生活污水处理过程中最佳絮凝微生物的筛选及其生物学特性研究
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摘要
微生物絮凝剂是一类由微生物产生的具有絮凝能力的代谢产物,主要有糖蛋白、多糖、蛋白质、纤维素和DNA等。相比无机絮凝剂铝盐的易发生老年痴呆,铁盐对设备的高要求,以及丙烯酰胺单体的毒性,高效、无毒、无二次污染、可生物降解等特点使得微生物絮凝剂在水处理、食品工业等方面具有广阔的应用前景,越来越受到研究者的重视。
1.本文考察了呼和浩特辛辛板污水处理厂处理污水过程中各阶段出水的微生物类群数量变化情况。各阶段微生物数量变化从多到少依次为活性污泥>曝气池>进水池>出水池>沉淀池。细菌、放线菌和真菌三大类微生物当中,各阶段出水都是细菌数量占绝对优势。
2.从污水处理厂的各处理阶段出水及活性污泥中初筛得到10株絮凝活性较高的菌株。复筛后选择了两株絮凝能力最强的菌株做了进一步研究。将它们命名为X-1和X-2。经生理生化特性研究和16SrDNA序列测定,确定菌株X-1和X-2都是克雷伯氏菌属(Klebsiella sp.)的细菌。
3.X-1的培养条件为葡萄糖20g/L,牛肉膏0.5g/L,磷酸氢二钾5g/L,磷酸二氢钾2g/L,氯化钠0.1g/L,硫酸镁为0.05g/／L,硝酸钾0.2g/L培养基初始pH值7.5,培养温度30℃,摇床转速160r/m。
菌株X-2的培养条件为葡萄糖15g/L,酵母膏1.0g/L,磷酸氢二钾为3g/L,磷酸二氢钾为2g/L,硫酸镁0.2g/L,氯化钠0.1g/L,氯化亚铁0.1g/L。培养基初始pH值7.5,培养温度30℃,摇床转速160r/m。
4.研究了X-1和X-2两菌株的部分生物学特性。X-1和X-2都是革兰氏阴性菌,菌株X-1菌体杆状,长2μm,宽1μm。菌落直径约3mm,圆形,边缘整齐,凸起,表面光滑,半透明,乳黄色,有光泽。有荚膜,无芽孢。在X-1的生长周期中,絮凝率最高的时期是在稳定期的后期,即培养32h时。
菌株X-2菌体长1.8μm,宽0.8μm。菌落直径3-5mm,圆形,边缘整齐,凸起,扁圆锥状,表面光滑,半透明,黄色,有光泽。有荚膜,无芽孢。X-2的生长周期中,絮凝率最高的时期是在对数生长期,即培养6h时。
5.在100mL 0.5%的高岭土悬液体系中,pH值为8,温度为25℃,加入800μL 10%的CaCl2溶液为助凝剂,培养32h的X-1发酵液600μL150r/m快搅1min后,50r/m慢搅2min,静置30min,菌株X-1能发挥最高絮凝活性,絮凝率达到93.57%。
对于菌株X-2来说,高岭土悬液pH值为7时,反应温度为25℃,助凝剂10%的CaCl2溶液600μL,培养6h的X-2发酵液用量500μL,搅拌方式与X-1相同等条件下,X-2絮凝率最高,达到92.82%。
同时加入培养适宜时间的X-1和X-2的发酵液,测得的絮凝率比单一菌株作用下的絮凝率高。
6.离心后发现X-1和X-2的絮凝活性物质都分布在上清液中。它们的热稳定性都比较高,随着温度的上升,絮凝率下降幅度不大。菌株传代次数和菌种的保存时间对两菌株的絮凝率有一定的影响。传代次数越多,絮凝活性越低。菌种保存时间越长,絮凝率也越低。
7.在净化实际生活污水的过程中发现,在X-1和X-2两菌株共同作用下对污水的净化程度较高。
Bioflocculant is a kind of biological metabolic substance produced by microorganism which has the ability of flocculating. Its main components are glycoprotein, amylum, protein, cellulose, DNA etc. Bioflocculant is efficient, nontoxic, biodegradable readily and without pollution to the environment. The studies of bioflocculants are gaining more and more attentions.
1. We detected the change of microbe quantity of sewage basin, aeration basin, sewage preliminary basin, sewage effluent and activated sludge in Hohhot sewage disposal plant. Microbe quantity was activated sludge, aeration basin, sewage basin, sewage effluent, sewage preliminary basin from more to less. Bacterium is dominant in Microorganism.
2. We selected 10 strains with high flocculating activity from Hohhot sewage disposal plant.Two strains of them with higher flocculating activity were do further research and named as X-1 and X-2. X-1 and X-2 were considered as Klebsiella sp. by assaying physiological and biochemical characteristics and 16SrDN A sequence.
3. Glucose was the best carbon source of X-1 and X-2, X-1 was 20g/L and X-2 was 15g/L. Beef extract was the best nitrogen source of X-1, when 0.5g/L it had the highest flocculating activity. Other optimum cultural conditions of X-1 were:K2HPO4 5g/L, KH2PO4 2g/L, NaCl 0.1g/L, MgSO4 0.05g/L, KNO3 0.2g/L.The initial pH value of the medium was adjusted to 7.5, and the strain X-1 was cultured at 30℃and shaken at 160 r/m in a rotary shaker for 32 hours.
Yeast extract was the best nitrogen source of X-2, the best dosage was 1.0g/L.Other optimum cultural conditions of the strain X-2 were:K2HPO4 3g,KH2PO4 2g,NaCl 0.1g, MgSO4 0.2g, FeCl2 0.1g in 1000mL distilled water. The best condition for X-2 were:the initial pH was 7.5, Temperature was 30℃, and shaken at 160 r/m in a rotary shaker for 6 hours.
4. We studied some biological characteristics of the two strains. X-1 and X-2 were gram-negative bacteria.
In the X-1's growth cycle, the highest rate of flocculation appeared in stable phase,32h cultured. In the X-2 growth cycle, the highest rate of flocculation appeared in logarithmic phase, that is, when cultured 6h.
5. X-1 could play the highest flocculating activity and its flocculating rate was 93.57%, when 0.5% kaolin suspension system in 100mL. And pH was 8, the temperature was 25℃, to add 800μL 10% CaCl2 as coagulant,32h cultured of the 600μL X-1 fermenting liquor,150r/m lmin,50r/m 2min, standing for 30min.
When pH of the kaolin suspension was 7, the reaction temperature was 25℃,10% CaCl2 was 600μL, the amount of X-2 liquid was 500μL 6h cultured, stirring the same way as X-1 for the X-2, flocculation rate of X-2 was to 92.82%,that was the highest flocculation rate of X-2.
6. After centrifugation, we found that the flocculating activity of substances of X-1 and X-2 both mainly existed in the supernatant. The flocculants produced by X-1 and X-2 relatively had high thermal stability, as the temperature increased the flocculation rate decreased slightly. To a certain extent, times of bacteria passages and storage time had effect on the flocculation rate. The more passages bacteria were, the lower flocculating activity was. The longer preservation species was, the lower flocculation rate was.
7. We found that X-1 and X-2 acted on sewage together, sewage disposal consequence was better than X-1 or X-2 acted alone.
1.本文考察了呼和浩特辛辛板污水处理厂处理污水过程中各阶段出水的微生物类群数量变化情况。各阶段微生物数量变化从多到少依次为活性污泥>曝气池>进水池>出水池>沉淀池。细菌、放线菌和真菌三大类微生物当中,各阶段出水都是细菌数量占绝对优势。
2.从污水处理厂的各处理阶段出水及活性污泥中初筛得到10株絮凝活性较高的菌株。复筛后选择了两株絮凝能力最强的菌株做了进一步研究。将它们命名为X-1和X-2。经生理生化特性研究和16SrDNA序列测定,确定菌株X-1和X-2都是克雷伯氏菌属(Klebsiella sp.)的细菌。
3.X-1的培养条件为葡萄糖20g/L,牛肉膏0.5g/L,磷酸氢二钾5g/L,磷酸二氢钾2g/L,氯化钠0.1g/L,硫酸镁为0.05g/／L,硝酸钾0.2g/L培养基初始pH值7.5,培养温度30℃,摇床转速160r/m。
菌株X-2的培养条件为葡萄糖15g/L,酵母膏1.0g/L,磷酸氢二钾为3g/L,磷酸二氢钾为2g/L,硫酸镁0.2g/L,氯化钠0.1g/L,氯化亚铁0.1g/L。培养基初始pH值7.5,培养温度30℃,摇床转速160r/m。
4.研究了X-1和X-2两菌株的部分生物学特性。X-1和X-2都是革兰氏阴性菌,菌株X-1菌体杆状,长2μm,宽1μm。菌落直径约3mm,圆形,边缘整齐,凸起,表面光滑,半透明,乳黄色,有光泽。有荚膜,无芽孢。在X-1的生长周期中,絮凝率最高的时期是在稳定期的后期,即培养32h时。
菌株X-2菌体长1.8μm,宽0.8μm。菌落直径3-5mm,圆形,边缘整齐,凸起,扁圆锥状,表面光滑,半透明,黄色,有光泽。有荚膜,无芽孢。X-2的生长周期中,絮凝率最高的时期是在对数生长期,即培养6h时。
5.在100mL 0.5%的高岭土悬液体系中,pH值为8,温度为25℃,加入800μL 10%的CaCl2溶液为助凝剂,培养32h的X-1发酵液600μL150r/m快搅1min后,50r/m慢搅2min,静置30min,菌株X-1能发挥最高絮凝活性,絮凝率达到93.57%。
对于菌株X-2来说,高岭土悬液pH值为7时,反应温度为25℃,助凝剂10%的CaCl2溶液600μL,培养6h的X-2发酵液用量500μL,搅拌方式与X-1相同等条件下,X-2絮凝率最高,达到92.82%。
同时加入培养适宜时间的X-1和X-2的发酵液,测得的絮凝率比单一菌株作用下的絮凝率高。
6.离心后发现X-1和X-2的絮凝活性物质都分布在上清液中。它们的热稳定性都比较高,随着温度的上升,絮凝率下降幅度不大。菌株传代次数和菌种的保存时间对两菌株的絮凝率有一定的影响。传代次数越多,絮凝活性越低。菌种保存时间越长,絮凝率也越低。
7.在净化实际生活污水的过程中发现,在X-1和X-2两菌株共同作用下对污水的净化程度较高。
Bioflocculant is a kind of biological metabolic substance produced by microorganism which has the ability of flocculating. Its main components are glycoprotein, amylum, protein, cellulose, DNA etc. Bioflocculant is efficient, nontoxic, biodegradable readily and without pollution to the environment. The studies of bioflocculants are gaining more and more attentions.
1. We detected the change of microbe quantity of sewage basin, aeration basin, sewage preliminary basin, sewage effluent and activated sludge in Hohhot sewage disposal plant. Microbe quantity was activated sludge, aeration basin, sewage basin, sewage effluent, sewage preliminary basin from more to less. Bacterium is dominant in Microorganism.
2. We selected 10 strains with high flocculating activity from Hohhot sewage disposal plant.Two strains of them with higher flocculating activity were do further research and named as X-1 and X-2. X-1 and X-2 were considered as Klebsiella sp. by assaying physiological and biochemical characteristics and 16SrDN A sequence.
3. Glucose was the best carbon source of X-1 and X-2, X-1 was 20g/L and X-2 was 15g/L. Beef extract was the best nitrogen source of X-1, when 0.5g/L it had the highest flocculating activity. Other optimum cultural conditions of X-1 were:K2HPO4 5g/L, KH2PO4 2g/L, NaCl 0.1g/L, MgSO4 0.05g/L, KNO3 0.2g/L.The initial pH value of the medium was adjusted to 7.5, and the strain X-1 was cultured at 30℃and shaken at 160 r/m in a rotary shaker for 32 hours.
Yeast extract was the best nitrogen source of X-2, the best dosage was 1.0g/L.Other optimum cultural conditions of the strain X-2 were:K2HPO4 3g,KH2PO4 2g,NaCl 0.1g, MgSO4 0.2g, FeCl2 0.1g in 1000mL distilled water. The best condition for X-2 were:the initial pH was 7.5, Temperature was 30℃, and shaken at 160 r/m in a rotary shaker for 6 hours.
4. We studied some biological characteristics of the two strains. X-1 and X-2 were gram-negative bacteria.
In the X-1's growth cycle, the highest rate of flocculation appeared in stable phase,32h cultured. In the X-2 growth cycle, the highest rate of flocculation appeared in logarithmic phase, that is, when cultured 6h.
5. X-1 could play the highest flocculating activity and its flocculating rate was 93.57%, when 0.5% kaolin suspension system in 100mL. And pH was 8, the temperature was 25℃, to add 800μL 10% CaCl2 as coagulant,32h cultured of the 600μL X-1 fermenting liquor,150r/m lmin,50r/m 2min, standing for 30min.
When pH of the kaolin suspension was 7, the reaction temperature was 25℃,10% CaCl2 was 600μL, the amount of X-2 liquid was 500μL 6h cultured, stirring the same way as X-1 for the X-2, flocculation rate of X-2 was to 92.82%,that was the highest flocculation rate of X-2.
6. After centrifugation, we found that the flocculating activity of substances of X-1 and X-2 both mainly existed in the supernatant. The flocculants produced by X-1 and X-2 relatively had high thermal stability, as the temperature increased the flocculation rate decreased slightly. To a certain extent, times of bacteria passages and storage time had effect on the flocculation rate. The more passages bacteria were, the lower flocculating activity was. The longer preservation species was, the lower flocculation rate was.
7. We found that X-1 and X-2 acted on sewage together, sewage disposal consequence was better than X-1 or X-2 acted alone.
引文
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