Stivaktakis Polychronis D, Kavvalakis Matthaios P, Tzatzarakis Manolis N, Alegakis Athanasios K, Panagiotakis Michael N, Fragkiadaki Persefoni, Vakonaki Elena, Ozcagli Eren, Hayes Wallace A, Rakitskii Valerii N, Tsatsakis Aristidis M
Center of Toxicology Science & Research, Division of Morphology, Medical School, University of Crete, Voutes Campus, Heraklion, 71003, Crete, Greece.
Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Istanbul University, 34116, Beyazit, Istanbul, Turkey.
Chemosphere. 2016 Apr;149:108-13. doi: 10.1016/j.chemosphere.2016.01.040. Epub 2016 Feb 5.
The present in-vivo study focuses on the genotoxic effect of the neonicotinoid pesticide imidacloprid (IMI) in rabbits. The purpose of the study was to establish a possible relationship between exposure to the pesticide (dose and duration) and genotoxicity. Furthermore, an analytical method for the simultaneous determination of IMI and its major metabolite 6-chloronicotinic acid (6-ClNA) in blood was developed and validated. The isolation of the two analytes from blood was performed by liquid-liquid extraction with dichloromethane. Analysis was performed by Liquid Chromatography - Atmospheric Pressure Chemical Ionization - Mass Spectrometry (LC-APCI-MS). The method was applied on the determination of IMI and 6-ClNA in serum samples obtained from rabbits fed with the insecticide at two low doses. Furthermore, parameters of genotoxicity and cytotoxicity were evaluated by measuring binucleated cells with micronuclei (BNMN), micronuclei (MN) and the Cytokinesis Block Proliferation Index (CBPI), in lymphocytes of exposed rabbits. The results revealed a genotoxic effect of IMI for both exposed groups. There were statistically significant differences in the frequencies of BNMN and MN between control and exposed groups but there was no dose-dependence, neither time-dependence of the genotoxic effect for the administered doses. This is the first time that long term exposure to IMI in rabbits was studied for the determination of its genotoxic effect. The genotoxic effect of IMI as it is depicted by the current study is in accordance with previous studies.
本体内研究聚焦于新烟碱类杀虫剂吡虫啉(IMI)对家兔的遗传毒性作用。该研究的目的是确定接触该杀虫剂(剂量和持续时间)与遗传毒性之间可能存在的关系。此外,还开发并验证了一种同时测定血液中IMI及其主要代谢物6-氯烟酸(6-ClNA)的分析方法。通过用二氯甲烷进行液-液萃取从血液中分离出这两种分析物。采用液相色谱-大气压化学电离-质谱联用(LC-APCI-MS)进行分析。该方法应用于测定从以两种低剂量喂食该杀虫剂的家兔获得的血清样本中的IMI和6-ClNA。此外,通过测量暴露家兔淋巴细胞中的含微核双核细胞(BNMN)、微核(MN)和胞质分裂阻滞增殖指数(CBPI)来评估遗传毒性和细胞毒性参数。结果显示,两个暴露组均出现了IMI的遗传毒性作用。对照组和暴露组之间BNMN和MN的频率存在统计学上的显著差异,但遗传毒性作用对于所施用剂量既不存在剂量依赖性,也不存在时间依赖性。这是首次对家兔长期接触IMI以确定其遗传毒性作用进行研究。当前研究所描述的IMI的遗传毒性作用与先前的研究一致。