Bees display limited acclimation capacity for heat tolerance.

Bees are essential pollinators and understanding their ability to cope with extreme temperature changes is crucial for predicting their resilience to climate change, but studies are limited. We measured the response of the critical thermal maximum (CTMax) to short-term acclimation in foragers of six bee species from the Greek island of Lesvos, which differ in body size, nesting habit, and level of sociality. We calculated the acclimation response ratio as a metric to assess acclimation capacity and tested whether bees' acclimation capacity was influenced by body size and/or CTMax. We also assessed whether CTMax increases following acute heat exposure simulating a heat wave. Average estimate of CTMax varied among species and increased with body size but did not significantly shift in response to acclimation treatment except in the sweat bee Lasioglossum malachurum. Acclimation capacity averaged 9% among species and it was not significantly associated with body size or CTMax. Similarly, the average CTMax did not increase following acute heat exposure. These results indicate that bees might have limited capacity to enhance heat tolerance via acclimation or in response to prior heat exposure, rendering them physiologically sensitive to rapid temperature changes during extreme weather events. These findings reinforce the idea that insects, like other ectotherms, generally express weak plasticity in CTMax, underscoring the critical role of behavioral thermoregulation for avoidance of extreme temperatures. Conserving and restoring native vegetation can provide bees temporary thermal refuges during extreme weather events.

Acute exposure to sublethal doses of neonicotinoid insecticides increases heat tolerance in honey bees.

The European honey bee, Apis mellifera L., is the single most valuable managed pollinator in the world. Poor colony health or unusually high colony losses of managed honey bees result from a myriad of stressors, which are more harmful in combination. Climate change is expected to accentuate the effects of these stressors, but the physiological and behavioral responses of honey bees to elevated temperatures while under simultaneous influence of one or more stressors remain largely unknown. Here we test the hypothesis that exposure to acute, sublethal doses of neonicotinoid insecticides reduce thermal tolerance in honey bees. We administered to bees oral doses of imidacloprid and acetamiprid at 1/5, 1/20, and 1/100 of LD50 and measured their heat tolerance 4 h post-feeding, using both dynamic and static protocols. Contrary to our expectations, acute exposure to sublethal doses of both insecticides resulted in higher thermal tolerance and greater survival rates of bees. Bees that ingested the higher doses of insecticides displayed a critical thermal maximum from 2 ˚C to 5 ˚C greater than that of the control group, and 67%-87% reduction in mortality. Our study suggests a resilience of honey bees to high temperatures when other stressors are present, which is consistent with studies in other insects. We discuss the implications of these results and hypothesize that this compensatory effect is likely due to induction of heat shock proteins by the insecticides, which provides temporary protection from elevated temperatures.

Sublethal imidacloprid effects on honey bee flower choices when foraging.

Neonicotinoids, systemic neuro-active pesticides similar to nicotine, are widely used in agriculture and are being investigated for a role in honey bee colony losses. We examined one neonicotinoid pesticide, imidacloprid, for its effects on the foraging behavior of free-flying honey bees (Apis mellifera anatoliaca) visiting artificial blue and white flowers. Imidacloprid doses, ranging from 1/5 to 1/50 of the reported LD50, were fed to bees orally. The study consisted of three experimental parts performed sequentially without interruption. In Part 1, both flower colors contained a 4 µL 1 M sucrose solution reward. Part 2 offered bees 4 µL of 1.5 M sucrose solution in blue flowers and a 4 µL 0.5 M sucrose solution reward in white flowers. In Part 3 we reversed the sugar solution rewards, while keeping the flower color consistent. Each experiment began 30 min after administration of the pesticide. We recorded the percentage of experimental bees that returned to forage after treatment. We also recorded the visitation rate, number of flowers visited, and floral reward choices of the bees that foraged after treatment. The forager return rate declined linearly with increasing imidacloprid dose. The number of foraging trips by returning bees was also affected adversely. However, flower fidelity was not affected by imidacloprid dose. Foragers visited both blue and white flowers extensively in Part 1, and showed greater fidelity for the flower color offering the higher sugar solution reward in Parts 2 and 3. Although larger samples sizes are needed, our study suggests that imidacloprid may not affect the ability to select the higher nectar reward when rewards were reversed. We observed acute, mild effects on foraging by honey bees, so mild that storage of imidacloprid tainted-honey is very plausible and likely to be found in honey bee colonies.

Feature-positive and feature-negative learning in honey bees.

Honey bees (Apis mellifera anatolica) were subjected to sequential trials where they were given the choice between a feature-positive and a feature-negative feeding plate. The 'feature' being manipulated is the presence of a single blue circle among three circles marking the location of a small sucrose reward. That is, a 'feature-negative' target had three white circles, while a 'feature-positive' target had two white circles and one blue one. Two experiments were performed. In both experiments, each bee was tested under two different reward scenarios (treatments). In the first experiment, during the feature-positive treatment bees received 4 µl of 2 mol l(-1) sucrose when choosing the feature-positive plate, but received 4 µl of saturated NaCl solution (saltwater) when choosing the feature-negative plate. During the feature-negative treatment, bees were rewarded when visiting the feature-negative plate, while visitation to the feature-positive plate only offered bees the saltwater. The second experiment was a repeat of the first except that pure water was offered instead of saltwater in the non-rewarding feeding plate. As an experimental control, a set of bees was offered sequential trials where both the feature-positive and feature-negative plates offered the sucrose reward. Bee feeding plate choice differed between the feature-positive and feature-negative treatments in both experiments. Bees favored the feeding plate type with the sucrose reward in each treatment, and never consumed the saltwater or pure water when encountered in either treatment. Further, behavior of bees during both the feature-positive and feature-negative treatments differed from that of control bees. However, neither feature-positive nor feature-negative learning reached high levels of success. Further, a feature-positive effect was seen when pure water was offered; bees learned to solve the feature-positive problem more rapidly. When we tested bees using simply the choice of blue versus white targets, where one color held the sucrose reward and the other the saltwater, a bee's fidelity to the color offering the sucrose reward quickly reached very high levels.

Ethanol increases HSP70 concentrations in honeybee (Apis mellifera L.) brain tissue.

Previous research on the honeybee ethanol model established how acute ethanol exposure altered function at different levels of organization: behavior and learning, ecology, and physiology. The purpose of this study was to evaluate whether ethanol doses that affect honeybee behavior also induce a significant stress response, measured by heat shock protein 70 (HSP70) concentrations, in honeybee brain tissues. Experiment 1 examined how pretreatment handling influenced brain HSP70 concentrations in three pretreatment groups of bees; immediately after being collected, after being harnessed and fed, and after 22-24h in a harness. HSP70 concentrations did not differ among pretreatment groups within replicates, although we observed significantly different HSP70 concentrations between the two replicates. Experiment 2 investigated the relationship between ethanol dose and brain HSP70 concentrations. Bees were placed in seven experimental groups, the three pretreatment groups as in Experiment 1 and four ethanol-fed groups. Bees in ethanol treatments were fed 1.5M sucrose (control) and 1.5M sucrose-ethanol solutions containing 2.5, 5, and 10% ethanol, allowed to sit for 4h, and dissected brains were assayed for HSP70. We observed ethanol-induced increases in honeybee brain HSP70 concentrations from the control group through the 5% ethanol group. Only bees in the 5% ethanol group had HSP70 concentrations significantly higher than the control group. The inverted U-shaped ethanol dose-HSP70 concentration response curve indicated that ingestion of 2.5% ethanol and 5% ethanol stimulated the stress response, whereas ingestion of 10% ethanol inhibited the stress response. Doses that show maximum HSP70 concentration (5% ethanol) or HSP70 inhibition (10% ethanol) correspond to those (> or =5% ethanol) that also impaired honeybees in previous studies. We conclude that acute ethanol intoxication by solutions containing > or =5% ethanol causes significant ethanol-induced stress in brain tissue that impairs honeybee behavior and associative learning.

Nest site selection influences mortality and stress responses in developmental stages of Megachile apicalis Spinola (Hymenoptera: Megachilidae).

We examined stress responses and survival in developmental stages of the invasive solitary bee Megachile apicalis Spinola during two nesting seasons in the Central Valley of California to consider whether abiotic stress tolerance of its offspring contributes to this species' successful colonization of the western United States. In 2001 and 2003, artificial nesting cavities were affixed to vertical plywood boards oriented to maximize nest cavity temperature and humidity differences: one side faced south (exposed to direct sun) and the other one faced north (shaded). After several weeks of nesting activity, we measured heat shock protein 70 (HSP70) concentrations in adults and offspring on 1 d in both years and offspring survival and mortality sources in 2003. In 2001, M. apicalis showed higher HSP70 concentrations in exposed nests than in shaded nests during all developmental stages, adults and their offspring. In 2003, overall survivorship was not significantly different between treatments because exposed nests experienced high offspring mortality caused by heat stress, whereas shaded nests suffered similarly high offspring mortality because of parasitoids. In both years of our study, females preferred shaded nests over exposed nests. M. apicalis successfully reproduces in grasslands of the Central Valley of California where offspring survive hot, dry nest sites and parasitoids in sufficient numbers to inoculate new grassland habitats, unpopulated by tolerance-limited native solitary bees, with incipient populations of this bee, M. apicalis.