N. americanus mortality.

Among the most immediate drivers of American burying beetle (Nicrophorus americanus Olivier) declines, nontarget toxicity to pesticides is poorly understood. Acute, episodic exposure to neonicotinoid insecticides at environmentally relevant concentrations is linked to negative impacts on beneficial terrestrial insect taxa. Beyond mortality, behavioral indicators of toxicity are often better suited to assess sublethal effects of residual concentrations in the environment. First, Nicrophorus spp. congeners were used to generate and identify a low-dose exposure rate (lethal dose 10%; LD10) from an acute, 24-hour exposure and the concentration-series was confirmed by LC–MS/MS. Next, we evaluated the effects of single and repeated low-dose (LD10 = 58.9 ng/beetle) imidacloprid exposure on N. americanus behavior (10 minutes post-dose) and mortality (10 days post-dose). Behavior parameters were analyzed using EthoVision-XT. Control N. americanus were significantly less mobile, demonstrating death-feigning, an anti-predator behavior. Single LD10 dosed N. americanus were hyperactive, traveling over 4 times farther (total distance; p = 0.03) and faster (mean velocity; p = 0.02) than controls. Single and repeated LD10 dosed N. americanus extended their wings without taking flight and flipped on their backs. All control N. americanus survived 10 days post-dose; single LD10 and repeated LD10 exhibited 30% and 50% mortality, respectively. A single LD10 exposure event was sufficient to significantly elicit greater movement and high predation risk behaviors, whereas repeated LD10 exposure did not worsen behavioral impairment but increased mortality over time. Collectively, generalized linear mixed effects models indicated that distance traveled, velocity, and extended wings were significant predictors of mortality. Recently reclassified, the federally threatened N. americanus may be at greater risk to insecticide exposure than previously thought and vulnerable to episodic, low-dose neonicotinoid exposure.