好氧有效微生物群反硝化的研究
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摘要
氮素是水体污染中的一类重要的污染物,对人体健康和环境有极大的危害,它主要以有机氮和无机氮两种形式存在。传统的生物脱氮是通过硝化和反硝化两个过程完成的,首先氨氮由自养的硝化细菌在好氧条件下转化为硝态氮,然后再通过异养的反硝化细菌在缺氧条件下将硝态氮还原为氮气,排出水体。因此,脱氮过程工艺环节较多,处理成本高,操作条件复杂等不足,极大地限制了生物脱氮技术的应用。好氧反硝化菌的发现,为生物脱氮提供了新的思路。本论文从土壤中筛选获得高效好氧反硝化有效微生物群,从而为生物脱氮技术提供较好菌源与技术。现将研究结果分述如下;
(1)通过反硝化培养基定向筛选,从土壤中分离获得好氧反硝化有效微生物群,该有效微生物群反硝化速率为143.71 mg/L·d,并命名为OAI-EMF。研究表明OAI-EMF在以淀粉为有机碳源时,其反硝化速率达到了144 mg/L·d,Cu~(2+)、Ni~(2+)和Hg~(2+)能抑制OAI-EMF反硝化作用,而Fe~(3+)、Zn~(2+)和Mn~(2+)能激活OAI-EMF反硝化作用。高浓度硝酸盐也能抑制OAI-EMF反硝化作用,而随着C/N的增加OAI-EMF反硝化速率和硝酸盐去除率都增加,在C/N为25∶1条件下,OAI-EMF的硝酸盐去除率达到100%,反硝化速率为184 mg/L·d。
(2)通过稀释涂布法从OAI-EMF中分离获得两株细菌,分别编号为NCDX—1和NCDX—2,根据菌株NCDX—1和菌株NCDX—2的生理生化实验,菌株NCDX—1和菌株NCDX—2分别符合动性球菌属(Planococcus sp.)和海球菌属(Marinococcus sp.)的特征,菌株NCDX—2初步鉴定为白色海球菌(Marhwcoccus albus)。通过传统分离纯化技术及其反硝化作用分析,该有效微生物群由两种以上细菌组成。
(3)选择聚乙烯醇作为EMF固定化载体,海藻酸钠为成型助剂,通过PVA—硼酸交联法制备了固定化EMF细胞。固定化EMF颗粒具有较好的脱氮生物活性、较好的机械稳定性和抗溶剂酸碱度,从而为固定化EMF在污水处理方面的应用提供了可能性。
(4)探讨了固定化EMF脱氮最佳工艺,研究发现固定化EMF在pH6.2-11.2,28℃～32℃,装料系数为20～40%左右的250 mL三角瓶中,培养48 h,固定化EM对硝酸盐的去除率最佳,达到了100%。与游离细胞比较,固定化EMF其对温度稳定性和对pH稳定性都有所提高,说明固定化EMF对外界不良环境的抗性增强了,能较好地适合工业化应用。
(5)在初始pH 8.2、空气流量为0.025 m~3/h、30℃、将活化的PVA固定化细胞颗粒(湿重)40 g放在装有300 mL脱氮试验培养基的流化床生物反应器(φ40×430mm)中、硝酸盐起始浓度为0.64 g/L条件下,其反硝化动力学规律为;
Nitrogen is one of the most important pollutants in the water.It is hazardous to people's health and environment.Two forms of nitrogen element in the water are organic nitrogen and inorganic nitrogen. Traditional biological denitrogenation involves two procedures—nitrification and denitrification. First of all, ammonia nitrogen is transformed, by autotrophic nitrobacteria in the presence of oxygen, into nitrate nitrogen which will then be reduced to nitrogen by heterotrophic denitrifyimg bacteria in the absence of oxygen, with nitrogen eliminated from the water. Therefore, biological denitrogenation procedures invovle many steps, making the processing cost very high. In addtion, there are other disadvantages, like the complication in the process, which to a great extent restrict the application of biological denitrogenation techniques. And the discovery of aerobic denitrifyimg bacteria offers a new perspective to biological denitrogenation. This reseach screened and isolated from soil possessing highly aerobic denitrification effective microorganisms flora(EMF), which then provides biological denitrogenation with a good source of bacteria and related technologies. All the results are descripted as follows:
(1) A newly OAI-EMF directed-screened by denitrification culture medium and isolated from soil possessing highly effective denitrification under aerobic condition. The denitrification rate of OAI-EMF is 143.71 mg/L·d. This research reveals that with the starch as its organic carbon source, the denitrification rate of OAI-EMF can reach as high as 144 mg/L·d. Though strongly inhibited by ions like Cu~(2+)、Ni~(2+) and Hg~(2+) , the process of denitrification is activated by ions of Fe~(3+),Zn~(2+) and Mn~(2+) . Nitrate with rich content also inhibits the denitrification of OAI-EMF, the increasing C/N, however, improves its denitrification rate as well as the removal rate of nitrate, which will reach 184 mg/L·d and 100% respectively when the C/N is 25:1.
(2) Two aerobic denitrifying bacterial strains named NCDX—1 and NCDX—2 were isolated from OAI-EMF . After physiological and biochemical experiments, the strain NCDX—1 was preliminary classified within the Planococcus sp. and the strain NCDX—2 was preliminary classified within the Marinococcus albus. It can be concluded that these effective microorganisms flora are composed of more than two kinds of bacteria, through traditional isolating and purifying technologies.
(3) Polyvinyl alcohol (PVA) was selected as the immobilized carrier of EMF and sodium alginate as the shaping assistant. The immobilized EMF bacteria were produced through PVA—boric acid crosslinking method. The immobolized EMF particles have good biological denitrogenation activity, mechanical stability and resistant to pH of sovlent, which then makes possible the application of immobilized EMF to waste water treatment.
(4) The thesis discussed the optical process of immobized EMF denitrogenation. And it's been found that in the triangular flasks of 250 mL with pH 6.2～11.2, the temperature 28℃～32℃, and the loading coefficient about 20～40 %, after 48 hours of culture, immobilzed EMF could realize the highest the removal rate of nitrate—100 %. Compared with free bacteria, immobolized EMF has better temperature consistency and PH stability, which shows it has stronger resistance against the outside unpleasant environment and it's more suitable for industrial application.
(5) Under the conditions of the initial pH 8.2, the air flow rate 0.025 m~3/h, the temperature 30℃, the activated PVA immobolized bacteria particles 40 g(natural weight), the fluidized bed bioreactor (φ40×430mm) of 300 mL of dinitrogenation experiment medium, and the initial concentration of nitrate 0.64 g/L, the rule ofdenitrification kinetics is:
(1)通过反硝化培养基定向筛选,从土壤中分离获得好氧反硝化有效微生物群,该有效微生物群反硝化速率为143.71 mg/L·d,并命名为OAI-EMF。研究表明OAI-EMF在以淀粉为有机碳源时,其反硝化速率达到了144 mg/L·d,Cu~(2+)、Ni~(2+)和Hg~(2+)能抑制OAI-EMF反硝化作用,而Fe~(3+)、Zn~(2+)和Mn~(2+)能激活OAI-EMF反硝化作用。高浓度硝酸盐也能抑制OAI-EMF反硝化作用,而随着C/N的增加OAI-EMF反硝化速率和硝酸盐去除率都增加,在C/N为25∶1条件下,OAI-EMF的硝酸盐去除率达到100%,反硝化速率为184 mg/L·d。
(2)通过稀释涂布法从OAI-EMF中分离获得两株细菌,分别编号为NCDX—1和NCDX—2,根据菌株NCDX—1和菌株NCDX—2的生理生化实验,菌株NCDX—1和菌株NCDX—2分别符合动性球菌属(Planococcus sp.)和海球菌属(Marinococcus sp.)的特征,菌株NCDX—2初步鉴定为白色海球菌(Marhwcoccus albus)。通过传统分离纯化技术及其反硝化作用分析,该有效微生物群由两种以上细菌组成。
(3)选择聚乙烯醇作为EMF固定化载体,海藻酸钠为成型助剂,通过PVA—硼酸交联法制备了固定化EMF细胞。固定化EMF颗粒具有较好的脱氮生物活性、较好的机械稳定性和抗溶剂酸碱度,从而为固定化EMF在污水处理方面的应用提供了可能性。
(4)探讨了固定化EMF脱氮最佳工艺,研究发现固定化EMF在pH6.2-11.2,28℃～32℃,装料系数为20～40%左右的250 mL三角瓶中,培养48 h,固定化EM对硝酸盐的去除率最佳,达到了100%。与游离细胞比较,固定化EMF其对温度稳定性和对pH稳定性都有所提高,说明固定化EMF对外界不良环境的抗性增强了,能较好地适合工业化应用。
(5)在初始pH 8.2、空气流量为0.025 m~3/h、30℃、将活化的PVA固定化细胞颗粒(湿重)40 g放在装有300 mL脱氮试验培养基的流化床生物反应器(φ40×430mm)中、硝酸盐起始浓度为0.64 g/L条件下,其反硝化动力学规律为;
Nitrogen is one of the most important pollutants in the water.It is hazardous to people's health and environment.Two forms of nitrogen element in the water are organic nitrogen and inorganic nitrogen. Traditional biological denitrogenation involves two procedures—nitrification and denitrification. First of all, ammonia nitrogen is transformed, by autotrophic nitrobacteria in the presence of oxygen, into nitrate nitrogen which will then be reduced to nitrogen by heterotrophic denitrifyimg bacteria in the absence of oxygen, with nitrogen eliminated from the water. Therefore, biological denitrogenation procedures invovle many steps, making the processing cost very high. In addtion, there are other disadvantages, like the complication in the process, which to a great extent restrict the application of biological denitrogenation techniques. And the discovery of aerobic denitrifyimg bacteria offers a new perspective to biological denitrogenation. This reseach screened and isolated from soil possessing highly aerobic denitrification effective microorganisms flora(EMF), which then provides biological denitrogenation with a good source of bacteria and related technologies. All the results are descripted as follows:
(1) A newly OAI-EMF directed-screened by denitrification culture medium and isolated from soil possessing highly effective denitrification under aerobic condition. The denitrification rate of OAI-EMF is 143.71 mg/L·d. This research reveals that with the starch as its organic carbon source, the denitrification rate of OAI-EMF can reach as high as 144 mg/L·d. Though strongly inhibited by ions like Cu~(2+)、Ni~(2+) and Hg~(2+) , the process of denitrification is activated by ions of Fe~(3+),Zn~(2+) and Mn~(2+) . Nitrate with rich content also inhibits the denitrification of OAI-EMF, the increasing C/N, however, improves its denitrification rate as well as the removal rate of nitrate, which will reach 184 mg/L·d and 100% respectively when the C/N is 25:1.
(2) Two aerobic denitrifying bacterial strains named NCDX—1 and NCDX—2 were isolated from OAI-EMF . After physiological and biochemical experiments, the strain NCDX—1 was preliminary classified within the Planococcus sp. and the strain NCDX—2 was preliminary classified within the Marinococcus albus. It can be concluded that these effective microorganisms flora are composed of more than two kinds of bacteria, through traditional isolating and purifying technologies.
(3) Polyvinyl alcohol (PVA) was selected as the immobilized carrier of EMF and sodium alginate as the shaping assistant. The immobilized EMF bacteria were produced through PVA—boric acid crosslinking method. The immobolized EMF particles have good biological denitrogenation activity, mechanical stability and resistant to pH of sovlent, which then makes possible the application of immobilized EMF to waste water treatment.
(4) The thesis discussed the optical process of immobized EMF denitrogenation. And it's been found that in the triangular flasks of 250 mL with pH 6.2～11.2, the temperature 28℃～32℃, and the loading coefficient about 20～40 %, after 48 hours of culture, immobilzed EMF could realize the highest the removal rate of nitrate—100 %. Compared with free bacteria, immobolized EMF has better temperature consistency and PH stability, which shows it has stronger resistance against the outside unpleasant environment and it's more suitable for industrial application.
(5) Under the conditions of the initial pH 8.2, the air flow rate 0.025 m~3/h, the temperature 30℃, the activated PVA immobolized bacteria particles 40 g(natural weight), the fluidized bed bioreactor (φ40×430mm) of 300 mL of dinitrogenation experiment medium, and the initial concentration of nitrate 0.64 g/L, the rule ofdenitrification kinetics is:
引文
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