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A family of major royal jelly proteins of the honeybee Apis mellifera L.

The characterization of major proteins of honeybee larval jelly (49-87 kDa) was performed by the sequencing of new complementary DNAs (cDNAs) obtained from a honeybee head cDNA library, by the determination of N-terminal sequences of the proteins, and by analyses of the newly obtained and known sequence data concerning the proteins. It was found that royal jelly (RJ) and worker jelly (WJ) contain identical major proteins and that all the proteins belong to one protein family designated MRJP (from Major Royal Jelly Proteins). The family consists of five main members (MRJP1, MRJP2, MRJP3, MRJP4, MRJP5). The proteins MRJP3 and MRJP5 are polymorphic. MRJPs account for 82 to 90% of total larval jelly protein, and they contain a relatively high amount of essential amino acids. These findings support the idea that MRJPs play an important role in honeybee nutrition.

Amino Acid Sequence↗

Foraging reactivation in the honeybee Apis mellifera L.: factors affecting the return to known nectar sources.

This paper addresses, what determines that experienced forager honeybees return to places where they have previously exploited nectar. Although there was already some evidence that dance and trophallaxis can cause bees to return to feed, the fraction of unemployed foragers that follow dance or receive food from employed foragers before revisiting the feeder was unknown. We found that 27% of the experienced foragers had no contact with the returning foragers inside the hive. The most common interactions were dance following (64%) and trophallaxis (21%). The great variability found in the amount of interactions suggests that individual bees require different stimulation before changing to the foraging mode. This broad disparity negatively correlated with the number of days after marking at the feeder, a variable that is closely related to the foraging experience, suggesting that a temporal variable might affect the decision-making in reactivated foragers.

Animals↗

Reassessing the role of the honeybee (Apis mellifera) Dufour's gland in egg marking.

Dufour's gland secretion may allow worker honeybees to discriminate between queen-laid and worker-laid eggs. To investigate this, we combined the chemical analysis of individually treated eggs with an egg removal bioassay. We partitioned queen Dufour's gland into hydrocarbon and ester fractions. The bioassay showed that worker-laid eggs treated with either whole gland extract, ester fraction or synthetic gland esters were removed more slowly than untreated worker-laid eggs. However, the effect only lasted up to 20 h. Worker-laid eggs treated with the hydrocarbon fraction were removed at the same rate as untreated eggs. The amount of ester which reduced the egg removal rate was far higher than that naturally found on queen-laid or worker-laid eggs, and at natural ester levels no effect was found. Our results indicate that esters or hydrocarbons probably do not function as the signal by which eggs can be discriminated.

Animals↗

Evidence for intra-colonial genetic variance in resistance to American foulbrood of honey bees ( Apis mellifera): further support for the parasite/pathogen hypothesis for the evolution of polyandry.

Explanations for the evolution of multiple mating by social insect (particularly honey bee) queens have been frequently sought. An important hypothesis is that multiple mating is adaptive because it increases intracolonial genetic diversity and thereby reduces the likelihood that parasites or pathogens will catastrophically infect a colony. We tested one assumption of this model: that honey bee worker patrilines should differ in disease resistance. We used American foulbrood (caused by the bacterium Paenibacillus larvae) as a model pathogen. We found that patrilines within colonies do indeed vary in their resistance to this disease.

Animals↗

Conservation of Bio synthetic pheromone pathways in honeybees Apis.

Social insects use complex chemical communication systems to govern many aspects of their life. We studied chemical changes in Dufour's gland secretions associated with ovary development in several genotypes of honeybees. We found that C28-C38 esters were associated only with cavity nesting honeybee queens, while the alcohol eicosenol was associated only with their non-laying workers. In contrast, both egg-laying anarchistic workers and all parasitic Cape workers from queenright colonies showed the typical queen pattern (i.e. esters present and eicosenol absent), while egg-laying wild-type and anarchistic workers in queenless colonies showed an intermediate pattern, producing both esters and eicosenol but at intermediate levels. Furthermore, neither esters nor eicosenol were found in aerial nesting honeybee species. Both esters and eicosenol are biosynthetically similar compounds since both are recognizable products of fatty acid biosynthesis. Therefore, we propose that in honeybees the biosynthesis of esters and eicosenol in the Dufour's gland is caste-regulated and this pathway has been conserved over evolutionary time.

Animals↗

Conditioning procedure and color discrimination in the honeybee Apis mellifera.

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.

Animals↗

Pheromone-modulated behavioral suites influence colony growth in the honey bee (Apis mellifera).

The success of a species depends on its ability to assess its environment and to decide accordingly which behaviors are most appropriate. Many animal species, from bacteria to mammals, are able to communicate using interspecies chemicals called pheromones. In addition to exerting physiological effects on individuals, for social species, pheromones communicate group social structure. Communication of social structure is important to social insects for the allocation of its working members into coordinated suites of behaviors. We tested effects of long-term treatment with brood pheromone on suites of honey bee brood rearing and foraging behaviors. Pheromone-treated colonies reared significantly greater brood areas and more adults than controls, while amounts of stored pollen and honey remained statistically similar. Brood pheromone increased the number of pollen foragers and the pollen load weights they returned. It appeared that the pheromone-induced increase in pollen intake was directly canalized into more brood rearing. A two-way pheromone priming effect was observed, such that some workers from the same age cohorts showed an increased and extended capacity to rear larvae, while others were recruited at significantly younger ages into pollen-specific foraging. Brood pheromone affected suites of nursing and foraging behaviors allocating worker and pollen resources associated with an important fitness trait, colony growth.

Animals↗

Larval salivary glands are a source of primer and releaser pheromone in honey bee (Apis mellifera L.).

A brood pheromone identified in honeybee larvae has primer and releaser pheromone effects on adult bees. Using gas chromatography-mass spectrometry (GC-MS) to evaluate fatty acid esters--the pheromonal compounds--in different parts of the larvae, we have localized the source of the esters as the larval salivary glands. A histochemical study describes the glands and confirms the presence of lipids in the glands. Epithelial cells of the gland likely secrete the fatty acids into the lumen of the gland. These results demonstrate the salivary glands to be a reservoir of esters, components of brood pheromone, in honeybee larvae.

Animals↗

Lower disease infections in honeybee (Apis mellifera) colonies headed by polyandrous vs monandrous queens.

We studied the relationship between genetic diversity and disease susceptibility in honeybee colonies living under natural conditions. To do so, we created colonies in which each queen was artificially inseminated with sperm from either one or ten drones. Of the 20 colonies studied, 80% showed at least one brood disease. We found strong differences between the two types of colonies in the infection intensity of chalkbrood and in the total intensity of all brood diseases (chalkbrood, sacbrood, American foulbrood, and European foulbrood) with both variables lower for the colonies with higher genetic diversity. Our findings demonstrate that disease can be an important factor in the ecology of honeybee colonies and they provide strong support for the disease hypothesis for the evolution of polyandry by social insect queens.

Animals↗

Do honeybees, Apis mellifera scutellata, regulate humidity in their nest?

Honeybees are highly efficient at regulating the biophysical parameters of their hive according to colony needs. Thermoregulation has been the most extensively studied aspect of nest homeostasis. In contrast, little is known about how humidity is regulated in beehives, if at all. Although high humidity is necessary for brood development, regulation of this parameter by honeybee workers has not yet been demonstrated. In the past, humidity was measured too crudely for a regulation mechanism to be identified. We reassess this issue, using miniaturised data loggers that allow humidity measurements in natural situations and at several places in the nest. We present evidence that workers influence humidity in the hive. However, there are constraints on potential regulation mechanisms because humidity optima may vary in different locations of the nest. Humidity could also depend on variable external factors, such as water availability, which further impair the regulation. Moreover, there are trade-offs with the regulation of temperature and respiratory gas exchanges that can disrupt the establishment of optimal humidity levels. As a result, we argue that workers can only adjust humidity within sub-optimal limits.

Animals↗

Does the spatial distribution of the parasitic mite varroa jacobsoni oud. (Mesostigmata: varroidae) in worker brood of honey bee apis mellifera L. (Hymenoptera: apidae) rely on an aggregative process?

Varroa jacobsoni is an ectoparasite of honey bees which reproduces in capped brood cells. Multi-infestation is frequently observed in worker brood and can be interpreted as an aggregative phenomenon. The aim of this study was to determine whether the distribution of V. jacobsoni in worker brood cells relies on a random or an aggregative process. We studied the distribution of Varroa females in capped worker brood at similar age by comparing, by a Monte Carlo test, the observed frequency distribution of mites per cell to simulated distributions based on a random process. A complementary approach, using the "nearest neighbor distances" (NND) with Monte Carlo tests, was investigated to study the spatial distribution (a) between mites in different cells and (b) between infested cells in brood. The observed distributions did not differ significantly from that expected by a random process, and we conclude that there is no aggregation during invasion of V. jacobsoni in worker brood.

Journal Article↗

Trophallaxis in filled-crop honeybees (Apis mellifera L.): food-loading time affects unloading behaviour.

Honeybees ingested 50% w/w (1.8 M) sucrose solution at a rate feeder offering either 16.5, 32.5 or 65 microliters/min. While the time spent ingesting solution at the feeder decreased significantly with increasing flow of solution, bees attained maximum crop loads with this range of flows. Different parameters related to mouth-to-mouth food exchange (trophallaxis) showed important modulations as the offered flow of solution was incremented. Trophallactic transfer rate, i.e. the speed at which liquid food is transferred from donor to recipient bee, was found to increase along with increasing profitability at the rate feeder. In the present case, food source profitability could have been evaluated by foragers either by measuring the time invested in ingesting the solution, or by direct assessment of the flow rate of the feeder. Thus it seems that perception of profitability conditions at the food source suffices for later representation in the hive through trophallactic contacts, independently of crop-filling state.

Animals↗

Effects of a chronic dietary exposure of the honeybee Apis mellifera (Hymenoptera: Apidae) to imidacloprid.

Previous studies have shown that imidacloprid and insecticidally active imidacloprid plant metabolites are rapidly metabolized by honeybees. Accordingly, no striking differences were expected between the acute and the chronic dietary toxicity of imidacloprid to honeybees. More recently, however, an unexpectedly high chronic dietary toxicity to honeybees was reported for imidacloprid and imidacloprid plant metabolites, and a novel pharmacologic mechanism unrelated to the parent toxophor was postulated. In an extensive literature survey, no further evidence was found for the reported high difference between the acute and the chronic dietary toxicity of imidacloprid and its plant metabolites to honeybees. The majority of data indicated a dietary no observed lethal-effect concentration >0.04 and 0.02 mg/L 50% sucrose solution, respectively, for an acute and a chronic dietary exposure of honeybees to either imidacloprid or its plant metabolites. Findings of chronic feeding studies with those plant metabolites where the toxophor had already been cleaved did not support the hypotheses of a novel pharmacologic mechanism unrelated to the parent toxophor. No increased treatment-related mortality or behavioral abnormalities were recorded in four independent research facilities during a 10-day dietary exposure of honeybees of different ages to sucrose solutions spiked with the respective metabolites at 0.0001, 0.001, and 0.010 mg/L 50% sucrose solution.

Administration, Oral↗

Effect of a new variety of Apis mellifera propolis on mutans Streptococci.

The effects of a new variety of propolis, from Northeastern Brazil (BA), on growth of mutans streptococci, cell adherence, and water-insoluble glucan (WIG) synthesis were evaluated. Propolis from Southeastern (MG) and Southern (RS) Brazil were also tested as an extension of our previous work. Ethanolic extracts of propolis (EEP) were prepared and analyzed by reversed-phase HPLC. For the antibacterial activity assays, minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of EEPs against Streptococcus mutans, S. sobrinus, and S. cricetus were determined. Cell adherence of S. mutans and S. sobrinus to a glass surface was measured spectrophotometrically at 550 nm. WIG synthesized from sucrose by glucosyltransferase (Gtf) was extracted and quantified by the phenol-sulfuric method. The HPLC profile of the new variety of propolis was entirely different from Southeastern and Southern propolis. Neither flavonoid aglycones nor p-coumaric acid were detected in EEP BA. All EEPs demonstrated biological activities against mutans streptococci; EEP BA showed the highest potency in all in vitro parameters evaluated in this study. The ranges of MIC values were 50 (EEP BA)-400 microg/ml (MG), for S. mutans; and 25 (BA)-400 microg/ml (MG), for S. sobrinus and S. cricetus. The bactericidal concentration of EEPs was four to eight times the MIC values. The adherence of S. mutans and S. sobrinus cells and WIG synthesis were markedly inhibited by EEPs, demonstrating significant inhibition at all concentrations compared with the control (80% ethanol) (p<0.05). EEP BA showed 80% inhibition of cell adherence and WIG synthesis at concentrations as low as 12.5 and 7.8 microg/ml, respectively. The results show that the new variety of propolis was exceptionally effective in all in vitro parameters tested against mutans streptococci; biological effects of propolis are likely not to be due solely to flavonoids and (hydroxy)cinnamic acid derivatives.

Animals↗