厌氧生物除磷及其微生物种群结构特性的研究
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摘要
通过厌氧培养基试验,在厌氧条件下,考察始末总磷的浓度,研究了不同泥源、不同磷源、不同起始磷浓度、不同氮源、不同初始氮磷比、不同碳源、不同初始碳磷比对厌氧除磷的影响,并对各影响因素稳定时期的微生物群落结构进行跟踪研究。试验结果表明:
以猪粪、鸡粪、SBR污泥、EGSB厌氧污泥、ASBR污泥和具有同步脱氮生物化学除磷的污泥为泥源的培养基试验中, ASBR污泥为泥源时对总磷去除量最高,其次为EGSB污泥为泥源的培养基。不同泥源稳定培养状态下微生物多样性最丰富,样品相似性不高,其中以ASBR污泥和EGSB污泥为泥源的厌氧培养基的相似性最高。
以磷酸氢二钾为磷源时的磷去除效果最佳其次为磷酸二氢钾,最差为六偏磷酸钠。采用磷酸二氢钾为磷源时,随着磷浓度的升高,总磷的去除量增加,当总磷含量高于50mg/L时,总磷去除量不随初始磷浓度的升高而升高,维持16mg/L左右。不同磷源及不同初始磷浓度培养基稳定培养状态下微生物多样性较丰富,磷源培养基样品相似性不高,磷浓度培养基样品相似性较高,磷源试验中以磷酸氢二钾和亚磷酸钠为磷源的厌氧培养基的相似性最低55.4%,磷浓度试验中初始磷浓度为50mg/L时与其它浓度的厌氧培养基的相似性均为70%以上。
以氯化铵为氮源时的总磷磷去除率最高,其次为乙酸氨,最差为硝酸钠。采用氯化铵为氮源时,初始氮磷比为1.0时总磷的去除率最高。不同氮源及不同初始氮磷比培养基稳定培养状态下微生物多样性较丰富,样品相似性不高,氮源试验中以氯化铵和蛋白胨为氮源的厌氧培养基的相似性最低44.3%,氮磷比试验中初始氮磷比为0.5和1.0情况下的厌氧培养基的相似性为最差仅为59.7%。
以甲醇为碳源时磷去除效果最佳,其次为啤酒和葡萄糖,最差为乙酸钠。不同碳磷比对厌氧培养基系统中总磷和COD去除的影响较大,随着碳磷比的增加,厌氧培养中总磷和COD的去除有所增加,但当达到一定程度时,随着碳磷比的增加,总磷去除率降低,COD去除率增高。5种不同碳源培养基稳定阶段的微生物多样性较丰富,样品相似性较高;8种不同初始碳磷比培养基稳定阶段的微生物多样性较丰富,样品相似性不是很高;初始碳磷比为8、12、16时厌氧培养基样品相似性较高,初始碳磷比为24、30、40时厌氧培养基样品相似性次之。
随着培养条件的优化,厌氧培养基总磷的去除率逐渐升高;培养基微生物多样性丰富,相似性相差整体上差异性很大;随着培养条件的优化微生物种群结构在逐渐的变化,微生物种群数逐渐减少,结合常规试验结果可知,条带4、7、12、13、17所代表的微生物种群是厌氧除磷产磷化氢的优势种群,可以用于后续测序种群鉴定。
In the condition of anaerobic envinronment, research the effects of different mud, different Phosphor and the original concentration of P, different Nitrogen and nitrogen-phosphorus ratio, different Carbon and carbon-phosphorus ratio on the anaerobic phosphorus removal through the anaerobic culture medium experiment, and combine to fall structure progress to follow a search to the microorganisms of each impact factor in stabilization time. And results are as follows.
The phosphorus removal efficiency is highest when the mud is EGSB anaerobic sludge or ASBR sludge, and it is better to remove P within the culture medium of ASBR sludge than with the EGSB sludge, compared with the shortly simultaneous nitrification and denitrification P removal sludge mixed by pig manure and fowl dung, SBR sludge and the simultaneously biochemical nitrogen and phosphorus removal sludge. Microbial diversity are of great abundance and its similarity is low when they are stably cultured in various mud source, among which the similarity between ASBR and EGSB is the most.
It has the best result to remove P when using the dipotassiun hydrogen phosphate, the monopotassiun phosphate is the second, and the sodium hexametaphosphate is the worst of the three. If using the monopotassium phosphate, as the concentration of P goes up, the P removal quantity increases. However, when the total P is more than 50mg/L, the P removal which keeps 20mg/L will not goes up as the concentration increases. Different phosphorus source and various initial concentrations of phosphorus culture media are cultured stably rendering rich microbial diversity, low similarity of phosphorus source culture media samples, and high similarity of concentration of phosphorus culture media samples. Among all experiments of phosphorus source, we found that anaerobic medium using dipotassiun hydrogen phosphate and sodium phosphite as phosphorus resource have the lowest similarity 55.4%, besides, in the test of initial phosphorus concentration, the initial phosphorus concentration is 50mg/L had the medium high similarity with the other initial phosphorus concentration which are more than 70%
It has the best results to remove P when using the ammonium chloride as N source, the ammonium acetate is the second, and the sodium nitrate is the worst of the three. If using the ammonium chloride as N source,it has the best results to remove P when the nitrogen-phosphorus ratio is 1.0.Different nitrogen source and initial Nitrogen-Phosphorus ratios are cultured stably rendering rich microbial diversity, low similarity of nitrogen source culture media samples. Among all experiments of nitrogen source, we found that anaerobic medium using ammonium chloride and peptone as nitrogen source have the lowest similarity 44.3%, besides, in the test of nitrogen-phosphorus ratio, the initial nitrogen-phosphorus ratio that are respectively 0.5 and 1.0 have the lowest anaerobic medium similarity which is just 59.7%.
It has the best results to remove P when using the methanol as C source, the beer and the glucose is the second, and the sodium acetate is the worst of the three. The different carbon-phosphorus ratio has a big impact on the total phosphorus removal and COD removal in the anaerobe culture system,as the carbon-phosphorus ratio increases, the total phosphorus removal and COD removal in the anaerobe culture system will increase until it reaches certain degree that the removement will decrease as the carbon-phosphorus ratio increases.Five kinds of various carbon sources cultured stably rendering rich microbial diversity, high similarity. Eight kinds of various carbon sources cultured stably rendering rich microbial diversity and the similarity is not high, similarity between the anaerobic culture mediums with the carbon- phosphorus ratio 8、12 and 1.5 is the higher than that of 24、30and 40.
As the condition of culture optimizes,in the condition of each best effect factor,the phosphorus removal in the anaerobic culture medium is raising;the microbialpopulation variety is abundant , the similarity is discrepancy as a whole;As the condition of culture optimizes,the microbialpopulation structure is changing gradually,the amount of the microbialpopulation are reducing gradually.Combine the result of routine experiments, the microbialpopulation which band 4 7 12 13 and 17 stands for are dominant microbialpopulation that remove P to produce phosphine in anaerobic condition, which can be used to the follow-up sequencing to identify the population.
以猪粪、鸡粪、SBR污泥、EGSB厌氧污泥、ASBR污泥和具有同步脱氮生物化学除磷的污泥为泥源的培养基试验中, ASBR污泥为泥源时对总磷去除量最高,其次为EGSB污泥为泥源的培养基。不同泥源稳定培养状态下微生物多样性最丰富,样品相似性不高,其中以ASBR污泥和EGSB污泥为泥源的厌氧培养基的相似性最高。
以磷酸氢二钾为磷源时的磷去除效果最佳其次为磷酸二氢钾,最差为六偏磷酸钠。采用磷酸二氢钾为磷源时,随着磷浓度的升高,总磷的去除量增加,当总磷含量高于50mg/L时,总磷去除量不随初始磷浓度的升高而升高,维持16mg/L左右。不同磷源及不同初始磷浓度培养基稳定培养状态下微生物多样性较丰富,磷源培养基样品相似性不高,磷浓度培养基样品相似性较高,磷源试验中以磷酸氢二钾和亚磷酸钠为磷源的厌氧培养基的相似性最低55.4%,磷浓度试验中初始磷浓度为50mg/L时与其它浓度的厌氧培养基的相似性均为70%以上。
以氯化铵为氮源时的总磷磷去除率最高,其次为乙酸氨,最差为硝酸钠。采用氯化铵为氮源时,初始氮磷比为1.0时总磷的去除率最高。不同氮源及不同初始氮磷比培养基稳定培养状态下微生物多样性较丰富,样品相似性不高,氮源试验中以氯化铵和蛋白胨为氮源的厌氧培养基的相似性最低44.3%,氮磷比试验中初始氮磷比为0.5和1.0情况下的厌氧培养基的相似性为最差仅为59.7%。
以甲醇为碳源时磷去除效果最佳,其次为啤酒和葡萄糖,最差为乙酸钠。不同碳磷比对厌氧培养基系统中总磷和COD去除的影响较大,随着碳磷比的增加,厌氧培养中总磷和COD的去除有所增加,但当达到一定程度时,随着碳磷比的增加,总磷去除率降低,COD去除率增高。5种不同碳源培养基稳定阶段的微生物多样性较丰富,样品相似性较高;8种不同初始碳磷比培养基稳定阶段的微生物多样性较丰富,样品相似性不是很高;初始碳磷比为8、12、16时厌氧培养基样品相似性较高,初始碳磷比为24、30、40时厌氧培养基样品相似性次之。
随着培养条件的优化,厌氧培养基总磷的去除率逐渐升高;培养基微生物多样性丰富,相似性相差整体上差异性很大;随着培养条件的优化微生物种群结构在逐渐的变化,微生物种群数逐渐减少,结合常规试验结果可知,条带4、7、12、13、17所代表的微生物种群是厌氧除磷产磷化氢的优势种群,可以用于后续测序种群鉴定。
In the condition of anaerobic envinronment, research the effects of different mud, different Phosphor and the original concentration of P, different Nitrogen and nitrogen-phosphorus ratio, different Carbon and carbon-phosphorus ratio on the anaerobic phosphorus removal through the anaerobic culture medium experiment, and combine to fall structure progress to follow a search to the microorganisms of each impact factor in stabilization time. And results are as follows.
The phosphorus removal efficiency is highest when the mud is EGSB anaerobic sludge or ASBR sludge, and it is better to remove P within the culture medium of ASBR sludge than with the EGSB sludge, compared with the shortly simultaneous nitrification and denitrification P removal sludge mixed by pig manure and fowl dung, SBR sludge and the simultaneously biochemical nitrogen and phosphorus removal sludge. Microbial diversity are of great abundance and its similarity is low when they are stably cultured in various mud source, among which the similarity between ASBR and EGSB is the most.
It has the best result to remove P when using the dipotassiun hydrogen phosphate, the monopotassiun phosphate is the second, and the sodium hexametaphosphate is the worst of the three. If using the monopotassium phosphate, as the concentration of P goes up, the P removal quantity increases. However, when the total P is more than 50mg/L, the P removal which keeps 20mg/L will not goes up as the concentration increases. Different phosphorus source and various initial concentrations of phosphorus culture media are cultured stably rendering rich microbial diversity, low similarity of phosphorus source culture media samples, and high similarity of concentration of phosphorus culture media samples. Among all experiments of phosphorus source, we found that anaerobic medium using dipotassiun hydrogen phosphate and sodium phosphite as phosphorus resource have the lowest similarity 55.4%, besides, in the test of initial phosphorus concentration, the initial phosphorus concentration is 50mg/L had the medium high similarity with the other initial phosphorus concentration which are more than 70%
It has the best results to remove P when using the ammonium chloride as N source, the ammonium acetate is the second, and the sodium nitrate is the worst of the three. If using the ammonium chloride as N source,it has the best results to remove P when the nitrogen-phosphorus ratio is 1.0.Different nitrogen source and initial Nitrogen-Phosphorus ratios are cultured stably rendering rich microbial diversity, low similarity of nitrogen source culture media samples. Among all experiments of nitrogen source, we found that anaerobic medium using ammonium chloride and peptone as nitrogen source have the lowest similarity 44.3%, besides, in the test of nitrogen-phosphorus ratio, the initial nitrogen-phosphorus ratio that are respectively 0.5 and 1.0 have the lowest anaerobic medium similarity which is just 59.7%.
It has the best results to remove P when using the methanol as C source, the beer and the glucose is the second, and the sodium acetate is the worst of the three. The different carbon-phosphorus ratio has a big impact on the total phosphorus removal and COD removal in the anaerobe culture system,as the carbon-phosphorus ratio increases, the total phosphorus removal and COD removal in the anaerobe culture system will increase until it reaches certain degree that the removement will decrease as the carbon-phosphorus ratio increases.Five kinds of various carbon sources cultured stably rendering rich microbial diversity, high similarity. Eight kinds of various carbon sources cultured stably rendering rich microbial diversity and the similarity is not high, similarity between the anaerobic culture mediums with the carbon- phosphorus ratio 8、12 and 1.5 is the higher than that of 24、30and 40.
As the condition of culture optimizes,in the condition of each best effect factor,the phosphorus removal in the anaerobic culture medium is raising;the microbialpopulation variety is abundant , the similarity is discrepancy as a whole;As the condition of culture optimizes,the microbialpopulation structure is changing gradually,the amount of the microbialpopulation are reducing gradually.Combine the result of routine experiments, the microbialpopulation which band 4 7 12 13 and 17 stands for are dominant microbialpopulation that remove P to produce phosphine in anaerobic condition, which can be used to the follow-up sequencing to identify the population.
引文
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