同步脱氮脱硫工艺生物强化及种群动态分析初探
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摘要
同步脱氮脱硫工艺是废水处理的研究热点之一,其目的是在废水处理过程中将硫化合物和硝酸盐同时去除,并产生单质硫和氮气,实现废物的资源化。将该工艺与硫酸盐废水处理工艺有机地结合起来可以降低硫酸盐废水处理所产生的出水硫化物浓度,同时与生物强化技术相结合,高效去除出水中的硫化物浓度。本课题从这一思路出发,进行了以下几个方面的研究:
采用平板夹层培养和Hungate厌氧培养技术相结合的方法,从土壤中分离到一株能够同时利用硫化物和硝酸盐的化能自养型兼性厌氧菌3-1F。鉴定和16S rDNA测序分析表明,该菌与红平红球菌的相似性达99%。菌株3-1F的代谢特征表现为接种后24h后进入生长对数期,60h达到生长高峰,此时,硫代硫酸盐和硝酸盐的利用率超过94%和75%,代谢产物以单质硫和氮气为主,含少量硫酸盐。目前,具备同步脱硫脱氮功能的红平红球菌尚未见报道。该菌株的获得为研究废水同步脱硫反硝化工艺并通过生物强化方式提高工艺的处理效能提供了微生物种源。
将分离得到的同步脱氮脱硫细菌3-1F投加到连续流反应器中进行生物强化试验,从对照试验来看,菌株投加后对反应器的pH、碱度、COD及硝酸盐的去除率没有明显的影响,对反应器硫酸盐去除的影响也是在2d内就恢复了,生物强化过程将出水硫化物浓度从130mg/L降低至30mg/L左右,硫酸根的去除率维持在60%以上,硝酸根去除率接近100%,并且利用DGGE技术分析了菌株3-1F投加后其在工艺内部的种群变化以及不同时期工艺中微生物种群之间的相似性分析,其相似性达到了82%。
采用FISH技术,分析同步脱氮脱硫工艺(厌氧膨胀床反应器)的功能微生物种群动态。自行设计了脱氮硫杆菌探针Thi-d,并采取PCR-测序技术确定了Thi-d探针的可行性,这在国内外的文献中尚无报道。选用SRB385作为硫酸盐还原菌的探针。通过杂交试验,确定了探针Thi-d和SRB385的最佳杂交温度为46℃。采用探针Thi-d和SRB385的双杂交试验没有得到理想的效果,分析认为,探针的杂交顺序、洗脱液配制方法等是存在的关键问题。
Simutaneous desulfurization and denitrfication process is one of hotspots in wastewater treatment research. Its goal is that realizes simultaneous removal of sulfide and nitrate while the elemental sulfur and the nitrogen gas are produced as metabolic products in the process of wastewater treatment. Combining simutaneous desulfurization and denitrfication process and the bioaugmentation technique with sulfide wastewater process could high effectively remove effluent concentration of sulfide in the process of sulfate wastewater process. On the basis of this research idea, the paper reported research works involving in the following several aspects.
A strain of facultative anaerobic bacterium, 3-1F, was isolated from soil using the method of double-layers plate and the Hungate anaerobic operation technique, for which, sulfide is adopted for its electron donor and nitrate for electron acceptor. Identification based on 16S rDNA sequence and physiological biochemical analysis indicated that the isolate has a homologous similarity of 99% with Rhodococcus erythropolis. The characterization of this strain showed that it could reach the logarithmic phase after inoculation for 24 h, and reach a peak after inoculation for 60 h. Under this conditions, the thiosulfate and nitrate removal rate over 94% and 75%, respectively. And the end products of this strain in mainly consist of elemental sulfur and nitrogen gas, as well as small amount of sulfate. Till now, no previous reports metioning the strain of Rhodococcus erythropolis has the function of simoutanous desulfurization and denitrification. The achievement of strain 3-1F provides a useful microbiology resource for the efficiency improvement of simutaneous desulfurization and denitrfication process throught the way of bioaugmentation.
In the process of bioaugmentation experiment, the strain 3-1F was put into the continuous flow bio-reactor. The result of bioaugmentation experiment showed that the operation had not obvious effect on pH, alkalinity, COD and nitrate removal rate. The bioaugmentation operation had produced a short term effect on sulfate removal rate, concentration of effluence sulfide decreased from 130mg/L to 30mg/L in the bioaugmentation experiment within 2d. Meanwhile,
采用平板夹层培养和Hungate厌氧培养技术相结合的方法,从土壤中分离到一株能够同时利用硫化物和硝酸盐的化能自养型兼性厌氧菌3-1F。鉴定和16S rDNA测序分析表明,该菌与红平红球菌的相似性达99%。菌株3-1F的代谢特征表现为接种后24h后进入生长对数期,60h达到生长高峰,此时,硫代硫酸盐和硝酸盐的利用率超过94%和75%,代谢产物以单质硫和氮气为主,含少量硫酸盐。目前,具备同步脱硫脱氮功能的红平红球菌尚未见报道。该菌株的获得为研究废水同步脱硫反硝化工艺并通过生物强化方式提高工艺的处理效能提供了微生物种源。
将分离得到的同步脱氮脱硫细菌3-1F投加到连续流反应器中进行生物强化试验,从对照试验来看,菌株投加后对反应器的pH、碱度、COD及硝酸盐的去除率没有明显的影响,对反应器硫酸盐去除的影响也是在2d内就恢复了,生物强化过程将出水硫化物浓度从130mg/L降低至30mg/L左右,硫酸根的去除率维持在60%以上,硝酸根去除率接近100%,并且利用DGGE技术分析了菌株3-1F投加后其在工艺内部的种群变化以及不同时期工艺中微生物种群之间的相似性分析,其相似性达到了82%。
采用FISH技术,分析同步脱氮脱硫工艺(厌氧膨胀床反应器)的功能微生物种群动态。自行设计了脱氮硫杆菌探针Thi-d,并采取PCR-测序技术确定了Thi-d探针的可行性,这在国内外的文献中尚无报道。选用SRB385作为硫酸盐还原菌的探针。通过杂交试验,确定了探针Thi-d和SRB385的最佳杂交温度为46℃。采用探针Thi-d和SRB385的双杂交试验没有得到理想的效果,分析认为,探针的杂交顺序、洗脱液配制方法等是存在的关键问题。
Simutaneous desulfurization and denitrfication process is one of hotspots in wastewater treatment research. Its goal is that realizes simultaneous removal of sulfide and nitrate while the elemental sulfur and the nitrogen gas are produced as metabolic products in the process of wastewater treatment. Combining simutaneous desulfurization and denitrfication process and the bioaugmentation technique with sulfide wastewater process could high effectively remove effluent concentration of sulfide in the process of sulfate wastewater process. On the basis of this research idea, the paper reported research works involving in the following several aspects.
A strain of facultative anaerobic bacterium, 3-1F, was isolated from soil using the method of double-layers plate and the Hungate anaerobic operation technique, for which, sulfide is adopted for its electron donor and nitrate for electron acceptor. Identification based on 16S rDNA sequence and physiological biochemical analysis indicated that the isolate has a homologous similarity of 99% with Rhodococcus erythropolis. The characterization of this strain showed that it could reach the logarithmic phase after inoculation for 24 h, and reach a peak after inoculation for 60 h. Under this conditions, the thiosulfate and nitrate removal rate over 94% and 75%, respectively. And the end products of this strain in mainly consist of elemental sulfur and nitrogen gas, as well as small amount of sulfate. Till now, no previous reports metioning the strain of Rhodococcus erythropolis has the function of simoutanous desulfurization and denitrification. The achievement of strain 3-1F provides a useful microbiology resource for the efficiency improvement of simutaneous desulfurization and denitrfication process throught the way of bioaugmentation.
In the process of bioaugmentation experiment, the strain 3-1F was put into the continuous flow bio-reactor. The result of bioaugmentation experiment showed that the operation had not obvious effect on pH, alkalinity, COD and nitrate removal rate. The bioaugmentation operation had produced a short term effect on sulfate removal rate, concentration of effluence sulfide decreased from 130mg/L to 30mg/L in the bioaugmentation experiment within 2d. Meanwhile,
引文
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