致命的甲壳类动物长期毒性试验卤虫franciscana</em
Summary
这项研究涉及了宝贵的方法论协议，以确定长期（14天）致死毒性化学物质，工业废水或污水和海水甲壳类的液体环境样品施加的发展和标准化，<em>卤虫franciscana</em>。
Abstract
我们的研究活动的目标环境质量评价的生物学方法的使用，特别是海水/苦咸水和泥沙。生物指标的选择必须基于可靠的科学知识，可能的话，对标准化程序的可用性。在这篇文章中，我们提出了一个标准化的协议，用于海洋甲壳动物卤虫评估毒性化学品和/或海洋环境矩阵。科学家建议，丰年虾（ 卤虫 ）是一个合适的人选标准为全球利用生物测定的发展。已发表 的论文数量，丰年虾（ 卤虫 ）的各种化学品和毒物的毒性作用。这种毒性研究的甲壳类动物的主要优势是干囊肿的整体可用性，这些都可以立即在测试中使用和培养困难不要求<suP> 1,2。囊肿的毒性检测价格便宜，连续可用，简单，可靠，因此，一个重要的答案毒性筛选的常规需求，工业监测要求或监管目的3。该方法涉及作为端点死亡率。幸存者的数量进行了清点和死亡的百分比计算。幼虫被认为是死亡，如果他们没有表现出任何内部或外部的运动，在几秒钟的观察4。这个过程是标准化测试参考物质（十二烷基硫酸钠）;在这项工作中的一些结果报告。本文附带的视频描述程序的毒性测试的性能，显示所有的有关协议的步骤。
Protocol
Discussion
卤虫是迄今所做的生态毒性测试和研究提供最有价值的测试生物体之一，让我们来说明它是可以维持有关卤虫使用毒理学和生态毒理学的几个选项。这个有机体变成一个合适的物种生物测定的特点是：广泛的地理分布，恶劣的环境条件和不同的营养素，相对简单的实验室培养和维护，抗操纵，生命周期短，大型苗种生产和高适应性大量存在的一些物种的信息。反对使用卤虫的…
Declarações
The authors have nothing to disclose.
Acknowledgements
Loredana Manfra环境的保护和研究研究所（伊斯普拉部环境质量监测）区域环境保护署（艾米利亚 – 罗马涅，意大利）和Federica Savorelli产生。
海水/苦咸水和水质的UNICHIM委员会（在化工行业的技术标准化工作的机构）的沉积物- 卤虫甲壳类生物方法集团内的子工作组议员参考。
录像和编辑由马尔科，伊斯普拉Pisapia（网络单位）。
由安娜·玛丽亚·西塞罗和Erika伊斯普拉Magaletti的的环境质量监测部生产协调。
我们要感谢集团的生物方法费拉拉科，艾米利亚 – 罗马涅，环境保护的区域代理 – 海水/苦咸水水和泥沙的UNICHIM委员会对水质和卢西亚娜米廖雷，Tor Vergata大学（罗马）的合作;洛舍Boscolo和马西莫加贝里尼，伊斯普拉（预防和减轻影响）提供财政支持的法比奥Matassa和伦敦在罗马的语言学校，他们的语言支持。
Materials
Produced by Loredana Manfra of Institute for environmental protection and research (ISPRA-Department of Environmental Quality Monitoring) and Federica Savorelli of Regional Agency for Environmental Protection (Emilia-Romagna, Italy).
Reference Members for the sub-working Group on the Artemia crustacean within the Biological Methods Group – Saltwater/Brackish Water and Sediment of the UNICHIM Commission on Water Quality (the institute for technical standardization operating in the chemical sector).
Videography and editing by Marco Pisapia of ISPRA (Scientific Advisory and Documentation Services).
Production Coordination by Anna Maria Cicero and Erika Magaletti of ISPRA (Department of Environmental Quality Monitoring).
We would like to thank the Regional Agency for Environmental Protection of Emilia-Romagna, Ferrara Branch and the Biological Methods Group – Saltwater /Brackish Water and Sediment of the UNICHIM Commission on Water Quality for their cooperation; Rossella Boscolo and Massimo Gabellini of ISPRA (Department of Prevention and Mitigation of Impacts) for their financial support; and the London School of Languages in Rome for their linguistic support.
| Name of the reagent | Company | Product number |
| Sodium Dodecyl Sulfate | Sigma-Aldrich | L4390 |
Appendix
The algal media has to be prepared using analytical quality reagents.
A liter of nutrient solution is prepared by dissolving 300 g of NaNO3, 30 g of KH2PO4 and 20 g of NH4Cl in distilled water.
Four initial solutions (A, B, C, D) of minerals are prepared as follows:
- A liter of solution “A” containing 30 g of ZnSO4 H2O, 25 g of CuSO4 5 H2O, 30 g of CoSO4 7 H2O and 20 g of MnSO4 H2O in distilled water;
- A liter of solution “B” containing 50 g of FeCl3 •6 H2O in distilled water;
- A liter of solution “C” containing 25 g of Na2MoO4 •2 H2O in distilled water;
- A liter of solution “D” containing 50 g di Na2EDTA •2 H2O in distilled water.
A liter of a final solution of minerals (solution “E”) is obtained adding aliquots of the above-mentioned solutions to distilled water. The following order of addition is recommended: 100 ml of solution “D”, 10 ml of solution “C”, 10 ml of solution “B” and 10 ml of solution “A”.
Three initial solutions (F, G, H) of vitamins are prepared as follows:
- A liter of solution “F” containing 100 mg of biotin in distilled water;
- A liter of solution “G” containing 10 g of thiamine in distilled water;
- A liter of solution “H” containing 100 mg of cobalamin in distilled water.
A liter of a final solution of vitamins (solution “I”) is obtained adding 10 ml of the three initial solutions to distilled water.
A liter of the algal media is prepared by dissolving 1 ml of the nutrient solution, 1 ml of the mineral final solution (solution “E”) and 1 ml of the vitamin final solution (solution “I”) in seawater. This latter may be obtained by dissolving recognized analytical quality reagents or a commercially available formulation in distilled or de-ionized water. Salt blends for creating the ideal saltwater are commercially available, e.g. the Instant Ocean mixtures. It is recommended the using of a seawater with salinity of 30 (±2) PSU and temperature of 20 (±1) °C.
Referências
- Togulga, M. The Short-Term Toxicity of Two Toxicants to Artemia Nauplii. Tr. J. of Zoology. 22, 259-266 (1998).
- Persoone, G., Wells, P. G., Sorgeloos, P. Artemia in aquatic toxicology: a review. Artemia Research and its Applications. Morphology, Genetics, Strain characterization Toxicology. , 259-275 (1987).
- Nunes, B. S., Carvalho, F. D., Guilhermino, L. M., Stappen, G. V. a. n. Use of the genus Artemia in ecotoxicity testing. Envir. Poll. 144, 453-462 (2006).
- APAT IRSA-CNR. . Metodi analitici per le acque Manuali e Linee guida 29/2003. Terzo Metodo, 8060 (2003).
- Sorgellos, P., Wielen, C. R. e. m. i. c. h. e. -. V. a. n. D. e. r., Persoone, G. The use of Artemia nauplii for toxicity tests – a critical analysis. Ecotox. Env. Saf. 2, 249-255 (1978).
- Savorelli, F., Palazzi, D., Gorbi, G., Invidia, M., Sei, S., Magaletti, E., Manfra, L., Gelli, F. Set up of a standard methodology for 14-day bioassay on Artemia franciscana and A. parthenogenetica. Biol. Amb. 21, 27-36 (2007).
- Brix, K. V., Cardwell, R. D., Adams, W. J. Chronic toxicity of arsenic to the Great Salt Lake brine shrimp, Artemia franciscana. Ecotox. Environ. Saf. 54, 169-175 (2003).
- Brix, K. V., Deforest, D. K., Cardwell, R. D., Adams, W. J. Derivation of a chronic site-specific water quality standard for selenium in the Great Salt Lake, Utah, USA. Environ. Toxicol. Chem. 23, 606-612 (2004).
- . Qualitá dell’acqua – Determinazione della tossicitá letale a 14 giorni con Artemia franciscana (Crustacea: Anostraca). Commissione UNICHIM “Qualitá dell’acqua” Gruppo di Lavoro “Metodi Biologici” Sottogruppo “Acque salate/salmastre e sedimenti”. , (2010).
- Varo, I., Navarro, J. C., Amat, F., Guilhermino, L. Characterisation of cholinesterases and evaluation of the inhibitory potential of chlorpyrifos and dichlorvos to Artemia salina and Artemia parthenogenetica. Chem. 48, 563-569 (2002).
