碳源种类对反硝化除磷系统的影响及反硝化聚磷菌(DPB)的分离
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摘要
反硝化除磷过程可以将生物除磷和脱氮两个原本相互独立的过程融为一体,被广泛认为是一个具有良好前景的生物除磷技术。论文通过对比试验,并采用连续运行方式研究了乙酸、丙酸、葡萄糖三种单碳源以及生活污水系统对反硝化除磷系统长期运行状态的影响,确定了不同碳源对系统反硝化除磷的作用与贡献;并使用ERIC-PCR指纹图谱技术研究分析了试验前后各反应器的菌群变化与脱氮除磷的关系以及各反应器的菌群特征,揭示了不同碳源下的菌群特征演变对反应器运行状态的影响。通过对分离的细菌进行反硝化与吸/释磷试验,并利用16SrDNA法对其进行初步鉴定。试验结果表明:
①运行良好的反硝化除磷系统,能在短时间内承受因碳源变化带来的冲击;三天后,单碳源系统出现了厌氧释磷和出水磷浓度的波动;两周后,乙酸、丙酸系统逐渐恢复稳定,葡萄糖系统则逐渐失去反硝化吸磷能力。乙酸可以提高生物除磷效果,可以作为富集反硝化聚磷菌DPB的有效碳源;丙酸虽然也可以提高生物除磷效果,但对反硝化除磷的促进作用不明显;葡萄糖的大量存在将使已成为优势菌群的PAOs和DPB逐渐被非聚磷菌取代。乙酸碳源基质的污泥产率系数低于丙酸和葡萄糖,葡萄糖碳源基质有利于获得更高的污泥产率。
②对试验前后的各反应器中菌群结构分析表明:生活污水系统和A2N-SBR脱氮除磷系统能稳定运行并取得良好的脱氮除磷效果,菌群结构呈现多样性分布状态;单碳源系统运行初期前三天,原系统稳定复杂的生态特征证明在短时间内能承受由于碳源变化带来的冲击。随着单碳源系统的运行,反应器中微生物在碳源竞争过程中的优胜劣汰,导致各系统在富集了不同优势菌的同时,某些原存的微生物也逐渐消亡。充分证明除温度、pH等因子外,碳源也是控制系统微生物生态位的重要因素之一。
③DPB是反硝化除磷系统中一类重要的功能菌群,对生物脱氮除磷起决定性的作用。文献报道的反硝化聚磷菌有:肠杆菌科细菌(Enterobacteriaceae colonies )、气单胞菌属( Aerimonas colonies )、假单胞菌属(Pseudomonas colonies)、莫拉氏菌属(Moraxella colonies)等。研究发现在富集了反硝化聚磷菌的乙酸系统存在两株兼具厌氧释磷、好氧吸磷典型聚磷菌特征和反硝化特征的细菌,编号为P1、P2,P1菌株好氧吸磷量约为厌氧释磷量的3倍,P2菌株好氧吸磷量为释磷量的2倍,而且两菌株反硝化过程不受产生的亚硝酸盐浓度的影响。对P1、P2菌株进行16SrDNA序列测定结果表明,P1菌株属于金黄杆菌属、P2菌株属于微杆菌属,是现有文献未见报道过的具有反硝化聚磷菌特征细菌类群。表明反硝化聚磷菌是一个宽泛的细菌类群,生物除磷系统中是由不同种属的细菌共同发挥脱氮除磷功能。
课题研究得到国家自然科学基金项目(50278101)与国家水体污染控制与治理重大科技专项(2008ZX07315)的资助。
The processes of denitrification and dephosphorization which were independent originally were integrated into Biological Denitrifying Phosphors and Nitrogen removal process. It was widely considered a bright technology of biological phosphorus removal. In this paper, compared with domestic sewage, the influence of different types of carbon sources on biological denitrifying phosphors and nitrogen removal system was researched. And three types of carbon sources, such as glucose, acetic acid and propionic acid were used. It determined how the different carbon sources made contribution to the systems. ERIC-PCR fingerprinting was used to investigate the relationship between variation of microbial community structure and nitrogen and phosphorus removal during the start and the end of the research, and microbial community structure of the reactors. Through denitrification test and phosphate uptake/release test, a new DPB species, which had seldom been reported in journals, was obtained from the acetic-acid system. And it was identified primarily by 16SrDNA method.
It was demonstrated that: When the biological denitrifying phosphors and nitrogen removal system performed perfectly, it would possess strong ability to resist the impacts of changing carbon sources in a short time; after three days, phosphate release in anaerobic conditions and fluctuation of phosphorus concentration were appeared; and two weeks later, the stability stabilization were restored in the systems, with acetic acid or propionic acid as carbon source, while with glucose as carbon source, the system lost the ability of denitrifying phosphorus uptake increasingly. Enhanced biological phosphorus removal, acetic acid was used as effective carbon source to enrich denitrifying phosphorus accumulating bacteria (DPB), while propionic acid had no significance on denitrifying phosphorus removal. Because of a great deal of glucose, the dominant bacteria PAOs and DPB were replaced by other bacteria which were unable to accumulate polyphosphate. The sludge yield coefficient of acetic acid was lower than that of propionic acid and glucose, and glucose produced higher sludge yield.
Analysis on the microbial community structure of activated sludge in the reactors during the research. The results indicated that microbial community structure in the domestic system and A2N-SBR system distributed a high level of diversity, and the two reactors could stable and effectively removing the nitrogen and phosphorous from wastewater. Because of the stable and complex microbial community, the single carbon reactors could resist the impacts of changing carbon sources in the first three days. As the operating of single carbon reactors, different dominant bacteria were enriched and bacteria which were survival in system were extinct because of the competition of carbon. It proved that carbon source included DO, pH value, were the important factors for controlling ecological niche of bacteria.
DPB (denitrifying phosphorus accumulating bacteria) is an important kind of bacteria in the process of biological nitrogen and phosphorus removal. Members of several different bacterial divisions of microorganisms had been proposed as DPB. These include the Enterobacteriaceae colonies, Aerimonas colonies, Pseudomonas colonies and Moraxella colonies. In this study, two different kinds of bacteria were isolated which had the characteristic of anaerobic phosphorus release, aerobic phosphorus absorb and denitrifying. They were named as P1 and P2. P1 has best quality of phosphorous removal, and P-uptake was three times amount of P-release, and P2’s P-uptake was twice amount of P-release. And nitrite concentration had no effect on them. Sequence analysis of 16SrDNA identified P1 and P2, P1 belonged to Chryseobacterium sp, and P2 belonged to Microbacterium sp. The bacteria of these genus hadn’t been reported as DPB. Data presented here add further supports to the view that may be mediated by different microbial populations under different conditions. The perfect of Biological Denitrifying Phosphors and Nitrogen removal system provide different kinds of DPB ecological environment to survive which exert function of removing phosphorous and nitrogen from wastewater efficiently.
This research was sponsored by the National Natural Science Foundation of China (50278101) and Grand science and technology special project in National Water Pollution Control and Management of China(2008ZX07315).
①运行良好的反硝化除磷系统,能在短时间内承受因碳源变化带来的冲击;三天后,单碳源系统出现了厌氧释磷和出水磷浓度的波动;两周后,乙酸、丙酸系统逐渐恢复稳定,葡萄糖系统则逐渐失去反硝化吸磷能力。乙酸可以提高生物除磷效果,可以作为富集反硝化聚磷菌DPB的有效碳源;丙酸虽然也可以提高生物除磷效果,但对反硝化除磷的促进作用不明显;葡萄糖的大量存在将使已成为优势菌群的PAOs和DPB逐渐被非聚磷菌取代。乙酸碳源基质的污泥产率系数低于丙酸和葡萄糖,葡萄糖碳源基质有利于获得更高的污泥产率。
②对试验前后的各反应器中菌群结构分析表明:生活污水系统和A2N-SBR脱氮除磷系统能稳定运行并取得良好的脱氮除磷效果,菌群结构呈现多样性分布状态;单碳源系统运行初期前三天,原系统稳定复杂的生态特征证明在短时间内能承受由于碳源变化带来的冲击。随着单碳源系统的运行,反应器中微生物在碳源竞争过程中的优胜劣汰,导致各系统在富集了不同优势菌的同时,某些原存的微生物也逐渐消亡。充分证明除温度、pH等因子外,碳源也是控制系统微生物生态位的重要因素之一。
③DPB是反硝化除磷系统中一类重要的功能菌群,对生物脱氮除磷起决定性的作用。文献报道的反硝化聚磷菌有:肠杆菌科细菌(Enterobacteriaceae colonies )、气单胞菌属( Aerimonas colonies )、假单胞菌属(Pseudomonas colonies)、莫拉氏菌属(Moraxella colonies)等。研究发现在富集了反硝化聚磷菌的乙酸系统存在两株兼具厌氧释磷、好氧吸磷典型聚磷菌特征和反硝化特征的细菌,编号为P1、P2,P1菌株好氧吸磷量约为厌氧释磷量的3倍,P2菌株好氧吸磷量为释磷量的2倍,而且两菌株反硝化过程不受产生的亚硝酸盐浓度的影响。对P1、P2菌株进行16SrDNA序列测定结果表明,P1菌株属于金黄杆菌属、P2菌株属于微杆菌属,是现有文献未见报道过的具有反硝化聚磷菌特征细菌类群。表明反硝化聚磷菌是一个宽泛的细菌类群,生物除磷系统中是由不同种属的细菌共同发挥脱氮除磷功能。
课题研究得到国家自然科学基金项目(50278101)与国家水体污染控制与治理重大科技专项(2008ZX07315)的资助。
The processes of denitrification and dephosphorization which were independent originally were integrated into Biological Denitrifying Phosphors and Nitrogen removal process. It was widely considered a bright technology of biological phosphorus removal. In this paper, compared with domestic sewage, the influence of different types of carbon sources on biological denitrifying phosphors and nitrogen removal system was researched. And three types of carbon sources, such as glucose, acetic acid and propionic acid were used. It determined how the different carbon sources made contribution to the systems. ERIC-PCR fingerprinting was used to investigate the relationship between variation of microbial community structure and nitrogen and phosphorus removal during the start and the end of the research, and microbial community structure of the reactors. Through denitrification test and phosphate uptake/release test, a new DPB species, which had seldom been reported in journals, was obtained from the acetic-acid system. And it was identified primarily by 16SrDNA method.
It was demonstrated that: When the biological denitrifying phosphors and nitrogen removal system performed perfectly, it would possess strong ability to resist the impacts of changing carbon sources in a short time; after three days, phosphate release in anaerobic conditions and fluctuation of phosphorus concentration were appeared; and two weeks later, the stability stabilization were restored in the systems, with acetic acid or propionic acid as carbon source, while with glucose as carbon source, the system lost the ability of denitrifying phosphorus uptake increasingly. Enhanced biological phosphorus removal, acetic acid was used as effective carbon source to enrich denitrifying phosphorus accumulating bacteria (DPB), while propionic acid had no significance on denitrifying phosphorus removal. Because of a great deal of glucose, the dominant bacteria PAOs and DPB were replaced by other bacteria which were unable to accumulate polyphosphate. The sludge yield coefficient of acetic acid was lower than that of propionic acid and glucose, and glucose produced higher sludge yield.
Analysis on the microbial community structure of activated sludge in the reactors during the research. The results indicated that microbial community structure in the domestic system and A2N-SBR system distributed a high level of diversity, and the two reactors could stable and effectively removing the nitrogen and phosphorous from wastewater. Because of the stable and complex microbial community, the single carbon reactors could resist the impacts of changing carbon sources in the first three days. As the operating of single carbon reactors, different dominant bacteria were enriched and bacteria which were survival in system were extinct because of the competition of carbon. It proved that carbon source included DO, pH value, were the important factors for controlling ecological niche of bacteria.
DPB (denitrifying phosphorus accumulating bacteria) is an important kind of bacteria in the process of biological nitrogen and phosphorus removal. Members of several different bacterial divisions of microorganisms had been proposed as DPB. These include the Enterobacteriaceae colonies, Aerimonas colonies, Pseudomonas colonies and Moraxella colonies. In this study, two different kinds of bacteria were isolated which had the characteristic of anaerobic phosphorus release, aerobic phosphorus absorb and denitrifying. They were named as P1 and P2. P1 has best quality of phosphorous removal, and P-uptake was three times amount of P-release, and P2’s P-uptake was twice amount of P-release. And nitrite concentration had no effect on them. Sequence analysis of 16SrDNA identified P1 and P2, P1 belonged to Chryseobacterium sp, and P2 belonged to Microbacterium sp. The bacteria of these genus hadn’t been reported as DPB. Data presented here add further supports to the view that may be mediated by different microbial populations under different conditions. The perfect of Biological Denitrifying Phosphors and Nitrogen removal system provide different kinds of DPB ecological environment to survive which exert function of removing phosphorous and nitrogen from wastewater efficiently.
This research was sponsored by the National Natural Science Foundation of China (50278101) and Grand science and technology special project in National Water Pollution Control and Management of China(2008ZX07315).
引文
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