Pesticide use is one of the main causes of pollinator declines in agricultural ecosystems. Traditionally, most laboratory studies on bee ecotoxicology test acute exposure to single compounds. However, under field conditions, bees are often chronically exposed to a variety of chemicals, with potential synergistic effects. We studied the effects of field-realistic concentrations of three pesticides measured in pollen and nectar of commercial melon fields on the solitary beeOsmia bicornisL. We orally exposed females of this species throughout their life span to 8 treatments combining two neonicotinoid insecticides (acetamiprid, imidacloprid) and a triazole fungicide (myclobutanil) via pollen and sugar syrup. We measured pollen and syrup consumption, longevity, ovary maturation and thermogenesis. Although bees consumed larger amounts of syrup than pollen, pesticide intake via syrup and pollen were similar. At the tested concentrations, no synergistic effects emerged, and we found no effects on longevity and ovary maturation. However, all treatments containing imidacloprid resulted in suppressed syrup consumption and drastic decreases in thoracic temperature and bee activity. Our results have important implications for pesticide regulation. If we had measured only lethal effects we would have wrongly concluded that the pesticide combinations containing imidacloprid were safe toO. bicornis. The incorporation of tests specifically intended to detect sublethal effects in bee risk assessment schemes should be an urgent priority. In this way, the effects of pesticide exposure on the dynamics of bee populations in agroecosystems will be better assessed.

Bee populations are exposed to multiple stressors, including land‐use change, biological invasions, climate change, and pesticide exposure, that may interact synergistically. We analyze the combined effects of climate warming and sublethal insecticide exposure in the solitary bee Osmia cornuta. Previous Osmia studies show that warm wintering temperatures cause body weight loss, lipid consumption, and fat body depletion. Because the fat body plays a key role in xenobiotic detoxification, we expected that bees exposed to climate warming scenarios would be more sensitive to pesticides. We exposed O. cornuta females to three wintering treatments: current scenario (2007–2012 temperatures), near‐future (2021–2050 projected temperatures), and distant‐future (2051–2080). Upon emergence in spring, bees were orally exposed to three sublethal doses of an insecticide (Closer, a.i. sulfoxaflor; 0, 4.55 and 11.64 ng a.i./bee). We measured the combined effects of wintering and insecticide exposure on phototactic response, syrup consumption, and longevity. Wintering treatment by itself did not affect winter mortality, but body weight loss increased with increasing wintering temperatures. Similarly, wintering treatment by itself hardly influenced phototactic response or syrup consumption. However, bees wintered at the warmest temperatures had shorter longevity, a strong fecundity predictor in Osmia. Insecticide exposure, especially at the high dose, impaired the ability of bees to respond to light, and resulted in reduced syrup consumption and longevity. The combination of the warmest winter and the high insecticide dose resulted in a 70% longevity decrease. Smaller bees, resulting from smaller pollen–nectar provisions, had shorter longevity suggesting nutritional stress may further compromise fecundity in O. cornuta. Our results show a synergistic interaction between two major drivers of bee declines, and indicate that bees will become more sensitive to pesticides under the current global warming scenario. Our findings have important implications for pesticide regulation and underscore the need to consider multiple stressors to understand bee declines.

Planting flower strips adjacent to crops is among the habitat-management practices employed to offer alternative floral resources to pollinators. However, more information is needed to understand their potential spill-over of pollinators on nearby insect-pollinated crops. Over the course of two consecutive years, the suitability of a flower mixture of 10 herbaceous plants for pollinators was evaluated on a weekly basis, in a randomized block design of two melon plots (10 × 10 m²) with or without 1 m-wide flower strips. Floral coverage and pollinator visits to the plant species, as well as pollinator visits and the yield and quality of the crop, were assessed. Additionally, the selected mixture was tested for 1 year in a commercial field in order to ascertain how far the flower strip could influence visitors in the crop. The most suitable species for a flower strip in central Spain based on their attractiveness, floral coverage and staggered blossom were Coriandrum sativum L., Diplotaxis virgata L., Borago officinalis L. and Calendula officinalis L. The flower strip can act as either pollinator competitor or facilitator to the crop, depending on their floral coverage and/or the predominant species during the crop bloom period. The concurrence of blooming of the rewarding plant C. officinalis with the melon crop should be avoided in our area. In the commercial field, the bee visitation rate in the melon flowers decreased with the distance to the flower strip. No influence of the specific flower strip evaluated on crop productivity or quality was found.

Residues detected in pollen collected by honey bees are often used to estimate pesticide exposure in ecotoxicological studies. However, for a more accurate assessment of pesticides effect on foraging pollinators, residues found directly on flowers are a more realistic exposure approximation. We conducted a multi-residue analysis of pesticides on pollen and nectar of melon flowers collected from five fields. The cumulative chronic oral exposure Risk Index (RI) was calculated for Apis mellifera, Bombus terrestris and Osmia bicornis to multiple pesticides. However, this index could underestimate the risk since sublethal or synergistic effects are not considered. Therefore, a mixture containing three of the most frequently detected pesticides in our study was tested for synergistic impact on B. terrestris micro-colonies through a chronic oral toxicity test. According to the result, pollen and nectar samples contained numerous pesticide residues, including nine insecticides, nine fungicides, and one herbicide. Eleven of those were not applied by farmers during the crop season, revealing that melon agroecosystems may be pesticide contaminated environments. The primary contributor to the chronic RI was imidacloprid and O. bircornis is at greatest risk for lethality resulting from chronic oral exposure at these sites. In the bumblebee micro-colony bioassay, dietary exposure to acetamiprid, chlorpyrifos and oxamyl at residue level concentration, showed no effects on worker mortality, drone production or drone size and no synergies were detected when pesticide mixtures were evaluated. In conclusion, our findings have significant implications for improving pesticide risk assessment schemes to guarantee pollinator conservation. In particular, bee pesticide risk assessment should not be limited to acute exposure effects to isolated active ingredients in honey bees. Instead, risk assessments should consider the long-term pesticide exposure effects in both pollen and nectar on a range of bees that reflect the diversity of natural ecosystems and the synergistic potential among pesticide formulations.

In agricultural environments, bees are routinely exposed to combinations of pesticides. For the most part, exposure to these pesticide mixtures does not result in acute lethal effects, but we know very little about potential sublethal effects and their consequences on reproductive success and population dynamics. In this study, we orally exposed newly emerged females of the solitary bee Osmia cornuta to environmentally-relevant levels of acetamiprid (a cyano-substituted neonicotinoid insecticide) singly and in combination with tebuconazole (a sterol-biosynthesis inhibitor (SBI) fungicide). The amount of feeding solution consumed during the exposure phase was lowest in bees exposed to the pesticide mixture. Following exposure, females were individually marked and released into oilseed rape field cages to monitor their nesting performance and assess their reproductive success. The nesting performance and reproductive success of bees exposed to the fungicide or the insecticide alone were similar to those of control bees and resulted in a 1.3-1.7 net population increases. By contrast, bees exposed to the pesticide mixture showed lower establishment, shortened nesting period, and reduced fecundity. Together, these effects led to a 0.5-0.6 population decrease. Female establishment and shortened nesting period were the main population bottlenecks. We found no effects of the pesticide mixture on nest provisioning rate, offspring body weight or sex ratio. Our study shows how sublethal pesticide exposure may affect several components of bee reproductive success and, ultimately, population growth. Our results calls for a rethinking of pollinator risk assessment schemes, which should target not only single compounds but also combinations of compounds likely to co-occur in agricultural environments.

The recent EU ban of the three most widely used neonicotinoids (imidacloprid, thiamethoxam and clothianidin) to all outdoors applications has stimulated the introduction of new insecticides into the market. Sulfoxaflor is a new systemic insecticide that, like neonicotinoids, acts as a modulator of nicotinic acetylcholine receptors. In agro-environments, bees can be exposed to this compound via contaminated pollen and nectar for long periods of time. Therefore, it is important to assess the potential effects of chronic exposure to sulfoxaflor, alone and in combination with fungicides, on pollinators. In this study, we tested the effects of chronic exposure to two field concentrations of sulfoxaflor (20 and 100 ppb) alone and in combination with four concentrations of the fungicide fluxapyroxad (7500, 15,000, 30,000 and 60,000 ppb) on syrup consumption and longevity in females of the solitary bee Osmia bicornis L. Exposure to 20 ppb of sulfoxaflor, alone and in combination with the fungicide, stimulated syrup consumption, but did not affect longevity. In contrast, syrup consumption decreased in bees exposed to 100 ppb, all of which died after 2-6 days of exposure. We found no evidence of synergism between the two compounds at any of the two sulfoxaflor concentrations tested. Comparison of our findings with the literature, confirms that O. bicornis is more sensitive to sulfoxaflor than honey bees. Our results highlight the need to include different bee species in risk assessment schemes.