Catae Aline Fernanda, Roat Thaisa Cristina, Pratavieira Marcel, Silva Menegasso Anally Ribeiro da, Palma Mario Sergio, Malaspina Osmar
Departamento de Biologia, Centro de Estudos de Insetos Sociais, UNESP - Univ. Estadual Paulista, campus de Rio Claro, Instituto de Biociências, Rio Claro, SP, Brazil.
Ecotoxicology. 2018 Mar;27(2):109-121. doi: 10.1007/s10646-017-1874-4. Epub 2017 Nov 10.
The use of insecticides has become increasingly frequent, and studies indicate that these compounds are involved in the intoxication of bees. Imidacloprid is a widely used neonicotinoid; thus, we have highlighted the importance of assessing its oral toxicity to Africanized bees and used transmission electron microscopy to investigate the sublethal effects in the brain, the target organ, and the midgut, responsible for the digestion/absorption of food. In addition, the distribution of proteins involved in important biological processes in the brain were evaluated on the 1st day of exposure by MALDI-imaging analysis. Bioassays were performed to determine the Median Lethal Concentration (LC) of imidacloprid to bees, and the value obtained was 1.4651 ng imidacloprid/μL diet. Based on this result, the sublethal concentration to be administered at 1, 4 and 8 days was established as a hundredth (1/100) of the LC. The results obtained from the ultrastructural analysis showed alterations in the midgut cells of bees as nuclear and mitochondrial damage and an increase of vacuoles. The insecticide caused spacing among the Kenyon cells in the mushroom bodies, chromatin condensation and loss of mitochondrial cristae. The MALDI-imaging analysis showed an increase in the expression of such proteins as vascular endothelial growth factor receptor, amyloid protein precursor and protein kinase C, which are related to oxygen supply, neuronal degeneration and memory/learning, and a decrease in the expression of the nicotinic acetylcholine receptor alpha 1, which is fundamental to the synapses. These alterations demonstrated that imidacloprid could compromise the viability of the midgut epithelium, as well as inhibiting important cognitive processes in individuals, and may be reflected in losses of the colony.
杀虫剂的使用越来越频繁,研究表明这些化合物与蜜蜂中毒有关。吡虫啉是一种广泛使用的新烟碱类杀虫剂;因此,我们强调了评估其对非洲化蜜蜂口服毒性的重要性,并使用透射电子显微镜研究其对大脑(靶器官)和中肠(负责食物消化/吸收)的亚致死效应。此外,在暴露的第1天通过基质辅助激光解吸电离成像分析评估大脑中参与重要生物学过程的蛋白质分布。进行生物测定以确定吡虫啉对蜜蜂的半数致死浓度(LC),得到的值为1.4651 ng吡虫啉/μL饲料。基于此结果,将在第1、4和8天施用的亚致死浓度确定为LC的百分之一(1/100)。超微结构分析结果显示,蜜蜂中肠细胞出现核损伤和线粒体损伤以及液泡增多等变化。该杀虫剂导致蘑菇体中的肯扬细胞之间出现间隙、染色质浓缩和线粒体嵴丢失。基质辅助激光解吸电离成像分析显示,与氧气供应、神经元变性和记忆/学习相关的血管内皮生长因子受体、淀粉样蛋白前体和蛋白激酶C等蛋白质的表达增加,而对突触至关重要的烟碱型乙酰胆碱受体α1的表达减少。这些变化表明,吡虫啉可能会损害中肠上皮的活力,以及抑制个体的重要认知过程,并可能反映在蜂群的损失上。