摘要
储油罐底油泥是一种废弃物,若能研究出一套经济实用的方法对其进行无公害处理,就会成为一种资源,并产生一定的经济社会效益。生物处理法以其无二次污染、操作简单、费用低等优点目前被认为是处理含油污泥的一种比较好的方法。目前的研究多集中于单一菌株对石油的降解作用。本研究从分离表面活性剂产生菌和石油降解菌出发,通过菌株组合试验,获得一个石油降解混合菌群Flo.2,并对其石油降解条件进行研究;通过摇床试验,利用混合菌群对储油罐底油泥进行无公害处理,以期为治理含油污泥造成的污染提供一条技术途径。
目的:从储油罐底泥及石油污染土壤分离表面活性剂产生菌和石油降解菌,完成石油降解混合菌群的组建,并研究其石油降解条件,利用混合菌群对储油罐底泥进行无公害处理。以期为实现含油污泥微生物治理工艺奠定理论基础。
方法:通过富集分离法,获得表面活性剂产生菌和石油降解菌;通过菌株组合试验,组建石油降解混合菌群;通过菌株、菌落形态观察试验对菌群中各菌株进行初步鉴定;设计单因素试验选择菌群降解石油的的最佳氮、磷营养盐、最适pH范围及摇床转速;并先后采用L_9(3~4)、L_(16)(4~5)正交试验对菌群中各菌株的最适接种比例、菌群的石油降解条件进行优化研究;通过摇床试验,利用混合菌群对储油罐底油泥进行无公害处理。
结果:分离得到2株石油降解菌(编号为PB10,SY-8),3株表面活性产生菌(编号为SY-3,PB2,GJ4),组合试验得到一个石油降解混合菌群Flo.2(由菌株PB2,PB10,SY-3和SY-8组成),初步鉴定4株菌SY-3、SY-8、PB2和PB10均为芽孢杆菌属(Bacillussp.),其适宜的生长和石油降解条件:pH值5～9,最适pH值7.5;温度25～50℃,最适温度35℃;盐浓度2～12g/L,最适盐浓度7g/L;
菌群Flo.2石油降解的最佳氮源为NaNO_3,磷源为K_2HPO_4,最适pH范围7～9,摇床转数180 r/min,优化后菌群Flo.2中各菌株的最适接种比例PB2:PB10:SY-3:SY-8为5:1:5:1,最佳石油降解条件为pH 8.0,初始油浓度12g/L,NaNO_34g/L,K_2HPO_43g/L,接种量7%,在此条件下,其石油降解率达73.19%,石油粘度降低61.83%。利用混合菌群Flo.2对储油罐底油泥进行处理,结果表明同时投加混合菌群Flo.2和无机盐营养物质能够提高罐底油泥中石油的降解速率,处理16d,石油降解率达71.4%。气相色谱检测分析表明混合菌群Flo.2对石油的不同组分有不同程度的降解作用,经菌群F10.2作用后,罐底油泥中油、泥沙分离效果相对较好。
结论:该研究为解决油田环境问题提供了一定的菌种资源和信息,同时为治理含油污泥造成的污染或其他石油污染提供了一条可行的途径。
Oily sludge is a waste,it will become a resource and bring certain economic benefits if we can search a set of economical and practical way to deal with it.Biological treatment method for its no secondary pollution,simple operation,low cost is considered a better method of treating oily sludge currently.The recent study focused on a single strain of the oil degradation.In this thesis,oil degrading and surfactant-producing strains were isolated from oily sludge of oil-tank firstly,and via combination,the mixd culture Flo.2 was obtained, further studying the optimizational conditions of oily sludge,by shaing flask test,the biodegradation experiment of oily sludge from tank bottom was finished with mixd culture. This will be providing a technical way to control pollution caused by oil-tank sludge.
Objectives:Oil degrading and surfactant-producing strains were isolated from oily sludge from oil-tank and oil-contaminantd soil firstly in order to construct a mixd culture; study on the optimizational conditions of oil degradation of mixd culture;through shaing flask test,the biodegradation experiment of oily sludge from tank bottom was finished with mixd culture.This will be laying a theoretical basis for the realization of oily sludge treatment. Methods:Through enrichment and separation,the oil degrading and surfactant-producing strains were obtained;and via combination,a mixd culture was constructed;by using morphological,physiological and biochemical methods of the various strains in the mixd culture;choice of single-factor experiment for the best oil degradation of nitrogen and phosphorus nutrients,the suitable pH value range and rotate speed;the optimal inoculation proportion of strains and the optimal conditions of oil degradation were screened by L_9(3~4) and L_(16)(4~5) orthogonal experiment;the biodegradation experiment of oily sludge from tank bottom was finished with mixd culture by shaing flask test.Results:two strains of oil degrading strains(numbered PB10,SY-8) and 3 strains of surfactant-producing strains (numbered SY-3,PB2,GJ4) were isolated from oil-tank sludge,the mixd culture composed of PB10,SY-8,SY-3 and PB2 was abtained via combination test,the strains SY-3,SY-8,PB2 and PB10 as Bacillus sp.,for the four strains the suitable pH value range is 5～9,the temperature range is 25℃～50℃,the range of salt concentration is 2～12 g/L,the optimal conditions:the temperature 35℃,the pH 7.5,the salt concentration 7 g/L;The optimal oil degrading ingredient of Flo.2 was:N sources and P sources were respectively NaNO_3 and K_2HPO_4,the optimal range of pH value was 7～9,rotate speed 180 r/min;the best inoculation proportion of each strain was PB2:PB10:SY-3:SY-8=5:1:5:1,the optimal conditions of oil degradation were that pH 8.0,initial oil concentration 12 g/L,NaNO_3 4 g/L,K_2HPO_4 3 g/L, inoculation volume7%,under the optimal conditions,the oil degradation rate reached 73.19%, the viscosity reduced 61.83%.The result of biodegradation experiment found that the oil degradation rate was increased rapidly when adding both nutrients and mixd culture Flo.2,in 16 days,it reached 71.4%.Gas chromatography detection analysis showed that Flo.2 can degraded saturated and aromatic hydrocarbon drastically,the oil and sand were separated effectively after degradation.Conclusions:This research provides some strain resource and information in solving oil field environment pollution,and also a feasible way in aspects of the biological control for future pollution caused by oil-tank sludge or other oil pollution.
引文
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