Melatonin enhances the antioxidant capacity to rescue the honey bee Apis mellifera from the ecotoxicological effects caused by environmental imidacloprid

Abstract

Imidacloprid severely poisons the nontarget insect honey bee Apis mellifera. Few treatments are available to mitigate the adverse effects of imidacloprid. The primary concern is that the molecular understanding of imidacloprid toxicity is not comprehensive enough. Oxidative stress is the primary pathophysiological mechanism by which pesticides cause high mortality. Our pilot study found for the first time that imidacloprid stimulates bee brains to secrete melatonin, a free radical scavenger. However, the molecular basis for imidacloprid toxicity and the role of melatonin in coping with imidacloprid have not been systematically investigated in bees. This study administered an environmental dose of imidacloprid (36 ng/bee) orally to A. mellifera. The detoxification gene cytochrome P450 CYP4G11 was significantly induced. However, potent cytotoxicity of imidacloprid suppressed the expression of the antioxidants catalase (CAT) and thioredoxin reductase (TrxR), and the activity of guaiacol peroxidase (GPX), superoxide dismutase (SOD), and reduced glutathione (GSH) was not induced. The levels of reactive oxygen species (ROS) and the lipid peroxidation marker malondialdehyde (MDA) were increased. The expression of the apoptotic genes cysteinyl aspartate specific proteinase (Caspase-3) and apoptosis inducing factor (AIF) increased, and the apoptotic features of midgut cells were prominently apparent. These results suggest that imidacloprid disrupts the bee antioxidant system, causing severe oxidative stress and tissue damage and ultimately leading to apoptosis. Significantly, however, imidacloprid exposure also stimulated bee brains to continuously secrete melatonin. Further preadministration of exogenous melatonin (200 ng/bee) orally to bees significantly reversed and enhanced the activity of the imidacloprid-suppressed antioxidants CAT, SOD, and GSH, which allowed imidacloprid-induced ROS accumulation to be effectively alleviated. The MDA content, apoptotic genes Caspase-3 and AIF, and detoxification gene CYPG411 expression were restored to normalization; midgut cell damage, apoptosis, and mortality were significantly reduced. These findings strongly suggest that melatonin enhanced bee antioxidant capacity, thus attenuating oxidative stress and apoptosis to confer imidacloprid tolerance to honey bees. Melatonin secretion may be a defense mechanism to mitigate imidacloprid toxicity.