稀有鮈鲫胚胎急性毒性试验的实验室间比对
Inter-laboratory Comparison for Fish Embryo Acute Toxicity Test with Chinese Rare Minnows
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摘要: 评价稀有鮈鲫胚胎急性毒性试验在各实验室的重现性，验证稀有鮈鲫胚胎是否适用于鱼类胚胎急性毒性试验，为相关化学品测试国家标准的制定提供依据。选择3，4-二氯苯胺和五水硫酸铜作为参比物质，按照《OECD化学品测试准则No.236鱼类胚胎急性毒性试验》，进行6家实验室间比对试验以评价试验结果的重现性。实验室间比对试验中，3，4-二氯苯胺和五水硫酸铜对稀有鮈鲫胚胎急性毒性96 h半数致死浓度（LC50）的总体平均值和标准差分别为（9.25±3.62） mg·L-1和（1.63±0.59） mg·L-1，变异系数分别为39.1%和36.0%。采用z分数方法评价各实验室间比对试验结果的差异大小，各实验室3，4-二氯苯胺和五水硫酸铜比对结果的z分数分别在-1.42~1.23之间和-0.898~1.56之间。虽然变异系数较大，但各实验室的96 h-LC50均在xpt±2σpt范围之内。稀有鮈鲫胚胎急性毒性试验结果具有良好的重现性，且稀有鮈鲫胚胎对3，4-二氯苯胺和五水硫酸铜的敏感性和重复性与斑马鱼（Danio rerio）类似，毒性值差异均未超过一个数量级，因此稀有鮈鲫可作为鱼类胚胎急性毒性试验的本土受试鱼种。
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关键词:
- 稀有鮈鲫 /
- 鱼类胚胎急性毒性试验 /
- 实验室间比对 /
- 重现性
Abstract: By comparing the results of Chinese rare minnows (Gobiocypris rarus) embryo acute toxicity test from different laboratories, this study aims to validate whether Gobiocypris rarus embryos are applicable for the fish embryo acute toxicity test as a test organism, and to lay a foundation for the national standards of chemical testing. According to "OECD Guidelines for the Testing of Chemicals No. 236:Fish Embryo Acute Toxicity (FET) Test", six domestic laboratories were involved in the ring test program to evaluate the reproducibility of the method. Gobiocypris rarus embryo were exposed to two reference substances (3,4-dichloroaniline and copper sulfate pentahydrate), and the overall means of the concentration that is estimated to be lethal to 50% of embryos after 96 h exposure relative to controls (LC50) were (9.25±3.62) mg·L-1 for 3,4-dichloroaniline and (1.63±0.59) mg·L-1 for copper sulfate pentahydrate, with coefficient of variation (CV) of 39.1% and 36.0%, respectively. The difference in the results of the inter-laboratory testing were evaluated by z-score method, and a range from -1.42 to 1.23 for 3,4-dichloroaniline and that from -0.898 to 1.56 for copper sulfate pentahydrate were obtained. Although the coefficients of variation of both datasets were significant, the 96 h-LC50 of the six laboratories were within the range of xpt±2σpt. The reproducibility of this testing method using Gobiocypris rarus embryos has presented a promising result, and the level of sensitivity and repeatability of this native fish species embryo is comparable to that of Danio rerio embryo in relation to the exposure of 3,4-dichloroaniline and copper sulfate pentahydrate, i.e., the differences of the two species less than an order of magnitude. Therefore, Gobiocypris rarus can serve as standard test organism for fish toxicity studies. -
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Organization for Economic Co-operation and Development (OECD). Guidelines for testing of chemicals, 203:Fish, acute toxicity testing[S]. Paris:OECD, 2019 European Union (EU). Directive 2010/63/EU of the European Parliament and Council of 22 September 2010 on the protection of animals used for scientific purposes[J]. Official Journal of the European Union, 2010, 276:33-79 European Commission. Regulation (EC) No 1907/2006 of the European Parliament and the Council of 18 December 2006 concerning the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), establishing a European Chemicals Agency, amending Directive 1999/45/EC and repealing Council Regulation (EEC) No 793/93 and Commission Regulation (EC) No 1488/94 as well as Council Directive 76/769/EEC and Commission Directives 91/155/EEC, 93/67/EEC, 93/105/EC and 2000/21/EC[J]. Official Journal of the European Union, 2006, 396:1-849 Organization for Economic Co-operation and Development (OECD). Short guidance on the threshold approach for acute fish toxicity testing. Series on testing and assessment No. 126[S]. Paris:OECD, 2016 Organization for Economic Co-operation and Development (OECD). OECD validation report (phase 1) for the zebrafish embryo toxicity test. Series on Testing and Assessment. No. 157[R]. Paris:OECD, 2011 Organization for Economic Co-operation and Development (OECD). OECD validation report (phase 2) for the zebrafish embryo toxicity test. Series on Testing and Assessment, No. 179[R]. Paris:OECD, 2012 Organization for Economic Co-operation and Development (OECD). Guidelines for the testing of chemicals. Section 2:Effects on biotic systems test No. 236:Acute embryo acute toxicity (FET) test[S]. Paris:OECD, 2013 Braunbeck T, Boettcher M, Hollert H, et al. Towards an alternative for the acute fish LC(50) test in chemical assessment:The fish embryo toxicity test goes multi-species-An update[J]. ALTEX, 2005, 22(2):87-102 曹文宣, 王剑伟. 稀有鮈鲫——一种新的鱼类实验动物[J]. 实验动物科学与管理, 2003, 20(S1):96-99 Cao W X, Wang J W. Rare minnow:A new laboratory animal in China[J]. Laboratory Animal Science and Administration, 2003, 20(S1):96-99(in Chinese)
王蕊, 殷浩文. 斑马鱼胚胎发育中适宜的毒理学指标分析[J]. 环境与职业医学, 2004, 21(2):88-89 , 93 Wang R, Yin H W. Analysis on feasible toxicologic endpoints in embryonic development of Brachydanio rerio[J]. Journal of Labour Medicine, 2004, 21(2):88-89, 93(in Chinese)
朱晓鸣, 解绶启, 崔奕波, 等. 摄食水平和性别对稀有鮈鲫生长和能量收支的影响[J]. 海洋与湖沼, 2001, 32(3):240-247 Zhu X M, Xie S Q, Cui Y B, et al. Effect of ration level on growth and energy budget of male and female rare minnow Gobiocypris rarus [J]. Oceanologia et Limnologia Sinica, 2001, 32(3):240-247(in Chinese)
王剑伟, 乔晔, 陶玉岭. 稀有鮈鲫仔鱼的摄食和耐饥饿能力[J]. 水生生物学报, 1999, 23(6):648-654 Wang J W, Qiao Y, Tao Y L. Feeding and starvation tolerance of the yolk-sac larvae of Gobiocypris rarus [J]. Acta Hydrobiologica Sinica, 1999, 23(6):648-654(in Chinese)
王剑伟. 稀有鮈鲫的繁殖生物学[J]. 水生生物学报, 1992, 16(2):165-174 , 195 Wang J W. Reproductive biology of Gobiocypris rarus [J]. Acta Hydrobiologica Sinica, 1992, 16(2):165-174, 195(in Chinese)
常剑波,王剑伟,曹文宣. 稀有鮈鲫胚胎发育研究[J]. 水生生物学报, 1995, 19(2):97-103 Chang J B, Wang J W, Cao W X. The embryonic development of Gobiocpris rarus[J]. Acta Hydrobiologica Sinica, 1995, 19(2):97-103(in Chinese)
ISO/IEC ISO 17043, Conformity assessment-General requirements for proficiency testing; International Organization for Standardization:Geneva, 2010 张京佶, 王绿平, 张琨. 稀有鮈鲫在鱼类胚胎急性毒性试验中的适用性研究[J]. 环境科学研究, 2019, 32(7):1162-1169 Zhang J J, Wang L P, Zhang K. Applicability of Gobiocypris rarus in fish embryo acute toxicity test[J]. Research of Environmental Sciences, 2019, 32(7):1162-1169(in Chinese)
International Organization for Standardization (ISO). ISO 5725-2 Accuracy (trueness and precision) of measurement methods and results:Part 2:Basic method for the determination of repeatability and reproducibility of a standard measurement method[S]. Geneva:ISO, 1994 International Organization for Standardization (ISO). ISO 13528:2015. Statistical methods for use in proficiency testing by interlaboratory comparison[S]. Geneva:ISO, 2015 International Organization for Standardization (ISO). ISO/IEC 17043:2010 Conformity assessment:General requirements for proficiency testing[S]. Geneva:ISO, 2010 U S Environmental Protection Agency (US EPA). Methods for measuring the acute toxicity of effluents and receiving waters to freshwater and marine organisms-Fifth Edition (EPA-821-R-02-012)[R].Washington DC:Environmental Protection Agency Office of Water (4303T), 2002 顾党恩, 于学颖, 王剑伟. 稀有鮈鲫封闭群Ihb:IHB生长和繁殖性能的监测[J]. 中国实验动物学报, 2014, 22(1):71-75 Gu D G, Yu X Y, Wang J W. Monitoring of the growth and reproductive performance of a rare closed colony of Gobiocypris rarus Ihb:IHB[J]. Acta Laboratorium Animalis Scientia Sinica, 2014, 22(1):71-75(in Chinese)
王绿平, 张京佶, 殷浩文. 不同实验条件下稀有鮈鲫胚胎的自然受精率和孵化率[J]. 生态毒理学报, 2020, 15(3):100-107 Wang L P, Zhang J J, Yin H W. Natural fertility and hatchability of Gobiocypris rarus embryos under different experimental conditions[J]. Asian Journal of Ecotoxicology, 2020, 15(3):100-107(in Chinese)
范博, 樊明, 刘征涛, 等. 稀有鮈鲫物种敏感性及其在生态毒理学与水质基准中的应用[J]. 环境科学研究, 2019, 32(7):1153-1161 Fan B, Fan M, Liu Z T, et al. Species sensitivity and application in ecotoxicology and water quality criterion for Gobiocypris rarus [J]. Research of Environmental Sciences, 2019, 32(7):1153-1161(in Chinese)
王绿平, 张京佶, 赵华清. 稀有鮈鲫作为鱼类胚胎急性毒性试验受试鱼种的敏感性研究[J]. 生态毒理学报, 2021, 16(5):102-112 Wang L P, Zhang J J, Zhao H Q. Sensitivity of Chinese rare minnows (Gobiocypris rarus) for fish embryo acute toxicity test[J]. Asian Journal of Ecotoxicology, 2021, 16(5):102-112(in Chinese)
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