基于发光细菌法和鱼类暴露实验评价6种化学消油剂的遗传毒性
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摘要
为评价海洋溢油应急处置中所使用的消油剂的环境安全性,分别以发光细菌Acinetobacter sp.Rec A和海水青鳉(Oryzias melastigma)幼鱼为受试生物,对6种化学消油剂的遗传毒性进行检测.结果显示,采用发光细菌法仅检测到HLD-501(常规型)在浓度较高(500 mg/L和1 000 mg/L)时表现出一定水平的遗传毒性,其等效应的丝裂霉素C(MMC)浓度分别为0.039 mg/L和0.032 mg/L.在彗星实验中,6种消油剂对海水青鳉幼鱼细胞的DNA损伤率从高到低依次为RS-II(浓缩型)、RS-I(浓缩型)、HLD-501(常规型)、富肯-2(常规型)、RS-I(常规型)和微普(常规型),然而HLD-501(常规型)处理的3级损伤率最高,其遗传毒性最大.综上,采用两种不同营养级的受试生物对6种化学消油剂遗传毒性检测结果的相关度较好;本研究可为消油剂用于处理实际海洋溢油污染的环境安全性评价提供参考.
Chemical dispersants are widely used in emergency response of marine oil spills. However, their genotoxicity information is relatively scarce. This research aimed to evaluate the environmental safety of chemical dispersants used for rapid removal and cleaning up of marine oil spills. We used luminescent bacteria test(LBT) based on Acinetobacter sp. Rec A combined with fish exposure experiment based on marine medaka(Oryzias melastigma) to detect the genotoxicity of 6 chemical dispersants. In LBT, the 500 mg/L and 1 000 mg/L of chemical dispersant HLD-501 exhibited genotoxicity of 0.039 mg/L and 0.032 mg/L of mitomycin C(MMC), respectively. In addition, the DNA damage ratio of O. melastigma by the 6 chemical dispersants in the comet assay was in the order of concentrate type RS-II > concentrate type RS-I > conventional type HLD-501 > conventional type Fuken-2 > conventional type RS-I > conventional type Weipu. However, HLD-501 resulted in the most serious DNA damage(level 3), being the most genotoxic among the 6 dispersants. The result of these two methods for genotoxicity detection fitted well with each other. This study could provide a useful reference for environmental safety evaluation of the chemical dispersants used in cleaning up of the marine oil spills.
Chemical dispersants are widely used in emergency response of marine oil spills. However, their genotoxicity information is relatively scarce. This research aimed to evaluate the environmental safety of chemical dispersants used for rapid removal and cleaning up of marine oil spills. We used luminescent bacteria test(LBT) based on Acinetobacter sp. Rec A combined with fish exposure experiment based on marine medaka(Oryzias melastigma) to detect the genotoxicity of 6 chemical dispersants. In LBT, the 500 mg/L and 1 000 mg/L of chemical dispersant HLD-501 exhibited genotoxicity of 0.039 mg/L and 0.032 mg/L of mitomycin C(MMC), respectively. In addition, the DNA damage ratio of O. melastigma by the 6 chemical dispersants in the comet assay was in the order of concentrate type RS-II > concentrate type RS-I > conventional type HLD-501 > conventional type Fuken-2 > conventional type RS-I > conventional type Weipu. However, HLD-501 resulted in the most serious DNA damage(level 3), being the most genotoxic among the 6 dispersants. The result of these two methods for genotoxicity detection fitted well with each other. This study could provide a useful reference for environmental safety evaluation of the chemical dispersants used in cleaning up of the marine oil spills.
引文
1钱国栋,赵宇鹏.消油剂对环境生物毒性评价研究[J].船海工程,2014,43(5):108-111[Qian GD,Zhao YP.Assessment on the toxicity of oil spill dispersant on marine organisms[J].Ship Ocean Eng,2014,43(5):108-111]
2 Temkin AM,Bowers RR,Magaletta ME,Holshouser S,Maggi A,Ciana P,Guillette LJ,Bowden JA,Kucklick JR,Baatz JE,Spyropoulos DD.Effects of crude oil/dispersant mixture and dispersant components on PPARγactivity in vitro and in vivo:identification of dioctyl sodium sulfosuccinate(DOSS;CAS#577-11-7)as a probable obesogen[J].Environ Heal Persp,2016,124(1):113
3赵玉慧,张友篪,孙培艳.化学消油剂在溢油污染控制中的应用及其今后发展方向[J].海洋环境科学,2006,25(A01):97-100[Zhao YH.Zhang YC,Sun PY.Application and further development of dispersants in pollution control of oll spill[J].Mar Environ Sci,2006,25(A01):97-100]
4 Singer MM,George S,Lee I,Jacobson S,Weetman LL,Blondina G,Tjeerdema RS,Aurand D,Sowby ML.Effects of dispersant treatment on the acute aquatic toxicity of petroleum hydrocarbons[J].Arch Environ Contam Toxicol,1998,34(2):177-187
5 Mitchell FM,Holdway DA.The acute and chronic toxicity of the dispersants Corexit 9527 and 9500,water accom modated f raction(WAF)of crude oil,and dispersant enhanced WAF(DEWAF)to Hydra viridissima(green hydra)[J].Water Res,2000,34(1):343-348
6高亚丽,吕昕璐,杨柏林,熊德琪,姚念娣.消油剂处理溢油对海洋浮游生物的毒性效应影响[J].大连海事大学学报:自然科学版,2014,40(1):78-82[Gao YL,Lu XL,Yang BL,Xiong DQ,Yao ND.Toxic effect of oil spill processed by the dispersant on marine planktons[J].J Dal Marit Univ,2014,40(1):78-82]
7 Gao J,Zhou W,Chen R.Acute toxicity by four kinds of oil dispersants in Cynoglossus semilaevis[M].Advan Appl Biotechnol,2015:219-222
8 Hemmer MJ,Barron MG,Greene RM.Comparative toxicity of eight oil dispersants,Louisiana sweet crude oil(LSC),and chemically dispersed LSC to two aquatic test species[J].Environ Toxicol Chem,2011,30(10):2244-2252
9姚念娣.消油剂处理燃料油对两种海洋生物的毒性效应影响研究[D].大连:大连海事大学,2012[Yao ND.Research on toxicity of fuel oil processed by the dispersant to two marine organisms[D].Dalian:Dalian Maritime University,2012]
10段美娜,杨柏林,丁光辉,熊德琪.消油剂处理溢油对海胆Cy IIa和SP-Runt基因突变效应[J].海洋环境科学,2015,34(3):468-472,480[Duan MN,Yang BL,Ding GH,Xiong DQ.Effects on mutation of Cy IIa and SP-Runt gene in sea urchins of dispersed oil[J].Mar Environ Sci,2015,34(3):468-472,480]
11 Flora DS,Renzi GPD,Camoirano A,Basso C,Zanacchi P,Bennicelli C.Genotoxicity assay of oil dispersants in bacteria(mutation,differential lethality,SOS DNA-repair)and yeast(mitotic crossing-over)[J].Mutat Res,1985,158(1):19-30
12宋一之,黄巍,张旭,李广贺.遗传毒性检测生物传感细胞的灵敏度及稳定性研究[J].清华大学学报自然科学版,2010,50(11):1880-1884[Song YZ,Huang W,Zhang X,Li GH.Sensitivity and stability study of a whole-cell biosensor for geno-toxicity assessment[J].J Tsinghua Univ(Sci Technol),2010,50(11):1880-1889]
13黄新新,何苗,罗虹.p UCD-rec A重组发光菌构建及对遗传毒性污染物响应作用[J].环境科学,2009,30(6):1722-1726[Huang XX,He M,Luo H.Construction of the pUCD-rec A recombinate luminescence bacterium to evaluate the genetoxicty of environment pollutants[J].Environ Sci,2009,30(6):1722-1726]
14 Horii T,Ogawa T,Ogawa H.Nucleotide sequence of the lex A gene of E.coli[J].Cell,1981,23(3):689-697
15农永光,胡刚.发光细菌法在国内水质监测中的应用[J].分析仪器,2012,1(1):79-81[Nong YG,Hu G.Applications ofluminous bacteria method in water quality monitoring in China[J].Anal Instr,2012,1(1):79-81]
16杜丽娜,杨帆,穆玉峰,余若祯;左剑恶,高俊发,余忻,滕丽君,汤薪瑶.某制药废水对发光细菌急性毒性的评价研究[J].环境科学,2014,35(1):286-291[Du LN,Yang F,Mu YF,Yu RZ,Zuo JE,Gao JF,Yu X,Teng LJ,Tang XY.Evaluation of the acute toxicity of pharmaceutical wastewater to luminescent bacteria[J].Environ Sci,2014,35(1):286-291]
17朱文杰,徐亚同,张秋卓,杨洁,丁国荣,王晓媛.发光细菌法在环境污染物监测中的进展与应用[J].净水技术,2010,29(4):54-59[Zhu WJ,Xu YT,Zhang QZ,Yang J,Ding GR,Wang XY.Applications ofluminous bacteria method in water quality monitoring in China[J].Water Purif Technol,2010,29(4):54-59]
18王春香,李媛媛,徐顺清.生物监测及其在环境监测中的应用[J].生态毒理学报,2010,5(5):628-638[Wang QX,Li YY,Xu SQ.Biological monitoring and its application in environmental monitoring[J].Asian JEcotoxicol,2010,5(5):628-638]
19 Sorensen SJ,Burmolle M,Hansen LH.Making bio-sense of toxicity:new developments in whole-cell biosensors[J].Curr Opin Biotechnol,2006,17(1):11-16
20崔志松,栾晓,高伟,李倩,杨官品,郑立.发光细菌法在环渤海污水样品遗传毒性检测中的应用[J].海洋与湖沼,2015,46(1):228-232[Cui ZS,Luan X,Gao W,Li Q,Yang GP,Zheng L.Using luminescent bacteria to detect genotoxicity of seawage in outfall of the bohai sea[J].Oceanol ET Limnol Sci,2015,46(1):228-232]
21邢彩虹,李桂兰.单细胞凝胶电泳法检测苯作业工人淋巴细胞DNA损伤[J].中华劳动卫生职业病杂志,2000,18(5):257-259[Xing CH,Li GL.study of DNA damage among workers exposed to benzene by alkaline single cell gel electrophoresis detection[J].Chin J Indud Hyg Occup Dis,2000,18(5):257-259]
22 Azqueta A,Collins AR.The essential comet assay:a comprehensive guide to measuring DNA damage and repair[J].Arch Toxicol,2013,87(6):949-968
23王丽,鲁志松,丁书茂,杨旭.四膜虫彗星实验在环境水质检测中的应用[J].环境科学与技术,2006,29(5):31-33[Wang L,Lu ZS,Ding SM,Yang X.Environmental water quality detection by comet assay using tetrahymena application[J].Environ Sci Technol,2006,29(5):31-33]
24文小龙,崔志松,穆景利,靳非,高伟,栾晓,郑立,孙承君,詹天荣.10种凝油剂的生物毒性评价[J].应用与环境生物学报,2015,21(5):798-804[Wen XL,Cui ZS,Mu JL,Jin F,Gao W,Luan X,Zheng L,Sun CJ,Zhan TR.Toxicity evaluation of 10 oil gelling agents[J].Chin JAppl Environ Biol,2015,21(5):798-804]
25 ASTM D3987-2012.Standard practice for shake extraction of solid waste with water[S]
26栾晓,崔志松,高伟,文小龙,徐广飞,李倩,孙承君,郑立.发光细菌法在黄岛溢油污染海水遗传毒性评价中的应用[J].应用与环境生物学报,2015,21(4):665-671[Luan X,Cui ZS,Gao W,Wen XL,Xu GF,Li Q,Sun CJ,Zheng L.Genotoxicity evaluation based on luminescent bacteria Acinetobacter sp.Rec A for seawater contaminated by Huangdao oil spill[J].Chin J Appl Environ Biol,2015,21(4):665-671]
27赵倩,段丽菊,刘英帅.彗星实验检测单细胞DNA损伤的方法[J].公共卫生与预防医学,2004,15(5):57-59[Zhao Q,Duan LJ,Liu YS.Comet assay to detect cell DNA damage[J].J Pub Heal Pre Med,2004,15(5):57-59]
28李兴暖,韩雅莉,邓瑞鹏,敖丽梅,周小朋.三丁基锡对牡蛎鳃细胞DNA损伤的研究[J].水生生物学报,2005,29(3):336-339[Li XN,Han YL,Deng RP,Ao LM,Zhou XP.Effects of tributyltin on DNA damage of gill cell from Saccostrea cucullata[J].Acta Hydr Sin,2005,29(3):336-339]
2 Temkin AM,Bowers RR,Magaletta ME,Holshouser S,Maggi A,Ciana P,Guillette LJ,Bowden JA,Kucklick JR,Baatz JE,Spyropoulos DD.Effects of crude oil/dispersant mixture and dispersant components on PPARγactivity in vitro and in vivo:identification of dioctyl sodium sulfosuccinate(DOSS;CAS#577-11-7)as a probable obesogen[J].Environ Heal Persp,2016,124(1):113
3赵玉慧,张友篪,孙培艳.化学消油剂在溢油污染控制中的应用及其今后发展方向[J].海洋环境科学,2006,25(A01):97-100[Zhao YH.Zhang YC,Sun PY.Application and further development of dispersants in pollution control of oll spill[J].Mar Environ Sci,2006,25(A01):97-100]
4 Singer MM,George S,Lee I,Jacobson S,Weetman LL,Blondina G,Tjeerdema RS,Aurand D,Sowby ML.Effects of dispersant treatment on the acute aquatic toxicity of petroleum hydrocarbons[J].Arch Environ Contam Toxicol,1998,34(2):177-187
5 Mitchell FM,Holdway DA.The acute and chronic toxicity of the dispersants Corexit 9527 and 9500,water accom modated f raction(WAF)of crude oil,and dispersant enhanced WAF(DEWAF)to Hydra viridissima(green hydra)[J].Water Res,2000,34(1):343-348
6高亚丽,吕昕璐,杨柏林,熊德琪,姚念娣.消油剂处理溢油对海洋浮游生物的毒性效应影响[J].大连海事大学学报:自然科学版,2014,40(1):78-82[Gao YL,Lu XL,Yang BL,Xiong DQ,Yao ND.Toxic effect of oil spill processed by the dispersant on marine planktons[J].J Dal Marit Univ,2014,40(1):78-82]
7 Gao J,Zhou W,Chen R.Acute toxicity by four kinds of oil dispersants in Cynoglossus semilaevis[M].Advan Appl Biotechnol,2015:219-222
8 Hemmer MJ,Barron MG,Greene RM.Comparative toxicity of eight oil dispersants,Louisiana sweet crude oil(LSC),and chemically dispersed LSC to two aquatic test species[J].Environ Toxicol Chem,2011,30(10):2244-2252
9姚念娣.消油剂处理燃料油对两种海洋生物的毒性效应影响研究[D].大连:大连海事大学,2012[Yao ND.Research on toxicity of fuel oil processed by the dispersant to two marine organisms[D].Dalian:Dalian Maritime University,2012]
10段美娜,杨柏林,丁光辉,熊德琪.消油剂处理溢油对海胆Cy IIa和SP-Runt基因突变效应[J].海洋环境科学,2015,34(3):468-472,480[Duan MN,Yang BL,Ding GH,Xiong DQ.Effects on mutation of Cy IIa and SP-Runt gene in sea urchins of dispersed oil[J].Mar Environ Sci,2015,34(3):468-472,480]
11 Flora DS,Renzi GPD,Camoirano A,Basso C,Zanacchi P,Bennicelli C.Genotoxicity assay of oil dispersants in bacteria(mutation,differential lethality,SOS DNA-repair)and yeast(mitotic crossing-over)[J].Mutat Res,1985,158(1):19-30
12宋一之,黄巍,张旭,李广贺.遗传毒性检测生物传感细胞的灵敏度及稳定性研究[J].清华大学学报自然科学版,2010,50(11):1880-1884[Song YZ,Huang W,Zhang X,Li GH.Sensitivity and stability study of a whole-cell biosensor for geno-toxicity assessment[J].J Tsinghua Univ(Sci Technol),2010,50(11):1880-1889]
13黄新新,何苗,罗虹.p UCD-rec A重组发光菌构建及对遗传毒性污染物响应作用[J].环境科学,2009,30(6):1722-1726[Huang XX,He M,Luo H.Construction of the pUCD-rec A recombinate luminescence bacterium to evaluate the genetoxicty of environment pollutants[J].Environ Sci,2009,30(6):1722-1726]
14 Horii T,Ogawa T,Ogawa H.Nucleotide sequence of the lex A gene of E.coli[J].Cell,1981,23(3):689-697
15农永光,胡刚.发光细菌法在国内水质监测中的应用[J].分析仪器,2012,1(1):79-81[Nong YG,Hu G.Applications ofluminous bacteria method in water quality monitoring in China[J].Anal Instr,2012,1(1):79-81]
16杜丽娜,杨帆,穆玉峰,余若祯;左剑恶,高俊发,余忻,滕丽君,汤薪瑶.某制药废水对发光细菌急性毒性的评价研究[J].环境科学,2014,35(1):286-291[Du LN,Yang F,Mu YF,Yu RZ,Zuo JE,Gao JF,Yu X,Teng LJ,Tang XY.Evaluation of the acute toxicity of pharmaceutical wastewater to luminescent bacteria[J].Environ Sci,2014,35(1):286-291]
17朱文杰,徐亚同,张秋卓,杨洁,丁国荣,王晓媛.发光细菌法在环境污染物监测中的进展与应用[J].净水技术,2010,29(4):54-59[Zhu WJ,Xu YT,Zhang QZ,Yang J,Ding GR,Wang XY.Applications ofluminous bacteria method in water quality monitoring in China[J].Water Purif Technol,2010,29(4):54-59]
18王春香,李媛媛,徐顺清.生物监测及其在环境监测中的应用[J].生态毒理学报,2010,5(5):628-638[Wang QX,Li YY,Xu SQ.Biological monitoring and its application in environmental monitoring[J].Asian JEcotoxicol,2010,5(5):628-638]
19 Sorensen SJ,Burmolle M,Hansen LH.Making bio-sense of toxicity:new developments in whole-cell biosensors[J].Curr Opin Biotechnol,2006,17(1):11-16
20崔志松,栾晓,高伟,李倩,杨官品,郑立.发光细菌法在环渤海污水样品遗传毒性检测中的应用[J].海洋与湖沼,2015,46(1):228-232[Cui ZS,Luan X,Gao W,Li Q,Yang GP,Zheng L.Using luminescent bacteria to detect genotoxicity of seawage in outfall of the bohai sea[J].Oceanol ET Limnol Sci,2015,46(1):228-232]
21邢彩虹,李桂兰.单细胞凝胶电泳法检测苯作业工人淋巴细胞DNA损伤[J].中华劳动卫生职业病杂志,2000,18(5):257-259[Xing CH,Li GL.study of DNA damage among workers exposed to benzene by alkaline single cell gel electrophoresis detection[J].Chin J Indud Hyg Occup Dis,2000,18(5):257-259]
22 Azqueta A,Collins AR.The essential comet assay:a comprehensive guide to measuring DNA damage and repair[J].Arch Toxicol,2013,87(6):949-968
23王丽,鲁志松,丁书茂,杨旭.四膜虫彗星实验在环境水质检测中的应用[J].环境科学与技术,2006,29(5):31-33[Wang L,Lu ZS,Ding SM,Yang X.Environmental water quality detection by comet assay using tetrahymena application[J].Environ Sci Technol,2006,29(5):31-33]
24文小龙,崔志松,穆景利,靳非,高伟,栾晓,郑立,孙承君,詹天荣.10种凝油剂的生物毒性评价[J].应用与环境生物学报,2015,21(5):798-804[Wen XL,Cui ZS,Mu JL,Jin F,Gao W,Luan X,Zheng L,Sun CJ,Zhan TR.Toxicity evaluation of 10 oil gelling agents[J].Chin JAppl Environ Biol,2015,21(5):798-804]
25 ASTM D3987-2012.Standard practice for shake extraction of solid waste with water[S]
26栾晓,崔志松,高伟,文小龙,徐广飞,李倩,孙承君,郑立.发光细菌法在黄岛溢油污染海水遗传毒性评价中的应用[J].应用与环境生物学报,2015,21(4):665-671[Luan X,Cui ZS,Gao W,Wen XL,Xu GF,Li Q,Sun CJ,Zheng L.Genotoxicity evaluation based on luminescent bacteria Acinetobacter sp.Rec A for seawater contaminated by Huangdao oil spill[J].Chin J Appl Environ Biol,2015,21(4):665-671]
27赵倩,段丽菊,刘英帅.彗星实验检测单细胞DNA损伤的方法[J].公共卫生与预防医学,2004,15(5):57-59[Zhao Q,Duan LJ,Liu YS.Comet assay to detect cell DNA damage[J].J Pub Heal Pre Med,2004,15(5):57-59]
28李兴暖,韩雅莉,邓瑞鹏,敖丽梅,周小朋.三丁基锡对牡蛎鳃细胞DNA损伤的研究[J].水生生物学报,2005,29(3):336-339[Li XN,Han YL,Deng RP,Ao LM,Zhou XP.Effects of tributyltin on DNA damage of gill cell from Saccostrea cucullata[J].Acta Hydr Sin,2005,29(3):336-339]