The use of heterospecific scent marks by the sweat bee <Emphasis Type="Italic">Halictus aerarius</Emphasis>

To forage effectively amongst flowers, some bee species utilize olfactory cues left by previous visitors in addition to direct assessment of visual cues to identify rewarding flowers. This ability can be more advantageous if the bees can recognize and use scent marks left by heterospecifics, not just marks left by members of their own species. We conducted field experiments to investigate whether the sweat bee Halictus aerarius avoids visiting flowers of trailing water willow Justicia procumbens emptied by other bee species. We found that H. aerarius rejected the flowers visited by both heterospecifics and conspecifics. They also rejected visited flowers artificially replenished with nectar. Our results demonstrate that social bees outside the Apidae can detect marks left on flowers by heterospecifics but that (on this plant species) they are unable to discriminate against flowers by directly detecting nectar volume. H. aerarius exhibited different rejection rates according to the identity of the previous bee species. We suggest that the frequency of rejection responses may depend on the amount of chemical substances left by the previous bee. In general, the use of scent marks left by previous visitors is almost certainly advantageous, enabling foragers to avoid flowers with depleted nectar levels and thereby improving their foraging efficiency.     

Background complexity affects colour preference in bumblebees

Flowers adapted for hummingbird pollination are typically red. This correlation is usually explained by the assertion that nectar- or pollen-stealing bees are “blind” to red flowers. However, laboratory studies have shown that bees are capable of locating artificial red flowers and often show no innate preference for blue over red. We hypothesised that these findings might be artefacts of the simplified laboratory environment. Using bumblebees (Bombus impatiens) that had been trained to visit red and blue artificial flowers, we tested whether colour preference was influenced by complexity of the background on which they were foraging. Many bees were indifferent to flower colour when tested using a uniform green background like those commonly used in laboratory studies, but all bees showed strong colour preferences (usually for blue) when flowers were presented against a photograph of real foliage. Overall, preference for blue flowers was significantly greater on the more realistic, complex background. These results support the notion that the red of “hummingbird syndrome” flowers can function to reduce bee visits despite the ability of bees to detect red and highlight the need to consider context when drawing inferences about pollinator preferences from laboratory data. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The innate responses of bumble bees to flower patterns: separating the nectar guide from the nectary changes bee movements and search time

Nectar guides can enhance pollinator efficiency and plant fitness by allowing pollinators to more rapidly find and remember the location of floral nectar. We tested if a radiating nectar guide around a nectary would enhance the ability of naïve bumble bee foragers to find nectar. Most experiments that test nectar guide efficacy, specifically radiating linear guides, have used guides positioned around the center of a radially symmetric flower, where nectaries are often found. However, the flower center may be intrinsically attractive. We therefore used an off-center guide and nectary and compared “conjunct” feeders with a nectar guide surrounding the nectary to “disjunct” feeders with a nectar guide separated from the nectary. We focused on the innate response of novice bee foragers that had never previously visited such feeders. We hypothesized that a disjunct nectar guide would conflict with the visual information provided by the nectary and negatively affect foraging. Approximately, equal numbers of bumble bees (Bombus impatiens) found nectar on both feeder types. On disjunct feeders, however, unsuccessful foragers spent significantly more time (on average 1.6-fold longer) searching for nectar than any other forager group. Successful foragers on disjunct feeders approached these feeders from random directions unlike successful foragers on conjunct feeders, which preferentially approached the combined nectary and nectar guide. Thus, the nectary and a surrounding nectar guide can be considered a combination of two signals that attract naïve foragers even when not in the floral center.     

Floral odor learning within the hive affects honeybees’ foraging decisions

Honeybees learn odor cues quickly and efficiently when visiting rewarding flowers. Memorization of these cues facilitates the localization and recognition of food sources during foraging flights. Bees can also use information gained inside the hive during social interactions with successful foragers. An important information cue that can be learned during these interactions is food odor. However, little is known about how floral odors learned in the hive affect later decisions of foragers in the field. We studied the effect of food scent on foraging preferences when this learning is acquired directly inside the hive. By using in-hive feeders that were removed 24 h before the test, we showed that foragers use the odor information acquired during a 3-day stimulation period with a scented solution during a food-choice situation outside the nest. This bias in food preference is maintained even 24 h after the replacement of all the hive combs. Thus, without being previously collected outside by foragers, food odors learned within the hive can be used during short-range foraging flights. Moreover, correct landings at a dual-choice device after replacing the storing combs suggests that long-term memories formed within the colony can be retrieved while bees search for food in the field.     

Recruits of the stingless bee Scaptotrigona pectoralis learn food odors from the nest atmosphere

The ability to learn food odors inside the nest and to associate them with food sources in the field is of essential importance for the recruitment of nestmates in social bees. We investigated odor learning by workers within the hive and the influence of these odors on their food choice in the field in the stingless bee Scaptotrigona pectoralis. During the experiments, recruited bees had to choose between two feeders, one with an odor that was present inside the nest during the recruitment process, and one with an unknown odor. In all experiments with different odor combinations (linalool/phenylacetaldehyde, geraniol/eugenol) a significant majority of bees visited the feeder with the odor they had experienced in their nest (χ ²-tests; p?<?0.05). By contrast, the bees showed no preference for one of two feeders when they were either baited with the same odor (linalool) or contained no odor. Our results clearly show that naïve workers of S. pectoralis can learn the odor of a food source during the recruitment process from the nest atmosphere and that their subsequent food search in the field is influenced by the learned odor.     

Chemical compounds of the foraging recruitment pheromone in bumblebees

When the frenzied and irregular food-recruitment dances of bumblebees were first discovered, it was thought that they might represent an evolutionary prototype to the honeybee waggle dance. It later emerged that the primary function of the bumblebee dance was the distribution of an alerting pheromone. Here, we identify the chemical compounds of the bumblebee recruitment pheromone and their behaviour effects. The presence of two monoterpenes and one sesquiterpene (eucalyptol, ocimene and farnesol) in the nest airspace and in the tergal glands increases strongly during foraging. Of these, eucalyptol has the strongest recruitment effect when a bee nest is experimentally exposed to it. Since honeybees use terpenes for marking food sources rather than recruiting foragers inside the nest, this suggests independent evolutionary roots of food recruitment in these two groups of bees.     

How floral odours are learned inside the bumblebee (Bombus terrestris) nest

Recruitment in social insects often involves not only inducing nestmates to leave the nest, but also communicating crucial information about finding profitable food sources. Although bumblebees transmit chemosensory information (floral scent), the transmission mechanism is unknown as mouth-to-mouth fluid transfer (as in honeybees) does not occur. Because recruiting bumblebees release a pheromone in the nest that triggers foraging in previously inactive workers, we tested whether this pheromone helps workers learn currently rewarding floral odours, as found in food social learning in rats. We exposed colonies to artificial recruitment pheromone, paired with anise scent. The pheromone did not facilitate learning of floral scent. However, we found that releasing floral scent in the air of the colony was sufficient to trigger learning and that learning performance was improved when the chemosensory cue was provided in the nectar in honeypots; probably because it guarantees a tighter link between scent and reward, and possibly because gustatory cues are involved in addition to olfaction. Scent learning was maximal when anise-scented nectar was brought into the nest by demonstrator foragers, suggesting that previously unidentified cues provided by successful foragers play an important role in nestmates learning new floral odours.     

Floral colour change in <Emphasis Type="Italic">Byrsonima variabilis</Emphasis> (Malpighiaceae) as a visual cue for pollen but not oil foraging by oil-collecting bees

Pollinators search for multiple flora resources throughout their life cycle. Most studies, however, only assess how bees discriminate floral cues in the context of nectar foraging. In the present study, we sought to elucidate whether oil-collecting bees discriminate flowers of Byrsonima variabilis (Malpighiaceae) with petals of different colours when foraging for pollen or oil. As the colour of the standard petal changes during anthesis, we characterised the spectral reflectance patterns of flowers throughout anthesis and modelled chromatic perceptual space to determine how these colour patterns are perceived by bees. Through the quantification of flower pollen in the different phases, we found that the colour of the standard petal is an honest cue of the presence of pollen. Centridine bees preferentially visited flowers with a yellow (bee’s green) colour when searching for pollen, but indiscriminately visited flowers with different petal colours when searching for floral oil. We suggest that standard petals, in the species studied and others of the genus, like nectar guides, act as pollen guides, which oil-collecting females use to detect pollen-rich flowers. Moreover, they use different floral clues during foraging for different resources in the same host plant.     

Pollen foraging: learning a complex motor skill by bumblebees (<Emphasis Type="Italic">Bombus terrestris</Emphasis>)

To investigate how bumblebees (Bombus terrestris) learn the complex motor skills involved in pollen foraging, we observed naïve workers foraging on arrays of nectarless poppy flowers (Papaver rhoeas) in a greenhouse. Foraging skills were quantified by measuring the pollen load collected during each foraging bout and relating this to the number of flowers visited and bout duration on two consecutive days. The pollen standing crop (PSC) in each flower decreased drastically from 0530 to 0900 hours. Therefore, we related foraging performance to the changing levels of pollen available (per flower) and found that collection rate increased over the course of four consecutive foraging bouts (comprising between 277 and 354 individual flower visits), suggesting that learning to forage for pollen represents a substantial time investment for individual foragers. The pollen collection rate and size of pollen loads collected at the start of day 2 were markedly lower than at the end of day 1, suggesting that components of pollen foraging behaviour could be subject to imperfect overnight retention. Our results suggest that learning the necessary motor skills to collect pollen effectively from morphologically simple flowers takes three times as many visits as learning how to handle the most morphologically complex flowers to extract nectar, potentially explaining why bees are more specialised in their choice of pollen flowers.     

Heat-balling wasps by honeybees

Defensiveness of honeybee colonies of Apis cerana and Apis mellifera (actively balling the wasps but reduction of foraging) against predatory wasps, Vespa velutina, and false wasps was assessed. There were significantly more worker bees in balls of the former than latter. Core temperatures in a ball around a live wasp of A. cerana were significantly higher than those of A. mellifera, and also significantly more when exposed to false wasps. Core temperatures of bee balls exposed to false wasps were significantly lower than those exposed to V. velutina for both A. cerana and for A. mellifera. The lethal thermal limits for V. velutina, A. cerana and A. mellifera were significantly different, so that both species of honeybees have a thermal safety factor in heat-killing such wasp predators. During wasps attacks at the hives measured at 3, 6 and 12 min, the numbers of Apis cerana cerana and Apis cerana indica bees continuing to forage were significantly reduced with increased wasp attack time. Tropical lowland A. c. indica reduced foraging rates significantly more than the highland A. c. cerana bees; but, there was no significant effect on foraging by A. mellifera. The latency to recovery of honeybee foraging was significantly greater the longer the duration of wasp attacks. The results show remarkable thermal fine-tuning in a co-evolving predator–prey relationship.     

Longer visits on familiar plants?: testing a regular visitor’s tendency to probe more flowers than occasional visitors

An individual pollinator may tend to consecutively probe more flowers on a plant to which it returns at shorter intervals than other plants. In a large net cage, I let individually marked bumble bees forage on flowering heads of red clovers arranged in 37 bottles (plants), each of which was monitored by an observer to record every visit and probe for 2.5 h on each of 3 days. The data of collective visits by marked individuals revealed that the bees had their own foraging areas, in which they visited a set of plants frequently and others less often, i.e., the same individual bee repeatedly returned to certain plants as a regular visitor while sampling others as an occasional visitor. I further found that as a regular visitor, an individual bee tended to probe more flowering heads on familiar plants while probing fewer on unfamiliar plants as an occasional visitor. The mean number of consecutive probes by a bee was also positively correlated with its activity (the total number of plant visits made during the observation period). The fact that each bee behaves differently on different plants indicates that the same individual pollinator can exert different influence on the reproductive success of each plant: apparently, a pollinator likely reduces the potential for geitonogamous self-pollination when foraging as an occasional visitor. Attracting occasional visitors therefore may be beneficial for plants to avoid geitonogamy. This study thus emphasizes the importance of paying attention to pollinator individuality in pollination ecology.     

Flower-visiting behavior of male bees is triggered by nectar-feeding insects

Bees are important pollinators for many flowering plants. Female bees are thought to be more effective pollinators than male bees because they carry much more pollen than males. Males of some solitary bee species are known to patrol near flowers that females visit. Because patrolling males visit flowers to mate or defend their territories, they may function as pollinators. However, the significance of patrolling males to pollination has not been studied. We studied males of a solitary bee, Heriades fulvohispidus (Megachilidae), patrolling near flowers and visiting flowers that attracted nectar-feeding insects, including conspecifics, on the Ogasawara (Bonin) Islands. To test the hypothesis that patrolling male bees may function as pollen vectors, we compared the frequency of visits by H. fulvohispidus to flowers of an endemic plant, Schima mertensiana (Theaceae); comparisons were made among flowers with a dead H. fulvohispidus, a dead beetle, a piece of plastic, and nothing (control flowers). Patrolling H. fulvohispidus more frequently visited flowers with a dead conspecific, a dead beetle, or a piece of plastic than the control flowers. Our experiment demonstrates that nectar-feeding insects (including conspecifics and other insects) enhance the flower-visiting frequency of patrolling H. fulvohispidus males on S. mertensiana flowers. Furthermore, we observed S. mertensiana pollen on patrolling males as well as females, suggesting that male bees may also function as pollen vectors.     

Multicomponent floral signals elicit selective foraging in bumblebees

Flower constancy, or the tendency of individual pollinators to visit sequentially a single flower type even when other equally rewarding types are available, has important implications for animal-pollinated plants. Yet, the proximal reason for the behaviour still remains poorly understood. Here I show that bumblebees visiting equally rewarding flowers that differ in size and odour are more flower constant and less efficient (visited fewer flowers per minute) than bees visiting flowers that differ in size only and odour only. These results are consistent with the view that flower constancy in pollinators is related to their inability to perceive, process or recall multicomponent floral signals. I discuss these findings in the context of pollinator behavioural mechanisms and the evolution of floral diversity.     

Individual lifetime pollen and nectar foraging preferences in bumble bees

Foraging specialization plays an important role in the ability of social insects to efficiently allocate labor. However, relatively little is known about the degree to which individual bumble bees specialize on collecting nectar or pollen, when such preferences manifest, and if individuals can alter their foraging preferences in response to changes in the colony workforce. Using Bombus impatiens, we monitored all foraging visits made by every bee in multiple colonies and showed that individual foragers exhibit consistent lifetime foraging preferences. Based upon the distribution of foraging preferences, we defined three forager types (pollen specialists, nectar specialists, and generalists). In unmanipulated colonies, 16–36?% of individuals specialized (≥90?% of visits) on nectar or pollen only. On its first day of foraging, an individual’s foraging choices (nectar only, pollen only, or nectar and pollen) significantly predicted its lifetime foraging preferences. Foragers that only collected pollen on their first day of foraging made 1.61- to 1.67-fold more lifetime pollen foraging visits (as a proportion of total trips) than foragers that only collected nectar on their first foraging day. Foragers were significantly larger than bees that stayed only in the nest. We also determined the effect of removing pollen specialists at early (brood present) or later (brood absent) stages in colony life. These results suggest that generalists can alter their foraging preferences in response to the loss of a small subset of foragers. Thus, bumble bees exhibit individual lifetime foraging preferences that are established early in life, but generalists may be able to adapt to colony needs.     

The effect of flower-like and non-flower-like visual properties on choice of unrewarding patterns by bumblebees

How do distinct visual stimuli help bumblebees discover flowers before they have experienced any reward outside of their nest? Two visual floral properties, type of a pattern (concentric vs radial) and its position on unrewarding artificial flowers (central vs peripheral on corolla), were manipulated in two experiments. Both visual properties showed significant effects on floral choice. When pitted against each other, pattern was more important than position. Experiment 1 shows a significant effect of concentric pattern position, and experiment 2 shows a significant preference towards radial patterns regardless of their position. These results show that the presence of markings at the center of a flower are not so important as the presence of markings that will direct bees there.     

Landmark memories are more robust when acquired at the nest site than en route: experiments in desert ants

Foraging desert ants, Cataglyphis fortis, encounter different sequences of visual landmarks while navigating by path integration. This paper explores the question whether the storage of landmark information depends on the context in which the landmarks are learned during an ant's foraging journey. Two experimental set-ups were designed in which the ants experienced an artificial landmark panorama that was placed either around the nest entrance (nest marks) or along the vector route leading straight towards the feeder (route marks). The two training paradigms resulted in pronounced differences in the storage characteristics of the acquired landmark information: memory traces of nest marks were much more robust against extinction and/or suppression than those of route marks. In functional terms, this result is in accord with the observation that desert ants encounter new route marks during every foraging run but always pass the same landmarks when approaching the nest entrance.     

Nutrient compensatory foraging in a free-living social insect

The geometric framework model predicts that animal foraging decisions are influenced by their dietary history, with animals targeting a combination of essential nutrients through compensatory foraging. We provide experimental confirmation of nutrient-specific compensatory foraging in a natural, free-living population of social insects by supplementing their diet with sources of protein- or carbohydrate-rich food. Colonies of the ant Iridomyrmex suchieri were provided with feeders containing food rich in either carbohydrate or protein for 6 days, and were then provided with a feeder containing the same or different diet. The patterns of recruitment were consistent with the geometric framework: while feeders with a carbohydrate diet typically attracted more workers than did feeders with protein diet, the difference in recruitment between the two nutrients was smaller if the colonies had had prior access to carbohydrate than protein. Further, fewer ants visited feeders if the colony had had prior access to protein than to carbohydrates, suggesting that the larvae play a role in worker foraging behaviour.     

Comparison of pollination and defensive buzzes in bumblebees indicates species-specific and context-dependent vibrations

Bees produce vibrations in many contexts, including for defense and while foraging. Buzz pollination is a unique foraging behavior in which bees vibrate the anthers of flowers to eject pollen which is then collected and used as food. The relationships between buzzing properties and pollen release are well understood, but it is less clear to what extent buzzing vibrations vary among species, even though such information is crucial to understanding the functional relationships between bees and buzz-pollinated plants. Our goals in this study were (1) to examine whether pollination buzzes differ from those produced during defense, (2) to evaluate the similarity of buzzes between different species of bumblebees (Bombus spp.), and (3) to determine if body size affects the expression of buzzing properties. We found that relative peak amplitude, peak frequency, and duration were significantly different between species, but only relative peak amplitude differed between pollination and defensive buzzes. There were significant interactions between species and buzz type for peak frequency and duration, revealing that species differed in their patterns of expression in these buzz properties depending on the context. The only parameter affected by body size was duration, with larger bees producing shorter buzzes. Our findings suggest that although pollination and defensive buzzes differ in some properties, variability in buzz structure also exhibits a marked species-specific component. Species differences in pollination buzzes may have important implications for foraging preferences in bumblebees, especially if bees select flowers best matched to release pollen for their specific buzzing characteristics.     

A novel mutualism between an ant-plant and its resident pollinator

Pollination systems in which the host plant provides breeding sites for pollinators, invariably within flowers, are usually highly specialized mutualisms. We found that the pollinating bee Braunsapis puangensis breeds within the caulinary domatia of the semi-myrmecophyte Humboldtia brunonis (Fabaceae), an unusual ant-plant that is polymorphic for the presence of domatia and harbours a diverse invertebrate fauna including protective and non-protective ants in its domatia. B. puangensis is the most common flower visitor that carries the highest proportion of H. brunonis pollen. This myrmecophyte is pollen limited and cross-pollinated by bees in the daytime. Hence, the symbiotic pollinator could provide a benefit to trees bearing domatia by alleviating this limitation. We therefore report for the first time an unspecialised mutualism in which a pollinator is housed in a plant structure other than flowers. Here, the cost to the plant is lower than for conventional brood-site pollination mutualisms where the pollinator develops at the expense of plant reproductive structures. Myrmecophytes housing resident pollinators are unusual, as ants are known to be enemies of pollinators, and housing them together may decrease the benefits that these residents could individually provide to the host plant.     

Convergent evolution: floral guides, stingless bee nest entrances, and insectivorous pitchers

Several recent hypotheses, including sensory drive and sensory exploitation, suggest that receiver biases may drive selection of biological signals in the context of sexual selection. Here we suggest that a similar mechanism may have led to convergence of patterns in flowers, stingless bee nest entrances, and pitchers of insectivorous plants. A survey of these non-related visual stimuli shows that they share features such as stripes, dark centre, and peripheral dots. Next, we experimentally show that in stingless bees the close-up approach to a flower is guided by dark centre preference. Moreover, in the approach towards their nest entrance, they have a spontaneous preference for entrance patterns containing a dark centre and disrupted ornamentation. Together with existing empirical evidence on the honeybee's and other insects’ orientation to flowers, this suggests that the signal receivers of the natural patterns we examined, mainly Hymenoptera, have spontaneous preferences for radiating stripes, dark centres, and peripheral dots. These receiver biases may have evolved in other behavioural contexts in the ancestors of Hymenoptera, but our findings suggest that they have triggered the convergent evolution of visual stimuli in floral guides, stingless bee nest entrances, and insectivorous pitchers.