Multimodal processing of noisy cues in bumblebees.

Multimodal cues can improve behavioral responses by enhancing the detection and localization of sensory cues and reducing response times. Across species, studies have shown that multisensory integration of visual and olfactory cues can improve response accuracy. However, in real-world settings, sensory cues are often noisy; visual and olfactory cues can be deteriorated, masked, or mixed, making the target cue less clear to the receiver. In this study, we use an associative learning paradigm (Free Moving Proboscis Extension Reflex, FMPER) to show that having multimodal cues may improve the accuracy of bees' responses to noisy cues. Adding a noisy visual cue improves the accuracy of response to a noisy olfactory cue, despite neither the clear nor noisy visual cue being sufficient when paired with a novel olfactory cue. This may provide insight into the neural mechanisms underlying multimodal processing and the effects of environmental change on pollination services.

Flying, nectar-loaded honey bees conserve water and improve heat tolerance by reducing wingbeat frequency and metabolic heat production.

Heat waves are becoming increasingly common due to climate change, making it crucial to identify and understand the capacities for insect pollinators, such as honey bees, to avoid overheating. We examined the effects of hot, dry air temperatures on the physiological and behavioral mechanisms that honey bees use to fly when carrying nectar loads, to assess how foraging is limited by overheating or desiccation. We found that flight muscle temperatures increased linearly with load mass at air temperatures of 20 or 30 °C, but, remarkably, there was no change with increasing nectar loads at an air temperature of 40 °C. Flying, nectar-loaded bees were able to avoid overheating at 40 °C by reducing their flight metabolic rates and increasing evaporative cooling. At high body temperatures, bees apparently increase flight efficiency by lowering their wingbeat frequency and increasing stroke amplitude to compensate, reducing the need for evaporative cooling. However, even with reductions in metabolic heat production, desiccation likely limits foraging at temperatures well below bees' critical thermal maxima in hot, dry conditions.

Autonomous Expansion of Grasshopper Wings Reveals External Forces Contribute to Final Adult Wing Shape.

Ecdysis, transformation from juvenile to adult form in insects, is time-consuming and leaves insects vulnerable to predation. For winged insects, the process of wing expansion during ecdysis, unfurling and expanding the wings, is a critical bottleneck in achieving sexual maturity. Internal and external forces play a role in wing expansion. Vigorous abdominal pumping during wing expansion allows insects to pressurize and inflate their wings, filling them with hemolymph. In addition, many insects adopt expansion-specific postures and, if inhibited, do not expand their wings normally, suggesting that external forces such as gravity may play a role. However, two previous studies over 40 years ago, reported that the forewings of swarming locusts can expand autonomously when removed from the emerging insect and laid flat on a saline solution. Termed "autoexpansion," we replicated previous experiments of autoexpansion on flat liquid media, documenting changes in both wing length and area over time while also focusing on the role of gravity in autoexpansion. Using the North American bird grasshopper Schistocerca americana, we tested four autoexpansion treatments of varying surface tension and hydrophobicity (gravity, deionized water, buffer, and mineral oil) while simultaneously observing and measuring intact, normal wing expansion. Finally, we constructed a simple model of a viscoelastic expanding wing subjected to gravity, to determine whether it could capture aspects of wing expansion. Our data confirmed that wing autoexpansion does occur in S. americana, but autoexpanding wings, especially hindwings, failed to increase to the same final length and area as intact wings. We found that gravity plays an important role in wing expansion, early in the expansion process. Combined with the significant mass increase we documented in intact wings, it suggests that hydraulic pumping of hemolymph into the wings plays an important role in increasing the area of expanding wings, especially in driving expansion of the large, pleated hindwings. Autoexpansion in a non-swarming orthopteran suggests that local cues driving wing autoexpansion may serve a broader purpose, reducing total expansion time and costs by shifting some processes from central to local control. Documenting wing autoexpansion in a widely studied model organism and demonstrating a mathematical model provides a tractable new system for exploring higher level questions about the mechanisms of wing expansion and the implications of autoexpansion, as well as potential bioinspiration for future technologies applicable to micro-air vehicles, space exploration, or medical and prosthetic devices.

Close encounters of three kinds: impacts of leg, wing and body collisions on flight performance in carpenter bees.

Flying insects often forage among cluttered vegetation that forms a series of obstacles in their flight path. Recent studies have focused on behaviors needed to navigate clutter while avoiding all physical contact and, as a result, we know little about flight behaviors that do involve encounters with obstacles. Here, we challenged carpenter bees (Xylocopa varipuncta) to fly through narrow gaps in an obstacle course to determine the kinds of obstacle encounters they experience, as well as the consequences for flight performance. We observed three kinds of encounters: leg, body and wing collisions. Wing collisions occurred most frequently (in about 40% of flights, up to 25 times per flight) but these had little effect on flight speed or body orientation. In contrast, body and leg collisions, which each occurred in about 20% of flights (1-2 times per flight), resulted in decreased flight speeds and increased rates of body rotation (yaw). Wing and body collisions, but not leg collisions, were more likely to occur in wind versus still air. Thus, physical encounters with obstacles may be a frequent occurrence for insects flying in some environments, and the immediate effects of these encounters on flight performance depend on the body part involved.

Going against the flow: bumblebees prefer to fly upwind and display more variable kinematics when flying downwind.

Foraging insects fly over long distances through complex aerial environments, and many can maintain constant ground speeds in wind, allowing them to gauge flight distance. Although insects encounter winds from all directions in the wild, most lab-based studies have employed still air or headwinds (i.e. upwind flight); additionally, insects are typically compelled to fly in a single, fixed environment, so we know little about their preferences for different flight conditions. We used automated video collection and analysis methods and a two-choice flight tunnel paradigm to examine thousands of foraging flights performed by hundreds of bumblebees flying upwind and downwind. In contrast to the preference for flying with a tailwind (i.e. downwind) displayed by migrating insects, we found that bees prefer to fly upwind. Bees maintained constant ground speeds when flying upwind or downwind in flow velocities from 0 to 2 m s-1 by adjusting their body angle, pitching down to raise their air speed above flow velocity when flying upwind, and pitching up to slow down to negative air speeds (flying backwards relative to the flow) when flying downwind. Bees flying downwind displayed higher variability in body angle, air speed and ground speed. Taken together, bees' preference for upwind flight and their increased kinematic variability when flying downwind suggest that tailwinds may impose a significant, underexplored flight challenge to bees. Our study demonstrates the types of questions that can be addressed with newer approaches to biomechanics research; by allowing bees to choose the conditions they prefer to traverse and automating filming and analysis to examine massive amounts of data, we were able to identify significant patterns emerging from variable locomotory behaviors, and gain valuable insight into the biomechanics of flight in natural environments.

Complex hemolymph circulation patterns in grasshopper wings.

An insect's living systems-circulation, respiration, and a branching nervous system-extend from the body into the wing. Wing hemolymph circulation is critical for hydrating tissues and supplying nutrients to living systems such as sensory organs across the wing. Despite the critical role of hemolymph circulation in maintaining healthy wing function, wings are often considered "lifeless" cuticle, and flows remain largely unquantified. High-speed fluorescent microscopy and particle tracking of hemolymph in the wings and body of the grasshopper Schistocerca americana revealed dynamic flow in every vein of the fore- and hindwings. The global system forms a circuit, but local flow behavior is complex, exhibiting three distinct types: pulsatile, aperiodic, and "leaky" flow. Thoracic wing hearts pull hemolymph from the wing at slower frequencies than the dorsal vessel; however, the velocity of returning hemolymph (in the hindwing) is faster than in that of the dorsal vessel. To characterize the wing's internal flow mechanics, we mapped dimensionless flow parameters across the wings, revealing viscous flow regimes. Wings sustain ecologically important insect behaviors such as pollination and migration. Analysis of the wing circulatory system provides a template for future studies investigating the critical hemodynamics necessary to sustaining wing health and insect flight.

An evaluation of common methods for comparing the scaling of vertical force production in flying insects.

Maximum vertical force production (Fvert) is an integral measure of flight performance that generally scales with size. Numerous methods of measuring Fvert and body size are accessible to entomologists, but we do not know whether method selection affects inter- and intraspecific comparisons of Fvert-size scaling. We compared two common techniques for measuring Fvert in bumblebees (Bombus impatiens) and mason bees (Osmia lignaria), and examined Fvert scaling using five size metrics. Fvert results were similar with incremental or asymptotic load-lifting, but scaling analyses were sensitive to the size metric used. Analyses based on some size metrics indicated similar scaling exponents and coefficients between species, whereas other metrics indicated coefficients that differed by up to 18%. Furthermore, Fvert showed isometry with body lengths and fed and starved masses, but negative allometry with dry mass. We conclude that Fvert can be measured using either incremental or asymptotic loading but choosing a size metric for scaling studies requires careful consideration.

Wind and route choice affect performance of bees flying above versus within a cluttered obstacle field.

Bees flying through natural landscapes frequently encounter physical challenges, such as wind and cluttered vegetation, but the influence of these factors on flight performance remains unknown. We analyzed 548 videos of wild-caught honeybees (Apis mellifera) flying through an enclosure containing a field of vertical obstacles that bees could choose to fly within (through open corridors, without maneuvering) or above. We varied obstacle field height and wind condition (still, headwinds or tailwinds), and examined how these factors affected bees' flight altitude, ground speed, and side-to-side casting motions (lateral excursions). When obstacle fields were short, bees flew at altitudes near the midpoint between the tunnel floor and ceiling. When obstacle fields approached or exceeded this midpoint, bees tended to increase their altitude, but they did not always avoid flying through obstacles, despite having the freedom to do so. Bees that flew above the obstacles exhibited 40% faster ground speeds and 36% larger lateral excursions than bees that flew within the obstacle fields. Wind did not affect flight altitude, but bees flew 12-19% faster in tailwinds, and their lateral excursions were 19% larger when flying in headwinds or tailwinds, as compared to still air. Our results show that bees flying through complex environments display flexibility in their route choices (i.e., flying above obstacles in some trials and through them in others), which affects their overall flight performance. Similar choices in natural landscapes could have broad implications for foraging efficiency, pollination, and mortality in wild bees.

Bumblebees perceive the spatial layout of their environment in relation to their body size and form to minimize inflight collisions.

Animals that move through complex habitats must frequently contend with obstacles in their path. Humans and other highly cognitive vertebrates avoid collisions by perceiving the relationship between the layout of their surroundings and the properties of their own body profile and action capacity. It is unknown whether insects, which have much smaller brains, possess such abilities. We used bumblebees, which vary widely in body size and regularly forage in dense vegetation, to investigate whether flying insects consider their own size when interacting with their surroundings. Bumblebees trained to fly in a tunnel were sporadically presented with an obstructing wall containing a gap that varied in width. Bees successfully flew through narrow gaps, even those that were much smaller than their wingspans, by first performing lateral scanning (side-to-side flights) to visually assess the aperture. Bees then reoriented their in-flight posture (i.e., yaw or heading angle) while passing through, minimizing their projected frontal width and mitigating collisions; in extreme cases, bees flew entirely sideways through the gap. Both the time that bees spent scanning during their approach and the extent to which they reoriented themselves to pass through the gap were determined not by the absolute size of the gap, but by the size of the gap relative to each bee's own wingspan. Our findings suggest that, similar to humans and other vertebrates, flying bumblebees perceive the affordance of their surroundings relative their body size and form to navigate safely through complex environments.

Wind and obstacle motion affect honeybee flight strategies in cluttered environments.

Bees often forage in habitats with cluttered vegetation and unpredictable winds. Navigating obstacles in wind presents a challenge that may be exacerbated by wind-induced motions of vegetation. Although wind-blown vegetation is common in natural habitats, we know little about how the strategies of bees for flying through clutter are affected by obstacle motion and wind. We filmed honeybees Apis mellifera flying through obstacles in a flight tunnel with still air, headwinds or tailwinds. We tested how their ground speeds and centering behavior (trajectory relative to the midline between obstacles) changed when obstacles were moving versus stationary, and how their approach strategies affected flight outcome (successful transit versus collision). We found that obstacle motion affects ground speed: bees flew slower when approaching moving versus stationary obstacles in still air but tended to fly faster when approaching moving obstacles in headwinds or tailwinds. Bees in still air reduced their chances of colliding with obstacles (whether moving or stationary) by reducing ground speed, whereas flight outcomes in wind were not associated with ground speed, but rather with improvement in centering behavior during the approach. We hypothesize that in challenging flight situations (e.g. navigating moving obstacles in wind), bees may speed up to reduce the number of wing collisions that occur if they pass too close to an obstacle. Our results show that wind and obstacle motion can interact to affect flight strategies in unexpected ways, suggesting that wind-blown vegetation may have important effects on foraging behaviors and flight performance of bees in natural habitats.

Wind drives temporal variation in pollinator visitation in a fragmented tropical forest.

Wind is a critical factor in the ecology of pollinating insects such as bees. However, the role of wind in determining patterns of bee abundance and floral visitation rates across space and time is not well understood. Orchid bees are an important and diverse group of neotropical pollinators that harvest pollen, nectar and resin from plants. In addition, male orchid bees collect volatile scents that they store in special chambers in their hind legs, and for which the wind-based dispersal of odours may play a particularly crucial role. Here, we take advantage of this specialized scent foraging behaviour to study the effects of wind on orchid bee visitation at scent sources in a fragmented tropical forest ecosystem. Consistent with previous work, forest cover increased orchid bee visitation. In addition, we find that temporal changes in wind speed and turbulence increase visitation to scent stations within sites. These results suggest that the increased dispersal of attractive scents provided by wind and turbulence outweighs any biomechanical or energetic costs that might deter bees from foraging in these conditions. Overall, our results highlight the significance of wind in the ecology of these important pollinators in neotropical forests.

Kinematic flexibility allows bumblebees to increase energetic efficiency when carrying heavy loads.

Foraging bees fly with heavy loads of nectar and pollen, incurring energetic costs that are typically assumed to depend on load size. Insects can produce more force by increasing stroke amplitude and/or flapping frequency, but the kinematic response of a given species is thought to be consistent. We examined bumblebees (Bombus impatiens) carrying both light and heavy loads and found that stroke amplitude increased in proportion to load size, but did not predict metabolic rate. Rather, metabolic rate was strongly tied to frequency, which was determined not by load size but by the bee's average loading state and loading history, with heavily loaded bees displaying smaller changes in frequency and smaller increases in metabolic rate to support additional loading. This implies that bees can increase force production through alternative mechanisms; yet, they often choose the energetically costly option of elevating frequency, suggesting associated performance benefits that merit further investigation.

Sonicating bees demonstrate flexible pollen extraction without instrumental learning.

Pollen collection is necessary for bee survival and important for flowering plant reproduction, yet if and how pollen extraction motor routines are modified with experience is largely unknown. Here, we used an automated reward and monitoring system to evaluate modification in a common pollen-extraction routine, floral sonication. Through a series of laboratory experiments with the bumblebee, Bombus impatiens, we examined whether variation in sonication frequency and acceleration is due to instrumental learning based on rewards, a fixed behavioral response to rewards, and/or a mechanical constraint. We first investigated whether bees could learn to adjust their sonication frequency in response to pollen rewards given only for specified frequency ranges and found no evidence of instrumental learning. However, we found that absence versus receipt of a pollen reward did lead to a predictable behavioral response, which depended on bee size. Finally, we found some evidence of mechanical constraints, in that flower mass affected sonication acceleration (but not frequency) through an interaction with bee size. In general, larger bees showed more flexibility in sonication frequency and acceleration, potentially reflecting a size-based constraint on the range over which smaller bees can modify frequency and acceleration. Overall, our results show that although bees did not display instrumental learning of sonication frequency, their sonication motor routine is nevertheless flexible.

Neonicotinoid exposure disrupts bumblebee nest behavior, social networks, and thermoregulation.

Neonicotinoid pesticides can negatively affect bee colonies, but the behavioral mechanisms by which these compounds impair colony growth remain unclear. Here, we investigate imidacloprid's effects on bumblebee worker behavior within the nest, using an automated, robotic platform for continuous, multicolony monitoring of uniquely identified workers. We find that exposure to field-realistic levels of imidacloprid impairs nursing and alters social and spatial dynamics within nests, but that these effects vary substantially with time of day. In the field, imidacloprid impairs colony thermoregulation, including the construction of an insulating wax canopy. Our results show that neonicotinoids induce widespread disruption of within-nest worker behavior that may contribute to impaired growth, highlighting the potential of automated techniques for characterizing the multifaceted, dynamic impacts of stressors on behavior in bee colonies.

Living in a trash can: turbulent convective flows impair Drosophila flight performance.

Turbulent flows associated with thermal convection are common in areas where the ground is heated by solar radiation, fermentation or other processes. However, it is unknown how these flow instabilities affect the locomotion of small insects, like fruit flies, that inhabit deserts and urban landscapes where surface temperatures can reach extreme values. We quantified flight performance of fruit flies (Drosophila melanogaster) traversing a chamber through still air and turbulent Rayleigh-Bénard convection cells produced by a vertical temperature gradient. A total of 34% of individuals were unable to reach the end of the chamber in convection, although peak flow speeds were modest relative to typical outdoor airflow. Individuals that were successful in convection were faster fliers and had larger wing areas than those that failed. All flies displayed higher pitch angles and lower mean flight speeds in convection. Successful individuals took longer to cross the chamber in convection, due to lower flight speeds and greater path sinuosity. All individuals displayed higher flapping frequencies in convection, and successful individuals also reduced stroke amplitude. Our results suggest that thermal convection poses a significant challenge for small fliers, resulting in increased travel times and energetic costs, or in some cases precluding insects from traversing these environments entirely.

Author Correction: Spatial fidelity of workers predicts collective response to disturbance in a social insect.

The original version of the Article contained incorrect citation information in reference 67. The reference should read "Russell, A. L., Morrison, S. J., Moschonas, E. H. & Papaj, D. R. Patterns of pollen and nectar foraging specialization by bumblebees over multiple timescales using RFID. Sci. Rep. 7, 1-13 (2017)." This error has now been corrected in the PDF and HTML versions of the Article.

Dispensing Pollen via Catapult: Explosive Pollen Release in Mountain Laurel (Kalmia latifolia).

The astonishing amount of floral diversity has inspired countless assumptions about the function of diverse forms and their adaptive significance, yet many of these hypothesized functions are untested. We investigated an often-repeated adaptive hypothesis about how an extreme floral form functions. In this study, we conducted four investigations to understand the adaptive function of explosive pollination in Kalmia latifolia, the mountain laurel. We first performed a kinematic analysis of anther movement. Second, we constructed a heat map of pollen trajectories in three-dimensional space. Third, we conducted field observations of pollinators and their behaviors while visiting K. latifolia. Finally, we conducted a pollination experiment to investigate the importance of pollinators for fertilization success. Our results suggest that insect visitation dramatically improves fertilization success and that bees are the primary pollinators that trigger explosive pollen release.

Spatial fidelity of workers predicts collective response to disturbance in a social insect.

Individuals in social insect colonies cooperate to perform collective work. While colonies often respond to changing environmental conditions by flexibly reallocating workers to different tasks, the factors determining which workers switch and why are not well understood. Here, we use an automated tracking system to continuously monitor nest behavior and foraging activity of uniquely identified workers from entire bumble bee (Bombus impatiens) colonies foraging in a natural outdoor environment. We show that most foraging is performed by a small number of workers and that the intensity and distribution of foraging is actively regulated at the colony level in response to forager removal. By analyzing worker nest behavior before and after forager removal, we show that spatial fidelity of workers within the nest generates uneven interaction with relevant localized information sources, and predicts which workers initiate foraging after disturbance. Our results highlight the importance of spatial fidelity for structuring information flow and regulating collective behavior in social insect colonies.

Wings as impellers: honey bees co-opt flight system to induce nest ventilation and disperse pheromones.

Honey bees (Apis mellifera) are remarkable fliers that regularly carry heavy loads of nectar and pollen, supported by a flight system - the wings, thorax and flight muscles - that one might assume is optimized for aerial locomotion. However, honey bees also use this system to perform other crucial tasks that are unrelated to flight. When ventilating the nest, bees grip the surface of the comb or nest entrance and fan their wings to drive airflow through the nest, and a similar wing-fanning behavior is used to disperse volatile pheromones from the Nasonov gland. In order to understand how the physical demands of these impeller-like behaviors differ from those of flight, we quantified the flapping kinematics and compared the frequency, amplitude and stroke plane angle during these non-flight behaviors with values reported for hovering honey bees. We also used a particle-based flow visualization technique to determine the direction and speed of airflow generated by a bee performing Nasonov scenting behavior. We found that ventilatory fanning behavior is kinematically distinct from both flight and scenting behavior. Both impeller-like behaviors drive flow parallel to the surface to which the bees are clinging, at typical speeds of just under 1 m s-1 We observed that the wings of fanning and scenting bees frequently contact the ground during the ventral stroke reversal, which may lead to wing wear. Finally, we observed that bees performing Nasonov scenting behavior sometimes display 'clap-and-fling' motions, in which the wings contact each other during the dorsal stroke reversal and fling apart at the start of the downstroke. We conclude that the wings and flight motor of honey bees comprise a multifunctional system, which may be subject to competing selective pressures because of its frequent use as both a propeller and an impeller.

Wing flexibility improves bumblebee flight stability.

Insect wings do not contain intrinsic musculature to change shape, but rather bend and twist passively during flight. Some insect wings feature flexible joints along their veins that contain patches of resilin, a rubber-like protein. Bumblebee wings exhibit a central resilin joint (1m-cu) that has previously been shown to improve vertical force production during hovering flight. In this study, we artificially stiffened bumblebee (Bombus impatiens) wings in vivo by applying a micro-splint to the 1m-cu joint, and measured the consequences for body stability during forward flight in both laminar and turbulent airflow. In laminar flow, bees with stiffened wings exhibited significantly higher mean rotation rates and standard deviation of orientation about the roll axis. Decreasing the wing's flexibility significantly increased its projected surface area relative to the oncoming airflow, likely increasing the drag force it experienced during particular phases of the wing stroke. We hypothesize that higher drag forces on stiffened wings decrease body stability when the left and right wings encounter different flow conditions. Wing splinting also led to a small increase in body rotation rates in turbulent airflow, but this change was not statistically significant, possibly because bees with stiffened wings changed their flight behavior in turbulent flow. Overall, we found that wing flexibility improves flight stability in bumblebees, adding to the growing appreciation that wing flexibility is not merely an inevitable liability in flapping flight, but can enhance flight performance.