洛克沙胂对鲫鱼生态毒理学指标的影响
摘要
本实验研究了洛克沙胂对鲫鱼组织Na~+-K~+-ATP酶活性影响及肾细胞DNA损伤程度。发现洛克沙胂对于鲫鱼组织Na~+-K~+-ATP酶活性和肾细胞DNA损伤程度具有直接的影响。
首先以4.0、16.0、64.0mg/L洛克沙胂暴露48、96、144h后检测鲫鱼的鳃、肾脏和肝中的Na~+-K~+-ATP酶活性。试验结果表明,在同一组织中,随着处理浓度的提高,洛克沙胂对鲫鱼各组织Na~+-K~+-ATP酶活性抑制作用更为明显,呈现出明显的浓度-效应关系(P<0.01)。对于不同组织,同一处理浓度的洛克沙胂对鲫鱼鳃中Na~+-K~+-ATP酶活性抑制作用更为明显,其次是肝和肾(P<0.01)。在同一浓度下,洛克沙胂对鲫鱼各组织Na~+-K~+-ATP酶活性抑制率呈明显的时间-效应关系,暴露时间越长,抑制率越高(P<0.01)。
单细胞凝胶电泳试验(SCGE)结果表明,鲫鱼暴露于0.5、1.0、2.0mg/L洛克沙胂中72h能引起鲫鱼肾细胞明显的DNA损伤,与空白对照相比有显著性差异(P<0.05),并在设定剂量范围内呈现一定的剂量-效应关系;同时,鲫鱼肾细胞于体外分别暴露于1、10、100、500、1000mg/L洛克沙胂3、6、12h后,可引起严重的DNA损伤。可得出结论,无论体内染毒还是体外染毒,洛克沙胂均可致鲫鱼肾细胞DNA明显的断裂和迁移,对肾细胞DNA造成损伤,各染毒组与阴性对照组有显著差异。在实验期间,实验剂量内呈现剂量-效应关系和时间-效应关系。此外,将离体鲫鱼肾细胞于体外分别暴露于1、10、100、500、1000mg/L洛克沙胂的代谢产物亚砷酸钠3、6、12h后,可引起更为严重的DNA损伤,实验剂量内同样呈现剂量-效应关系和时间-效应关系。
In order to provide a reference that the influence of Roxarsone on Na~+-K~+-ATPase in crap’s tissue and DNA damage. Firstly, liver, kidney and gill were collected from craps at 48, 96, and 144 hours from Roxarsone exposure group in which craps were exposed to solution containing Roxarsone at the dose of 4.0,16.0,64.0mg/L.After that, the activities of Na~+-K~+-ATPase was analyzed. The result showed that Roxarsone can significantly inhibit the activation of Na~+-K~+-ATPase of crap in liver, kidney and gill. The inhibition of Na~+-K~+-ATPase increased with the concentrateion and the exposure time of Roxarsone,showing dose-response relationship and time-response relationship. Secondly, the results of single-cell gel eiectrophoresi(sSCGE)showed that Roxarsone at the dose of 0.5、1.0、2.0mg/L, could induce DNA damage in crap kidney cells in vivo, especially at high dose doses of exposure, and might have a potentiall genotoxicity. The damage caused by Roxarsone was obviously great and the differentce was significant,compared with those of the control(P<0.05). The extent of DNA damage increased with the concentrateion of Roxarsone, showing a striking dose-response relationship.At the same time, Roxarsone at the dose of 1.0,10,100,500,1000mg/L and exposuring for 3h,6h,12h respectively, could induce DNA damage on crap kidney cells in vitro, The damage caused by Roxarsone was obviously great and the differentce was significant,compared with those of the control(P<0.05). The extent of DNA damage showed a striking dose-response relationship and time-response relationship.
首先以4.0、16.0、64.0mg/L洛克沙胂暴露48、96、144h后检测鲫鱼的鳃、肾脏和肝中的Na~+-K~+-ATP酶活性。试验结果表明,在同一组织中,随着处理浓度的提高,洛克沙胂对鲫鱼各组织Na~+-K~+-ATP酶活性抑制作用更为明显,呈现出明显的浓度-效应关系(P<0.01)。对于不同组织,同一处理浓度的洛克沙胂对鲫鱼鳃中Na~+-K~+-ATP酶活性抑制作用更为明显,其次是肝和肾(P<0.01)。在同一浓度下,洛克沙胂对鲫鱼各组织Na~+-K~+-ATP酶活性抑制率呈明显的时间-效应关系,暴露时间越长,抑制率越高(P<0.01)。
单细胞凝胶电泳试验(SCGE)结果表明,鲫鱼暴露于0.5、1.0、2.0mg/L洛克沙胂中72h能引起鲫鱼肾细胞明显的DNA损伤,与空白对照相比有显著性差异(P<0.05),并在设定剂量范围内呈现一定的剂量-效应关系;同时,鲫鱼肾细胞于体外分别暴露于1、10、100、500、1000mg/L洛克沙胂3、6、12h后,可引起严重的DNA损伤。可得出结论,无论体内染毒还是体外染毒,洛克沙胂均可致鲫鱼肾细胞DNA明显的断裂和迁移,对肾细胞DNA造成损伤,各染毒组与阴性对照组有显著差异。在实验期间,实验剂量内呈现剂量-效应关系和时间-效应关系。此外,将离体鲫鱼肾细胞于体外分别暴露于1、10、100、500、1000mg/L洛克沙胂的代谢产物亚砷酸钠3、6、12h后,可引起更为严重的DNA损伤,实验剂量内同样呈现剂量-效应关系和时间-效应关系。
In order to provide a reference that the influence of Roxarsone on Na~+-K~+-ATPase in crap’s tissue and DNA damage. Firstly, liver, kidney and gill were collected from craps at 48, 96, and 144 hours from Roxarsone exposure group in which craps were exposed to solution containing Roxarsone at the dose of 4.0,16.0,64.0mg/L.After that, the activities of Na~+-K~+-ATPase was analyzed. The result showed that Roxarsone can significantly inhibit the activation of Na~+-K~+-ATPase of crap in liver, kidney and gill. The inhibition of Na~+-K~+-ATPase increased with the concentrateion and the exposure time of Roxarsone,showing dose-response relationship and time-response relationship. Secondly, the results of single-cell gel eiectrophoresi(sSCGE)showed that Roxarsone at the dose of 0.5、1.0、2.0mg/L, could induce DNA damage in crap kidney cells in vivo, especially at high dose doses of exposure, and might have a potentiall genotoxicity. The damage caused by Roxarsone was obviously great and the differentce was significant,compared with those of the control(P<0.05). The extent of DNA damage increased with the concentrateion of Roxarsone, showing a striking dose-response relationship.At the same time, Roxarsone at the dose of 1.0,10,100,500,1000mg/L and exposuring for 3h,6h,12h respectively, could induce DNA damage on crap kidney cells in vitro, The damage caused by Roxarsone was obviously great and the differentce was significant,compared with those of the control(P<0.05). The extent of DNA damage showed a striking dose-response relationship and time-response relationship.
引文
[1] Garbarino JR,Bednar AJ,Rutherford DW,et al.Environmental fate of roxarsone in poultry litter.I.Degradation ofroxarsone during composting.Environmental Sc-ience and Technology,2003,37:1509-1514
[2]王涛,衡正昌.彗星试验检测环境致癌物的敏感性评价.卫生毒理学杂志. 2001,15(1):24-26
[3] Lemiere, C., Malo, J. L., and Gautrin, D.:Nonsensitizing causes of occupational. asthma. Medical Clinics of North America 1996
[4]李兆利,陈海刚,徐韵等. 3种兽药及饲料添加剂对鱼类的毒理效应.生态与农村环境学报,2006,22(1):4-8
[5]杨永嘉,王国忠,王雷.高效液相色谱法测定洛克沙胂预混剂含量方法的研究.中国兽药杂志,2001, 35(6):23-26
[6] Momplaisir G M,Rosal C G,Heithma E M. Arsenic speciation methods for studying the environmental fate of organoarsenic animal-feed additives.U.S.EPA,NERL-LasVegas, 1991, 14(4): 1-11
[7]孙冬菊,孙志贤,聂继池等.铅对家兔脑组织血管内皮物质及ATP酶活力的影响.职业与健康,2007,23(10):25-27
[8]张纪亮,李瑞霞,王重刚.三丁基锡对罗非鱼稚鱼生长和ATP酶活力的影响.厦门大学学报,自然科学版,2006,45(6):28-30
[9] Hart M G,Hutchinson S,Hawkins L E.The effects of sediment-associated triorganotin compounds on the gills of the European flounder,Platichthys flesus(L.)[J].Exp.Mar.Bio1.Eco1.2001, 261(1):75-91
[10] Garbarino JR,Rutherford DW,Wershaw RL.Degradation of Roxarsone in Poult-ry Litter: In the proceedings of Arsenic in the Environment Workshop. Denver,Clorado;FebruaryU.S.Geological,Survey.2001, 8(12)21-22
[11] Overby LR,Lilian Straube.Metabolism of Arsanilic Acid,I.Metabolism Stability of Doubly Labeled Arsanilic Acid in chickens[J].Toxicology and Applied Phar- macology,1965, 7:850-854
[12] Moody JP,Williams RT.The fate of arsanilic acid and acetyl arsanilic acid in hens.Food Cosmet Toxicol,1964, 2:678-693
[13] Ostling O ,Johanson KJ . Microelectrophoretic study of radiation-induced DNA damage in individual mammalian cells [R] .Biochem. Biophys. Res. Commun ,1984 , (123) : 291-298
[14] Singh N P ,McCoy M T ,Tice R R ,et al. A simple technique for quantitation of low level of DNA damage in individual cells[J]. Exp. Cells Res ,1988 ,(175) :184-191
[15] http://www.geol.vt.edu/resesearch/gssrs/gssrs/absracts.2002
[16] Bednar AJ,Garbarino JR,Ferrer I,et al.Photodegradation of roxasone in poultry litter leachates. Sci Total Environ, 2003, 302: 237-245
[17]李银生,曾振灵,陈杖榴等.洛克沙砷的作用、毒性及环境行为.上海畜牧兽医通讯,2003, 1:10-12
[18] Richard F. Lee, Luis Felipe H. Niencheski, Karrie Brinkley.DNA strand breaks in grass shrimp embryos exposed to highway runoff sediments and sediments with coal fly ash. Marine Environmental Research, 2008, 27:22-35
[19]李芳柏,钟继洪,谭军.广东集约化养猪业的环境影响及其防治对策.土壤与环境,1999, 8(4):245-249
[20] Wershaw RL,Garbarino JR,Burkhardt MR.Roxarsone in Natural Water Systems,Effects of animal feeding operations on water resources and the environment-proceedings of the technical meeting,Fort Collins, Colorado, Geological Survey Open-File Report .2001, 8:189-204
[21] Awerkiew, Martin D?umer, Marcel Reiser, Ulrike C. Wend, Herbert Pfister, Rolf Kaiser, Wulf R. Willems, Wolfram H. Gerlich Multipliers on matrix ordered operator spaces and some K-groups. Journal of Clinical Virology, 2007,1:83-86
[22] Kotaro Aoyama, Keisuke Iwahori, Naoyuki Miyata. Application of Euglena gracilis cells to comet assay: evaluation of DNA damage and repair. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 2003,7(12) 155-162 .
[23]孟紫强,张连珍.单细胞凝胶电泳技术在人血淋巴细胞DNA损伤研究中的应用[J ].遗传学报,1998 ,8:294～300.
[24] Calvert CC.Arsenicals in animal feeds and waste.In E.A. Woolson(ed.)Arsenical pesticides.Am.Chem.Soc,1975,6:71-80
[25] Isabelle Manière, Thierry Godard, Daniel R. Doerge, Mona I. Churchwell, Magali Guffroy, Michel Laurentie, Jean-Michel Poul .DNA damage and DNA adduct formation in rat tissues following oral administration of acrylamide. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 2005, 1: 119-129.
[26]徐三元,徐仁达,陈仁尔等.经济高效多功能饲料添加剂—洛克沙胂.粮食与饲料工业,1996,7:32-33
[27]张宏欣,单永利,王双同.有机胂对鸡的营养作用及应用前景.饲料营养.2001,5:20-22
[28]李凤学,吴占福,张鹤亮.砷制剂在猪日粮中的添加效果.河北畜牧兽医.1997,3:162-163
[29]梁俊荣,闫贵龙,崔红玉等.砷制剂促生长机理研究.中国饲料,1998,9:15-16
[30] Scott A. Steinert, Rebecca Streib-Montee, James M. Leather, David B. Chadwick. DNA damage in mussels at sites in San Diego Bay. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 1998,3(13) 65-85 .
[31]徐韵,陈海刚,李兆利等.蚯蚓体腔细胞彗星试验检测阿散在泥浆体系中降解前后的遗传毒性变化.应用与环境生物学报, 2007,13(1): 46-49
[32]汪梦萍,陈培赛.洛克沙生和阿散酸对肉用仔鸡前期饲喂效果.粮食与饲料工业.1998,2:37-39
[36]易敢峰,李德发,万熙卿等.洛克沙生和阿散酸对肉仔鸡饲养效果的研究.粮食与饲料工业.1998,9:36
[37] Kerr KB,Cavett JW,Owen L,et al.The toxicity of an organic arsenical 3-nitro-4- hydroxyphenylarsonic acid I.Acute and subacute toxicity. Toxicology and Appl- ied Pharmacology,1963,5(4):507-525
[38]孙永学,陈杖榴,陈展飞等.洛克沙胂对鲫鱼暴露胁迫的毒性效应.华南农业大学学报,2004,3:101-104
[39] Kennedy S,Rice DA,Cush PF.Neuropathology of Experimental 3-nitro-4- hydro- xyphenylarsonic acid Toxicosis in pigs.Vet.Pathol,1986,23:454-461
[40] Brown BL,Slaugher AD,Schreiber ME.Controls on roxarsone transport in agric- ultural watersheds.Appl Geochem,2005,20:123-133
[2]王涛,衡正昌.彗星试验检测环境致癌物的敏感性评价.卫生毒理学杂志. 2001,15(1):24-26
[3] Lemiere, C., Malo, J. L., and Gautrin, D.:Nonsensitizing causes of occupational. asthma. Medical Clinics of North America 1996
[4]李兆利,陈海刚,徐韵等. 3种兽药及饲料添加剂对鱼类的毒理效应.生态与农村环境学报,2006,22(1):4-8
[5]杨永嘉,王国忠,王雷.高效液相色谱法测定洛克沙胂预混剂含量方法的研究.中国兽药杂志,2001, 35(6):23-26
[6] Momplaisir G M,Rosal C G,Heithma E M. Arsenic speciation methods for studying the environmental fate of organoarsenic animal-feed additives.U.S.EPA,NERL-LasVegas, 1991, 14(4): 1-11
[7]孙冬菊,孙志贤,聂继池等.铅对家兔脑组织血管内皮物质及ATP酶活力的影响.职业与健康,2007,23(10):25-27
[8]张纪亮,李瑞霞,王重刚.三丁基锡对罗非鱼稚鱼生长和ATP酶活力的影响.厦门大学学报,自然科学版,2006,45(6):28-30
[9] Hart M G,Hutchinson S,Hawkins L E.The effects of sediment-associated triorganotin compounds on the gills of the European flounder,Platichthys flesus(L.)[J].Exp.Mar.Bio1.Eco1.2001, 261(1):75-91
[10] Garbarino JR,Rutherford DW,Wershaw RL.Degradation of Roxarsone in Poult-ry Litter: In the proceedings of Arsenic in the Environment Workshop. Denver,Clorado;FebruaryU.S.Geological,Survey.2001, 8(12)21-22
[11] Overby LR,Lilian Straube.Metabolism of Arsanilic Acid,I.Metabolism Stability of Doubly Labeled Arsanilic Acid in chickens[J].Toxicology and Applied Phar- macology,1965, 7:850-854
[12] Moody JP,Williams RT.The fate of arsanilic acid and acetyl arsanilic acid in hens.Food Cosmet Toxicol,1964, 2:678-693
[13] Ostling O ,Johanson KJ . Microelectrophoretic study of radiation-induced DNA damage in individual mammalian cells [R] .Biochem. Biophys. Res. Commun ,1984 , (123) : 291-298
[14] Singh N P ,McCoy M T ,Tice R R ,et al. A simple technique for quantitation of low level of DNA damage in individual cells[J]. Exp. Cells Res ,1988 ,(175) :184-191
[15] http://www.geol.vt.edu/resesearch/gssrs/gssrs/absracts.2002
[16] Bednar AJ,Garbarino JR,Ferrer I,et al.Photodegradation of roxasone in poultry litter leachates. Sci Total Environ, 2003, 302: 237-245
[17]李银生,曾振灵,陈杖榴等.洛克沙砷的作用、毒性及环境行为.上海畜牧兽医通讯,2003, 1:10-12
[18] Richard F. Lee, Luis Felipe H. Niencheski, Karrie Brinkley.DNA strand breaks in grass shrimp embryos exposed to highway runoff sediments and sediments with coal fly ash. Marine Environmental Research, 2008, 27:22-35
[19]李芳柏,钟继洪,谭军.广东集约化养猪业的环境影响及其防治对策.土壤与环境,1999, 8(4):245-249
[20] Wershaw RL,Garbarino JR,Burkhardt MR.Roxarsone in Natural Water Systems,Effects of animal feeding operations on water resources and the environment-proceedings of the technical meeting,Fort Collins, Colorado, Geological Survey Open-File Report .2001, 8:189-204
[21] Awerkiew, Martin D?umer, Marcel Reiser, Ulrike C. Wend, Herbert Pfister, Rolf Kaiser, Wulf R. Willems, Wolfram H. Gerlich Multipliers on matrix ordered operator spaces and some K-groups. Journal of Clinical Virology, 2007,1:83-86
[22] Kotaro Aoyama, Keisuke Iwahori, Naoyuki Miyata. Application of Euglena gracilis cells to comet assay: evaluation of DNA damage and repair. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 2003,7(12) 155-162 .
[23]孟紫强,张连珍.单细胞凝胶电泳技术在人血淋巴细胞DNA损伤研究中的应用[J ].遗传学报,1998 ,8:294～300.
[24] Calvert CC.Arsenicals in animal feeds and waste.In E.A. Woolson(ed.)Arsenical pesticides.Am.Chem.Soc,1975,6:71-80
[25] Isabelle Manière, Thierry Godard, Daniel R. Doerge, Mona I. Churchwell, Magali Guffroy, Michel Laurentie, Jean-Michel Poul .DNA damage and DNA adduct formation in rat tissues following oral administration of acrylamide. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 2005, 1: 119-129.
[26]徐三元,徐仁达,陈仁尔等.经济高效多功能饲料添加剂—洛克沙胂.粮食与饲料工业,1996,7:32-33
[27]张宏欣,单永利,王双同.有机胂对鸡的营养作用及应用前景.饲料营养.2001,5:20-22
[28]李凤学,吴占福,张鹤亮.砷制剂在猪日粮中的添加效果.河北畜牧兽医.1997,3:162-163
[29]梁俊荣,闫贵龙,崔红玉等.砷制剂促生长机理研究.中国饲料,1998,9:15-16
[30] Scott A. Steinert, Rebecca Streib-Montee, James M. Leather, David B. Chadwick. DNA damage in mussels at sites in San Diego Bay. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 1998,3(13) 65-85 .
[31]徐韵,陈海刚,李兆利等.蚯蚓体腔细胞彗星试验检测阿散在泥浆体系中降解前后的遗传毒性变化.应用与环境生物学报, 2007,13(1): 46-49
[32]汪梦萍,陈培赛.洛克沙生和阿散酸对肉用仔鸡前期饲喂效果.粮食与饲料工业.1998,2:37-39
[36]易敢峰,李德发,万熙卿等.洛克沙生和阿散酸对肉仔鸡饲养效果的研究.粮食与饲料工业.1998,9:36
[37] Kerr KB,Cavett JW,Owen L,et al.The toxicity of an organic arsenical 3-nitro-4- hydroxyphenylarsonic acid I.Acute and subacute toxicity. Toxicology and Appl- ied Pharmacology,1963,5(4):507-525
[38]孙永学,陈杖榴,陈展飞等.洛克沙胂对鲫鱼暴露胁迫的毒性效应.华南农业大学学报,2004,3:101-104
[39] Kennedy S,Rice DA,Cush PF.Neuropathology of Experimental 3-nitro-4- hydro- xyphenylarsonic acid Toxicosis in pigs.Vet.Pathol,1986,23:454-461
[40] Brown BL,Slaugher AD,Schreiber ME.Controls on roxarsone transport in agric- ultural watersheds.Appl Geochem,2005,20:123-133