应用黑斑蛙胚胎测试化学品发育毒性的方法Developmental Toxicity Assay for Chemicals Using Pelophylaxnigromaculatus Embryos
苏红巧;付旭锋;赵亚娴;楼钦钦;秦占芬;
摘要(Abstract):
选择合适的尤其是本土生物种进行化学品生态毒性评估,对于各国化学品的环境管理十分重要。本文选择我国本土两栖种黑斑蛙的胚胎为试验生物,以半数致死浓度LC50、半数致畸浓度TC50、致畸指数TI及最小抑制生长浓度MCIG为终点指标,建立了黑斑蛙胚胎发育毒性试验方法。以五氯酚、视磺酸、氯化镉、重铬酸钾为测试物研究了方法的敏感性,并以五氯酚为测试物研究了方法的可重复性。结果显示:五氯酚、视磺酸、氯化镉和重铬酸钾对黑斑蛙胚胎的LC50分别为572.3μg·L-1、54.8μg·L-1、6.8 mg·L-1和97.5 mg·L-1。五氯酚、视磺酸和氯化镉对黑斑蛙胚胎的TC50分别为246.5μg·L-1、18.7μg·L-1和3.4 mg·L-1,TI分别为2.3、2.9和2.0,MCIG分别为160.0μg·L-1、10.1μg·L-1和2.4 mg·L-1。重复性试验发现,五氯酚的LC50、TC50、TI及MCIG的变异系数分别为12.6%、18.0%、23.0%和18.6%。与文献中非洲爪蟾胚胎试验的数据比较,这些数据显示黑斑蛙胚胎与非洲爪蟾胚胎对测试物的敏感性存在一定的差异,各个终点指标的变异系数相当。因此,本文建立的黑斑蛙胚胎发育毒性试验可以用于化学品的发育毒性评价,为我国化学品环境管理提供技术支持。
关键词(KeyWords): 黑斑蛙胚胎;发育毒性;化学品管理
基金项目(Foundation): “863”计划课题(2012AA06A302);; 环保公益专项(201109048)
作者(Author): 苏红巧;付旭锋;赵亚娴;楼钦钦;秦占芬;
Email:
DOI:
参考文献(References):
- [1]Colborn T,Vom Saal F S,Soto A M.Developmental effects of endocrine-disrupting chemicals in wildlife and humans[J].Environmental Health Perspectives,1993,101(5):378-384
- [2]American Society for Testing and Materials(ASTM).ASTM E1439-98 Standard Guide for Conducting the Frog Embryo Teratogenesis Aaasy-Xenopus(FETAX)[S].Philadelphia:Annual Book of ASTM Standards,2004
- [3]杨安峰,动物学.脊椎动物学[M].北京大学出版社:1992:109
- [4]应浩,陈晓萍,陈晓虹.河南新乡段卫河水对黑斑侧褶蛙早期胚胎发育的影响[J].四川动物,2008,27(5):878-880Ying H,Chen X P,Chen X H.Effect of Weihe River on early embryonic development of Pelophylax nigromaculata[J].Sichuan Journal of Zoology,2008,27(5):878-880(in Chinese)
- [5]汪学英,卢祥云,李春梅,等.重金属离子对黑斑蛙胚胎及蝌蚪的毒性影响[J].四川动物,2001,20(2):59-61Wang X Y,Lu X Y,Li C M,et.al.Toxicity of heavy metal irons to embryos and larvae of Rana nigromaculata[J].Sichuan Journal of Zoology,2001,20(2):59-61(in Chinese)
- [6]卢祥云,陈海波,韩曜平.几种农药对黑斑蛙胚胎及蝌蚪的毒性[J].四川动物,2002,21(2):84-85Lu X Y,Chen H B,Han Y P.Poisonous effect of some common pesticides on embryo and tadpole of Rana nigromaculata[J].Sichuan Journal of Zoology,2002,21(2):84-85(in Chinese)
- [7]任东凯,苏红巧,刘芃岩,等.全氟辛烷磺酸盐(PFOS)及其替代品对两栖类胚胎的发育毒性[J].生态毒理学报,2012,7(5):561-564Ren D K,Su H Q,Liu P Y,et al.Developmental toxicity of perfluorooctane sulfonate(PFOS)and its substitutes to amphibian embryos[J].Asian Journal of Ecotoxicology,2012,7(5):561-564(in Chinese)
- [8]沈英娃,王宏,卢玲.化学品环境管理与水生实验动物[J].中国比较医学杂志,2004,14(1):53-56Shen Y W,Wang H,Lu L.Environmental management of chemicals and aquatic laboratory animals[J].Chinese Journal of Comparative Medicine,2004,14(1):53-56(in Chinese)
- [9]苏红巧,张银凤,秦占芬,等.黑斑蛙与非洲爪蟾蝌蚪急性毒性试验方法敏感性的比较[J].生态毒理学报,2013,8(5):785-790Su H Q,Zhang Y F,Qin Z F,et al.Comparison of the sensitivity of Rana nigromaculata with Xenopus laevis in tadpole acute toxicity test[J].Asian Journal of Ecotoxicology,2013,8(5):785-790(in Chinese)
- [10]Gosner K L.A simplified table for staging anuran embryos and larvae with notes on identification[J].Herpetologica,1960,16(3):183-190
- [11]Prati M,Biganzoli E,Boracchi P,et al.Ecotoxicological soil evaluation by FETAX[J].Chemosphere,2000,41(10):1621-1628
- [12]Vrskova D,Modra H.Evaluation of the developmental toxicity of2-phenoxyethanol and clove oil anaesthetics using the Frog Embryo Teratogenesis Assay:Xenopus(FETAX)[J].Veterinarni Medicina,2012,57(5):245-250
- [13]Mortensen A S,Kortner T M,Arukwe A.Thyroid hormone-dependent gene expression as a biomarker of short-term 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene(DDE)exposure in European common frog(Rana temporaria)tadpoles[J].Biomarkers,2006,11(6):524-537
- [14]Oka T,Miyahara M,Yamamoto J,et al.Application of metamorphosis assay to a native Japanese amphibian species,Rana rugosa,for assessing effects of thyroid system affecting chemicals[J].Ecotoxicology and Environmental Safety,2009,72(5):1400-1405
- [15]国家环境保护部.环境保护部令第7号新化学物质环境管理办法[EB/OL].(2010-01-19)[2014-05-27].http://www.mep.gov.cn/gkml/hbb/bl/201002/t20100201_185231.htm
- [16]刘楚吾,何晓晓,肖亚梅.过量的维生素A对黑斑蛙早期胚胎发育的影响[J].湖南师范大学自然科学学报,1996,19(4):77-83Liu C W,He X X,Xiao Y M.Effects of vitamin A excess on early embryonic development of Rana nigromaculata[J].Journal of Natural Science of Hunan Normal University,1996,19(4):77-83(in Chinese)
- [17]Li X,Zhou J,Yu M,et al.Toxic effects of 1-methyl-3-octylimidazolium bromide on the early embryonic development of the frog Rana nigromaculata[J].Ecotoxicology and Environmental Safety,2009,72(2):552-556
- [18]王坤英,李卫国,赵艳红,等.重金属铬对黑斑蛙胚胎和蝌蚪生长发育的影响及血细胞DNA损伤效应[J].河南师范大学学报:自然科学版,2007,35(4):199-202Wang K Y,Li W G,Zhao Y H,et al.Effect of potassium dichromate on the development of embryo and tadpole of frog,Rana nigromaculata,and on DNA Damage[J].Journal of Henan Normal University(Natural Science),2007,35(4):199-202(in Chinese)
- [19]赵尔宓.介绍一种蛙类胚胎及蝌蚪发育的分期[J].生物学通报,1990(1):13-15
- [20]张彤,金洪钧.鱼类和两栖类胚胎幼体慢性毒性短期试验[J].应用与环境生物学报,1996,2(3):294-298Zhang T,Jin H J.Short-term chronic toxicity test for embryo-larval fish and amphibians[J].Chinese Journal of Applied and Environmental Biology,1996,2(3):294-298(in Chinese)
- [21]Osano O,Admiraal W,Otieno D.Developmental disorders in embryos of the frog Xenopus laevis induced by chloroacetanilide herbicides and their degradation products[J].Environmental Toxicology and Chemistry,2002,21(2):375-379
- [22]Collins M D,Mao G E.Teratology of retinoids[J].Annual Review of Pharmacology and Toxicology,1999,39(1):399-430
- [23]曹庆珍,钱丽娟,张小利,等.视黄酸对热带爪蟾胚胎致畸效应的表型特征[J].华东师范大学学报(自然科学版),2011,(2):134-141Cao Q Z,Qian L J,Zhang X L,et al.Teratogenic phenotypes induced in Xenopus tropicalis embryos by all-trans retinoic acid[J].Journal of East China Normal University:Natural Science,2011,(2):134-141(in Chinese)
- [24]Schwetz B A,Keeler P A,Gehring P J.The effect of purified and commercial grade pentachlorophenol on rat embryonal and fetal development[J].Toxicology and Applied Pharmacology,1974,28(1):151-161
- [25]Fort D J,Stover E L.Effect of low-level copper and pentachlorophenol exposure on various early life stages of Xenopus laevis[J].ASTM Special Technical Publication,1996,1306:188-203
- [26]熊力,马永鹏,毛思予,等.五氯酚对稀有鮈鲫胚胎毒性效应研究[J].中国环境科学,2012,32(2):337-344Xiong L,Ma Y P,Mao S Y,et al.Toxic effect of pentachlorophenol on the Chinese rare minnow embryos[J].China Environmental Science,2012,32(2):337-344(in Chinese)
- [27]Domingo J L.Metal‐induced developmental toxicity in mammals:A review[J].Journal of Toxicology and Environmental Health,Part A Current Issues,1994,42(2):123-141
- [28]Jezierska B,ugowska K,Witeska M.The effects of heavy metals on embryonic development of fish(a review)[J].Fish Physiology and Biochemistry,2009,35(4):625-640
- [29]Iijima S,Spindle A,Pedersen R A.Developmental and cytogenetic effects of potassium dichromate on mouse embryos in vitro[J].Teratology,1983,27(1):109-115
- [30]Thompson J,Bannigan J.Cadmium:toxic effects on the reproductive system and the embryo[J].Reproductive Toxicology,2008,25(3):304-315
- [31]Rayburn J R,Fort D J,Mcnew R,et al.Synergism and antagonism induced by three carrier solvents with t-retinoic acid and 6-aminonicotinamide using FETAX[J].Bulletin of Environmental Contamination and Toxicology,1991,46(4):625-632
- [32]Sunderman F W,Plowman M C,Hopfer S M.Embryotoxicity and teratogenicity of cadmium chloride in Xenopus laevis,assayed by the FETAX procedure[J].Annals of Clinical&Laboratory Science,1991,21(6):381-391
- [33]Bantle J A,Rayburn J R,Burton D T,et al.Initial interlaboratory validation study of FETAX:Phase I testing[J].Journal of Applied Toxicology,1994,14(3):213-223