Bees have a crucial role in pollination; therefore, it is important to determine the causes of their recent decline. Fipronil and imidacloprid are insecticides used worldwide to eliminate or control insect pests. Because they are broad-spectrum insecticides, they can also affect honeybees. Many researchers have studied the lethal and sublethal effects of these and other insecticides on honeybees, and some of these studies have demonstrated a correlation between the insecticides and colony collapse disorder in bees. The authors investigated the effects of fipronil and imidacloprid on the bioenergetic functioning of mitochondria isolated from the heads and thoraces of Africanized honeybees. Fipronil caused dose-dependent inhibition of adenosine 5'-diphosphate-stimulated (state 3) respiration in mitochondria energized by either pyruvate or succinate, albeit with different potentials, in thoracic mitochondria; inhibition was strongest when respiring with complex I substrate. Fipronil affected adenosine 5'-triphosphate (ATP) production in a dose-dependent manner in both tissues and substrates, though with different sensitivities. Imidacloprid also affected state-3 respiration in both the thorax and head, being more potent in head pyruvate-energized mitochondria; it also inhibited ATP production. Fipronil and imidacloprid had no effect on mitochondrial state-4 respiration. The authors concluded that fipronil and imidacloprid are inhibitors of mitochondrial bioenergetics, resulting in depleted ATP. This action can explain the toxicity of these compounds to honeybees.

The use of pesticides on crops contributes to the decline of bee populations, and in this sense, bioactive nutrients have been studied to counteract this effect. We suppose that caffeine might be one of these nutrients. We exposed honey bees (Apis mellifera L.) to 0.7 or 2.0 ng/mL imidacloprid, 5.0 μg/mL caffeine in syrup, or 5.0 μg/mL caffeine in syrup plus 0.7 or 2.0 ng/mL imidacloprid. After 72 h, the oxidative status and the food intake were verified. Imidacloprid increased glutathione peroxidase and catalase activities. Caffeine alone or with 2.0 ng/mL imidacloprid also stimulated the activity of glutathione peroxidase but did not alter the effect of the insecticide on the catalase activity. A significant reduction in the concentration of the thiol group of proteins was observed in the two imidacloprid-fed groups, and the addition of caffeine protected these groups. Imidacloprid increased the malondialdehyde concentration while the addition of caffeine partially decreased this effect. Food intake was higher for bees treated with 2.0 ng/mL imidacloprid. Our results show that imidacloprid increased the food intake resulting in oxidative damage, which was partially reversed by caffeine. From these findings, it is inferred that caffeine treatments can be used to mitigate the sublethal effects of this insecticide on honey bees.

Silkworm cocoon production has been reduced due to a number of problems other than those inherent in sericulture, such as diseases, malnutrition, and inappropriate management. The use of pesticides in areas surrounding mulberry fields can contaminate these plants and consequently harm caterpillars. The aim of this study was to evaluate whether the application of the fungicide pyraclostrobin in mulberry plants interferes with the mitochondrial bioenergetics and the productive performance of silkworms. Mulberry plants were treated with pyraclostrobin (0, 100, 200, and 300 g ha-1). After 30 d of fungicide application, fifth instar caterpillars were fed with leaves from the treated plants. We evaluated in vitro and in vivo mitochondrial bioenergetics of mitochondria from the head and intestines, as well as the feed intake and mortality rate of the caterpillars and the weight of fresh cocoons and cocoons shells. At doses of 50 µM (in vitro) and 200 g ha-1 (in vivo), pyraclostrobin inhibited oxygen consumption in state 3, dissipated membrane potential, and inhibited ATP synthesis in mitochondria. Pyraclostrobin acted as a respiratory chain inhibitor, affecting mitochondrial bioenergetics. The fungicide did not interfere with food consumption but negatively affected mortality rate and weight of cocoons. Mulberry leaves contaminated with pyraclostrobin negatively impact the mitochondrial bioenergetics of silkworms and cocoon production.