanogenesis and denitrification inoculated with the granular sludge from EGSB reactor, in which anaerobic ammonium oxidation, methanogenesis and denitrification were integrated, were investigated by batch experiment,respectively. The kinetic parameters of anaerobic ammonium oxidation were determined, where the maximum anaerobic ammonium oxidation rate was 6.65×10~(-3) mg NH+4-N/(mgMLSS.h), the half saturate coefficient of ammonium nitrogen and nitrite nitrogen were 87.1 mg/L and 15.39 mg/L,respectively;inhibition coefficient of ammonium nitrogen and nitrite nitrogen were 1123 mg/L and 159.5 mg/L,respectively.The kinetic parameters of methanogenesis were determined, where the maximum specific degradation rate, half saturation constant and yield coefficient were 0.158 mgCOD/(mgVSS·h), 464 mgCOD/L and 0.254 mL CH4/mgCOD respectively. The kinetic parameters of denitrification were determined, where the maximum denitrification rate, half saturation constant of NO-2-N and COD were 0.0069 mg NO_2~--N/(mgMLSS.h), 2.26 mg NO_2~--N/L and 4.72 mgCOD/L respectively.The rates of anaerobic ammonium oxidation, methanogenesis and denitrification in coupling experiments of them all decreased.
⑤The effect of trace NO_2 on anaerobic ammonium oxidation, methanogenesis and denitrification was investigated by batch experiment. Trace NO_2 apparently enhanced the anaerobic ammonium oxidation, the kinetic parameters of anaerobic ammonium oxidation were kept invariable, the function for NO_2 apparently to enhance ammonium oxidation was suggested using the Haldane based model. The parameters were estimated, where the half saturate coefficient of NO_2 was 2570 mg/m~3, inhibition coefficient of NO_2 was 3.9 mg/m~3, the maximum enhanced rate coefficient was 48.79, the basic rate coefficient was 0.0188.The methanogenesis and denitrification kinetics could be described using uncompetitive inhibition model. The maximum NaAc removal rate, NaAc half saturation constant and NO_2 inhibition coefficient in methanogenesis were 0.149 mgCOD/mgVSS·h, 396 mgCOD/L and 231 mg NO_2/ m~3,respectively. The maximum nitrite removal rate, half saturation constant of NO_2~--N and NO_2 inhibition coefficient in denitrification were 0.006865/h, 0.1 mg NO_2~--N/L and 1530 mg NO_2/m~3,respectively. The most of NOX in the experiment was lost.
⑥The effect of trace NO_2 on integrating anaerobic ammonium oxidation, methanogenesis and denitrification was investigated by batch experiment. The kinetic parameters of anaerobic ammonium oxidation were kept invariable, the function for NO_2 apparently to enhance ammonium oxidation was suggested using the Haldane based model. The parameters were estimated, where the half saturate coefficient of NO_2 was 1960 mg/m~3, inhibition coefficient of NO_2 was 8.2 mg/m~3, the maximum enhanced rate coefficient was 30.55, the basic rate coefficient was 0.0314 . The methanogenesis and denitrification kinetics the integration system could be described using uncompetitive inhibition model. The maximum NaAc removal rate, NaAc half saturation constant and NO_2 inhibition coefficient in methanogenesis were 0.15 mgCOD/mgVSS·h, 395 mgCOD/L and 623 mg NO_2/m~3 ,respectively. The maximum nitrite removal rate, half saturation constant of NO_2~--N and NO_2 inhibition coefficient in denitrification were 0.00685 /h, 0.214 mg NO_2~--N /L and 22400 mg NO_2/m~3,respectively. Most of the NOX in the experiment was lost.The rates of methanogenesis and denitrification in the coupling experiments of them were decreased. When NO_2 was lower than 50 ppm, the rate of anaerobic ammonium oxidation in the coupling experiments of them was smaller than it in the separate experiment, but larger when NO_2 was higher than 50 ppm.
⑦The mixed ammonium oxidizer culture was enriched from normal activated sludge in SBR reactor, in which the pH was controlled between 7.8 and 8.2 ,DO between 0.8 and 1.0 mg/L and temperature at 30±2℃. The experimentation was carried out through synthetic wastewater,in which the initial NH_4~+-N was 50 mg/L and was increased up to 250 mg/L gradually by the step of 50 mg/L.The MPN test was run every 15 days to characterize the enrichment of ammonium oxidizer.The concentration of the ammonium oxidizer culture increased by 300 times after the cultivation for 120 days. The results of batch experiment indicated that pH, DO and temperature had significant influence on the enrichment of aerobic ammonia oxidation bacteria. During the enrichment cultivation, pH, DO and temperature were the key factors, FA and nitrous acid could be properly controlled to inhibit the growth of nitrite oxidation bacteria, but not the ammonia oxidation bacteria.
⑧The biomass yield coefficient of 0.2119 mg COD produced/mg NH4+-NOD (or 0.7268 mg COD/mg NH_4~+-N)and the maximum specific ammonium nitrogen consumption rate of 0.1 mg NOD/(mg COD.h)(or 0.0292 mg N/(mg COD.h)) for autotrophic aerobic ammonia oxidation bacteria were obtained from batch respirometry. The kinetics of aerobic ammonia oxidation was established by adding 24μM NaN3 to the sequence batch reactor. The kinetic parameters were determined, where the half saturate coefficient of NH4+-N was 18.38 mg NOD/L (or 5.36 mg NH_4~+-N/L) and the half saturate coefficient of DO was 0.494 mg/L.
⑨The kinetics of the NO_2-dependent ammonia oxidation was developed for ammonia oxidizer mixed culture when there was no molecular oxygen in the batch tests. The kinetic parameters were determined, where the half saturate coefficient of NO_2 was 0.821μmol/L, inhibition coefficient of NO_2 concentration was 1.721μmol/L, the maximum ammonium oxidation rate was 0.144 mgNOD/(mgCOD·h). After adding 2%O_2 to trace NO_2, the ammonium oxidation rates increased obviously. The maximum ammonium oxidation rate, 0.198 mgNOD/(mgCOD·h) occurred under the condition of the mixed gasses containing 2%O_2 and 50 ppm NO_2. Under the condition of mixed gasses containing 21%O_2 and trace NO_2, the ammonium oxidation rates further increased greatly. The maximum ammonium oxidation rate, 0.477 mgNOD/(mgCOD·h) occurred under the condition of 21%O_2 and 100 ppm NO_2 in the mixed gas, which is 3 times higher than the general aerobic ammonium oxidation rate. The function for NO_2 apparently to enhance ammonium oxidation was suggested. The kinetic models of ammonium oxidation under the conditions of O_2 and trace NO_2 mixed gasses was developed. The model was validated by the results of ammonium oxidation experiments under the conditions of the mixed gasses containing 2%O_2 and trace NO_2. The mechanism for NO_2 to enhance ammonium oxidation under the conditions of O_2 and trace NO_2 mixed gasses was discussed.
⑩COD removal and aerobic ammonium oxidation were performed in BAF reactor filled with expanded clay.After the start-up operation of 20d, the effluent COD concentration below 30 mg/L ,ammonium nitrogen below 0.1 mg/L ,nitrite nitrogen 45.55 mg/L and nitrate nitrogen 4.18 mg/L were obtained with the influent ammonium nitrogen concentration about 50 mg/L, COD concentration about 100 mg/L and inner recycle ratio 200% in BAF reactor. Bigger or smaller inner recycle ratio was unsuitable for running of BAF reactor.
⑾The aerobic ammonium oxidation,anaerobic ammonium oxidation, methanogensis and denitrification were coupled in the integration of EGSB reactor technique and BAF reactor technique. The removal efficiency of total nitrogen was 80.6% and COD concentration was below 40 mg/L, ammonium nitrogen below 1 mg/L, nitrite nitrogen 4.3 mg/L, nitrate nitrogen 2.56 mg/L in the effluent under the performance conditions of influent flux 13 mL/min,ammonium nitrogen concentration 50 mg/L , COD concentration 500 mg/L and outer recycle ratio 200%. Because of application for EGSB-BAF system, the removal rate of total COD, total nitrogen and total ammonium nitrogen were 1.7122 (kg/m~3.d), 0.1363 (kg/m~3.d) and 0.1632 (kg/m~3.d) ,respectively; it can saved 0.2405 (kg O_2/m~3.d) and 0.1775 (kg COD /m~3.d), received 554.94(L CH4 /m~3.d).
引文
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