许玉洁^1,2,3,4,
高亮^1,2,3,4,
陈进林^1,2,3,4,
余明喧^1,2,3,4,
冯宝欣^1,2,3,4,
黄叶梅^1,2,3,4,
梅承芳^1,2,3,4,
曾国驱^1,2,3,4,,
1. 广东省科学院广东省微生物研究所, 广州 510070;
2. 华南应用微生物国家重点实验室, 广州 510070;
3. 广东省菌种保藏与应用重点实验室, 广州 510070;
4. 广东省微生物应用新技术公共实验室, 广州 510070

作者简介: 许玉洁(1988-),女,硕士,研究方向为生态毒理学,E-mail:xuyj@gddcm.com.

通讯作者: 曾国驱,zenggq@gddcm.com

基金项目: 国家重点研发计划项目“化学品健康危害快速分级与确证技术研究”项目(2017YFF0211200)；广州市科技计划项目“生活污水厂出水中OMPs毒害风险的同步多级快速检测与评估”(201707010162)


中图分类号: X171.5

Effects of Test Medium on the Growth, Reproduction and Offspring of Daphnia magna

Xu Yujie^1,2,3,4,
Gao Liang^1,2,3,4,
Chen Jinlin^1,2,3,4,
Yu Mingxuan^1,2,3,4,
Feng Baoxin^1,2,3,4,
Huang Yemei^1,2,3,4,
Mei Chengfang^1,2,3,4,
Zeng Guoqu^1,2,3,4,,
1. Guangdong Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, China;
2. State Key Laboratory of Applied Microbiology Southern China, Guangzhou 510070, China;
3. Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangzhou 510070, China;
4. Guangdong Open Laboratory of Applied Microbiology, Guangzhou 510070, China

Corresponding author: Zeng Guoqu,zenggq@gddcm.com

CLC number: X171.5

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摘要:大型溞(Daphnia magna)急性活动抑制试验和繁殖试验是研究和评价新化学物质对水生生物危害性的重要手段之一。由于经济合作与发展组织(Organization for Economic Co-operation and Development, OECD)测试导则推荐的Elendt M4培养基中含有的乙二胺四乙酸(EDTA)可能会影响金属离子的活性，不适合用于含有金属离子样品的毒理学测试。为选出合适的替代培养基，采用Elendt M4培养基、美国材料与试验协会(American Society for Testing and Materials, ASTM)推荐的ASTM培养基和国际标准化组织(International Organization for Standardization, ISO)推荐的ISO培养基进行大型溞繁殖试验，并收集繁殖试验中亲溞产下的非头胎幼溞进行重铬酸钾的急性活动抑制试验。结果表明，3种培养基对大型溞的生长、存活均无抑制作用，ASTM培养基和ISO培养基中大型溞的繁殖量分别为111±10和70±13，显著低于Elendt M4培养基的132±4 (P<0.05)。3种培养基中大型溞的繁殖量均能满足OECD 211-2012大型溞繁殖试验标准中繁殖量大于60的要求，但在ISO培养基中有死胎出现。重铬酸钾对Elendt M4培养基、ASTM培养基和ISO培养基中繁殖幼溞的24 h半数效应浓度(24 h-EC₅₀)值分别为1.29、1.84和0.772 mg·L^-1，均在OECD 202-2004大型溞急性活动抑制试验标准要求的0.6～2.1 mg·L^-1范围内，说明3种培养基中繁殖的大型溞均能用于大型溞急性活动抑制试验的标准测试中。综上所述，进行含有金属离子样品的毒理测试时，ASTM培养基和ISO培养基均可用于大型溞的短期驯养和急性活动抑制试验，ASTM培养基还可以用于长期驯养和繁殖试验。以上结果为大型溞毒性试验中试验培养基的选择提供理论依据。
关键词: 重铬酸钾/
大型溞/
繁殖试验/
培养基

Abstract:The acute immobilization test and reproduction test of Daphnia magna are important technical means to study and evaluate the hazard of chemicals on aquatic organisms. As ethylene diamine tetraacetic acid (EDTA) in the Elendt M4 medium recommended by the Organization for Economic Cooperation and Development (OECD) testing guidelines may affect the activity of metal ions, Elendt M4 medium is not suitable for toxicological testing of samples containing metals. In order to select the appropriate medium, three different test media of Elendt M4 medium, ASTM medium recommended by American Society for Testing and Materials (ASTM) and ISO medium recommended by International Organization for Standardization (ISO) were adopted to perform the reproduction test of Daphnia magna and the young daphnids, which were not first brood progeny, were collected and used in the acute immobilization test of reference substance K₂Cr₂O₇. The results showed that the three test media had no inhibitory effect on the growth and survival of Daphnia magna, and the number of offspring in ASTM medium and ISO medium was 111±10 and 70±13, respectively, which was significantly lower than the value of 132±4 of Elendt M4 medium (P<0.05). The number of offspring in the three test media could all reach the requirement of OECD 211-2012 “Daphnia magna Reproduction Test Guideline” which should be over 60, while dead offspring occurred in ISO medium. The results of acute immobilization test showed that 24 h-EC₅₀ values of K₂Cr₂O₇ in Elendt M4 medium, ASTM medium and ISO medium were 1.29, 1.84 and 0.772 mg·L^-1, respectively. The 24 h-EC₅₀ values in three test media were all in the range of 0.6～2.1 mg·L^-1, which met the requirement of OECD 202-2004 “Daphnia sp., Acute Immobilization Test Guideline”. It is indicated that the offspring in three test media can be used in the standard test of daphnia activity inhibition. In conclusion, both ASTM medium and ISO medium can be used for short-term domestication and acute toxicity test when test samples containing metals, and ASTM medium can also be used for long-term domestication or reproduction test. Such results provide a theoretical basis for the selection of test medium in daphnia toxicity tests.
Key words:K₂Cr₂O₇/
Daphnia magna/
reproduction test/
test medium.