基于A/O/A运行模式的SBR工艺脱氮除磷效能及其微生物特性研究
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摘要
随着“水体富营养化”问题的日渐突出,污水排放标准的日趋严格,如何经济、高效的去除氮磷污染成为水体污染防治领域的研究热点。生物脱氮除磷工艺因其经济、环保等优点而得到了广泛应用,但其本身也存在一些问题亟待解决,如脱氮和除磷过程存在底物竞争和污泥龄等矛盾,使得生物脱氮除磷工艺的去除效能难以提高。一些新的脱氮除磷理论,如反硝化除磷、同步硝化反硝化等为解决生物脱氮除磷工艺中存在的矛盾提供了新思路。
以反硝化除磷理论为基础,采用A/O/A脱氮除磷工艺为研究主体,分别以人工合成污水和实际生活污水为处理对象,全面系统的研究了A/O/A工艺启动过程中的去除效能及污泥形态的变化,详细的考察了两个系统内颗粒污泥特性的差异。对比分析了环境因子对A/O/A工艺运行效果的冲击和持续影响。探讨了启动期系统内微生物的群落演替,及环境因子对微生物种群结构的影响。在此基础上,对筛选出的反硝化聚磷菌株进行鉴定,并研究了其除磷性能。
A/O/A脱氮除磷工艺启动过程中,随着系统去除效能的提高,污泥形态逐渐由絮体转化为颗粒、原生动物从无到有。稳定运行过程中,运行模式相同,进水水质不同的两个系统内的颗粒污泥性质存在较大差异。以实际生活污水培养的颗粒污泥与人工配水培养的颗粒污泥相比较,具有粒径小、沉淀速率慢、孔径大、比表面积小、比重轻、含水率高、活性高、胞外聚合物含量高、胞内聚合物含量低,释磷量小、反硝化效果好等特点。
影响因子对A/O/A脱氮除磷工艺去除性能的冲击影响显著。乙酸钠和丙酸钠为最佳碳源;丁二酸钠略差;乙醇和葡萄糖较差;异戊酸为最差碳源。进水pH值在6.0~8.0的范围内,对系统去除COD无明显影响。进水pH值为7.0时,系统的脱氮除磷效果最佳。进水C/N比在0.5~8.3的范围内,对系统去除COD的影响不大。随着进水C/N比的升高,系统对TN的去除有增加趋势。进水C/N比在0.5~6.1的范围内,系统对PO_4~(3-)-P的去除效果较好。进水中缺少钙、镁和钾离子,对系统去除有机物和脱氮没有明显的影响。钙、镁和钾离子皆参与释磷和吸磷反应,与磷的摩尔比分别为6.1×10~(-2),3.8×10~(-1),2.8×10~(-1)。缺少钙、镁和钾离子会引起聚磷菌超量释磷,释磷量顺序依次为:缺钾>缺钙>缺镁>正常系统。环境温度在10℃~30℃范围内,对系统去除COD的影响较小,释磷和吸磷速率随温度的升高而升高。温度为10℃时,系统硝化效果较差。浓度低于30mg/L时,NO_3~--N和NO_2~--N皆可替代O_2作为电子受体,进行过量吸磷反应。吸磷速率顺序依次为:O_2> NO_3~--N> NO_2~--N。
影响因子对A/O/A脱氮除磷工艺的持续影响与冲击影响结果并不完全一致。较长时间以乙酸钠和丙酸钠为进水碳源,系统皆可保持良好的脱氮除磷效果;而丁二酸钠为碳源则可导致脱氮效果下降,且除磷效果也随之恶化。系统pH值在7.5~8.5的范围内运行,可取得良好的去除效果;而长期在pH值为6.5的条件下运行,系统的处理效果会逐渐恶化。系统C/N长期在2.6~3.3的范围内运行,可取得理想的去除效果。但C/N进一步升高,系统的出水中磷浓度会逐渐升高,导致处理效能逐渐下降。
启动期,A/O/A脱氮除磷工艺内的微生物种群结构发生了动态变化。人工配水系统中部分优势菌如Candidatus Accumulibacter phosphatis在系统中始终存在;部分优势菌如Rhodospirillales随着系统的运行逐渐消失,取而代之出现新的优势菌如Rhodocyclaceae和Candidatus Accumulibacter phosphates。实际生活污水系统内亦发生了微生物的群落演替,优势菌M1和M3(β- proteobacteria)始终存在,随着系统的运行出现新的优势菌M14(β- proteobacteria)。环境因子对系统内的微生物的种群结构影响显著。系统内始终存在的优势菌如Denitrifying bacterium和Uncultured Chloroflexi bacterium耐受力较强,为常驻菌;而部分优势菌如Ammonia oxidizing bacterium则随着运行条件的改变,逐渐被系统淘汰。
从稳定运行的系统中分离纯化出25株菌,通过初筛和复筛试验选取除磷效率较高的4株菌:JL4、FH6、FQ3和FQ4。采用生理生化试验、脂肪酸鉴定及16S rDNA基因测序等手段进行鉴定:菌株JL4 Pseudomonas sp.,菌株FH6、FQ3和FQ4均为Bacillus sp.。环境因子pH值、碳源和温度对菌株生长及除磷效果影响显著。在pH值为5的条件下,菌株几乎停止生长,除磷效果极差;最适pH值为中性。菌株JL4、FH6和FQ3的最适温度为30℃,菌株FQ4的最适温度为25℃。在不同碳源的条件下,菌株的生长速率及除磷率顺序依次为:乙酸钠>丙酸钠>丁二酸钠>葡萄糖。
As the issue of eutrophication grows more severely and the regulatory requirements of wastewater discharge become increasingly stricter, economical and efficient removal of nitrogen and phosphorus has been a focus of wastewater treatment research. Though biological nitrogen and phosphorus removal process has been applied widely as one of economical and sustainable processes, it has some inherent problems that need to be solved. For example, the contradictions of substrates competition and SRT between the nitrogen and phosphorus removal, have inhibited the enhancement of removal efficiency. Some novel theories of nitrogen and phosphorus removal, such as denitrifying phosphorus removal and simultaneous nitrification and denitrification supply new ideas in solving theses contradictions.
Based on the theory of denitrifying phosphorus removal, A/O/A simultaneous nitrogen and phosphorus removal process was adopted as research subject. Dynamic changes in the removal efficiencies and morphologies of activated sludge of two A/O/A processes which treated with synthetic wastewater and domestic wastewater respectively were investigated systematically, and the differences of granular sludge characteristics in the two systems were analyzed comprehensively. Short and long -term influences of environmental factors on the removal efficiency of the A/O/A process were analyzed compar comparatively. Microbial community succession in the systems during the start-up period and the effect of environmental factors on the microbial population structure were investigated. Isolated denitrifying phosphorus removing bacteria were identified, and their phosphorus removal efficiency were further studied.
During the start-up period, flocculent sludge transformed into granular sludge gradually with the enhancement of removal efficiency, and lots of protozoa appeared in the A/O/A nitrogen and phosphorus removal processes. Granular sludge in the two systems treating different wastewater were quite distinct under the same operational condition in the stable phase. Granular sludge cultivated with real domestic wastewater was smaller in particle size, slower in sedimentation, larger in pore size, smaller in specific surface area, lighter in specific weight, higher in water content, higher in activity, more in extracellular polymeric content, lower in intracellular polymeric content, less in phosphorus release, and better in dintrification, compared with that cultivated with synthetic wastewater.
Environmental factors had vital influence on the performance of A/O/A nitrogen and phosphorus removal process in short-term. Sodium acetate and sodium propionate were optimal carbon source, and sodium succinate was slightly worse than them. Ethanol and glucose were unsuitable carbon source, and isovalerate was the worst carbon source. As pH value of influent changed from 6.0 to 8.0, no significant effect on the COD removal efficiency of the system were found. Influent pH at 7.0, perfect performance was achived. Value of C/N in the range of 0.5 to 8.3 had little effect on the COD removal efficiency. Total nitrogen removal efficiency increased with the increase of influent C/N. phosphorus removal efficiency of the system was perfect while C/N was in the range of 0.5 to 6.1, but it deteriorated when C/N was larger than 6.1. Value of C/N in the range of 0.5 to 8.3 had little effect on the COD removal efficiency. Total nitrogen removal efficiency increased with the increase of influent C/N. phosphorus removal efficiency of the system was perfect while C/N was in the range of 0.5 to 6.1, but it deteriorated when C/N was larger than 6.1. Effect of calcium, magnesium and potassium cations absence on the COD and nitrogen removal efficiency of the system was invisible. Calcium, magnesium and potassium cations were all involved in phosphorus release and uptake reactions, and the molar ratio of them to phosphorus were 6.1×10~(-2),3.8×10~(-1),2.8×10~(-1), respectively. Absence of calcium, magnesium and potassium cations in the influent led to surplus phosphorus release, and the order of the amounts of phosphorus release was obtained as follow: potassium absence >calcium absence >magnesium absence >normal system. In the range of 10oC to 30oC, changes in temperature had slightly effect on the COD removal efficiency, and the rate of phosphorus release and uptake both rose with the increase of temperature. Nitrification of the system got worse at temperature 10oC. Below 30mg/L, both nitrate and nitrite could be used as electron acceptor instead of oxygen for phosphorus uptake, and the order of phosphorus uptake rate was observed as follow: oxygen > nitrate > nitrite.
The effects of environmental factors on the A/O/A nitrogen and phosphorus removal procss in long-term and short-term were not entirely consistent. When sodium acetate and sodium propionate were used as influent carbon source in long-term, the system could keep favorable performance. But sodium succinate as carbon source led decrease in nitrogen removal efficiency, and the phosphorus removal deteriorated with the reduction of nitrogen removal efficiency. Favorable removal performance was achieved as pH in the range of 7.5 to 8.5. The performance got worse gradually, when the system was operated at pH 6.5 in long-term. As C/N in the range of 2.6~3.3, the performance of the system was excellent. However, with the increase of influent C/N, the phosphorus concentration in the effluent increased and removal efficiency of the system decreased gradually. In the start-up phase, Dynamic changes were observed in microbial species structure the A/O/A nitrogen and phosphorus removal process. In the synthetic wastewater system, partial dominant bacteria such as Candidatus Accumulibacter phosphatis always existed, partial dominant bacteria such as Rhodospirillales was eliminated and replaced by new strains, e.g Rhodocyclaceae and Candidatus Accumulibacter phosphates. Microbial commuities succession also occured in the domestic wastewater. Bacteria M1 and M3 (β- proteobacteria ) were always dominant, bacteria M14 (β- proteobacteria ) appeared with the operation of the system. Environmental factors had extraordinary influences on the microbial community structure. Dominant bacteria such as Denitrifying bacterium and Uncultured Chloroflexi bacterium with strong tolerance were resident bacteria, and some dominant bacteria such as Ammonia oxidizing bacterium were gradually washed out from the system with the transformation of the operational condition.
Through primary and secondary screening tests, four efficient phosphorus removing strains named as JL4, FH6, FQ3 and FQ4 were chosen from twenty-five strains isolated from the system with stable performance. Strain JL4 was identified as Pseudomonas sp., and strains FH6, FQ3 and FQ4 were identified as Bacillus sp. By the methods of physiological-biochemical tests, fatty acid identification and phylogenetic analysis of 16S rDNA clone. Environmental factors such as pH value, carbon source and temperature had distinguished effects on the growth and phosphorus removal efficiency of strains. The growth of strains had almost ceased and phosphorus removal performance deteriorated under the condition of pH 5, and the optimal pH was neutral. The optimum temperature was 30oC for strains JL4, FH6 and FQ3, but 25oC for strain FQ4. Cultivated with different carbon source, the sequence of the growth rate and phosphorus removal efficiency of the strains was concluded as follow: Sodium acetate > sodium propionate > sodium succinate >glucose.
以反硝化除磷理论为基础,采用A/O/A脱氮除磷工艺为研究主体,分别以人工合成污水和实际生活污水为处理对象,全面系统的研究了A/O/A工艺启动过程中的去除效能及污泥形态的变化,详细的考察了两个系统内颗粒污泥特性的差异。对比分析了环境因子对A/O/A工艺运行效果的冲击和持续影响。探讨了启动期系统内微生物的群落演替,及环境因子对微生物种群结构的影响。在此基础上,对筛选出的反硝化聚磷菌株进行鉴定,并研究了其除磷性能。
A/O/A脱氮除磷工艺启动过程中,随着系统去除效能的提高,污泥形态逐渐由絮体转化为颗粒、原生动物从无到有。稳定运行过程中,运行模式相同,进水水质不同的两个系统内的颗粒污泥性质存在较大差异。以实际生活污水培养的颗粒污泥与人工配水培养的颗粒污泥相比较,具有粒径小、沉淀速率慢、孔径大、比表面积小、比重轻、含水率高、活性高、胞外聚合物含量高、胞内聚合物含量低,释磷量小、反硝化效果好等特点。
影响因子对A/O/A脱氮除磷工艺去除性能的冲击影响显著。乙酸钠和丙酸钠为最佳碳源;丁二酸钠略差;乙醇和葡萄糖较差;异戊酸为最差碳源。进水pH值在6.0~8.0的范围内,对系统去除COD无明显影响。进水pH值为7.0时,系统的脱氮除磷效果最佳。进水C/N比在0.5~8.3的范围内,对系统去除COD的影响不大。随着进水C/N比的升高,系统对TN的去除有增加趋势。进水C/N比在0.5~6.1的范围内,系统对PO_4~(3-)-P的去除效果较好。进水中缺少钙、镁和钾离子,对系统去除有机物和脱氮没有明显的影响。钙、镁和钾离子皆参与释磷和吸磷反应,与磷的摩尔比分别为6.1×10~(-2),3.8×10~(-1),2.8×10~(-1)。缺少钙、镁和钾离子会引起聚磷菌超量释磷,释磷量顺序依次为:缺钾>缺钙>缺镁>正常系统。环境温度在10℃~30℃范围内,对系统去除COD的影响较小,释磷和吸磷速率随温度的升高而升高。温度为10℃时,系统硝化效果较差。浓度低于30mg/L时,NO_3~--N和NO_2~--N皆可替代O_2作为电子受体,进行过量吸磷反应。吸磷速率顺序依次为:O_2> NO_3~--N> NO_2~--N。
影响因子对A/O/A脱氮除磷工艺的持续影响与冲击影响结果并不完全一致。较长时间以乙酸钠和丙酸钠为进水碳源,系统皆可保持良好的脱氮除磷效果;而丁二酸钠为碳源则可导致脱氮效果下降,且除磷效果也随之恶化。系统pH值在7.5~8.5的范围内运行,可取得良好的去除效果;而长期在pH值为6.5的条件下运行,系统的处理效果会逐渐恶化。系统C/N长期在2.6~3.3的范围内运行,可取得理想的去除效果。但C/N进一步升高,系统的出水中磷浓度会逐渐升高,导致处理效能逐渐下降。
启动期,A/O/A脱氮除磷工艺内的微生物种群结构发生了动态变化。人工配水系统中部分优势菌如Candidatus Accumulibacter phosphatis在系统中始终存在;部分优势菌如Rhodospirillales随着系统的运行逐渐消失,取而代之出现新的优势菌如Rhodocyclaceae和Candidatus Accumulibacter phosphates。实际生活污水系统内亦发生了微生物的群落演替,优势菌M1和M3(β- proteobacteria)始终存在,随着系统的运行出现新的优势菌M14(β- proteobacteria)。环境因子对系统内的微生物的种群结构影响显著。系统内始终存在的优势菌如Denitrifying bacterium和Uncultured Chloroflexi bacterium耐受力较强,为常驻菌;而部分优势菌如Ammonia oxidizing bacterium则随着运行条件的改变,逐渐被系统淘汰。
从稳定运行的系统中分离纯化出25株菌,通过初筛和复筛试验选取除磷效率较高的4株菌:JL4、FH6、FQ3和FQ4。采用生理生化试验、脂肪酸鉴定及16S rDNA基因测序等手段进行鉴定:菌株JL4 Pseudomonas sp.,菌株FH6、FQ3和FQ4均为Bacillus sp.。环境因子pH值、碳源和温度对菌株生长及除磷效果影响显著。在pH值为5的条件下,菌株几乎停止生长,除磷效果极差;最适pH值为中性。菌株JL4、FH6和FQ3的最适温度为30℃,菌株FQ4的最适温度为25℃。在不同碳源的条件下,菌株的生长速率及除磷率顺序依次为:乙酸钠>丙酸钠>丁二酸钠>葡萄糖。
As the issue of eutrophication grows more severely and the regulatory requirements of wastewater discharge become increasingly stricter, economical and efficient removal of nitrogen and phosphorus has been a focus of wastewater treatment research. Though biological nitrogen and phosphorus removal process has been applied widely as one of economical and sustainable processes, it has some inherent problems that need to be solved. For example, the contradictions of substrates competition and SRT between the nitrogen and phosphorus removal, have inhibited the enhancement of removal efficiency. Some novel theories of nitrogen and phosphorus removal, such as denitrifying phosphorus removal and simultaneous nitrification and denitrification supply new ideas in solving theses contradictions.
Based on the theory of denitrifying phosphorus removal, A/O/A simultaneous nitrogen and phosphorus removal process was adopted as research subject. Dynamic changes in the removal efficiencies and morphologies of activated sludge of two A/O/A processes which treated with synthetic wastewater and domestic wastewater respectively were investigated systematically, and the differences of granular sludge characteristics in the two systems were analyzed comprehensively. Short and long -term influences of environmental factors on the removal efficiency of the A/O/A process were analyzed compar comparatively. Microbial community succession in the systems during the start-up period and the effect of environmental factors on the microbial population structure were investigated. Isolated denitrifying phosphorus removing bacteria were identified, and their phosphorus removal efficiency were further studied.
During the start-up period, flocculent sludge transformed into granular sludge gradually with the enhancement of removal efficiency, and lots of protozoa appeared in the A/O/A nitrogen and phosphorus removal processes. Granular sludge in the two systems treating different wastewater were quite distinct under the same operational condition in the stable phase. Granular sludge cultivated with real domestic wastewater was smaller in particle size, slower in sedimentation, larger in pore size, smaller in specific surface area, lighter in specific weight, higher in water content, higher in activity, more in extracellular polymeric content, lower in intracellular polymeric content, less in phosphorus release, and better in dintrification, compared with that cultivated with synthetic wastewater.
Environmental factors had vital influence on the performance of A/O/A nitrogen and phosphorus removal process in short-term. Sodium acetate and sodium propionate were optimal carbon source, and sodium succinate was slightly worse than them. Ethanol and glucose were unsuitable carbon source, and isovalerate was the worst carbon source. As pH value of influent changed from 6.0 to 8.0, no significant effect on the COD removal efficiency of the system were found. Influent pH at 7.0, perfect performance was achived. Value of C/N in the range of 0.5 to 8.3 had little effect on the COD removal efficiency. Total nitrogen removal efficiency increased with the increase of influent C/N. phosphorus removal efficiency of the system was perfect while C/N was in the range of 0.5 to 6.1, but it deteriorated when C/N was larger than 6.1. Value of C/N in the range of 0.5 to 8.3 had little effect on the COD removal efficiency. Total nitrogen removal efficiency increased with the increase of influent C/N. phosphorus removal efficiency of the system was perfect while C/N was in the range of 0.5 to 6.1, but it deteriorated when C/N was larger than 6.1. Effect of calcium, magnesium and potassium cations absence on the COD and nitrogen removal efficiency of the system was invisible. Calcium, magnesium and potassium cations were all involved in phosphorus release and uptake reactions, and the molar ratio of them to phosphorus were 6.1×10~(-2),3.8×10~(-1),2.8×10~(-1), respectively. Absence of calcium, magnesium and potassium cations in the influent led to surplus phosphorus release, and the order of the amounts of phosphorus release was obtained as follow: potassium absence >calcium absence >magnesium absence >normal system. In the range of 10oC to 30oC, changes in temperature had slightly effect on the COD removal efficiency, and the rate of phosphorus release and uptake both rose with the increase of temperature. Nitrification of the system got worse at temperature 10oC. Below 30mg/L, both nitrate and nitrite could be used as electron acceptor instead of oxygen for phosphorus uptake, and the order of phosphorus uptake rate was observed as follow: oxygen > nitrate > nitrite.
The effects of environmental factors on the A/O/A nitrogen and phosphorus removal procss in long-term and short-term were not entirely consistent. When sodium acetate and sodium propionate were used as influent carbon source in long-term, the system could keep favorable performance. But sodium succinate as carbon source led decrease in nitrogen removal efficiency, and the phosphorus removal deteriorated with the reduction of nitrogen removal efficiency. Favorable removal performance was achieved as pH in the range of 7.5 to 8.5. The performance got worse gradually, when the system was operated at pH 6.5 in long-term. As C/N in the range of 2.6~3.3, the performance of the system was excellent. However, with the increase of influent C/N, the phosphorus concentration in the effluent increased and removal efficiency of the system decreased gradually. In the start-up phase, Dynamic changes were observed in microbial species structure the A/O/A nitrogen and phosphorus removal process. In the synthetic wastewater system, partial dominant bacteria such as Candidatus Accumulibacter phosphatis always existed, partial dominant bacteria such as Rhodospirillales was eliminated and replaced by new strains, e.g Rhodocyclaceae and Candidatus Accumulibacter phosphates. Microbial commuities succession also occured in the domestic wastewater. Bacteria M1 and M3 (β- proteobacteria ) were always dominant, bacteria M14 (β- proteobacteria ) appeared with the operation of the system. Environmental factors had extraordinary influences on the microbial community structure. Dominant bacteria such as Denitrifying bacterium and Uncultured Chloroflexi bacterium with strong tolerance were resident bacteria, and some dominant bacteria such as Ammonia oxidizing bacterium were gradually washed out from the system with the transformation of the operational condition.
Through primary and secondary screening tests, four efficient phosphorus removing strains named as JL4, FH6, FQ3 and FQ4 were chosen from twenty-five strains isolated from the system with stable performance. Strain JL4 was identified as Pseudomonas sp., and strains FH6, FQ3 and FQ4 were identified as Bacillus sp. By the methods of physiological-biochemical tests, fatty acid identification and phylogenetic analysis of 16S rDNA clone. Environmental factors such as pH value, carbon source and temperature had distinguished effects on the growth and phosphorus removal efficiency of strains. The growth of strains had almost ceased and phosphorus removal performance deteriorated under the condition of pH 5, and the optimal pH was neutral. The optimum temperature was 30oC for strains JL4, FH6 and FQ3, but 25oC for strain FQ4. Cultivated with different carbon source, the sequence of the growth rate and phosphorus removal efficiency of the strains was concluded as follow: Sodium acetate > sodium propionate > sodium succinate >glucose.
引文
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