Ozone exposure induces metabolic stress and olfactory memory disturbance in honey bees.

Human activities, urbanization, and industrialization contribute to pollution that affects climate and air quality. A main atmospheric pollutant, the tropospheric ozone (O3), can damage living organisms by generating oxidative radicals, causing respiratory problems in humans and reducing yields and growth in plants. Exposure to high concentrations of O3 can result in oxidative stress in plants and animals, eventually leading to substantial ecological consequences. Plants produce volatile organic compounds (VOCs) emitted in the environment and detected by pollinators (mainly by their antennae), foraging for nutritious resources. Several pollinators, including honey bees, recognize and discriminate flowers through olfactory cues and memory. Exposure to different concentrations of O3 was shown to alter the emission of floral VOCs by plants as well as their lifetime in the atmosphere, potentially impacting plant-pollinator interactions. In this report, we assessed the impacts of exposure to field-realistic concentrations of O3 on honey bees' antennal response to floral VOCs, on their olfactory recall and discriminative capacity and on their antioxidant responses. Antennal activity is altered depending on VOCs structure and O3 concentrations. During the behavioral tests, we first check consistency between olfactory learning rates and memory scores after 15 min. Then bees exposed to 120 and 200 ppb of ozone do not exert specific recall responses with rewarded VOCs 90 min after learning, compared to controls whose specific recall responses were consistent between time points. We also report for the first time in honey bees how the superoxide dismutase enzyme, an antioxidant defense against oxidative stress, saw its enzymatic activity rate decreases after exposure to 80 ppb of ozone. This work tends to demonstrate how hurtful can be the impact of air pollutants upon pollinators themselves and how this type of pollution needs to be addressed in future studies aiming at characterizing plant-insect interactions more accurately.

Acute ozone exposure impairs detection of floral odor, learning, and memory of honey bees, through olfactory generalization.

Air pollution stemming from human activities affects the environment in which plant and animal species live and interact. Similar to primary air pollutants which are emitted, secondary air pollutants, such as tropospheric ozone (O3) formed from nitrogen oxides, are also harmful to human health and plant physiology. Yet, few reports studied the effects of O3 on pollinators' physiology, despite that this pollutant, with its high oxidative potential, likely affects pollinators behaviors, especially the perception of signals they rely on to navigate their environment. Volatile Organic Compounds (VOCs) released by plants are used as signals by different animals. For pollination services, VOCs attract different insects to the flowers and strengthen these interactions. Here, we used the honey bee Apis mellifera as a model to characterize the effects of acute exposure to different realistic mixing ratios of O3 (80-, 120-, and 200-ppb) on two crucial aspects: first, how exposed honey bees detect VOCs; and second, how O3 affects these pollinators' learning and memory processes. With electroantennogram (EAG) recordings, we showed that increasing O3 mixing ratios had a biphasic effect: an initial 25% decrease of the antennal activity when bees were tested directly after exposure (O3 direct effect), followed by a 25% increase in activity and response when bees were allowed a two-hour rest after exposure (O3 delayed effect). In parallel, during olfactory conditioning, increasing O3 mixing ratios in both exposure protocols scarcely affected olfactory learning, followed by a decrease in recall of learned odors and an increase of response to new odors, leading to a higher generalization rate (i.e., discrimination impairment). These results suggest a link between O3-related oxidative stress and olfactory coding disturbance in the honey bee brain. If ozone affects the pollinators' olfaction, foraging behaviors may be modified, in addition with a possible long-term harmful effect on pollination services.