Toxicological studies have shown that the cockroach Periplaneta americana is a classical model for studying the mode of action of commonly used insecticides. In a previous study, we demonstrated that thiamethoxam and clothianidin decreased locomotor activity and decreased the locomotor activity in an open-field-like apparatus. Here, we tested the effect of the neonicotinoid acetamiprid when applied orally, topically, or injected into the haemolymph. We found that acetamiprid decreased locomotor activity in the open-field-like apparatus. When treated with acetamiprid, a strong alteration in locomotor activity was observed 1 h, 24 h and 48 h after haemolymph and topical applications. Oral application of acetamiprid demonstrated that it impaired locomotor activity at 24 h and 48 h. A comparison of the present data with our previously published results showed that neonicotinoids were more active when injected into the haemolymph compared to oral and topical applications. These findings increased our understanding of the effect of neonicotinoid insecticides on insect locomotor activity, and demonstrated that the cyano-substituted neonicotinoid, acetamiprid, was able to alter cockroach locomotor activity.

Neonicotinoids, such as dinotefuran (DF), have caused a variety of problems, such as massive loss and winter failure of the bee colony, as a price for the benefit of reducing farm work, because it continues to maintain a high insecticide activity over a long period of time. In this study, a field experiment was conducted for about six months to investigate the effects of DF on bee colonies damaged by Varroa mites. This study examined the long-term changes in such as the size of bee colonies, the intake of sugar syrup (SS), intake of pollen paste (PP), which is a vehicle for administering DF, the intake of DF, the mite-prevalence of bees and the inside and outside temperatures of hive-boxes. The variation width of the inner temperature of the hive-box is less than that of the ambient temperature (Ta). The inner temperature of the hive-box is adjusted with about 30 ℃ of Ta as the boundary. If Ta is lower than 30 ℃, the inner temperature of the box is higher than Ta, and if Ta is higher than 30 ℃, it is lower than Ta. The temperature variation width of the DF-exposed colony is greater than that of the control colony. The average intake of SS per bee per day of the DF-exposed colony is more than that of the control colony. The average intake of PP per bee per day of the DF-exposed colony is almost equal to that of the control colony. These results suggest that bees do not avoid DF, and ingest PP without distinction between toxic and pesticide-free. In the period from the start of DF administration to the colony extinction, the intake of DF per colony is about 865 µg/colony, the intake per bee is 14 ng/bee, and the intake per bee per day is less than 0.1 ng/bee/day in this work. These intakes are much lower than the previous ones (60-65 ng/bee, 0.27-2.32 ng/bee/day). These discrepancies may be because attacks of mites and Japanese giant hornets hastened the colony collapse. Seasonal changes in mite-prevalence of honeybees is approximately the same regardless of the bee colonies. At the end of August (the start of attacks by Japanese giant hornets), the mite-prevalence will increase rapidly. Even if the number of bees damaged by mites turns to decrease, the mite-prevalence will continue to increase, with approaching 100% before bee colonies become extinct. In this study, it was found that the bee colony was collapsed by the intake of a smaller amount of DF due to the synergistic effect of DF and mite-damage. To prevent a bee colony collapse, not only to make an effort to minimalize the adverse effect on the bee colony of neonicotinoids such as DF with long-term residual effect and high insecticide properties, it is necessary to reduce the damage of mites as much as possible, while considering the synergistically adverse effects of neonicotinoids and miticides.