硫酸铜和马度米星铵联合暴露对鲫鱼的毒性和效应标记物研究
Toxicity and Toxic Effect Markers of Combined Exposure of Copper Sulphate and Maduramicin on Carassius auratus
-
摘要: 为探究硫酸铜和马度米星铵对水产动物的生态毒性，本研究以鲫鱼为模型，从急性毒性、亚急性毒性和遗传毒性3个方面研究硫酸铜和马度米星铵单一与联合暴露对鲫鱼的毒性效应。结果表明，硫酸铜对鲫鱼96 h的半数致死浓度（50% lethal concentration，LC50）为3.6 mg·L-1，硫酸铜和马度米星铵联合暴露时，硫酸铜对鲫鱼的96 h-LC50为1.4 mg·L-1，马度米星铵对鲫鱼的96 h-LC50为4.2 mg·L-1，硫酸铜对鲫鱼为高等毒性；硫酸铜和马度米星铵对鲫鱼的联合毒性表现为协同作用。染毒第7天，硫酸铜和马度米星铵均可影响过氧化氢酶（catalase，CAT）和谷胱甘肽过氧化物酶（glutathione peroxidase，GPx）的活性，随着剂量和染毒时间的增加，CAT和GPx的活性逐渐下降。硫酸铜和马度米星铵单一或联合暴露7 d或21 d显著下调cat基因表达，对sod和gpx基因表达无显著影响。染毒后期，染毒组谷丙转氨酶（alanine aminotransferase，ALT）、谷草转氨酶（aspartate transaminase，AST）和碱性磷酸酶（alkaline phosphatase，ALP）活性及丙二醛（malondialdehyde，MDA）含量均高于对照组。硫酸铜和马度米星铵单一及联合染毒均能诱导鲫鱼红细胞产生微核，微核数与硫酸铜和马度米星铵的浓度呈正相关；随着硫酸铜和马度米星铵浓度的增加，肝细胞的脱氧核糖核酸（deoxyribonucleic acid，DNA）出现断裂，彗星尾部所占比例逐渐升高；高浓度联合染毒组红细胞微核率和肝细胞尾部DNA百分比显著高于单一染毒组。硫酸铜和马度米星铵联合暴露可导致鲫鱼血液毒性、肝细胞氧化损伤及遗传毒性的加和性，为评估Cu2+与马度米星铵复合污染对鲫鱼的生态毒性提供基础数据和理论依据。Abstract: To explore the ecotoxicity of copper sulphate and maduramicin on aquatic animals, Carassius auratus was used as an animal model to understand the acute toxicity, sub-acute toxicity and genotoxicity of single and combined exposure of copper sulphate and maduramicin. The results indicated that the 96 h 50% lethal concentration (LC50) of copper sulphate on Carassius auratus was 3.6 mg·L-1, when combined exposure, the 96 h-LC50 of copper sulphate and maduramicin were 1.4 mg·L-1 and 4.2 mg·L-1, respectively, suggesting high toxicity to Carassius auratus. The joint toxic effect of copper sulphate and maduramicin on Carassius auratus was synergistic. When copper sulphate and maduramicin were exposed for 7 d, activities of catalase (CAT) and glutathione peroxidase (GPx) in liver of Carassius auratus were elevated; with the increasing of exposure concentration and time, the activities of CAT and GPx were gradually decreased. Single and combined exposure of copper sulphate and maduramicin for 7 d and 21 d significantly decreased the expression of cat but no significant effect on sod and gpx. At 21 d, the activities of alanine aminotransferase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) as well as the content of malondialdehyde (MDA) in liver of Carassius auratus were significantly increased by copper sulphate and maduramicin treatment. Single and combined exposure of copper sulphate and maduramicin all induced micronucleus generation of red blood cells in Carassius auratus in a concentration-dependent manner. With increasing of copper sulphate and maduramicin concentration, deoxyribonucleic acid (DNA) of hepatocytes in Carassius auratus were broken, and the percentage of comet tails was increased gradually. Furthermore, the micronucleus rate and the percentages of comet tails of combined exposure group were significantly higher than single exposure group. In summary, combined exposure of copper sulphate and maduramicin induce additivity of hematotoxicity, oxidative damage of hepatocytes, and genotoxicity on Carassius auratus, which together provides foundation data and the theoretical basis for the evaluation of ecotoxicity of combined contamination of copper ions and maduramicin in Carassius auratus.
-
Key words:
- copper /
- maduramicin /
- Carassius auratus /
- joint toxicity /
- ecotoxicity /
- oxidative stress /
- DNA damage
-
-
Breitholtz M, Nyholm J R, Karlsson J, et al. Are individual NOEC levels safe for mixtures? A study on mixture toxicity of brominated flame-retardants in the copepod Nitocra spinipes[J]. Chemosphere, 2008, 72(9):1242-1249 Yang W W, Tang Z P, Zhou F Q, et al. Toxicity studies of tetracycline on Microcystis aeruginosa and Selenastrum capricornutum[J]. Environmental Toxicology and Pharmacology, 2013, 35(2):320-324 张新宇, 刘晗璐, 张海华, 等. 微量元素铜在毛皮动物生产中的应用及对环境的影响[J]. 饲料研究, 2019, 42(12):105-107 Zhang X Y, Liu H L, Zhang H H, et al. Application of trace element copper in production of fur-bearing animals and effect on environment[J]. Feed Research, 2019, 42(12):105-107(in Chinese)
钱璟, 耿冉, 张耀武, 等. 铜离子对中国大鲵幼体的胁迫效应[J]. 广西科学院学报, 2019, 35(3):200-205 Qian J, Geng R, Zhang Y W, et al. The response of juvenile Chinese giant salamander on the stresses by copper ions[J]. Journal of Guangxi Academy of Sciences, 2019, 35(3):200-205(in Chinese)
姚志峰, 章龙珍, 庄平, 等. 铜对中华鲟幼鱼的急性毒性及对肝脏抗氧化酶活性的影响[J]. 中国水产科学, 2010, 17(4):731-738 Yao Z F, Zhang L Z, Zhuang P, et al. Effects of antioxidant enzyme in liver and acute toxicity of Cu2+ on juvenile Chinese sturgeon[J]. Journal of Fishery Sciences of China, 2010, 17(4):731-738(in Chinese)
谭树华, 欧阳铭. 铜对水生动物生理及毒性作用研究进展[J]. 江西农业学报, 2010, 22(11):139-142 Tan S H, Ouyang M. Research advance in physiological and toxicological effects of copper on aquatic animals[J]. Acta Agriculturae Jiangxi, 2010, 22(11):139-142(in Chinese)
Yap C K, Ismail A, Tan S G. Effects of total soft tissue and shell thickness on the accumulation of heavy metals (Cd, Cu, Pb, And Zn) in the green-lipped mussel Perna viridis[J]. Russian Journal of Marine Biology, 2003, 29(5):323-327 Johnson A, Carew E, Sloman K A. The effects of copper on the morphological and functional development of zebrafish embryos[J]. Aquatic Toxicology, 2007, 84(4):431-438 Sun Y, Zhang G, He Z Z, et al. Effects of copper oxide nanoparticles on developing zebrafish embryos and larvae[J]. International Journal of Nanomedicine, 2016, 11:905-918 Buckley J T, Roch M, McCarter J A, et al. Chronic exposure of coho salmon to sublethal concentrations of copper-I. Effect on growth, on accumulation and distribution of copper, and on copper tolerance[J]. Comparative Biochemistry and Physiology Part C:Comparative Pharmacology, 1982, 72(1):15-19 van Aardt W J, Hough M. The effect of short-term Cu exposure on the oxygen consumption and Cu accumulation of mudfish (Labeo capensis) and the largemouth bass (Micropteris salmoides) in hard water[J]. Water SA, 2019, 33(5):701-708 Folz S D, Lee B L, Nowakowski L H, et al. Anticoccidial evaluation of halofuginone, lasalocid, maduramicin, monensin and salinomycin[J]. Veterinary Parasitology, 1988, 28(1-2):1-9 Bastianello S S, Fourie N, Prozesky L, et al. Cardiomyopathy of ruminants induced by the litter of poultry fed on rations containing the ionophore antibiotic, maduramicin. I. Epidemiology, clinical signs and clinical pathology[J]. Onderstepoort Journal of Veterinary Research, 1991, 58(4):291-296 刘晓晓. 马度米星铵对克氏原螯虾的毒性研究[D]. 南京:南京农业大学, 2018:36 Liu X X. Research of toxic effects of maduramicin ammonium on the freshwater crayfish, Procambarus clarkia[D]. Nanjing:Nanjing Agricultural University, 2018:36(in Chinese) Gao X G, Liu X X, Song X H, et al. Effect of maduramicin on crayfish (Procambius clarkii):Hematological parameters, oxidative stress, histopathological changes and stress response[J]. Ecotoxicology and Environmental Safety, 2021, 211:111896 Ni H, Peng L, Gao X G, et al. Effects of maduramicin on adult zebrafish (Danio rerio):Acute toxicity, tissue damage and oxidative stress[J]. Ecotoxicology and Environmental Safety, 2019, 168:249-259 Yan L Q, Feng M B, Liu J Q, et al. Antioxidant defenses and histological changes in Carassius auratus after combined exposure to zinc and three multi-walled carbon nanotubes[J]. Ecotoxicology and Environmental Safety, 2016, 125:61-71 Xie Z X, Luan H Y, Zhang Y X, et al. Interactive effects of diclofenac and copper on bioconcentration and multiple biomarkers in crucian carp (Carassius auratus)[J]. Chemosphere, 2020, 242:125141 王晓峰, 蔡建民, 任丽华, 等. 电子垃圾拆解地区多氯联苯经口暴露健康风险评价[J]. 中国公共卫生, 2015, 31(6):734-738 Wang X F, Cai J M, Ren L H, et al. Health risk assessment of oral exposure to PCBs among residents in an e-waste dismantling area[J]. Chinese Journal of Public Health, 2015, 31(6):734-738(in Chinese)
王宗保, 张元凯, 李一琴. 黄曲霉毒素B1诱发鲫鱼肝细胞癌的实验研究[J]. 衡阳医学院学报, 1996, 24(1):18-20 Wang Z B, Zhang Y K, Li Y Q. Hepatocellular carcinoma of Carassius auratus L. induced by aflatoxin B1[J]. Journal of Hengyang Medical College, 1996, 24(1):18-20(in Chinese)
王慧珠, 罗义, 徐文青, 等. 四环素和金霉素对水生生物的生态毒性效应[J]. 农业环境科学学报, 2008, 27(4):1536-1539 Wang H Z, Luo Y, Xu W Q, et al. Ecotoxic effects of tetracycline and chlortetracycline on aquatic organisms[J]. Journal of Agro-Environment Science, 2008, 27(4):1536-1539(in Chinese)
Xiao X, Zhao Y, Zhou Y, et al. Plasma pharmacokinetics and muscle residue dynamics of mebendazole in Carassius auratus[J]. Journal of Veterinary Pharmacology and Therapeutics, 2017, 40(6):670-674 Yang F, Yang F, Wang G, et al. Pharmacokinetics of orbifloxacin in crucian carp (Carassius auratus) after intravenous and intramuscular administration[J]. Journal of Veterinary Pharmacology and Therapeutics, 2018, 41(4):599-604 Guo T, Lou C L, Zhai W W, et al. Increased occurrence of heavy metals, antibiotics and resistance genes in surface soil after long-term application of manure[J]. Science of the Total Environment, 2018, 635:995-1003 中华人民共和国国家质量监督检验检疫总局, 中国国家标准化管理委员会. 化学品鱼类急性毒性试验:GB/T 27861-2011[S]. 北京:中国标准出版社, 2011 General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China, China National Standardization Management Committee. Chemicals. Fish acute toxicity test:GB/T 27861-2011[S]. Beijing:Standards Press of China, 2011 (in Chinese)
杨丽华, 方展强, 郑文彪. 重金属对鲫鱼的急性毒性及安全浓度评价[J]. 华南师范大学学报:自然科学版, 2003, 35(2):101-106 Yang L H, Fang Z Q, Zheng W B. Safety assessment and acute toxicity of heavy metals to crucian Carassius auratus[J]. Journal of South China Normal University:Natural Science, 2003, 35(2):101-106(in Chinese)
马莉芳. 水体铜对黄河鲤毒性的研究[D]. 郑州:河南农业大学, 2013:12 Ma L F. The study of copper exposure on Yellow River carp (Cyprinus carpio)[D]. Zhengzhou:Henan Agricultural University, 2013:12(in Chinese) Ebrahimpour M, Alipour H, Rakhshah S. Influence of water hardness on acute toxicity of copper and zinc on fish[J]. Toxicology and Industrial Health, 2010, 26(6):361-365 孔令云, 田大勇, 石恬恬, 等. 混合化合物联合毒性研究进展[J]. 中国科技论文, 2014, 9(6):663-668 Kong L Y, Tian D Y, Shi T T, et al. Research progress in joint toxicity of mixed compounds[J]. China Sciencepaper, 2014, 9(6):663-668(in Chinese)
杨蓉, 李娜, 饶凯锋, 等. 环境混合物的联合毒性研究方法[J]. 生态毒理学报, 2016, 11(1):1-13 Yang R, Li N, Rao K F, et al. Review on methodology for environmental mixture toxicity study[J]. Asian Journal of Ecotoxicology, 2016, 11(1):1-13(in Chinese)
Hermesz E, Nemcsók J, Ábrahám M. Stress response in fish[J]. Pathophysiology, 1998, 5:96 徐奇友, 王常安, 许红, 等. 大豆分离蛋白替代鱼粉对哲罗鱼稚鱼生长、体成分和血液生化指标的影响[J]. 水生生物学报, 2008, 32(6):941-946 Xu Q Y, Wang C A, Xu H, et al. Effects of replacing fish meal with soy protein isolated on the growth performance, body composition and biochemical indexes of juvenile Hucho taimen[J]. Acta Hydrobiologica Sinica, 2008, 32(6):941-946(in Chinese)
Banaee M, Ahmadi K. Sub-lethal toxicity impacts of endosulfan on some biochemical parameters of the freshwater crayfish (Astacus leptodactylus)[J]. Research Journal of Environmental Sciences, 2011, 5(11):827-835 Butterfield D A, Koppal T, Howard B, et al. Structural and functional changes in proteins induced by free radical-mediated oxidative stress and protective action of the antioxidants N-tert-butyl-alpha-phenylnitrone and vitamin E[J]. Annals of the New York Academy of Sciences, 1998, 854:448-462 Zhang C, Wang J, Zhang S, et al. Acute and subchronic toxicity of pyraclostrobin in zebrafish (Danio rerio)[J]. Chemosphere, 2017, 188:510-516 刘冰, 梁婵娟. 生物过氧化氢酶研究进展[J]. 中国农学通报, 2005, 21(5):223-224 , 232 Liu B, Liang C J. Recent advances of catalase in organism[J]. Chinese Agricultural Science Bulletin, 2005, 21(5):223-224, 232(in Chinese)
Knapen M F C M, Zusterzeel P L M, Peters W H M, et al. Glutathione and glutathione-related enzymes in reproduction:A review[J]. European Journal of Obstetrics & Gynecology and Reproductive Biology, 1999, 82(2):171-184 王咏梅. 自由基与谷胱甘肽过氧化物酶[J]. 解放军药学学报, 2005, 21(5):369-371 Jin Y X, Zhang X X, Shu L J, et al. Oxidative stress response and gene expression with atrazine exposure in adult female zebrafish (Danio rerio)[J]. Chemosphere, 2010, 78(7):846-852 中华人民共和国环境保护部. 渔业水质标准:GB 11607-89[S]. 北京:中国标准出版社, 1990 Lutnicka H, Bojarski B, Witeska M, et al. Exposure to herbicide linuron results in alterations in hematological profile and stress biomarkers of common carp (Cyprinus carpio)[J]. Ecotoxicology (London, England), 2019, 28(1):69-75 史延萍. 微核技术监测污水养鱼鱼体污染率[J]. 农业环境与发展, 1998, 15(3):11-12 邢卫平, 赵恒奎, 姜宗荣. 微核及微核试验在遗传毒理学中的应用[J]. 安徽预防医学杂志, 2002, 8(5):317-320 Bojarski B, Witeska M. Blood biomarkers of herbicide, insecticide, and fungicide toxicity to fish-A review[J]. Environmental Science and Pollution Research, 2020, 27(16):19236-19250 王世锋, 徐萍, 古丽米热·安外尔, 等. 甘草对铜胁迫下鲫鱼肝胰脏的抗氧化酶活性及血液红细胞中微核数量的影响[J]. 水产科学, 2014, 33(11):713-717 Wang S F, Xu P, Anwaier G, et al. Effect of licorice on antioxidant enzyme activity in hepatopancreas and micronucleus number in red blood cell of crucian carp Carassius auratus exposed to Cu2+ stress[J]. Fisheries Science, 2014, 33(11):713-717(in Chinese)
秦椿华, 沈建英, 黄仕和, 等. DNA断裂检测方法——单细胞凝胶电泳法[J]. 生物化学与生物物理进展, 1995, 22(6):517-522 Qin C H, Shen J Y, Huang S H, et al. Single cell gel electrophoresis assay:A DNABreakage detection technique[J]. Progress in Biochemistry and Biophysics, 1995, 22(6):517-522(in Chinese)
-
点击查看大图
计量
- 文章访问数: 1539
- HTML全文浏览数: 1539
- PDF下载数: 29
- 施引文献: 0
下载:
