厌氧氨氧化性能的研究
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摘要
氮素污染已危及人类健康和生态安全,为了控制氮素污染,环保工作者求知创新,不辱使命,推出了众多新型废水脱氮技术。其中,厌氧氨氧化技术具有高效、稳定、经济等优点,一经问世便博得大家青睐。然而,厌氧氨氧化菌生长缓慢,工程启动时间很长,严重制约了推广应用。根据电子受体的不同,厌氧氨氧化可分为亚硝酸盐型厌氧氨氧化(NH_4~++N0O_2~-→N_2+H_2O,N-anammox)和硫酸盐型厌氧氨氧化(NH_4~++SO_4~(-2)→N_2+S+H_2O,S-anammox)。对于亚硝酸盐型厌氧氨氧化,本课题从优化营养条件和改进反应器构型着手,攻坚克难,实现了对菌体的“开源截流”(促进细胞生长,高效持留菌体)。对于硫酸盐型厌氧氨氧化,本课题从创建自养条件切入,开拓进取,证实了硫酸盐型厌氧氨氧化的存在,为氮、硫地球生物化学循环提供了新的认识。主要研究结果如下:
1)发明了亚硝酸盐型厌氧氨氧化生物膜膨胀床反应器。试验发现:①以反硝化污泥启动亚硝酸盐型厌氧氨氧化工艺,可依次呈现菌体自溶阶段、活性停滞阶段和活性提高阶段。②反应器容积总氮去除率可高达3.02 kgN·m~(-3)·d~(-1)。③反应器内的接种污泥逐渐从深黄色絮状污泥转变成棕灰色颗粒污泥,再进一步转变成红色颗粒污泥。④红色颗粒污泥中的优势亚硝酸盐型厌氧氨氧化菌为“Candidatus Brocadia”,它是反应器亚硝酸盐型厌氧氨氧化功能的主要承载者。
2)探明了亚铁和高铁离子对亚硝酸盐型厌氧氨氧化菌的影响。试验发现:①亚铁和高铁离子对亚硝酸盐型厌氧氨氧化菌的基质转化具有促进作用,这种促进作用随亚铁或高铁离子浓度升高而增强;当Fe~(2+)离子浓度和Fe~(3+)离子浓度分别为4.60 mg·L~(-1)时,试验反应器对NH_4~+-N和NO_2~--N的去除速率分别为对照反应器的1.95倍和1.71倍以及2.18倍和2.84倍。②Fe~(2+)离子和Fe~(3+)离子能够缓解基质亚硝酸对亚硝酸盐型厌氧氨氧化菌的抑制作用。③常用的亚硝酸盐型厌氧氨氧化菌培养基中铁离子含量不足,提高亚硝酸盐型厌氧氨氧化菌培养基中Fe~(2+)离子或Fe~(3+)离子浓度,可促进亚硝酸盐型厌氧氨氧化菌生长;在高浓度亚铁作用下,试验反应器的VS和ATP分别是对照反应器的2.16倍和3.53倍;在高浓度高铁作用下,试验反应器的VS和ATP依次是对照反应器的4.15倍和3.37倍。④提高铁离子浓度可增大亚硝酸盐型厌氧氨氧化菌对总溶解性铁的转化量,改变亚硝酸盐型厌氧氨氧化菌铁和血红素含量;提高Fe~(2+)离子浓度,亚硝酸盐型厌氧氨氧化菌的铁含量和血红素C含量同时增加;提高Fe~(3+)离子浓度,亚硝酸盐型厌氧氨氧化菌的铁含量增加,血红素C含量下降。⑤高浓度Fe~(2+)离子的长期作用可改变亚硝酸盐型厌氧氨氧化菌的细胞结构,细胞内产生不明灰色区域;但高浓度Fe~(3+)离子的长期作用不影响亚硝酸盐型厌氧氨氧化菌细胞结构。⑥Fe~(2+)离子和Fe~(3+)离子的长期作用可改变反应器内的微生物菌群结构,其中许多微生物属于非培养细菌,潜在菌源丰富。
3)证明了自养型硫酸盐型厌氧氨氧化的存在。NH_4~+和SO_4~(2-)的化学性质稳定,两者间不发生化学反应。在厌氧反应器中接种厌氧消化污泥,利用含有NH_4~+和SO_4~(2-)的无机培养基可驯化厌氧消化污泥,使其产生硫酸盐型厌氧氨氧化功能。在高基质浓度下,NH_4~+-N和SO_4~(2-)-S浓度平均降低71.67 mg·L~(-1和18.94 mg·L~(-1)。高基质浓度和低氧化还原电位可促进硫酸盐型厌氧氨氧化。
Nitrogenous pollution has posed hazardous effects on human health and ecological safety.Environmentalists have tried their best to develop several novel nitrogen removal technologies to solve the problem.Anaerobic ammonium oxidation(anammox) has attracted much attention because it was highly efficient,stable and cost-effective. However,the long start-up period of the process due to extremely slow growth rate of anammox bacteria retards its application.Anammox process can be categoried into two kinds,including nitrite-dependent anammox(N-anammox) and sulfate-dependent anammox(S-anammox) based on different electron receptors.As for N-anammox process,endevours were made to enhance bacteria growth and cell retention in by medium optimization and reactor configuration improvement.As for S-anammox,the experiment was conducted in autotrophic conditions,and the existence of autotrophic S-anammox process was confirmed.The study provided new knowledge for biogeochemical cycle of nitrogenous and sulfurous compounds.Major research results are as follows:
1) A new N-anammox expanded-bed reactor was invented.It was discovered that:①The start-up course of N-anammox process inoculated with denitrifying sludge could be divided into three stages including autolysis phase,activity lag phase and activity elevation phase.②The maximum total nitrogen removal rate reached 3.02kg N·m~(-3)·d~(-1).③The seeding sludge in the reactor turned from khaki flocs to sandybrown granules and finally to red granules.④The predominant N-anammox bacteria in red granules was "Candidatus Brocadia", which was the major contributor to the N-anammox process in the reactor.
2) The effects of ferrous and ferric ion on N-anammox bacteria were investigated. It was found that:①Ferrous and ferric ion stimulated substrate removal of N-anammox bacteria and the stimulatory effect was enhanced with increased ferrous or ferric ion concentration.NH_4~+-N and NO_2~--N removal rates in the test reactor were 1.95- and 1.71-fold of those in the control reactor when ferrous ion concentration was 4.60mg·L~(-1);NH_4~+-N and NO_2~--N removal rates in the test reactor were 2.18- and 2.84-fold of those in the control reactor when ferric ion concentration was 4.60mg·L~(-1).②The inhibitory effect of substrate NO_2~--N to N-anammox bacteria could be relieved by ferrous and ferric ion.③The fequently used medium for N-anammox bacteria was lack of iron.The growth of N-anammox bacteria was enhanced by increment of ferrous or ferric ion in the medium.The VS and ATP of the test reactor in ferrous ion experiment was 2.16- and 3.53-fold of those in the control reactor;the VS and ATP of the test reactor in ferric ion experiment was 4.15- and 3.37-fold of those in the control reactor.④Conversion of total dissolved iron was increased by extra addition of ferrous or ferric ion.Ferrous ion addition resulted in iron and Heme C content increment,and ferric ion addition lead to iron content increment but Heme C decrease.⑤Long-term effect of ferrous-ion-replete condition resulted in cellular structure alteration of N-anammox bacteria.An unknown grey area appeared,but no similar area appeared in N-anammox bacteria under ferric-ion-replete condition.⑥The microbial community altered with extra addition of ferrous or ferric ion.Most of the microorganisms were uncultred bacteria,which implies that the potential microbial resource is abundant.
3) The existence of autotrophic S-anammox was confirmed.The results showed that both SO_4~(2-) and NH_4~+ were chemically stable.They did not react with each other in the absence of biological catalyst(sludge).Anaerobic digested sludge cultivated in an anaerobic reactor for three years took on the ability of oxidizing ammonium with sulfate anaerobically.The average reduction of NH_4~+-N and SO_4~(2-)-S were 71.67mg·L~(-1) and 18.94mg·L~(-1) at high subtrate concentrations. The experiment demonstrated that high substrate concentrations and low oxidation-reduction potential(ORP) were favourable for the biological reaction.
1)发明了亚硝酸盐型厌氧氨氧化生物膜膨胀床反应器。试验发现:①以反硝化污泥启动亚硝酸盐型厌氧氨氧化工艺,可依次呈现菌体自溶阶段、活性停滞阶段和活性提高阶段。②反应器容积总氮去除率可高达3.02 kgN·m~(-3)·d~(-1)。③反应器内的接种污泥逐渐从深黄色絮状污泥转变成棕灰色颗粒污泥,再进一步转变成红色颗粒污泥。④红色颗粒污泥中的优势亚硝酸盐型厌氧氨氧化菌为“Candidatus Brocadia”,它是反应器亚硝酸盐型厌氧氨氧化功能的主要承载者。
2)探明了亚铁和高铁离子对亚硝酸盐型厌氧氨氧化菌的影响。试验发现:①亚铁和高铁离子对亚硝酸盐型厌氧氨氧化菌的基质转化具有促进作用,这种促进作用随亚铁或高铁离子浓度升高而增强;当Fe~(2+)离子浓度和Fe~(3+)离子浓度分别为4.60 mg·L~(-1)时,试验反应器对NH_4~+-N和NO_2~--N的去除速率分别为对照反应器的1.95倍和1.71倍以及2.18倍和2.84倍。②Fe~(2+)离子和Fe~(3+)离子能够缓解基质亚硝酸对亚硝酸盐型厌氧氨氧化菌的抑制作用。③常用的亚硝酸盐型厌氧氨氧化菌培养基中铁离子含量不足,提高亚硝酸盐型厌氧氨氧化菌培养基中Fe~(2+)离子或Fe~(3+)离子浓度,可促进亚硝酸盐型厌氧氨氧化菌生长;在高浓度亚铁作用下,试验反应器的VS和ATP分别是对照反应器的2.16倍和3.53倍;在高浓度高铁作用下,试验反应器的VS和ATP依次是对照反应器的4.15倍和3.37倍。④提高铁离子浓度可增大亚硝酸盐型厌氧氨氧化菌对总溶解性铁的转化量,改变亚硝酸盐型厌氧氨氧化菌铁和血红素含量;提高Fe~(2+)离子浓度,亚硝酸盐型厌氧氨氧化菌的铁含量和血红素C含量同时增加;提高Fe~(3+)离子浓度,亚硝酸盐型厌氧氨氧化菌的铁含量增加,血红素C含量下降。⑤高浓度Fe~(2+)离子的长期作用可改变亚硝酸盐型厌氧氨氧化菌的细胞结构,细胞内产生不明灰色区域;但高浓度Fe~(3+)离子的长期作用不影响亚硝酸盐型厌氧氨氧化菌细胞结构。⑥Fe~(2+)离子和Fe~(3+)离子的长期作用可改变反应器内的微生物菌群结构,其中许多微生物属于非培养细菌,潜在菌源丰富。
3)证明了自养型硫酸盐型厌氧氨氧化的存在。NH_4~+和SO_4~(2-)的化学性质稳定,两者间不发生化学反应。在厌氧反应器中接种厌氧消化污泥,利用含有NH_4~+和SO_4~(2-)的无机培养基可驯化厌氧消化污泥,使其产生硫酸盐型厌氧氨氧化功能。在高基质浓度下,NH_4~+-N和SO_4~(2-)-S浓度平均降低71.67 mg·L~(-1和18.94 mg·L~(-1)。高基质浓度和低氧化还原电位可促进硫酸盐型厌氧氨氧化。
Nitrogenous pollution has posed hazardous effects on human health and ecological safety.Environmentalists have tried their best to develop several novel nitrogen removal technologies to solve the problem.Anaerobic ammonium oxidation(anammox) has attracted much attention because it was highly efficient,stable and cost-effective. However,the long start-up period of the process due to extremely slow growth rate of anammox bacteria retards its application.Anammox process can be categoried into two kinds,including nitrite-dependent anammox(N-anammox) and sulfate-dependent anammox(S-anammox) based on different electron receptors.As for N-anammox process,endevours were made to enhance bacteria growth and cell retention in by medium optimization and reactor configuration improvement.As for S-anammox,the experiment was conducted in autotrophic conditions,and the existence of autotrophic S-anammox process was confirmed.The study provided new knowledge for biogeochemical cycle of nitrogenous and sulfurous compounds.Major research results are as follows:
1) A new N-anammox expanded-bed reactor was invented.It was discovered that:①The start-up course of N-anammox process inoculated with denitrifying sludge could be divided into three stages including autolysis phase,activity lag phase and activity elevation phase.②The maximum total nitrogen removal rate reached 3.02kg N·m~(-3)·d~(-1).③The seeding sludge in the reactor turned from khaki flocs to sandybrown granules and finally to red granules.④The predominant N-anammox bacteria in red granules was "Candidatus Brocadia", which was the major contributor to the N-anammox process in the reactor.
2) The effects of ferrous and ferric ion on N-anammox bacteria were investigated. It was found that:①Ferrous and ferric ion stimulated substrate removal of N-anammox bacteria and the stimulatory effect was enhanced with increased ferrous or ferric ion concentration.NH_4~+-N and NO_2~--N removal rates in the test reactor were 1.95- and 1.71-fold of those in the control reactor when ferrous ion concentration was 4.60mg·L~(-1);NH_4~+-N and NO_2~--N removal rates in the test reactor were 2.18- and 2.84-fold of those in the control reactor when ferric ion concentration was 4.60mg·L~(-1).②The inhibitory effect of substrate NO_2~--N to N-anammox bacteria could be relieved by ferrous and ferric ion.③The fequently used medium for N-anammox bacteria was lack of iron.The growth of N-anammox bacteria was enhanced by increment of ferrous or ferric ion in the medium.The VS and ATP of the test reactor in ferrous ion experiment was 2.16- and 3.53-fold of those in the control reactor;the VS and ATP of the test reactor in ferric ion experiment was 4.15- and 3.37-fold of those in the control reactor.④Conversion of total dissolved iron was increased by extra addition of ferrous or ferric ion.Ferrous ion addition resulted in iron and Heme C content increment,and ferric ion addition lead to iron content increment but Heme C decrease.⑤Long-term effect of ferrous-ion-replete condition resulted in cellular structure alteration of N-anammox bacteria.An unknown grey area appeared,but no similar area appeared in N-anammox bacteria under ferric-ion-replete condition.⑥The microbial community altered with extra addition of ferrous or ferric ion.Most of the microorganisms were uncultred bacteria,which implies that the potential microbial resource is abundant.
3) The existence of autotrophic S-anammox was confirmed.The results showed that both SO_4~(2-) and NH_4~+ were chemically stable.They did not react with each other in the absence of biological catalyst(sludge).Anaerobic digested sludge cultivated in an anaerobic reactor for three years took on the ability of oxidizing ammonium with sulfate anaerobically.The average reduction of NH_4~+-N and SO_4~(2-)-S were 71.67mg·L~(-1) and 18.94mg·L~(-1) at high subtrate concentrations. The experiment demonstrated that high substrate concentrations and low oxidation-reduction potential(ORP) were favourable for the biological reaction.
引文
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