Search

Bilateral symmetry has been considered as an indicator of phenotypic and genotypic quality supporting innate preferences for highly symmetric partners. Insect pollinators preferentially visit flowers of a particular symmetry type, thus leading to the suggestion that they have innate preferences for symmetrical flowers or flower models. Here we show that flower-naïve bumblebees (Bombus terrestris), with no experience of symmetric or asymmetric patterns and whose visual experience was accurately controlled, have innate preferences for bilateral symmetry. The presence of color cues did not influence the bees' original preference. Our results thus show that bilateral symmetry is innately preferred in the context of food search, a fact that supports the selection of symmetry in flower displays. Furthermore, such innate preferences indicate that the nervous system of naïve animals may be primed to respond to relevant sensory cues in the environment.

Generalization is a cognitive ability that allows similar stimuli along a given dimension to be treated as equivalents. Insects exhibit high levels of visual generalization. Honeybees trained to recognize complex visual stimuli on the basis of a single feature generalize their choice to novel stimuli sharing that common feature with the trained stimuli. The demonstration of this kind of performance has been limited to the use of a single visual feature, and the possibility that bees link different features in learning a visual pattern has been denied. Here we show that honeybees trained with a series of complex patterns sharing a common layout comprising four edge orientations remember these orientations simultaneously in their appropriate positions, and generalize their response to novel stimuli that preserve the trained layout. Honeybees also generalize their response to patterns with fewer correct orientations, depending on their match with the trained layout. Stimulation of the achromatic L-photoreceptor input is necessary for this task. The mini-brain of the honeybee can thus extract regularities in its environment and establish correspondences among correlated features. It can thus generate a large set of object descriptions from a finite set of elements.

We studied the influence of the conditioning procedure on color discrimination by free-flying honeybees. We asked whether absolute and differential conditioning result in different discrimination capabilities for the same pairs of colored targets. In absolute conditioning, bees were rewarded on a single color; in differential conditioning, bees were rewarded on the same color but an alternative, non-rewarding, similar color was also visible. In both conditioning procedures, bees learned their respective task and could also discriminate the training stimulus from a novel stimulus that was perceptually different from the trained one. Discrimination between perceptually closer stimuli was possible after differential conditioning but not after absolute conditioning. Differences in attention inculcated by these training procedures may underlie the different discrimination performances of the bees.

We asked whether honeybees, Apis mellifera, could solve a transitive inference problem. Individual free-flying bees were conditioned with four overlapping premise pairs of five visual patterns in a multiple discrimination task (A+ vs. B-, B+ vs. C-, C+ vs. D-, D+ vs. E-, where + and - indicate sucrose reward or absence of it, respectively). They were then tested with the nonadjacent pairs A vs. E and B vs. D. Preference of B to D is consistent with the use of the implicit hierarchy A > B > C > D > E. Equal choice of B and D supports choice based on the associative strength of the stimuli. The bees' choice was determined by their memory constraints: experience with the last premise pair (D+ vs. E-) predominated. In the tests, bees preferred A to E and chose equally B and D. An analysis of the performance in terms of a reward/penalty ratio showed that B had a higher associative strength than D. Thus, bees do not establish transitive inferences but, rather, guide their choices by the joint action of a recency effect and the associative strength of the stimuli. The former supports choice of D, whereas the latter supports choice of B, thus determining equal choice of B and D in the tests.

The present work introduces a form of associative mechanosensory conditioning of the proboscis extension reflex (PER) in honeybees. In our paradigm, harnessed honeybees learn the elemental association between mechanosensory, antennal stimulation and a reward of sucrose solution delivered to the proboscis. Thereafter, bees extend their proboscis to the antennal mechanosensory stimulation alone. We show that bees can learn such an association in a side-specific manner, that is, they learn the association on the antennal side that was rewarded and not on the side that was not rewarded. Responding produced by the paired training does likely contain a substantial Pavlovian component. Responding is only elicited by mechanosensory stimulation and not by spurious cues such as olfactory, visual, and contextual ones. The interstimulus interval (ISI) affects one-trial mechanosensory learning: a bell-shaped curve with a maximum of responding approximately 4 sec ISI was obtained. Mechanosensory memory is still operative 24 h after conditioning. Apart from absolute conditioning in which mechanosensory stimulation of one antenna is paired with sucrose, differential, side-specific, mechanosensory conditioning using two mechanosensory stimulations, one rewarded and the other not, is also possible. This paradigm constitutes, therefore, a new standard procedure for further learning studies in honeybees.

The brain of a honeybee contains only 960,000 neurons and its volume represents only 1 mm3. However, it supports impressive behavioral capabilities. Honeybees are equipped with sophisticated sensory systems and have well developed learning and memory capacities, whose essential mechanisms do not differ drastically from those of vertebrates. Here, I focus on non-elemental forms of learning by honeybees. I show that bees exhibit learning abilities that have been traditionally ascribed to a restricted portion of vertebrates, as they go beyond simple stimulus-stimulus or response-stimulus associations. To relate these abilities to neural structures and functioning in the bee brain we focus on the antennal lobes and the mushroom bodies. We conclude that there is a fair chance to understand complex behavior in bees, and to identify the potential neural substrates underlying such behavior by adopting a cognitive neuroethological approach. In such an approach, behavioral and neurobiological studies are combined to understand the rules and mechanisms of plastic behavior in a natural context.

We addressed the question of whether the amount of individual experience determines the use of elemental or configural visual discrimination strategies in free-flying honeybees Apis mellifera. We trained bees to fly into a Y-maze to collect sucrose solution on a rewarded stimulus presented in one of the arms of the maze. Stimuli were colour disks, violet (V), green (G) or yellow (Y), which were of equal psychophysical salience for honeybees. Training followed an A+, BC+ design, followed by an AC versus BC test. Training consisted of 6 (3 A+ and 3 BC+), 20 (10 A+ and 10 BC+) or 40 (20 A+ and 20 BC+) acquisition trials. Elemental models of compound processing predict a preference for the non-trained stimulus AC while configural models predict a preference for the trained stimulus BC. Our results show that increasing the number of acquisition trials results in a change of the internal representation of stimuli. After six training trials, bees favoured an elemental strategy and preferred AC to BC during the tests. Generally, increasing the number of training trials resulted in an increase of the choice of BC. Thus, short training favoured processing of the compound as the sum of its elements (elemental account) while long training favoured its processing as being different from the sum of its elements (configural account). Additionally, we observed that the change in stimulus processing was also influenced by stimulus similarity. Colour perceptual similarity favoured configural processing with increasing experience.

In patterning discriminations, animals have to differentiate a compound stimulus AB from each of its elements A and B. In positive patterning (PP), the compound is reinforced whilst the single elements are non-reinforced. In negative patterning (NP), single elements are reinforced whilst the compound is non-reinforced. Using olfactory conditioning of the proboscis extension response (PER), we asked whether honeybees (Apis mellifera) can solve these patterning problems when odorants are given unilaterally as well as bilaterally to the antennae. Separating the olfactory input space of bees into two independent zones using plastic walls placed between the antennae, we conditioned bees in PP and NP procedures, with input on one side, on both sides, or in an ambiguous problem where bees had to solve PP on one side and NP on the other side. We found that bees with simultaneous bilateral input solve both patterning tasks efficiently. In contrast, PP but not NP was learned by bees receiving unilateral olfactory input. Bees subjected to the ambiguous NP/PP problem only solved PP. As PP can be solved through mere elemental processes, but NP is critically dependent on the use of non-elemental learning processes, our results suggest that bilateral olfactory input is necessary for non-elemental processing to take place in the bee brain.

We investigated the capability of honeybees to discriminate between single odorants, binary olfactory mixtures, and ternary olfactory mixtures in olfactory conditioning of the proboscis extension reflex. In Experiment 1, three single odorants (A+, B+, and C+) and three binary mixtures of these odors (AB+, AC+, and BC+) were reinforced while the ternary compound, consisting of all three odors (ABC-), was nonreinforced. In Experiment 2, only one single odorant (A+) and one binary olfactory compound (BC+) were reinforced while the ternary compound (ABC-) consisting of the single odor and the binary compound was nonreinforced. We studied whether bees can solve these problems and whether the course of differentiation can be predicted by the unique cue theory, a modified unique cue theory, or Pearce's configural theory. Honeybees were not able to differentiate reinforced from nonreinforced stimuli in Experiment 1. However, summation to ABC observed at the beginning of training contradicts the predictions of Pearce's configural theory. In Experiment 2, differentiation between the single odorant A and the ternary compound developed more easily than between the binary compound BC and ABC. This pattern of differentiation is in line with a modified unique cue theory and Pearce's configural theory. Summation to ABC at the beginning of training, however, again was at odds with Pearce's configural theory. Thus, olfactory compound processing in honeybees can best be explained by a modified unique cue theory.

The mushroom bodies (MBs), a paired structure in the insect brain, play a major role in storing and retrieving olfactory memories. We tested whether olfactory learning and odor processing is impaired in honeybees in which MB subunits were partially ablated. Using hydroxyurea (HU) to selectively kill proliferating cells, we created honeybees with varying degrees of MB lesions. Three-dimensional reconstructions of brains were generated to analyze the drug-induced morphological changes. These reconstructions show that, with few exceptions, only the MBs were affected by the drug, while other brain areas remained morphometrically intact. Typically, lesions affected only the MB in one hemisphere of the brain. To preclude HU-induced physiologic deficits in the antennal lobe (AL) affecting olfactory learning, we measured the responses to odors in the AL using an in vivo calcium imaging approach. The response patterns did not differ between the AL of intact versus ablated brain sides within respective specimens. We, therefore, carried out side-specific classical discriminative olfactory conditioning of the proboscis extension reflex (PER) with control bees and with HU-treated bees with or without MB ablations. All experimental groups learned equally to discriminate and respond to a rewarded (CS+) but not to an unrewarded (CS-) conditioned stimulus during acquisition and retention tests. Thus, our results indicate that partial MB lesions do not affect this form of elemental olfactory learning., (Copyright 2002 Wiley Periodicals, Inc.)

Two human Pavlovian conditioning experiments investigated the impact of reinforcement density (the number of reinforced trials divided by the total number of trials) on discrimination learning. Experiment 1 used a negative patterning problem (A+, B+, AB-) and Experiment 2 used a positive patterning problem (A-, B-, AB+). In both experiments, reinforcement density varied across four levels. Response differentiation between reinforced and non-reinforced stimuli was linearly related to the decrease in reinforcement density. Responses to nonreinforced stimuli did not differ between the four groups in either experiment. In contrast to this, responses to reinforced stimuli were considerably more pronounced in conditions with lower reinforcement density. For negative patterning, this replicates and extends similar observations from other species. For positive patterning, this is a finding that has not yet been reported in other experiments. The results are in agreement with the comparator hypothesis (Miller & Matzel, 1988) and with Wagner's (1981) "standard operating procedures" (SOP) model.

Honeybees Apis mellifera can associate an originally neutral odor with a reinforcement of sucrose solution. Forward pairings of odor and reinforcement enable the odor to release the proboscis extension reflex in consecutive tests. Bees can also be conditioned differentially: They learn to respond to a reinforced odor and not to a nonreinforced one. They can also learn to reverse their choice. Here we ask whether honeybees can learn successive olfactory differential conditioning tasks involving different overlapping pairs of odors. The conditioning schedules were established in order to train the animals with 3, 2, 1, or 0 reversals previous to a last differential conditioning phase in which two additional reversals were present. We studied whether or not successive reversal learning is possible and whether or not learning olfactory discrimination reversals affects the solving of subsequent discrimination reversals. Therefore we compared the responses of bees that had experienced reversals with those of bees that had not experienced such reversals when both are confronted with a new reversal situation. In experiment 1 we showed that bees that had experienced three previous reversals were better in solving the final reversal task than bees with no previous reversal experience. In experiment 2, we showed that one reversal learning is enough for bees to perform better in the final reversal task. The successive different reversals trained in our experiments resemble the natural foraging situation in which a honeybee forager has to switch successively from an initial floral species to different ones. The fact that experiencing such changes seems to improve a bee's performance in dealing with further new exploited food sources has therefore an adaptive impact for the individual and for the colony as a whole.

We studied the ability of honeybees to discriminate between single odorants and binary olfactory mixtures. We analyzed the effect of the number of common elements between these two stimulus classes on olfactory discrimination. We used olfactory conditioning of the honeybees' proboscis extension reflex (PER), a paradigm in which odors can be associated with a reinforcement of sucrose solution. Bees were asked to discriminate reinforced from nonreinforced olfactory stimuli. They were trained with two elemental odors (A and B) versus a binary olfactory mixture. The mixture was either AB (group 2CE, two common elements), AC (group 1CE, one common element A), or CD (group 0CE, no common element). Three groups followed a positive patterning schedule (mixture reinforced and elements nonreinforced: groups 2CE+, 1CE+, and 0CE+) and three other groups a negative patterning schedule (mixture nonreinforced and elements reinforced: groups 2CE-, 1CE-, and 0CE-). We showed that a reduction of similarity (number of common elements) between elemental odors and compounds enhanced the ability to discriminate elements from compounds and that the kind of compound processing used by the bees supports theories that assume nonelemental compound processing (i.e., that exclude the mere summation of the elemental associative strengths upon compound presentation).

Hydroxyurea (HU) treatment of first instar honeybee larvae was previously shown to cause mushroom body (MB) ablations. Predominantly, either one or both median MB subunits were ablated. This prompted us to analyze the effects of asymmetrical or symmetrical HU-induced MB ablation on both the morphology of the brain and on the level of three proteins (synapsin, PKA RII, and PKC), which are considered to play a role in synaptic plasticity, learning, and memory. In brains with one median MB subunit missing the volume of the overall MB calyx neuropil in the lesioned side was diminished by 35%. This strong reduction occurred although the remaining lateral MB calyx of the lesioned brain side was found to be significantly larger than that of the intact side. Accordingly, in brains with both median MB subunits missing the size of the remaining lateral calyces increased. The various types of MB ablation differentially affected the amounts of synapsin, PKA RII, and PKC expressed in the central brain. In animals with bilateral and thus symmetrical MB ablation (both median calyces ablated) the protein amount was found to be similar to that in control animals. However, unilateral MB ablation causes an increase in the amounts of the tested proteins in the intact brain side, while the levels in the ablated side were the same as in control animals. These findings not only show that HU-induced ablation of MB subunits is accompanied by volume changes and by changes in protein expression, but also suggest that these processes are highly regulated between the brain sides. The latter is of general importance in understanding the potential contribution of the MB subunits to learning and memory and their interaction between the brain sides., (Copyright 2002 John Wiley & Sons, Inc.)

Insects process and learn information flexibly to adapt to their environment. The honeybee Apis mellifera constitutes a traditional model for studying learning and memory at behavioural, cellular and molecular levels. Earlier studies focused on elementary associative and non-associative forms of learning determined by either olfactory conditioning of the proboscis extension reflex or the learning of visual stimuli in an operant context. However, research has indicated that bees are capable of cognitive performances that were thought to occur only in some vertebrate species. For example, honeybees can interpolate visual information, exhibit associative recall, categorize visual information and learn contextual information. Here we show that honeybees can form 'sameness' and 'difference' concepts. They learn to solve 'delayed matching-to-sample' tasks, in which they are required to respond to a matching stimulus, and 'delayed non-matching-to-sample' tasks, in which they are required to respond to a different stimulus; they can also transfer the learned rules to new stimuli of the same or a different sensory modality. Thus, not only can bees learn specific objects and their physical parameters, but they can also master abstract inter-relationships, such as sameness and difference.

We asked whether the detection range of two-coloured centre-surround patterns differs from that of single-coloured targets. Honeybees Apis mellifera were trained to distinguish between the presence and absence of a single-coloured disc or a coloured pattern at different visual angles. The patterns presented colours which were either different in chromatic and L-receptor contrasts to the background, equal in chromatic but different in L-receptor contrasts, or vice-versa. Patterns with colours presenting only chromatic contrast were also tested. Patterns with higher L-receptor contrast in its outer than in its inner element were better detected than patterns with a reversed L-contrast distribution. However, both were detected worse than single-coloured discs of the respective colours. When the L-receptor contrast was the same for both elements, the detection range of the two-coloured and single-coloured targets was the same. Patterns whose colours lacked L-receptor contrast were detected just as single-coloured targets of the same colours. These results demonstrate that both chromatic and L-receptor contrasts mediate the detection of coloured patterns and that particular distributions of L-receptor contrast within a target are better detected than others. This finding is consistent with the intervention of neurons with centre-surround receptive fields in the detection of coloured patterns.

Honeybees have small brains, but their behavioural repertoire is impressive. In this article we focus on the extent to which adaptive behaviour in honeybees exceeds elementary forms of learning. We use the concept of modularity of cognitive functions to characterize levels of complexity in the honeybee brain. We show that behavioural complexity in the honeybee cannot be explained by independent functions of vertically arranged, domain-specific processing modules, but requires horizontal integration in a central state, and we identify neural mechanisms that may underlie domain-specific processing and central integration. The honeybee may serve as a useful model for the study of intermediate levels of complexity in cognitive functions and the search for their neural substrates.

We analysed how the assembling behaviour of Triatoma infestans is modulated by the convergence of chemical cues released by their faeces and the spectral quality of the light associated with refuges. Second-instar larvae were confronted with refuges associated with a visual stimulus (either blue, green or red lights having the same intensity, or darkness) and a chemical cue (presence or absence of faeces). In this context, faeces constitute a major attractant for bugs. In the absence of faeces, bugs always assembled in dark places. Green light was always rejected despite the presence of faeces, i.e. the assembling behaviour was controlled by a photonegative reaction to this light. In the presence of red light, orientation towards the chemical cue dominated over the bugs' photonegative reaction to this light. Such a light was avoided in the absence of faeces but not in their presence. Also, negative phototaxis to blue light could be counteracted by the presence of the chemical cue. Thus, a concrete interaction between visual and olfactory cues occurred in these experiments: the bugs' response changed depending on the specific combination of spectral light and faeces. Finally, bugs responded differentially to blue, red and green lights of the same intensity. They assembled preferentially on red, followed by blue and then by green. Thus, discrimination between lights of different spectral quality is possible, probably through an achromatic mechanism.

Floral symmetry has a relevant status in the study of both pollination biology and animal behavior. In this work, a brief review and classification of symmetry types in flowers is provided as a basis for understanding the role of floral symmetry in pollination phenomena. We focus on insects as a fundamental group of pollinators, and we discuss symmetry from the perspective of insect perception. We conclude that symmetry is a specific cue with a signal value that is perceived by insect pollinators. A simple nervous system, such as that of honeybees, is capable of an extremely flexible and adaptive processing of symmetry. Performances consistent with categorization and concept building may be observed, provided that appropriate learning paradigms are employed. Perfectly symmetrical flowers might signal a high quality and/or quantity of nectar or pollen to pollinators that, in turn, might exert strong selection pressure on symmetric features. However, coadaptation arguments in the strict sense are not adequate because it is impossible to determine whether the insect's capacity to perceive symmetry is younger or older than is the origin of flower symmetry.

In recognizing a pattern, honeybees Apis mellifera, may focus either on its ventral frontal part, or on the whole frontal image. We asked whether the conditioning procedure used to train the bees to a pattern determines the recognition strategy employed. Bees were trained with the same patterns presented vertically on the back walls of a Y maze. Conditioning was either absolute, that is, bees should learn to choose a rewarded pattern when there is no alternative, or differential, that is, bees should learn to choose a rewarded pattern that is paired with a different, nonrewarded one. Bees used different pattern recognition strategies depending on the conditioning procedure: absolute conditioning restricted recognition to the lower half whilst differential conditioning extended it to the whole pattern. Bees trained with absolute conditioning saw and learned the features of the upper part of the trained patterns, but assigned more weight to the lower part. Bees trained with differential conditioning learned not only the features of the reinforced stimulus in an excitatory way, but also those of the nonreinforced one in an inhibitory way. Thus, conditioning tasks that involve not only excitatory acquisition of the conditioned stimulus per se, but also discrimination of nonreinforced stimuli, result in an increase in the visual field assigned to the recognition task. Conditioning tasks that involve only excitatory acquisition of the rewarded stimulus result in a higher weighting of the lower pattern half and thus in a more reduced field assigned to the recognition task. This difference may reflect that existing between a conditioned and an incidental behavioural modification. Copyright 1999 The Association for the Study of Animal Behaviour.

Changes of the stinging response threshold of Apis mellifera scutellata were measured on foragers fixed on a holder and stimulated with an electric shock as a noxious stimulus. The threshold of responsiveness to the noxious stimulus increased when bees were previously stimulated with isopentyl acetate, which is a main component of the alarm pheromone of the sting chamber. This effect is antagonised by previous injection of naloxone-hydrochloride (Endo Laboratories Inc.). Results suggest that in the honeybee an endogenous opioid system activated by isopentyl acetate is responsible for modulation of perception for nociceptive stimuli. The resulting stress-induced analgesia in the defender bee would reduce its probability of withdrawal thus increasing its efficiency against enemies.

Despite their relatively simple nervous systems, insects display a rich behavioural repertoire, in which vision plays a major role. In the past two years, much knowledge has been gained about how insects are capable of a variety of flexible, visually guided tasks that involve a high level of complexity. From long-range navigation to median-range orientation and close-up recognition, insects apply different strategies that complement each other, that are used sequentially during their approach flight towards their goals, and that may replace each other, depending on the salience of, and the attention towards, particular visual cues.

Symmetrical visual patterns have a salient status in human perception, as evinced by their prevalent occurrence in art, and also in animal perception, where they may be an indicator of phenotypic and genotypic quality. Symmetry perception has been demonstrated in humans, birds, dolphins and apes. Here we show that bees trained to discriminate bilaterally symmetrical from non-symmetrical patterns learn the task and transfer it appropriately to novel stimuli, thus demonstrating a capacity to detect and generalize symmetry or asymmetry. We conclude that bees, and possibly flower-visiting insects in general, can acquire a generalized preference towards symmetrical or, alternatively, asymmetrical patterns depending on experience, and that symmetry detection is preformed or can be learned as perceptual category by insects, because it can be extracted as an independent visual pattern feature. Bees show a predisposition for learning and generalized symmetry because, if trained to it, they choose it more frequently, come closer to and hover longer in front of the novel symmetrical stimuli than the bees trained for asymmetry do for the novel asymmetrical stimuli. Thus, even organisms with comparatively small nervous systems can generalize about symmetry, and favour symmetrical over asymmetrical patterns.

Experimental evidence is reported for flower-marking by honeybees (Apis mellifera ligustica) while they were foraging on an artificial patch of flowers yielding a continuous and equal flow of sucrose solution. Honeybees marked with scent and rejected all recently visited and nectar-depleted flowers. The short fade-out time of this scent allowed discrimination of flowers that temporarily provided no food. The repellent nature of this scent mark was demonstrated by the use of an air extractor connected to the patch; when the apparatus was turned on, the rejection behaviour disappeared. The movement pattern of foraging bees also contributed to foraging efficiency, as the probability of an immediate return to the flower just abandoned was very low. However, when a quick repeat visit took place, the presence of the repellent scent-mark promoted rapid rejection.