A micronucleus assay detects genotoxic effects of herbicide exposure in a protected butterfly species

Lycaena disparHawort (Lepidoptera: Lycaenidae), a protected butterfly, is declining in Europe, but it thrives in rice fields in northern Italy. Here, agrochemical usage could threaten its long-term survival. We investigated, by micronucleus (MN) assay, the genotoxic effect of glyphosate, a common herbicide, onL. disparlarvae. Micronuclei (MNi) are DNA fragments separated from the main nucleus and represent the result of genomic damage that has been transmitted to daughter cells. In a control/treatment experiment, we extracted epithelial cells from last-instar larvae fed withRumexspp. plants sprayed with a solution containing 3.6 g/L of glyphosate, and from larvae fed with unsprayed plants. MNi and other chromosomal aberrations-nuclear buds (NBUDs) and bi-nucleated cells-were then scored in 1000 cells/subject. Significant differences were found between glyphosate-exposed and control groups in terms of MNi and total genomic damage, but not in terms of NBUDs or bi-nucleated cells. We reported a possible genomic damage induced by glyphosate on larvae ofL. dispar. For the first time, a MN assay was used in order to evaluate the genomic damage on a phytophagous invertebrate at the larval stage. Increased levels of MNi reflect a condition of genomic instability that can result in reduced vitality and in an increased risk of local extinction. Therefore, farmland management compatible with wildlife conservation is needed.