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Incentive contrast in honey bees.

Bees trained to come to the laboratory for a 20% sucrose solution accept it readily, but bees trained with a 40% sucrose solution and tested with the 20% solution show a pattern of interrupted feeding that may last for several minutes. Bees trained with 20% and tested with 40% sucrose are undisturbed. When the animals are offered two samples of the 20% solution simultaneously, they drink to repletion from whichever they first taste on each visit, but if both a 20% and a 40% drop are offered the 20% solution is rejected after a single experience of the 40% solution. Although these results are analogous in many respects to incentive contrast effects found in mammals, they can be understood in sensory terms and do not require the assumption of learning about reward.

Animals↗

Honey bee recruitment to food sources: olfaction or language?

Honey bee recruits locate food sources by olfaction and not by use of distance and direction information contained in the recruitment dance. Recruitment efficiency increases as odor of the food source accumulates in the hive, from hour to hour and from day to day. Flight patterns, landing patterns, bee odor, and Nassanoff secretion apparently do not aid in recruitment of bees.

Animal Communication↗

Crystal structure of bee-venom phospholipase A2 in a complex with a transition-state analogue.

The 2.0 angstroms crystal structure of a complex containing bee-venom phospholipase A2 (PLA2) and a phosphonate transition-state analogue was solved by multiple isomorphous replacement. The electron-density map is sufficiently detailed to visualize the proximal sugars of the enzyme's N-linked carbohydrate and a single molecule of the transition-state analogue bound ot its active center. Although bee-venom PLA2 does not belong to the large homologous Class I/II family that encompasses most other well-studied PLA2s, there is segmental sequence similarity and conservation of many functional substructures. Comparison of the bee-venom enzyme with other phospholipase structures provides compelling evidence for a common catalytic mechanism.

Amino Acid Sequence↗

Bee venom enhances guanylate cyclase activity.

Bee venom and phospholipase A2 extracted from bee venom enhanced guanylate cyclase (E.C. 4.6.1.2) activity two- to threefold in rat liver, lung, heart, kidney, ileum, and cerebellum. Dose-response relationships revealed that bee venom at concentrations as low as 1 microgram per milliliter and phospholipase A2 at 1 microunit per milliliter caused a maximal enhancement of guanylate cyclase.

Animals↗

Helical, motile mycoplasmas associated with flowers and honey bees in California.

Spiroplasmas were cultured from nonsurface-sterilized flowers of magnolia tree (Magnolia grandiflora L.) and tulip tree (Liriodendron tulipfera L.) in Alameda, Solano, and Yolo counties in California. Spiroplasmas were also isolated from honey bees (Apis mellifera L.) collected in the vicinity of Davis and Berkeley, CA. Most of the isolates grew relatively slowly at 31 or 37 degrees C reaching maximum population at 31 degrees C in 8-10 days. The flower isolates were serologically indistinguishable by deformation and growth inhibition tests of spiroplasma strain AS-576 from honey bee from Maryland. Isolates from honey bees were serologically closely related to spiroplasma strain SE-3 from tulip tree flower from Connecticut.

Bees↗

Properties of a free and a solubilized form of bound alpha,alpha-trehalase purified from honey bee thorax.

The free and bound forms of alpha,alpha-trehalase (EC 3.2.1.28) of the honey bee thorax were separated and the bound enzyme was solubilized by raising the pH to 8.0 for 10 h. Both enzymes were purified. They were homogeneous as determined by several electrophoretic criteria. It was found that the two enzymes had very similar Km's (each about 0.89 mM), Vm's (53.2 and 54.3 U/mg for free and solubilized, respectively), inhibition characteristics, specificities (both only hydrolyzed alpha,alpha-trehalose), pH maxima (each had maxima at about 3.5 and 6.5), molecular weights (65,000), isoelectric points (5.1), reactivities to sulfhydryl reagents, electrophoretic mobilities, activation energies (about 12.8 kcal/mol), and similar stabilities to heat, pH, and urea. Some significant differences between the two enzymes were, however, found: the solubilized alpha,alpha-trehalase floated at 70% saturation of ammonium sulfate while the free alpha,alpha-trehalase did not; the solubilized alpha,alpha-trehalase did not dissociate into subunits as readily as did the free one; and the solubilized alpha,alpha-trehalase was found to bind more readily to a hydrophobic grouping than the free enzyme. In addition to these comparisons, three new findings relating to thorax alpha,alpha-trehalases are reported. (1) Thorax alpha,alpha-trehalases are strongly inhibited by beta-glucosides (Ki values of about 8 x 10(-4) M); (2) under certain conditions thorax alpha,alpha-trehalases from honey bees dissociated into subunits of one-half the normal molecular weight; (3) honey bee thorax alpha,alpha-trehalases have unusual biphasic pH activity profiles.

Animals↗

The localization of honey bee thorax trehalase.

Differential and sucrose gradient centrifugation of honey bee thoraces, disrupted by gentle methods and using mannitol-triethanolamine-EDTA buffer at pH 6.5, showed that in the honey bee thorax 92-94.8% of the trehalase was mitochondrial. Since only 92-95% of the cytochrome c oxidase, a known mitochondrial enzyme, was found in the mitochondrial fraction by these methods, it was concluded that honey bee trehalase is totally mitochondrial. Significant amounts of 'microsomal' or 'soluble' trehalase were formed only by harsh methods of thorax disruption and similar 'microsomal' or 'soluble' trehalases were also formed by harsh treatment of purified whole mitochondria. They thus seem to be artifacts of the isolation procedure. Studies (using marker enzymes) with purified intact mitochondria which were dispersed by various chemical, enzymatic, and physical methods showed that the trehalase in the mitochondria was membrane bound and that it was bound to either the outside of the inner membrane or to one of the sides of the outer membrane.

Animals↗

The use of bee venom melittin to assess the topography of membrane vesicles derived from Paracoccus denitrificans.

There exists considerable controversy regarding membrane topography in vesicles derived by osmotic lysis of spheroplasts of Gram-negative bacteria. It has been reported by others that bee venom can be used to quantitate the portion of a heterogeneous vesicle population with an inside-out orientation by determining the degree of loss of crypticity of NADH dehydrogenase activity. We have demonstrated that a major component of bee venom, melittin, causes an increase in the activity of several different respiratory enzymes in isolated membrane vesicles of Paracoccus denitrificans. The degree of stimulation produced by melittin is dependent upon (i) the nature of the respiratory substrates, (ii) the pH, (iii) the presence of Mg2+, (iv) the melittin: membrane protein ratio, and (v) the growth history of the cells from which the membrane vesicles were derived. Melittin-induced enhancement of TMPD:ascorbate and cytochrome c oxidase activities cannot be accounted for by increased accessibility of nonpermeant substrate to the interior of the vesicle. The stimulatory effect of melittin may rely in part on its ability to alter the proton permeability of the membrane thereby abolishing respiratory control. Collectively these observations call into question the usefulness of bee venom melittin in quantitative analyses of membrane topography. These results are consistent with the postulated existence of a homogeneous vesicle population in which the topography of the NADH dehydrogenase is different from that of the intact cell.

Bee Venoms↗

Changing paradigms in insect social evolution: insights from halictine and allodapine bees.

Until the 1980s theories of social insect evolution drew strongly on halictine and allodapine bees. However, that early work suffered from a lack of sound phylogenetic inference and detailed information on social behavior in many critical taxa. Recent studies have changed our understanding of these bee groups in profound ways. It has become apparent that forms of social organization, caste determination, and sex allocation are more labile and complex than previously thought, although the terminologies for describing them are still inadequate. Furthermore, the unexpected complexity means that many key parameters in kin selection and reproductive skew models remain unquantified, and addressing this lack of information will be formidable. At the same time, phylogenetic questions have become more tractable, and DNA sequence-based studies have resolved questions that earlier studies could not resolve, radically changing our understanding of the number of origins and losses of sociality in these bees.

Animals↗

The water-soluble fraction (<10 kD) of bee venom (Apis mellifera) produces inhibitory effect on apical transporters in renal proximal tubule cells.

Human envenomation caused by bee stings has been reported to cause acute renal failure and the pathogenetic mechanisms of these renal functional changes are still unclear. Bee venom is also a complex mixture of enzymes and proteins. Thus, this study was conducted to examine the effects of bee venom (BV, Apis mellifera) fractions on apical transporters' activity and its related signal pathways in primary cultured renal proximal tubule cells. Whole BV was extracted into three fractions according to solubility [a water-soluble fraction (BVA), an ethylacetate-soluble fraction (BVE), and a hexane-soluble fraction (BVH)]. BVA fraction was further separated to three portions according to molecular weights: BF1 (>20 kD), BF2 (10-20 kD), and BF3 (<10 kD). Each fraction was treated to the PTCs to the ratio of BV (1 microg/ml). BVA (930 ng/ml) significantly decreased cell viability, but BVH (27 ng/ml) and BVE (43 ng/ml) did not. BF3 (710 ng/ml) among BVA fractions predominantly decreased cell viability and inhibited alpha-methyl-D-glucopyranoside (alpha-MG), phosphate (Pi), and Na(+) uptake. In addition, BF3 increased [(3)H] arachidonic acid release, lipid peroxide formation, and Ca(2+) uptake. These effects of BF3 were blocked by mepacrine and AACOCF(3) (phospholipase A(2) inhibitors) or N-acetylcysteine, vitamin C, and vitamin E (antioxidants). In conclusion, BF3 (<10 kD) among BV fractions is the most effective portion in BV-induced inhibition of alpha-MG, P(i), and Na(+) uptake and these effects of BF3 are associated with phospholipase A(2)-oxidative stress-Ca(2+) signal cascade in the primary cultured rabbit renal proximal tubule cells.

Animals↗

Characterization of nucleic acids associated with Arkansas bee virus.

Arkansas bee virus (ABV) is a 30-nm isometric virus composed of one major species of polypeptide (mol. wt. 43 X 10(3] and one species of single-stranded RNA (mol. wt. 1.8 X 10(6]. The size of the genomic RNA and the lack of evidence for encapsidation or synthesis of a second smaller RNA species exclude further consideration of this virus for inclusion with the physicochemically similar viruses of the Nodaviridae. Several independent isolations of ABV were made from bees, and in each case it was associated in mixed infection with a picorna-like virus termed Berkeley bee picornavirus (BBPV). No evidence for replicative dependence, serological relationship, or genome relatedness by complementary DNA hybridization was obtained between ABV and BBPV.

Animals↗

IgG and IgE antibodies after immunotherapy with bee and wasp venom.

IgG and IgE antibody levels have been followed for a period of 2 years in patients receiving immunotherapy with bee and wasp venom. 106 adult patients who had had anaphylactic reactions to wasp stings had initially low IgG antibody levels to wasp venom which rose with therapy (p less than 0.001). IgE antibody levels also showed an initial rise but subsequently fell (p less than 0.001). The pattern was similar to that previously reported in children who had had anaphylactic reactions to bee stings, but who, after a course of immunotherapy, were able to tolerate stings with impunity. 60 adults who had had anaphylactic reactions to bee stings showed a different pattern, with initially high IgG antibody levels which did not rise further. Since two thirds of this group were beekeepers or members of beekeeper's families, the high initial IgG antibody levels could have been a response to the frequent stings to which such individuals are prone. The fact that high levels did not protect against anaphylaxis shows, however, that the classical concept of 'blocking antibody' is in need of revision.

Bee Venoms↗

Appearance of macrophage migration inhibition factor in patients with systemic reactions to bee venom.

Cellular responses in bee venom (BV) allergy is a controversial issue. Previous studies could not reach an agreement whether this mechanism is activated as a result of allergic sensitization to bee venom. All previous works have used lymphocyte proliferation as their method to analyze cell-mediated immunity. In the present work, we tried to explore whether the production of macrophage migration inhibition factor (MIF), which is another in vitro correlate with cellular responses, is increased in these patients. We also examined which of the major antigenic components of BV played a significant role in the cellular response. Peripheral blood lymphocytes from 10 patients with systemic allergic reactions to bee sting and 9 healthy volunteers were examined for their ability to induce positive MIF responses. Macrophage inhibition was significantly increased in allergic patients when tested with BV, phospholipase A2 (PLA2) and with melittin. Positive MIF responses to other components were also more common in allergic patients than in the control group. Our results indicate that cellular response to BV is expressed in patients with systemic allergic reaction to BV. When major antigenic components of BV are examined, PLA2 seems to play the major role in inducing this response.

Acid Phosphatase↗

Mechanism of action of honey bee (Apis mellifera L.) venom on different types of muscles.

1. The effect of crude honeybee (Apis mellifera) venom on the skeletal, smooth as well as cardiac muscles were studied in this investigation. 2. Perfusion of gastrocnemius-sciatic nerve preparation of frogs with 1 microgram/ml venom solution has weakened the mechanical contraction of the muscle without recovery. Blocking of nicotinic receptors with 3 micrograms/ml flaxedil before bee venom application sustained normal contraction of gastrocnemius muscle. 3. The electrical activity of duodenum rabbits was recorded before and after the application of 1 microgram/ml venom solution. The venom has depressed the amplitude of the muscle contraction after 15 min pretreatment with atropine nearly abolished the depressor effect of the venom on smooth muscle. 4. In concentrations from 0.5-2 micrograms/ml, bee venom caused decrease of heart rate of isolated perfused toad heart. This bradycardia was accompanied by elongation in the P-R interval. A gradual and progressive increase in the R-wave amplitude reflected a positive inotropism of the venom. Application of 5 micrograms/ml verapamil, a calcium channels blocking agent, abolished the noticed effect of the venom. 5. Marked electrocardiographic changes were produced within minutes of the venom application on the isolated perfused hearts, like marked injury current (elevation or depression of the S-T segment), atrioventricular conduction disturbances and sinus arrhythmias. Atropine and nicotine could decrease the toxic effect of the venom on the myocardium. 6. Results of the present work lead to the suggestion that bee venom is mediated through the peripheral cholinergic neurotransmitter system. General neurotoxicity of an inhibitory nature involving the autonomic as well as neuromuscular system are established as a result of the venom, meanwhile a direct effect on the myocardium membrane stabilization has been suggested.

Animals↗

Visual resolution of gratings by the compound eye of the bee Apis mellifera.

Early measurements of the resolution of horizontal versus vertical gratings were confirmed, with a limit near a period of 2.5 degrees, and the resolution is similar when vertical or horizontal gratings are tested separately against grey. Bees were next trained to discriminate from a distance between gratings at 45 degrees versus 135 degrees, with no green contrast, on targets presented in a vertical plane at a fixed distance. As expected, they fail to learn; however, with green contrast but no modulation difference the resolution limit is near 3.5 degrees. With vertical and horizontal gratings with no green contrast they discriminate but do not learn an orientation cue. In order to eliminate the orientation cue altogether, new bees were then trained with alternating vertical and horizontal gratings versus grey, or with a black and white checkerboard versus grey. Tests of these trained bees with horizontal or with vertical gratings separately against grey again show a resolution down to a period near 2.5 degrees. These results, taken together, show that when edge orientation alone is the cue, the limit of resolution is near 3.5 degrees, but when receptor modulation is the cue, the limit is near 2.5 degrees.

Animals↗

Allometry of maximum vertical force production during hovering flight of neotropical orchid bees (Apidae: Euglossini).

The ability of orchid bees to generate vertical forces was evaluated using a load-lifting method that imposed asymptotically increasing loads during ascending flight, ultimately eliciting maximum forces while hovering. Among 11 orchid bee species varying by approximately an order of magnitude in body mass, the capacity to produce vertical forces expressed relative either to body weight or to flight muscle weight declined linearly with increased body mass. Allometric analysis of log-transformed data, by contrast, found maximum vertical force to scale isometrically with body mass, but also to exhibit a slightly negative allometry with respect to flight muscle mass. Maximum stroke amplitude at limiting loads averaged 140 degrees and was remarkably constant among species, a result consistent with anatomical constraints of the hymenopteran thorax on wing motions. By contrast, wing-beat frequencies during maximum performance declined with increasing body mass. Repeated lifting by individual bees reduced performance only when the number of consecutive lifts exceeded 15. Variation in linear mass density of the lifted load did not systematically alter performance estimates, although measurements on one species in two consecutive years at different thermal environments yielded significant differences in estimates of maximum force production. These findings suggest an adverse scaling of vertical force production at greater body mass even if flight muscle mass scales isometrically.

Animals↗

Thermal stability and muscle efficiency in hovering orchid bees (Apidae: Euglossini).

To test whether variation in muscle efficiency contributes to thermal stability during flight in the orchid bee, Euglossa imperialis, we measured CO2 production, heat loss and flight kinematics at different air temperatures (Ta). We also examined the relationship between wingbeat frequency (WBF) and Ta in five additional species of orchid bees. Mean thoracic temperature (Tth) for Eg. imperialis hovering in a screened insectary and in the field was 39.3+/-0.77 degrees C (mean +/- 95% C.I.), and the slope of Tth on Ta was 0.57. Head and abdominal temperature excess ratios declined with Ta, indicating that Eg. imperialis were not increasing heat dissipation from the thorax at high Ta. Elevation of Tth above Ta was correlated with WBF, but Tth alone was not. Estimates of heat production from both respirometry and heat loss experiments decreased 33% as Ta rose from 24 to 34 degrees C. Mean muscle efficiency over this temperature range was 18% assuming perfect elastic energy storage and 22% assuming zero elastic energy storage. Both efficiency estimates increased significantly as Ta rose from 24 to 34 degrees C. In all six species examined, WBF declined significantly with Ta. These data indicate that hovering orchid bees regulate heat production through changes in wingbeat kinematics and consequent changes in energy conversion by the flight motor. Temperature-dependent variation in elastic energy storage or muscle contraction efficiency or both may contribute to the observed trends.

Animals↗

Energy metabolism in orchid bee flight muscles: carbohydrate fuels all.

The widely accepted idea that bees fuel flight through the oxidation of carbohydrate is based on studies of only a few species. We tested this hypothesis as part of our research program to investigate the size-dependence of flight energetics in Panamanian orchid bees. We succeeded in measuring rates of O(2) consumption and CO(2) production in vivo during hovering flight, as well as maximal activities (V(max) values) in vitro of key enzymes in flight muscle energy metabolism in nine species belonging to four genera. Respiratory quotients (ratios of rates of CO(2) production to O(2) consumption) in all nine species are close to 1.0. This indicates that carbohydrate is the main fuel used for flight. Trehalase, glycogen phosphorylase and hexokinase activities are sufficient to account for the glycolytic flux rates estimated from rates of CO(2) production. High activities of other glycolytic enzymes, as well as high activities of mitochondrial oxidative enzymes, are consistent with the estimated rates of carbohydrate-fueled oxidative metabolism. In contrast, hydroxyacylCoA dehydrogenase, an enzyme involved in fatty acid oxidation, was not detectable in any species. Thoracic homogenates displayed ADP-stimulated oxidition of pyruvate + proline, but did not oxidize palmitoyl l-carnitine + proline as substrates. A metabolic map, based on data reported herein and information from the literature, is presented. The evidence available supports the hypothesis that carbohydrate serves as the main fuel for flight in bees.

3-Hydroxyacyl CoA Dehydrogenases↗