原油降解菌的筛选及降油率比较
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摘要
经牛肉膏蛋白胨液体培养基驯化、固体培养基分离纯化,从长庆油田措施废液集中处理后残渣中筛选出了5株具有较强降解石油能力的微生物菌株,将其分别编号为D1、D2、D3、D4、D5。通过形态学和生理生化实验对分离得到的纯种菌株进行鉴定,结果表明D1属于芽孢杆菌属(Bacillus sp.),D2属于链球菌属(Streptococcus sp.),D3属于黄杆菌属(Flavobacterium Bergey sp.),D4属于微球菌属(Micrococcus Cohn sp.),D5属于产碱菌属(Alcaligenes sp.)。将得到的5株纯菌株在牛肉膏蛋白胨液体培养基中培养3 d,分别按单株菌株5 mL、2种菌株以1:1的比例各5 mL、3种菌株以1:1:1的比例各5 mL的接种量,分别接种到原油液体培养基中培养,7 d后,按照《CJ/T 57-1999》标准测定原油降解率。比较原油降油率,得到2种菌株组合接种的降油率高于3种菌株组合高于单菌菌株。其中D1、D4按照1:1比例,各5 mL的接种量接种到原油液体培养基中得到的降油率最高,为89.39%。
Through cultivation and acclimation in the beef paste peptone liquid culture medium, separation and purification, 5 strains of microorganisms with strong petroleum degradation capacity were selected from the residue after concentrated treatment of Changqing oilfield wastewater, and they were numbered as D1, D2,, D3, D4, D5. The isolated strains were identified by morphology and physiological and biochemical tests. The results showed that D1 belonged to Bacillus sp., D2 belonged to Streptococcus sp., and D3 belonged to Flavobacterium sp., D4 belongs to Micrococcus Cohn sp. and D5 belongs to Alcaligenes sp. The obtained 5 strains of pure strain were cultured for 3 days in the beef paste peptone liquid culture medium. 5 m L of each strain, two strains of each strain at 1:1 ratio of 5 m L, and three strains at 1:1:1 ratio of 5 m L were inoculated in crude oil liquid medium. After 7 days, the degradation rate of crude oil was measured according to "CJ/T 57-1999" standard. Comparing the crude oil degrading rate, the oil degradation rate of the 2 strains inoculated was higher than that of the 3 strains, and was also higher than that of the single strain. Among them, the oil degradation rate of D1 and D4 inoculated into crude oil liquid medium in the ratio of1:1 and the inoculation amount of each 5 m L was the highest, 89.39%.
Through cultivation and acclimation in the beef paste peptone liquid culture medium, separation and purification, 5 strains of microorganisms with strong petroleum degradation capacity were selected from the residue after concentrated treatment of Changqing oilfield wastewater, and they were numbered as D1, D2,, D3, D4, D5. The isolated strains were identified by morphology and physiological and biochemical tests. The results showed that D1 belonged to Bacillus sp., D2 belonged to Streptococcus sp., and D3 belonged to Flavobacterium sp., D4 belongs to Micrococcus Cohn sp. and D5 belongs to Alcaligenes sp. The obtained 5 strains of pure strain were cultured for 3 days in the beef paste peptone liquid culture medium. 5 m L of each strain, two strains of each strain at 1:1 ratio of 5 m L, and three strains at 1:1:1 ratio of 5 m L were inoculated in crude oil liquid medium. After 7 days, the degradation rate of crude oil was measured according to "CJ/T 57-1999" standard. Comparing the crude oil degrading rate, the oil degradation rate of the 2 strains inoculated was higher than that of the 3 strains, and was also higher than that of the single strain. Among them, the oil degradation rate of D1 and D4 inoculated into crude oil liquid medium in the ratio of1:1 and the inoculation amount of each 5 m L was the highest, 89.39%.
引文
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[19]朱妍,赵敏,杨琴,等.电动力强化对微生物修复石油污染土壤的影响[J].油气田环境保护,2016,26(03):21-25+61.
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[2]张帆,郝春博,王丽华,等.华北某油田石油降解菌的筛选及降解特性[J].环境工程学报,2014,8(10):4533-4539.
[3]刘明,孙鹏,张祥胜.原油降解菌YSL28的分离鉴定及降解特性研究[J].东北农业大学学报,2013,44(05):47-50.
[4]韦朝海,廖建波,刘浔,等. PBDEs来源特征、环境分布及污染控制[J].环境科学学报,2015,35(10):3025-3041.
[5]王晓玉,葛树生.利用电化学技术处理含油污泥[J].油气田地面工程,2014,3(6):62.
[6]Xu J, Pancras T, Grotenhuis T. Chemical oxidation of cable insulating oilcontaminatedsoil[J]. Chemosphere, 2011,84(2):272-277.
[7] Yang G, Kang D. Enhancing the biodegradation of total petroleumpollutants in oily sludge by a modified bioau-ugmentation strategy[J].International Biodeterioration&Bio-degradation,2011,65(1):130-134.
[8]刘宇程,王姗镒,马丽丽,等.石油降解菌筛选鉴定及耐受性分析[J].东北农业大学学报,2017,48(01):49-57.
[9]祝威.石油污染土壤和油泥生物处理技术[M].北京:中国石化出版社,2010,33-34.
[10]孙先锋,杨波波,朱欣洁.不同生物修复技术处理油污土壤的效果研究[J].生物技术通报,2016,32(03):68-72.
[11]杨波波,孙先锋,朱欣洁,等.堆置法处理油污土壤参数的优化[J].环境工程学报,2016,10(08):4523-4528.
[12]张闻,陈贯虹,高永超,等.石油和重金属污染土壤的微生物修复研究进展[J].环境科学与技术,2012(12):174-181.
[13]王悦明,王继富,李鑫,等.石油污染土壤微生物修复技术研究进[J].环境工程, 2014,32(08):157-161+130.
[14]王悦明,王继富,李鑫,等.石油污染土壤微生物修复技术研究进展[J].环境工程,2014,32(08):157-161+130.
[15]姚瑶,郑青松,刘兆普,等. 2株石油降解菌生长和降解石油条件的优化[J].生态与农村环境学报,2011,27(5):48-53.
[16]金芬.电场强化复合微生物菌种修复油污土壤的研究[D].西安工程大学,2015.
[17]刘舒.陕北地区石油污染土壤微生物修复菌种选育研究[D].西安:西安工程大学,2013.
[18]刘宇程,王姗镒,马丽丽,等.石油降解菌筛选鉴定及耐受性分析[J].东北农业大学学报,2017,48(01):49-57.
[19]朱妍,赵敏,杨琴,等.电动力强化对微生物修复石油污染土壤的影响[J].油气田环境保护,2016,26(03):21-25+61.
[20]周力超.应用于油脂废水处理的全细胞脂肪酶的构建及其应用[D].湖北大学,2012.
[21]王庆莲.大庆油田典型含油固废热解处理及资源化探讨[D].大庆石油学院,2009.
[22]李乐,周飞,孙先锋.石油降解菌的筛选及复合菌群的构建[J].当代化工,2018,47(04):672-676.