Huska Dalibor, Krizkova Sona, Beklova Miroslava, Havel Ladislav, Zehnalek Josef, Diopan Vaclav, Adam Vojtech, Zeman Ladislav, Babula Petr, Kizek Rene
Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic.
Department of Veterinary Ecology and Environmental Protection, Faculty of Veterinary Hygiene and Ecology,Palackeho 1-3, CZ-612 42 Brno, Czech Republic.
Sensors (Basel). 2008 Feb 19;8(2):1039-1047. doi: 10.3390/s8021039.
Metallothioneins belong to a group of intracellular, high molecular andcysteine-rich proteins whose content in an organism increase with increasing concentrationof a heavy metal. The aim of this work was to apply the electrochemical analysis for theanalysis of metallothioneins in earthworms exposed to cadmium ions and brewery sludge.Here we utilized adsorptive transfer technique coupled with differential pulse voltammetryBrdicka reaction to determine metallothionein in different biological samples. By meansthis very sensitive technique it was possible to analyze metallothionein in concentrationsbelow 1 μmol.l⁻1 with the standard deviation of 4-5%. We found out that the average MTlevel in the non-treated earthworms oscillated between 19 and 48 μmol.l. When weanalysed samples of earthworms treated by cadmium, we observed that the MT contentincreased with the exposition length and increase dose of cadmium ions. Finally, weattempted to study and compare the toxicity of the raw sludge and its leach by using ofearthworms. The raw brewery sludge caused the death of the earthworms quickly.Earthworms held in the presence of leach from brewery sludge increased their weight of147 % of their original weight because they ingested the nutrients from the sludge. Themetallothionein level changes markedly with increasing time of exposition and applieddose of toxic compound. It clearly follows from the obtained results that the MT synthesisis insufficient in the first hours of the exposition and increases after more than 24 h.
金属硫蛋白属于一类细胞内、高分子量且富含半胱氨酸的蛋白质,其在生物体中的含量会随着重金属浓度的增加而升高。本研究旨在应用电化学分析方法,对暴露于镉离子和啤酒厂污泥中的蚯蚓体内的金属硫蛋白进行分析。在此,我们采用吸附转移技术结合差分脉冲伏安法的布氏反应来测定不同生物样品中的金属硫蛋白。通过这种非常灵敏的技术,可以分析浓度低于1μmol·L⁻¹的金属硫蛋白,标准偏差为4 - 5%。我们发现,未处理蚯蚓体内的金属硫蛋白平均水平在19至48μmol·L之间波动。当我们分析镉处理过的蚯蚓样本时,观察到金属硫蛋白的含量随着镉离子暴露时间的延长和剂量的增加而增加。最后,我们试图通过蚯蚓来研究和比较原生污泥及其浸出液的毒性。原生啤酒厂污泥会迅速导致蚯蚓死亡。置于啤酒厂污泥浸出液中的蚯蚓体重增加了其原始体重的147%,因为它们从污泥中摄取了营养物质。金属硫蛋白水平会随着暴露时间的延长和有毒化合物施用剂量的增加而显著变化。从获得的结果可以明显看出,在暴露的最初几个小时内金属硫蛋白的合成不足,而在超过24小时后会增加。