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Dancing bees tune both duration and rate of waggle-run production in relation to nectar-source profitability.

For more than 50 years, investigators of the honey bee's waggle dance have reported that richer food sources seem to elicit longer-lasting and livelier dances than do poorer sources. However, no one had measured both dance duration and liveliness as a function of food-source profitability. Using video analysis, we found that nectar foragers adjust both the duration (D) and the rate (R) of waggle-run production, thereby tuning the number of waggle runs produced per foraging trip (W, where W= DR) as a function of food-source profitability. Both duration and rate of waggle-run production increase with rising food-source profitability. Moreover, we found that a dancing bee adjusts the rate of waggle-run production (R) in relation to food-source profitability by adjusting the mean duration of the return-phase portion of her dance circuits. This finding raises the possibility that bees can use return-phase duration as an index of food-source profitability. Finally, dances having different levels of liveliness have different mean durations of the return phase, indicating that dance liveliness can be quantified in terms of the time interval between consecutive waggle runs.

Adaptation, Physiological↗

The effect of genotype, age, sex, and caste on response thresholds to sucrose and foraging behavior of honey bees (Apis mellifera L.).

Bees derived from artificially selected high- and low-pollen-hoarding strains were tested for their proboscis extension reflex response to water and varying sucrose concentrations. High-strain bees had a lower response threshold to sucrose than low-strain bees among pre-foragers, foragers, queens and drones. Pre-foraging low-strain workers showed ontogenetic changes in their response threshold to sucrose which was inversely related to age. High-strain foragers were more likely to return with loads of water compared to low-strain foragers. Whereas low-strain foragers were more likely to return with loads of nectar. Low-strain nectar foragers collected nectar with significantly higher sucrose concentrations than did the high-strain nectar foragers. Alternatively, low-strain foragers were more likely to return empty compared to high-strain foragers. These studies demonstrate how a genotypically varied sensory-physiological process, the perception of sucrose, are associated with a division of labor for foraging.

Aging↗

Acute disseminated encephalomyelitis after bee sting.

Acute disseminated encephalomyelitis (ADEM) usually follows a viral infection or an immunization and is thought to be an immunomediated disease. We describe a patient with ADEM after multiple yellow jacket bee stings. The patient recovered after treatment with a high dose of methylprednisolone. Although the pathologic mechanism exact remains unclear, potential cross-reactivity between bee toxins and the central nervous system myelin could induce demyelination. ADEM should be considered a rare complication of bee stings.

Animals↗

Observations on the honey bee tracheal mite Acarapis woodi (Acari: Tarsonemidae) using low-temperature scanning electron microscopy.

Observations were made of cryo-preserved honey bee tracheal mites Acarapis woodi (Rennie) using scanning electron microscopy. We describe various new morphological attributes of A. woodi based on the ability of the cryo-technique to capture live mites in natural positions and observe the Low-Temperature Scanning Electron Microscopy (LT-SEM) photographs under a 3-D viewer. Most striking was the observation that each leg has the ability to independently twist its segments with the ambulacrum rotating a minimum of 180 degrees during locomotion; this is a more sophisticated form of locomotion than has been proposed for the Acari. Adult daughter mites are known to be the dispersal instar moving from the tracheal tube to the thoracic hairs of the bee and then transferring to a new bee. We hypothesize that adult tarsal claws and setae on the legs play a role in attachment to hairs during dispersal. However, our evidence is that none of the life stases use their tarsal claws within the tracheal tubes. Larvae were observed to be 'freely moving' within the tracheal system, their tarsal claws rendered inoperative due to an enlarged swollen pulvillar pad. The solenidia of leg I are now known to have striations and the famulus is bifurcated. The bifurcated famulus, solenidial striations, and segmentation of leg IV of females may have taxonomic implications in the family Tarsonemidae. The body and leg setae of adults appear to be used as a tactile tool to sense the amount of space within the tracheal tubes; most of the setae are oriented distally and may help the mite to measure the space or radius of the tracheal tubes. The modified caudal region of the male revealed remnants of the h1 and h2 setae and a smooth clean surface, void of a film, supporting that pharate nymphs are not attached in this species.

Animals↗

Cuticular hydrocarbons as sex pheromone of the bee Colletes cunicularius and the key to its mimicry by the sexually deceptive orchid, Ophrys exaltata.

Male Colletes cunicularius bees pollinate the orchid, Ophrys exaltata, after being sexually deceived by the orchid's odor-mimicry of the female bee's sex pheromone. We detected biologically active volatiles of C. cunicularius by using gas chromatographic-electroantennographic detection (GC-EAD) with simultaneous flame ionization detection. After identification of the target compounds by coupled gas chromatography mass spectrometry (GC-MS), we performed behavioral tests using synthetic blends of the active components. We detected 22 EAD active compounds in cuticular extracts of C. cunicularius females. Blends of straight chain, odd-numbered alkanes and (Z)-7-alkenes with 21-29 carbon atoms constituted the major biologically active compounds. Alkenes were the key compounds releasing mating behavior, especially those with (Z)-7 unsaturation. Comparison of patterns of bee volatiles with those of O. exaltata subsp. archipelagi revealed that all EAD-active compounds were also found in extracts of orchid labella. Previous studies of the mating behavior in C. cunicularius showed linalool to be an important attractant for patrolling males. We confirmed this with synthetic linalool but found that it rarely elicited copulatory behavior, in accordance with previous studies. A blend of active cuticular compounds with linalool elicited both attraction and copulation behavior in patrolling males. Thus, linalool appears to function as a long-range attractant, whereas cuticular hydrocarbons are necessary for inducing short-range mating behavior.

Animals↗

1,4-Dimethoxybenzene, a floral scent compound in willows that attracts an oligolectic bee.

Many bees are oligolectic and collect pollen for their larvae only from one particular plant family or genus. Here, we identified flower scent compounds of two Salix species important for the attraction of the oligolectic bee Andrena vaga, which collects pollen only from Salix. Flower scent was collected by using dynamic-headspace methods from Salix caprea and S. atrocinerea, and the samples were subsequently analyzed by coupled gas chromatographic-electroantennographic detection (GC-EAD) to detect possible attractants of A. vaga. EAD active compounds were identified by gas chromatography coupled to mass spectrometry. Both Salix species had relatively similar scent profiles, and the antennae of male and female bees responded to at least 16 compounds, among them different benzenoids as well as oxygenated monoterpenoids and sesquiterpenoids. The strongest antennal responses were triggered by 1,4-dimethoxybenzene, and in field bioassays, this benzenoid attracted females of A. vaga at the beginning of its flight period, but not at the end.

Animals↗

Hexyl decanoate, the first trail pheromone compound identified in a stingless bee, Trigona recursa.

Foragers of many species of stingless bees guide their nestmates to food sources by means of scent trails deposited on solid substrates between the food and the nest. The corresponding trail pheromones are generally believed to be produced in the mandibular glands, although definitive experimental proof has never been provided. We tested the trail following behavior of recruits of Trigona recursa in field experiments with artificial scent trails branching off from natural scent trails of this stingless bee. First-time recruits (newcomers) did not follow these trails when they were laid with pure solvent or mandibular gland extract. However, they did follow trails made with labial gland extract. Chemical analyses of labial gland secretions revealed that hexyl decanoate was the dominant component (72.4 +/- 1.9% of all volatiles). Newcomers were significantly attracted to artificial trails made with synthetic hexyl decanoate, demonstrating its key function in eliciting scent-following behavior. According to our experiments with T. recursa, the trail pheromone is produced in the labial glands and not in the mandibular glands. Hexyl decanoate is the first component of a trail pheromone identified and proved to be behaviorally active in stingless bees.

Animal Communication↗

Characteristics of neuron activity in the honey bee (Apis mellifera L.) in conditions of kynurenine deficiency.

Neuron activity was studied electrophysiologically in the antennal lobes, mushroom bodies, and cervical connective during ontogenesis (days 1, 3, 7, and 25) in snow(laranija) mutant bees, which suffer kynurenine deficiency, and in wild-type bees. Neuron activity recorded from the cervical connective was found to show the greatest dependence on kynurenine content. Mutation in the structural gene for tryptophan oxygenase, the first and key enzyme in the kynurenine pathway for tryptophan metabolism (KPTM) and which leads to kynurenine deficiency, decreased the frequency of neuron spike activity recorded from the cervical connective. This would appear to be associated with a possible deficiency of glutamic acid; it was independent of the deficiency of kynurenic acid and kynurenine, a point subject to further studies. Clear manifestation of the effect of the mutation requires the presence of two mutant alleles in the bee genome (homozygosity). The effect of the mutation, to inhibit neuron activity in the cerebral ganglion, corresponds to the effect seen previously at the behavioral level. Less clear effects were obtained from recordings of neuron activity from the antennal lobes and mushroom bodies. This may be associated with the complexity of the spectrum of biochemical lesions in different parts of the brain resulting from the mutation.

Action Potentials↗

In vitro biosynthesis of juvenile hormone in larval honey bees: comparison of six media.

We have formulated a tissue culture medium based on the components of larval honey bee hemolymph. Using an in vitro radiochemical assay to measure juvenile hormone biosynthesis, we compared our larval-based medium to four commercially available media (Grace's, Medium-199; Shields and Sang M3, and Minimum Essential Medium), and a medium based on adult honey bee hemolymph. All media were formulated without methionine. There was no significant difference in the amounts of juvenile hormone produced by the larval medium and Grace's; both of these media, however, were more suitable than the remaining four. Our larval-based tissue culture medium should prove useful in studies aimed at elucidating the underlying hormonal mechanism(s) of caste development in honey bees.

Animals↗

High-level expression in Escherichia coli and rapid purification of enzymatically active honey bee venom phospholipase A2.

Bee venom phospholipase A2 (BV-PLA2) is a hydrolytic enzyme that specifically cleaves the sn-2 acyl bond of phospholipids at the lipid/water interface. The same enzyme is also believed to be responsible for some systemic anaphylactic reactions in bee venom sensitized individuals. To study the structure/function relationships of this enzyme and to define the molecular determinants responsible for its allergenic potential, a synthetic gene encoding the mature form of BV-PLA2 was expressed in Escherichia coli. This enzyme was produced as a fusion protein with a 6xHis-tag on its amino-terminus yielding 40-50 mg of fusion protein per 1 of culture after metal ion affinity chromatography. A kallikrein protease recognition site was engineered between the 6xHis-tag and the amino-terminus of the enzyme allowing isolation of the protein with its correct N-terminus. Recombinant affinity purified BV-PLA2 was refolded, purified to homogeneity, and cleaved with kallikrein, resulting in a final yield of 8-9 mg of active enzyme per 1 of culture. The enzymatic and immunological properties of the recombinant BV-PLA2 are identical to enzyme isolated from bee venom indicating a native-like folding of the protein.

Amino Acid Sequence↗

Two-step inactivation of bee venom phospholipase A2 by scalaradial.

Scalaradial (SLD), a marine natural product isolated from the sponge (Cacospongia sp., possesses anti-inflammatory properties in vivo and in vitro (Pharmacologist 32: 168, 1990). In this study we characterize its effects against bee venom phospholipase A2 (PLA2; EC 3.1.1.4). SLD is a potent inactivator of bee venom PLA2 with an IC50 value of 0.07 microM. Inactivation of bee venom PLA2 occurred in a time-dependent, irreversible manner. The rate of inactivation followed first-order reaction kinetics and was dependent on the concentration of SLD. Kinetic analysis suggested a two-step mechanism of inactivation: an initial apparent noncovalent binding (Ki = 4.5 x 10(-5) M) followed by covalent modification. The rate of inactivation was reduced markedly in the presence of excess phosphatidylcholine, suggesting that modification of the enzyme occurs at or near the substrate binding site.

Anti-Inflammatory Agents↗

A method to generate antigen-specific suppressor T cells in vitro from peripheral blood T cells of honey bee venom-sensitive, allergic patients.

Peripheral blood mononuclear cells of patients allergic to honey bee venom were stimulated with denatured bee venom phospholipase A2, and the antigen-activated T cells were propagated for 4 days by human IL-2 in the presence or absence of recombinant human lipocortin I. Upon antigenic stimulation with the denatured phospholipase A2 and autologous monocytes or by cross-linking of CD3 by anti-CD3 antibody, the activated T cells, which had propagated by IL-2 alone, formed N-glycosylated IgE-binding factors and glycosylation enhancing factor (GEF), while those propagated in the presence of lipocortin formed unglycosylated IgE-binding factors and glycosylation inhibiting factor (GIF). The GEF and GIF formed by the antigen- or anti-CD3-stimulated T cells had affinity for bee venom phospholipase A2 and could be purified by using anti-lipomodulin Sepharose. In the mouse lymphocyte system, the major cell source of GIF is antigen-specific suppressor T cells, and the antigen-binding GIF from the cells suppressed the in vivo antibody response in an antigen (carrier)-specific manner. In view of the findings in the mouse system, the present results may provide an immunological maneuver to generate allergen-specific suppressor T cells, and to obtain allergen-specific suppressor factor from T cell populations in the peripheral blood of allergic patients.

Annexins↗

The glycoprotein nature of phospholipase A2, hyaluronidase and acid phosphatase from honey-bee venom.

Experiments with immobilized concanavalin A strongly suggest a glycoprotein nature of three honey-bee venom enzymes, phospholipase A2, hyaluronidase and acid phosphatase. The electrophoretically and chromatographically detectable heterogeneity of phospholipase A2 results from absence of carbohydrate in a subfraction. Mannose, fucose and N-acetylglucosamine, but not galactose nor N-acetylgalactosamine, are present in the con A-binding fraction of bee venom. It is therefore concluded that only N-glycosidically linked carbohydrate occurs in bee venom glycoproteins.

Acid Phosphatase↗

Hydrolysis of short-chain phosphatidylcholines by bee venom phospholipase A2.

In order to find out the aggregation state of the substrate, preferred by bee venom phospholipase A2 (EC 3.1.1.4), its action on short-chain phosphatidylcholines with two identical (C6-C10) fatty acids has been tested. The rate of hydrolysis as a function of acyl chain length showed a maximum at dioctanoylphosphatidylcholine. The effects of alcohols, NaCl and Triton X-100, which affect the aggregation state of phospholipids in water, were also studied. The addition of n-alcohol led to a significant inhibition of the hydrolysis of the substrates present in micellar form and activated the hydrolysis of substrates which form liposomes. The inhibitory effect increased with increasing length of the aliphatic carbon chain of the alcohol. Triton X-100 at low Triton/phospholipid molar ratios enhanced enzyme activity. These results do not agree with the accepted idea that bee venom phospholipase A2 hydrolyzes short-chain lecithins in their molecularly dispersed form and that micelles cannot act as substrates. The data indicate that short-chain lecithins in the aggregated state are hydrolyzed and that the requirements of bee venom phospholipase A2 for the aggregation state of the substrate are not strict.

Alcohols↗

Contribution of bee venom phospholipase A2 contamination in melittin fractions to presumed activation of tissue phospholipase A2.

Melittin from bee venom has been suggested to activate tissue phospholipase A2 (PLA2) activity, and subsequently has been used as a specific PLA2 probe. The melittin in most cases was obtained commercially and used without further purification or treatment. To test the hypothesis that commercially obtained melittin specifically activates tissue PLA2, we radiolabeled the lipids of immortalized epithelial cells by incubating the cells for 22 hr with 14C-linoleic acid. The cells were then incubated with 2 microM melittin, 2nM bee venom PLA2, 2 microM melittin treated with p-bromophenacyl bromide (p-BPB) or PLA2 plus p-BPB-treated melittin. Lipids were extracted and separated by thin-layer chromatography. The radioactivity in each lipid fraction was then quantitated. The melittin-stimulated PLA2 activity observed in cells was primarily associated with phosphatidylcholine. Fatty acid release was decreased by 75% when the melittin fraction was pretreated with p-BPB to reduce contaminating venom PLA2 activity. Adding PLA2 to the p-BPB-treated melittin at an amount about equal to the original contamination (0.1%) resulted in the same PLA2 activity in cell as observed with the untreated melittin fraction. These findings suggest that bee venom PLA2 contamination, even at very low levels, can account for approximately 75% of the PLA2 activity in cells treated with commercial melittin fractions.

Acetophenones↗

Allergens in bee venom II. Two new high molecular weight allergenic specificities.

Two new allergenic specificites were detected in honeybee venom and the two corresponding protein substances isolated by gel filtration, immunoadsorption, and ion exchange chromatography. The first of these, allergen B, has a molecular weight ranging from 49,000 to more than 200,00 d and can be recognized by rabbit and guinea pig antisera as well as by human reaginic sera using the radioallergosorbent test (RAST). Allergen B gives a single line in immunodiffusion distinct from hyaluronidase, phospholipase A, melittin, and the other high molecular weight substances described and gives a single band at 49,000 d in sodium dodecyl sulfate (SDS) polyacrylamide gel. The second substance, allergen C, has a molecular weight of 105,000 d and was separated from allergen B by immunoadsorption with insoluble antibody. Allergen C was shown to be distinct from the other sustances in bee venom by immunodiffusion with animal antisera. One human reaginic serum was monospecific for allergen C. Two other minor components of 86,000 and 71,000 d are present in bee venom; their allergenic activities are unknown. The two specifities, B and C, comprise most of the reactivity of the previously described Sephadex G-75 fraction 1 and clearly are important allergens, reacting with 98% of sera from bee venom-allergic individuals.

Allergens↗

Partitioning non-linearities in the response of honey bee olfactory receptor neurons to binary odors.

In many organisms, of which honey bees are one example, a general (i.e., non-pheromonal) olfactory receptor neuron may respond to some odorants by increasing its firing rate and to others by decreasing its firing rate. In the latter case, this decrease will be with respect to a background firing rate determined by intrinsic (internal noise) and extrinsic (background odors) factors. To analyse receptor neurons of this complexity, we extend Beidler's model of receptor protein activation dynamics to account for the competition between depolarizing and hyperpolarizing pathways and couple the model to a phenomenological description of the non-linear relationship between the proportion of activate membrane receptors and the receptor cell spike generation rates. We then examine the implications of this theory for predicting the response of receptor neurons to odor mixtures based on their response to pure odorants at concentrations matched to the mixture. We derive inequalities that must be satisfied under our normative model, and propose that deviations from the model be designated as synergisms and inhibitions, depending on the direction in which various equalities and inequalities are violated. We then apply our inequalities to identifying synergisms and inhibitions in data analysed in a different way elsewhere (Akers, R.P. and Getz, W.M. Response of olfactory receptor neurons in honey bees to odorants and their binary mixtures. J. Comp. Physiol. (in press)). In these data regarding the response of honey bee placode sensilla to a number of odorants and their binary combinations, we demonstrate the presence of synergisms and inhibitions--that is, elevated or repressed responses that are not due to competitive interactions of mixture component odorants for receptor sites or Beidler (Beidler, L.M., 1962. Taste receptor stimulation. Prog. Biophys. Biophys. Chem. 12, 107-151) saturation mechanisms.

Animals↗

Phototactic behaviour correlates with gustatory responsiveness in honey bees (Apis mellifera L.).

The response threshold hypothesis of division of labour in honey bees assumes that individuals differ in their responsiveness to different stimulus modalities. However, previous experiments have shown that responsiveness to gustatory stimuli correlates with responsiveness to odours, pollen and tactile stimuli. Evaluation of these stimuli involves sensory receptors on the antenna. We tested whether responsiveness to gustatory stimuli correlates with responsiveness to visual stimuli in a phototaxis experiment, which is independent of antennal input. Gustatory responsiveness was measured using the proboscis extension response to antennal stimulation with water and different sucrose concentrations. Phototaxis was quantified by measuring the walking times a bee needed to reach light sources of different intensities. Walking behaviour in the darkness was measured to test for differences in locomotor behaviour. The walking time towards a light stimulus, the path length, and the walking speed depended on the intensity of the light stimulus. Responsiveness to visual stimuli correlated significantly with gustatory responsiveness. Bees displaying a high gustatory responsiveness were also very sensitive to light. Locomotor activity did not correlate with gustatory responsiveness. This shows that gustatory responsiveness is a good indicator of sensitivity for visual stimuli, which are not perceived by the antenna.

Analysis of Variance↗