Comparing Insect Predation by Birds and Insects in an Apple Orchard and Neighboring Unmanaged Habitat: Implications for Ecosystem Services.

Preserving ecosystem services, such as natural enemies that can provide pest control, can positively impact crops without compromising agricultural yield. Even though controlling pests by natural enemies has been suggested to reduce pests in agriculture, growers continue using conventional pesticides that kill beneficial predators. Here we studied whether the predation of avian and insect-beneficial predators varies in an apple orchard with conventional insecticide use compared to a bordering tree stand without insecticides. We studied the predation rates of mealworm pupae as a proxy to coddling moth pupae at 42 stations in both an apple orchard and a Eucalyptus stand at three distances (0 m, 50 m, and 100 m) from the border. Half of the stations were netted to prevent bird predation but were accessible to insects. The other half were non-netted and accessible to birds. We conducted six trials, each lasting two weeks, during which we recorded the predation of 504 stations with 5040 pupae. To validate which species predated the pupae, we added video cameras that took RGB videos during the day and IR videos at night in 45 stations and found that in net-free stations, birds preyed in 94.1% of stations in the orchard and 81.8% in the Eucalyptus stand. However, ants predated 70% of the pupae in stations with nets in the orchards and 100% in stations in the Eucalyptus strands. In addition, we found a significant rise in predation by birds as the distance into the orchard increased. Conversely, insect predation declined within the orchard but escalated in the adjacent unmanaged area. These findings suggest that the orchard's environment negatively affects beneficial insect activity, specifically predatory ants. This study demonstrates that birds can play an essential role in predating insect pests inside the orchard. In addition, we believe that the decreased predation of ants within the orchard was due to intense insecticide use.

Bumble Bee Workers Give Up Sleep to Care for Offspring that Are Not Their Own.

Sleep is ubiquitous in vertebrates and invertebrates, and its loss is typically associated with reduced performance, health, or survival, for reasons that are yet unclear [1-3]. Nevertheless, some animals can reduce sleep for increasing foraging time [4], under predation risk [5-8], during seasonal migration [9-11], or for having greater mating opportunities [12, 13]. Here, we tested the hypothesis that social bumble bee (Bombus terrestris) workers give up sleep for improving brood care. We combined video recordings, detailed behavioral analyses, sleep-deprivation experiments, and response-threshold assessments to characterize the sleep behavior of worker bees and showed that immobility bouts of ≥5 min provide a reliable proxy for sleep. We next used this index to study sleep with an automated video-based activity monitoring system. We found that isolated workers severely reduce sleep time in the presence of both larvae that need to be fed and pupae that do not. Reduced sleep was also correlated with around-the-clock activity and wax-pot building, which are typical for nest-founding mother queens. Cocoons, from which we removed the pupae, elicited a similar but transient sleep loss in tending workers, suggesting that the pupa effect on sleep is mediated by pheromonal signals. Sleep time increased following brood removal but remained lower compared to control bees, suggesting that the brood modulated sleep need. This first evidence for brood modulation of sleep in an insect suggests that plasticity in sleep can evolve as a mechanism to improve care for dependent juveniles, even in social insect workers that do not care for their own offspring.

Task-Related Phasing of Circadian Rhythms in Antennal Responsiveness to Odorants and Pheromones in Honeybees.

The insect antennae receive olfactory information from the environment. In some insects, it has been shown that antennal responsiveness is dynamically regulated by circadian clocks. However, it is unknown how general this phenomenon is and what functions it serves. Circadian regulation in honeybee workers is particularly interesting in this regard because they show natural task-related chronobiological plasticity. Forager bees show strong circadian rhythms in behavior and brain gene expression, whereas nurse bees tend brood around-the-clock and have attenuated circadian rhythms in activity and whole-brain gene expression. Here, we tested the hypothesis that there is task-related plasticity in circadian rhythms of antennal responsiveness to odorants in worker honeybees. We used electroantennogram (EAG) to measure the antennal responsiveness of nurses and foragers to general odorants and pheromones around the day. The capacity to track 10-Hz odorant pulses varied with time of day for both task groups but with different phases. The antennal pulse-tracking capacity was higher during the subjective day for the day-active foragers, whereas it was better during the night for around-the-clock active nurses. The task-related phases of pulse-tracking rhythms were similar for all the tested stimuli. We also found evidence for circadian rhythms in the EAG response magnitude of foragers but not of nurses. To the best of our knowledge, these results provide the first evidence for circadian regulation of antennal olfactory responsiveness and odorant pulse-tracking capacity in bees or any other hymenopteran insect. Importantly, our study shows for the first time that the circadian phase of olfactory responsiveness may be socially regulated.

Nurse honeybee workers tend capped brood, which does not require feeding, around the clock.

'Nurse' honeybees tend brood around the clock with attenuated or no circadian rhythms, but the brood signals inducing this behavior remain elusive. We first tested the hypothesis that worker circadian rhythms are regulated by brood pheromones. We monitored locomotor activity of individually isolated nurse bees that were exposed to either various doses of larval extract or synthetic brood ester pheromone (BEP). Bees orally treated with larval extract showed attenuated circadian rhythms in one of four tested colonies; a similar but statistically non-significant trend was seen in two additional colonies. Nurse bees treated with synthetic BEP showed rhythm attenuation in one of three tested colonies. Next, we tested the hypothesis that capped brood, which does not require feeding, nevertheless induces around-the-clock activity in nurses. By combining a new protocol that enables brood care by individually isolated nurse bees, detailed behavioral observations and automatic high-resolution monitoring of locomotor activity, we found that isolated nurses tended capped brood around the clock with attenuated circadian rhythms. Bees individually isolated in similar cages but without brood showed strong circadian rhythms in locomotor activity and rest. This study shows for the first time that the need to feed hungry larvae is not the only factor accounting for around-the-clock activity in nurse bees. Our results further suggest that the transition between activity with and without circadian rhythms is not a simple switch triggered by brood pheromones. Around-the-clock tending may enhance brood development and health in multiple ways that include improved larval feeding, thermoregulation or hygienic behavior.

Potent social synchronization can override photic entrainment of circadian rhythms.

Circadian rhythms in behaviour and physiology are important for animal health and survival. Studies with individually isolated animals in the laboratory have consistently emphasized the dominant role of light for the entrainment of circadian rhythms to relevant environmental cycles. Although in nature interactions with conspecifics are functionally significant, social signals are typically not considered important time-givers for the animal circadian clock. Our results challenge this view. By studying honeybees in an ecologically relevant context and using a massive data set, we demonstrate that social entrainment can be potent, may act without direct contact with other individuals and does not rely on gating the exposure to light. We show for the first time that social time cues stably entrain the clock, even in animals experiencing conflicting photic and social environmental cycles. These findings add to the growing appreciation for the importance of studying circadian rhythms in ecologically relevant contexts.

The involvement of the antennae in mediating the brood influence on circadian rhythms in "nurse" honey bee (Apis mellifera) workers.

Age-related division of labor in honey bees is associated with plasticity in circadian rhythms. Forager bees that are typically older than 3 weeks of age show strong behavioral and molecular circadian rhythms with higher activity during the day. Younger bees that typically care for ("nurse") the brood are active around the clock with similar brain clock gene levels throughout the day. However, nurses that are caged on brood-less combs inside or outside the hive show robust circadian rhythms with higher activity during the day, suggesting that direct contact with the brood mediates the plasticity in the circadian system. The nature of the brood signals affecting the workers' circadian system and the modalities by which they are detected are unknown. Given that the antennae are pivotal sensory organs in bees, we hypothesized that they are involved in mediating the brood influence on the plasticity in circadian rhythms. The flagella of the antennae are densely covered with diverse sensory structures able to detect a wide range of signals. To test our hypothesis, we removed the flagella of nurses and observed their behavior in isolation and in free-foraging colonies. We found that individually-isolated flagella-less bees under constant laboratory conditions show robust circadian rhythms in locomotor activity. In observation hives, flagella-less bees cared for the brood, but were more active during the day. By contrast, sham-treated bees were active around the clock as typical of nurses. Detailed video recordings showed that the brood-tending behavior of flagella-less and sham-treated bees is similar. These observations suggest that the difference in the patterns of brood care activity is not because the flagella-less bees did not contact the brood. Our results suggest that nurses are able to find the brood in the dark environment of the hive without their flagella, perhaps by using other sensory organs. The higher activity of flagella-less bees during the day further suggests that the flagella are involved in mediating the brood signals modulating plasticity in the circadian system.