Cytotoxic and genotoxic effects of a pendimethalin-based herbicide in Apis mellifera

Public concern about the effects of pesticides on non-target organisms has increased in the recent years. Nevertheless, there is a limited number of studies that address the actual toxic effects of herbicides on insects. This study investigated the side effects of herbicides on non-target organisms inhabiting agroecosystems and performing essential ecological and economic functions such as crop pollination. We analysed morphological alterations in the gut, Malpighian tubules and circulating haemocytes of Apis mellifera workers as markers of exposure effects. A commercial formulation of a pendimethalin-based herbicide (PND) was administered orally under laboratory conditions at a realistic concentration admitted in the field (330gL^-1 of active ingredient., 4 L ha^-1 for cereal and vegetable crops). The worker bees were exposed to a single application of PND for a period of one week, to simulate the exposure that can occur when foraging bees accidentally drink drops of contaminated water upon treatments. Histopathological analyses of the midgut, ileum and Malpighian tubules showed alterations over time (from 24 to 72 h after the beginning of exposure) such as loss of epithelial organisation, cellular vacuolisation and altered pyknotic nuclei as well as disruption of the peritrophic membrane over time. Semiquantitative analyses of the midgut showed a significant increase in the organ injury index 24 and 72 h after the initial exposure in PND-exposed bees compared to control bees. In addition, a change in positivity to Gram staining was observed in the midgut histological sections. A recovery of cytotoxic effects was observed one week after the initial exposure, which was favoured by the periodic renewal of the intestinal epithelium and the herbicide dissipation time. Cytochemical staining with Giemsa of haemocytes from PND-treated workers over 24 and 72 h showed significant nuclear alterations such as lobed or polymorphic nuclei and micronuclei compared to bees in the control group. These results show that the dose of PND used to protect crops from weeds can lead to significant cytotoxic and genotoxic effects in non-target organisms such as honey bees. In croplands, the sublethal effects on cell morphology can impair vital physiological processes such as nutrition, osmoregulation, and resistance to pathogens, contributing to the decline in biodiversity and abundance of species that play a prominent ecological role, such as pollinators.