Varó I, Amat F, Navarro J C
Departamento de Biología Funcional y Antropología Física, Facultad de Ciencias Biológicas, Universidad de Valencia, C/ Doctor Moliner s/n, Burjasot, Valencia, Spain.
Aquat Toxicol. 2008 Jun 2;88(1):53-61. doi: 10.1016/j.aquatox.2008.03.004. Epub 2008 Mar 14.
This study evaluates the toxic effects of the organophosphate pesticide (OP) dichlorvos to the endangered Iberian toothcarp (Aphanius iberus). To this end, the lethal toxicity of dichlorvos based on 96h-LC50 bioassays was determined in saline water (50g/L), and in vivo effects of dichlorvos on cholinesterase (ChE) activity were investigated in adult female and male specimens. The 96h-LC50 value determined by probit analysis was 3.17mg/L (95% confidence limits: 1.34-3.97). The characterisation of the ChE using different substrates and specific inhibitors was also carried out in head and muscle tissues. Acetylthiocholine was the substrate preferred by both head and muscle ChE in males and females. Eserine sulphate and BW284C51 significantly inhibited both head and muscle enzyme activity at low concentrations (muM range), and iso-OMPA had no significant effect. These results indicate that in the head and muscle the predominant ChE form is acetylcholinesterase (AChE) for both sexes. The kinetic parameters for ChE activity (Km and Vmax) were similar in both sexes. The 96h-LC50 value obtained for adult specimens of Iberian toothcarp was 3.17mg/L. ChE activity in head and body tissues of both sexes was significantly inhibited in all concentrations tested (0.5, 1, 2 and 4mg/L) after "in vivo" dichlorvos exposure. However, Iberian toothcarp was able to tolerate high concentrations of dichlorvos, and resist high levels of brain and muscle ChE inhibition without mortality. Both ChE inhibition and recovery followed a similar time-course pattern in response to sub-lethal exposure to dichlorvos (1mg/L), and the enzyme activity did not return to control levels after 96h in clean water. The results of this study show that ChE activity is a good biomarker of exposure to OP in the Iberian toothcarp adults.
本研究评估了有机磷农药敌敌畏对濒危物种伊比利亚鳉(Aphanius iberus)的毒性作用。为此,在盐水(50g/L)中通过96小时半数致死浓度(LC50)生物测定法测定了敌敌畏的致死毒性,并在成年雌性和雄性样本中研究了敌敌畏对胆碱酯酶(ChE)活性的体内影响。通过概率分析确定的96小时LC50值为3.17mg/L(95%置信区间:1.34 - 3.97)。还对头和肌肉组织中使用不同底物和特异性抑制剂的ChE进行了表征。乙酰硫代胆碱是雄性和雌性头部及肌肉ChE均偏好的底物。硫酸毒扁豆碱和BW284C51在低浓度(微摩尔范围)时能显著抑制头部和肌肉的酶活性,而异丙基甲基膦酸酯无显著影响。这些结果表明,在头部和肌肉中,两性的主要ChE形式均为乙酰胆碱酯酶(AChE)。两性的ChE活性动力学参数(Km和Vmax)相似。伊比利亚鳉成年样本的96小时LC50值为3.17mg/L。在“体内”接触敌敌畏后,所有测试浓度(0.5、1、2和4mg/L)下,两性头部和身体组织中的ChE活性均受到显著抑制。然而,伊比利亚鳉能够耐受高浓度的敌敌畏,并在脑和肌肉ChE受到高水平抑制的情况下存活而不死亡。在亚致死剂量(1mg/L)接触敌敌畏后,ChE抑制和恢复遵循相似的时间进程模式,并且在清洁水中96小时后酶活性未恢复到对照水平。本研究结果表明,ChE活性是伊比利亚鳉成体接触有机磷农药的良好生物标志物。