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Developmental expression of a tyramine receptor gene in the brain of the honey bee, Apis mellifera.

This study reveals that the tyramine receptor gene, Amtyr1, is expressed in the developing brain, as well as in the brain of the adult worker honey bee. Changes in levels of Amtyr1 expression were examined using Northern analysis. Age-related increases in Amtyr1 transcript levels were observed not only during metamorphic adult development, but also in the brain of the adult worker bee. RNA in situ hybridization revealed the pattern of Amtyr1 expression. Cell bodies staining intensely for tyramine receptor-gene transcript were observed throughout the somata rind, with well-defined clusters of cells associated with developing mushroom bodies, optic lobes, and antennal lobes of the brain. Staining for Amtyr1 transcript was particularly intense within the three major divisions of mushroom body intrinsic neurons (outer compact, noncompact, and inner compact cells), suggesting that Amtyr1 is highly expressed in these structures. Activation of AmTYR1 receptors heterologously expressed in insect (Spodoptera frugiperda) cells led to a reduction in intracellular levels of cAMP similar to that reported for AmTYR1 receptors expressed in mammalian (HEK 293) cells (Blenau et al. [2000] J Neurochem 74:900-908). Taken together, these results suggest that AmTYR1 receptors may play a role in the developing brain as well as in the brain of the adult worker bee. The actions of tyramine are likely to be mediated, at least in part, via the cAMP-signaling pathway.

Animals↗

Biogenic amines in the antennal lobes and the initiation and maintenance of foraging behavior in honey bees.

Previous findings showed that high levels of octopamine and serotonin in the antennal lobes of adult worker honey bees are associated with foraging behavior, and octopamine treatment induces precocious foraging. To better characterize the relationship between amines and foraging behavior in honey bees, we performed a detailed correlative analysis of amine levels in the antennal lobes as a function of various aspects of foraging behavior. Flight activity was measured under controlled conditions in a large outdoor flight cage. Levels of octopamine in the antennal lobes were found to be elevated immediately subsequent to the onset of foraging, but they did not change as a consequence of preforaging orientation flight activity, diurnal pauses in foraging, or different amounts of foraging experience, suggesting that octopamine helps to trigger and maintain the foraging behavioral state. In contrast, levels of serotonin and dopamine did not show changes that would implicate them as either causal agents of foraging, or as neurochemical systems affected by the act of foraging. Serotonin treatment had no effect on the likelihood of foraging. These results provide further support for the hypothesis that an increase in octopamine levels in the antennal lobes plays a causal role in the initiation and maintenance of the behavioral state of foraging, and thus is involved in the regulation of division of labor in honey bees.

5-Hydroxytryptophan↗

Use of RAPD analysis for in situ identification of Ascosphaera aggregata and Ascosphaera larvis in larval cadavers of the alfalfa leafcutting bee, Megachile rotundata.

Chalkbrood of the alfalfa leafcutting bee, Megachile rotundata, is caused by the fungus Ascosphaera aggregata. We used random amplified polymorphic DNA (RAPD) analysis for the in situ identification of A. aggregata and a related species, Ascosphaera larvis, in larval cadavers of M. rotundata. A simple DNA extraction method was developed to preferentially isolate DNA from fungal spores on the cadaver surface, or from ascocysts beneath the cuticle. Similar banding patterns were obtained in A. aggregata-infected larval cadavers from different sources and geographic areas. The RAPD banding pattern of cadavers infected with A. aggregata differed from that of healthy leafcutting bee prepupae. RAPD analyses of cadavers infected with A. aggregata and A. larvis resulted in similar banding profiles as those obtained from corresponding pure fungal cultures of the two species. This suggests that the RAPD bands of infected cadavers were amplified from fungal DNA, rather than from other DNA associated with the leafcutting bee cadaver. The banding patterns of "sporulating" and "non-sporulating" chalkbrood cadavers exhibited no differences; this provides the first definitive evidence that both forms of the disease result from infection with A. aggregata.

Animals↗

Social inhibition and the regulation of temporal polyethism in honey bees.

Honey bee division of labor is characterized by temporal polyethism, in which young workers remain in the hive and perform tasks there, whereas old workers perform more risky outside tasks, mainly foraging. We present a model of honey bee division of labor based on (1) an intrinsic process of behavioral development and (2) inhibition of development through social interactions among the workers in a colony. The model shows that these two processes can explain the main features of honey bee temporal polyethism: the correlation between age and task performance; the age at which a worker first forages and how this age varies among hives; the balanced allocation of workers to hive tasks and foraging; the recovery of a colony from demographic perturbations; and the differentiation of workers into different behavioral roles. The model provides a baseline picture of individual and colony behavior that can serve as the basis for studies of more fine-grained regulation of division of labor.

Aging↗

The major opsin in bees (Insecta: Hymenoptera): A promising nuclear gene for higher level phylogenetics.

We report the phylogenetic utility of the nuclear gene encoding the long-wavelength opsin (LW Rh) for tribes of bees. Aligned nucleotide sequences were examined in multiple taxa from the four tribes comprising the corbiculate bees within the subfamily Apinae. Phylogenetic analyses of sequence variation in a 502-bp fragment (approx 40% of the coding region) strongly supported the monophyly of each of the four tribes, which are well established from previous studies of morphology and DNA. Trees estimated from parsimony and maximum likelihood analyses of LW Rh sequences show a strongly supported relationship between the tribes Meliponini and Bombini, a relationship that has been found uniformly in studies of other genes (28S, 16S, and cytochrome b). All of the tribal clades as well as relationships among the tribes are supported by high bootstrap values, suggesting the utility of LW Rh in estimating tribal and subfamily rank for these bees. The sequences exhibit minimal base composition bias. Both 1st + 2nd and 3rd position sites provide information for estimating a reliable tree topology. These results suggest that LW Rh, which has not been reported previously in studies of organismal phylogenetics, could provide important new data from the nuclear genome for phylogeny reconstruction.

Animals↗

Phylogenetic relationships among bumble bees (Bombus, latreille) inferred from mitochondrial cytochrome b and cytochrome oxidase I sequences.

We conducted a molecular study intending to derive an estimate of the relationships within the genus Bombus (bumble bees) by comparing the mitochondrial cytochrome b and cytochrome oxidase I (COI) genes from 19 species, spanning 10 of approximately 16 European subgenera and 3 subgenera from North and South America. Our trees differ from the most recent classifications of bumble bees. Although bootstrap values for deep branches are low, our sequences show significant data structure and low homoplasy, and all trees share some groups and patterns. In all cases, the subgenus Bombus s. str. clusters among the most derived bumble bees, contrary to other molecular studies. In all trees, B. funebris is the sister taxon of B. robustus, and in five of the six trees, B. wurflenii is the sister taxon to this clade. B. nevadensis is basal to the other species in the analysis of the cytochrome b gene, but appears to be among the most derived according to the analysis of the COI region. The species representing the subgenera Thoracobombus and Fervidobombus are consistently among the earliest diverged. Species that appear in very different positions in different trees are B. nevadensis, B. mesomelas, B. balteatus, and B. hyperboreus. All subgenera with two representatives in our analysis are apparently monophyletic except Fervidobombus, Melanobombus, and Pyrobombus. The groups formed by pocket makers and non-pocket makers within Bombus also appear to be paraphyletic, and therefore some subgenera may not accurately reflect phylogeny.

Animals↗

Linkage analysis of sex determination in the honey bee (Apis mellifera).

A colony-level phenotype was used to map the major sex determination locus (designated X) in the honey bee (Apis mellifera). Individual queen bees (reproductive females) were mated to single drones (fertile males) by instrumental insemination. Haploid drone progeny of an F1 queen were each backcrossed to daughter queens from one of the parental lines. Ninety-eight of the resulting colonies containing back-cross progeny were evaluated for the trait 'low brood-viability' resulting from the production of diploid drones that were homozygous at X. DNA samples from the haploid drone fathers of these colonies were used individually in polymerase chain reactions (PCR) with 10-base primers. These reactions generated random amplified polymorphic DNA (RAPD) markers that were analyzed for cosegregation with the colony-level phenotype. One RAPD marker allele was shared by 22 of 25 drones that fathered low brood-viability colonies. The RAPD marker fragment was cloned and partially sequenced. Two primers were designed that define a sequence-tagged site (STS) for this locus. The primers amplified DNA marker fragments that cosegregated with the original RAPD marker. In order to more precisely estimate the linkage between X and the STS locus, another group of bees consisting of progeny from one of the low-brood viability colonies was used in segregation analysis. Four diploid drones and 181 of their diploid sisters (workers, nonfertile females) were tested for segregation of the RAPD and STS markers. The cosegregating RAPD and STS markers were codominant due to the occurrence of fragment-length alleles. The four diploid drones were homozygous for these markers but only three of the 181 workers were homozygotes (recombinants). Therefore the distance between X and the STS locus was estimated at 1.6 cM. An additional linked marker was found that was 6.6 cM from the STS locus.

Animals↗

Allozyme variation in bumble bees (Hymenoptera: Apidae).

Allozyme variation at an average of 37.3 loci was assessed in queens of 16 Bombus and 2 Psithyrus bumble bee species from North America. The mean expected heterozygosity (H) for the Bombus species was 0.008 +/- 0.006 (95% confidence limits) and that for the Psithyrus was 0.007 +/- 0.007. These levels are significantly lower than found in other Hymenoptera but are comparable to those found in previous studies of bumble bees based on far fewer loci. Neutral mutation and random genetic drift can account for the observed variation, but this implies a very small effective population size for species of bumble bees.

Alleles↗

Genetic component in learning ability in bees.

Twenty-five bees, five from each of five hives, were trained to collect food at a table. When the bee reached the table, time was recorded for 12 visits. Then a blue and yellow pan was substituted for the original metal pan, and time and correct responses were recorded for 30 trips (discrimination phase). Finally, food was taken from the pan and extinction was recorded as incorrect responses for 20 visits. Variance analysis was carried out, and genetic variance was undetected for discrimination, but was detected for extinction. It is concluded that learning is very important for bees, so that any impairment in such ability affects colony survival.

Animals↗

Heat stress induced enhancement of heat shock protein gene activity in the honey bee (Apis mellifera).

We employed in vitro translation of mRNA and product separation using SDS-PAGE to examine the heat-shock response of the worker honey bee. Increases in the levels of 6 translatable RNA populations were observed following heat stress. The greatest response was observed among bees aged 9 days. Slight levels of induction of 70 and 82 kDa heat shock proteins were evident among bees taken directly from the colony.

Age Factors↗

[Death after bee-sting].

A ten-year-old girl collapsed on the street with the signs of acute anaphylaxis. Cardiopulmonary resuscitation was unsuccessful. At the autopsy a thorn-like object was found sticking in the tongue and was later identified as the stinger of a honey bee. Witnesses reported that the girl had drunk some ice tea from a box and then had complained about pains in the mouth. Postmortem blood tests revealed increased total IgE level as well as increased levels of specific IgE against bee and wasp poisons. Cause of death was a bronchospasm caused by an allergic reaction due to a sting of a honey bee.

Age Factors↗

African bees to control African elephants.

Numbers of elephants have declined in Africa and Asia over the past 30 years while numbers of humans have increased, both substantially. Friction between these two keystone species is reaching levels which are worryingly high from an ecological as well as a political viewpoint. Ways and means must be found to keep the two apart, at least in areas sensitive to each species' survival. The aggressive African bee might be one such method. Here we demonstrate that African bees deter elephants from damaging the vegetation and trees which house their hives. We argue that bees can be employed profitably to protect not only selected trees, but also selected areas, from elephant damage.

Aggression↗

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

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

Animals↗

Highly controlled nest homeostasis of honey bees helps deactivate phenolics in nectar.

Honey bees have a highly developed nest homeostasis, for example, maintaining low CO2 levels and stable nest temperatures at 35 degrees C.We investigate the role of nest homeostasis in deactivating phenolic compounds present in the nectar of Aloe littoralis. We show that the phenolic content in nectar was reduced (from 0.65% to 0.49%) after nectar was incubated in a nest of Apis cerana, and that it was reduced still more (from 0.65% to 0.37%) if nectar was mixed with hypopharyngeal gland proteins (HGP) of worker bees before being placed inside a nest. HGP had little effect on samples outside a nest, indicating that nest conditions are necessary for HGP to deactivate phenolics in nectar. Consequently, the highly controlled nest homeostasis of honey bees facilitates direct deactivation of phenolics in nectar, and plays a role in the action of HGP as well.

Aloe↗

Worker piping in honey bee swarms and its role in preparing for liftoff.

Worker piping, previously reported only in hives, was observed in swarms as they prepared to liftoff to fly to a new home. Pipers are excited bees which scramble through the swarm cluster, pausing every second or so to emit a pipe. Each pipe consists of a sound pulse which lasts 0.82 +/- 0.43 s and rises in fundamental frequency from 100-200 Hz to 200-250 Hz. Many. if not all, of the pipers are nest-site scouts. The scouts pipe when it is time to stimulate the non-scouts to warm themselves to a flight-ready temperature (35 degrees C) in preparation for liftoff. The time-course of worker piping matches that of swarm warming, both start at a low level, about an hour before liftoff, and both build to a climax at liftoff. When we excluded pipers from bees hanging in the cool, outermost layer of a swarm cluster, we found that these bees did not warm up. The form of worker piping that we have studied in swarms differs from the form of worker piping that others have studied in hives. We call the two forms "wings-together piping" (in swarms) and "wings-apart piping" (in hives).

Animals↗

How long will honey bees (Apis mellifera L.) be stimulated by scent to revisit past-profitable forage sites?

Honey bees utilise floral food sources that vary temporally in their relative and absolute quality. Via a sophisticated colony organisation, a honey bee colony allocates its foragers such that the colony focuses on the most profitable forage sites while keeping track of changes within its foraging environment. One important mechanism of the allocation of foragers is the ability of experienced foragers to revisit past-profitable forage sites after a period of temporary dearth caused by, for example, inclement weather. The scent of past-profitable forage within the colony brought back by other foragers is sufficient to reactivate these experienced foragers. Here I determine for how long bees react to the scent of a past-profitable forage site. I show that the ability of foragers to revisit the location of a past-profitable food source diminishes rapidly over a period of 10 days, until no forager reacts to the cue (scent). I discuss the implications of these findings with respect to the colony's ability to react rapidly to changing foraging conditions.

Animals↗

Selective modulation of task performance by octopamine in honey bee (Apis mellifera) division of labour.

Octopamine treatment has previously been shown to increase honey bee foraging behaviour. We determined the effects of octopamine on other tasks to learn how octopamine affects division of labour in honey bee colonies. Octopamine treatment did not increase the rate of corpse removal from the hive, suggesting that elevated brain levels of octopamine do not act to increase the performance of all flight-related tasks. Octopamine treatment also did not increase attendance in the queen's retinue, suggesting that elevated brain levels of octopamine do not act to increase responsiveness to all olfactory stimuli. Consistent with these findings, octopamine treatment enhanced the foraging response to brood pheromone but not the cell capping response, a component of brood care. These results demonstrate a relatively specific form of neuromodulation by octopamine in the regulation of division of labour in honey bee colonies.

Animals↗

Manipulating the light/dark cycle: effects on dopamine levels in optic lobes of the honey bee (Apis mellifera) brain.

This study examines the relationship between cyclical variations in optic-lobe dopamine levels and the circadian behavioural rhythmicity exhibited by forager bees. Our results show that changing the light-dark regimen to which bees are exposed has a significant impact not only on forager behaviour, but also on the levels of dopamine that can be detected in the optic lobes of the brain. Consistent with earlier reports, we show that foraging behaviour exhibits properties characteristic of a circadian rhythm. Foraging activity is entrained by daily light cycles to periods close to 24 h, it changes predictably in response to phase shifts in light, and it is able to free-run under constant conditions. Dopamine levels in the optic lobes also undergo cyclical variations, and fluctuations in endogenous dopamine levels are influenced significantly by alterations to the light/dark cycle. However, the time course of these changes is markedly different from changes observed at a behavioural level. No direct correlation could be identified between levels of dopamine in the optic lobes and circadian rhythmic activity of the honey bee.

Animals↗