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The dice of fate: the csd gene and how its allelic composition regulates sexual development in the honey bee, Apis mellifera.

Perhaps 20% of known animal species are haplodiploid: unfertilized haploid eggs develop into males and fertilized diploid eggs into females. Sex determination in such haplodiploid species does not rely on a difference in heteromorphic sex chromosome composition but the genetic basis has been elucidated in some hymenopteran insects (wasps, sawflies, ants, bees). In these species, the development into one sex or the others depends on an initial signal whether there is only one allele or two different alleles of a single gene, the complementary sex determiner (csd), in the zygotic genome. The gene has been most-recently identified in the honey bee and has been found to encode an arginine serine-rich (SR) type protein. Heterozygosity generates an active protein that initiates female development while hemizygosity/homozygosity results in a non-active CSD protein and default male development. I will discuss plausible models of how the molecular decision of male and female is made and implemented. Comparison to hierarchies of dipteran insects suggests that SR-type protein has facilitated the differentiation of sex-determining systems and hierarchies.

Alleles↗

DNA methylation in stingless bees with low and high heterochromatin contents as assessed by restriction enzyme digestion and image analysis.

BACKGROUND: The stingless bee genus Melipona has been divided into two groups, based on their heterochromatin content. Melipona quadrifasciata and Melipona rufiventris have low and high levels of heterochromatin, respectively. Since condensed chromatin may be rich in methylated DNA sequences, M. quadrifasciata and M. rufiventris nuclei may contain different amounts of methylated CpG. These differences could be assessed by comparing Feulgen-DNA values obtained by image analysis of cells treated with the restriction enzymes Msp I and Hpa II that distinguish between methylated and unmethylated DNA. Msp I and Hpa II cleave the sequence -CCGG-, but there is no cleavage by Hpa II if the cytosine of the central CG dinucleotide is methylated. METHODS: Malpighian tubules of M. quadrifasciata and M. rufiventris were treated with Msp I and Hpa II prior to the Feulgen reaction, and analyzed by automatic scanning microspectrophotometry. RESULTS: The Feulgen-DNA values for the heterochromatin of M. rufiventris and for the small heterochromatin and some euchromatin domains of M. quadrifasciata mostly decreased after treatment with Msp I, but were unchanged after treatment with Hpa II. CONCLUSION: CpG methylation, although detected in diverse chromatin compartments in different bee species, may induce silencing effects required for the same cell physiology.

Animals↗

Bee venom phospholipase A2-specific T cell clones from human allergic and non-allergic individuals: cytokine patterns change in response to the antigen concentration.

Protein antigens with both allergenic and immunoprotective properties represent appropriate molecules to study IgE and IgG regulation. We have established a panel of T cell clones specific to bee venom phospholipase A2 (PLA) from human individuals allergic, hyposensitized or immune (protected) to bee sting. All clones obtained were CD3+, CD4+ and expressed alpha, beta T cell receptor. Depending on the T cell clone, maximal stimulation required 1 to 100 micrograms/ml of PLA, and the addition of interleukin (IL)-2 and/or IL-4 increased their antigen-dependent proliferation. Following antigen stimulation, the clones produced IL-4, interferon-gamma (IFN-gamma) and granulocyte-macrophage colony-stimulating factor. Most clones also produced tumor necrosis factor alpha (TNF-alpha) and tumor necrosis factor beta (TNF-beta), and some produced IL-5 and/or IL-2. Both absolute and relative amounts of secreted cytokines depended on the antigen concentration. At low antigen doses, IL-4 was produced but little or not IFN-gamma, whereas at higher PLA concentrations significant amounts of both IL-4 and IFN-gamma were obtained. Thus, these PLA-specific T cell clones could be classified according to the changes in the ratio of IL-4/IFN-gamma production in response to increasing antigen concentrations. Clones derived from allergic and hyposensitized individuals required higher critical amounts of antigen for IFN-gamma induction, and expressed increasing IL-4/IFN-gamma ratios with increasing concentrations of PLA. Modulation of cytokine patterns by the dose of the antigen may be a driving force for IgE or IgG formation resulting in allergy or immunoprotection.

Bee Venoms↗

An infarct-like myocardial lesion experimentally induced in Wistar rats with Africanized bee venom.

There is evidence in the literature that death following a bee or wasp sting may result from cardiac involvement. This study describes acute cardiac lesions experimentally induced in Wistar Rats submitted to intravenous inoculation of Africanized bee venom (ABV) and killed 1, 4, and 24 h after inoculation. Significant increases in serum enzyme levels were detected; light microscopy showed necrosis of the myocardium; and enzyme histochemistry showed inactivation of enzymes in and around the areas of necrosis. This is the first report of an acute necrotizing cardiac lesion, similar to human myocardial infarction, produced by the inoculation of ABV.

Animals↗

Molecular phylogeny of the large carpenter bees, genus Xylocopa (Hymenoptera: apidae), based on mitochondrial DNA sequences.

Carpenter bees, genus Xylocopa Latreille, a group of bees found on all continents, are of particular interest to behavioral ecologists because of their utility for studies of the evolution of mating strategies and sociality. This paper presents phylogenetic analyses based on sequences of two mitochondrial genes cytochrome oxidase 1 and cytochrome b for 22 subgenera of Xylocopa. Maximum-parsimony and maximum-likelihood methods were used to infer phylogenetic relationships. The analyses resulted in three resolved clades of subgenera: a South American group (including the subgenera Stenoxylocopa, Megaxylocopa, and Neoxylocopa), a group including the subgenera Xylocopa s.s. and Ctenoxylocopa, and an Ethiopean group (including the subgenera Afroxylocopa, Mesotrichia, Alloxylocopa, Platynopoda, Hoploxylocopa, and Koptortosoma). The relationships between the 11 other subgenera and the resolved clades are unclear. Within the Ethiopian group we found a clear separation of the African and the Oriental taxa and apparent polyphyly of the subgenus Koptortosoma. Using an evolutionary rate for ants, we investigated whether Gondwana vicariance or more recent dispersal events could best explain the present-day distribution of subgenera. Although some taxa show divergences that approach Gondwanan breakup times, most divergences between geographic groups are too recent to support a vicariance hypothesis.

Animals↗

Phylogenetic relationships within Diadasia, a group of specialist bees.

We estimated phylogenetic relationships among species of the bee genus Diadasia, a group of new world, specialist bees. We sequenced approximately 2 kb of the mitochondrial genes cytochrome oxidase subunit I and II and tRNA leucine and approximately 1 kb of the nuclear gene elongation factor 1-alpha for 24 North American Diadasia species, 4 South American species, and five outgroup genera. Parsimony analyses of the two data sets were highly congruent. A combined analysis produced a well-resolved phylogenetic hypothesis that supported the monophyly of Diadasia, but not that of traditional subgenera: Diadasia s. str. was paraphyletic in all analyses. With one exception, the North and South American species formed separate clades, supporting previous hypotheses that two lineages of Diadasia have dispersed from South to North America: a more recent dispersal of D. ochracea and an older dispersal of the ancestor to all other North American species. Different species of Diadasia specialize on pollen from at least five different plant families; the phylogeny presented here, along with known host affinities, indicates that host-switching has been rare.

Animals↗

Structural specialization in the dorsal retina of the bee, Apis mellifera.

Electron microscopic investigations on the eye of the worker bee showed that the ommatidia located in the uppermost part of the dorsal half of the eye are characterized by a distinct structural specialization: Nine visual cells contribute microvilli to the rhabdom over its full length. Within these rhabdoms the microvilli are arranged in at least three different directions. This specialization affects an area of at least 60 ommatidia. The most dorsal eye region differs, therefore, structurally from all other regions which have been investigated to date. Because the ommatidia in question are oriented skyward, their peculiar structure is discussed with respect to several concepts of polarized light detection by the bee.

Animals↗

The first optic ganglion of the bee. IV. Synaptic fine structure and connectivity patterns of receptor cell axons and first order interneurones.

The synaptic relationships between and within receptor-cell axons (RCAs), first-order interneurones (L-fibres) and accessory fibres (acc) in the first optic ganglion (the lamina) of the worker bee were studied in serial sections with Golgi-EM and routine transmission electron microscopy. The ommatidium contains nine retinular (photoreceptor) cells all of which project as RCAs to a single optical cartridge in the lamina. Six of the RCAs end as short visual fibres (svf) in the lamina, while the remaining three, the so-called long visual fibres (lvf), pass the lamina and end in the second optic ganglion, the medulla. In addition to the RCAs and an unknown number of accessory fibres, the cartridge also contains four L-fibres (L1--4). The spatial arrangement of the RCAs and L-fibres within a cartridge is constant throughout the depth of the lamina. Serial sections reveal a great number of chemical synapses interconnecting RCAs, L-and ace fibres. Double T-shaped presynaptic dense projections are surrounded and in close association with either spherical or flattened synaptic vesicles. The finding of gap junctions between and within identified RCAs and L-fibres suggest that these axons may be electronically coupled. A model for information processing in the lamina of the bee is suggested from observations of synaptic connectivity between and within fibres of one cartridge.

Animals↗

Glycerol-3-phosphate dehydrogenase (G-3-PDH; EC 1.1.1.8) variation in Brazilian stingless bees and in wasp species.

In only 1 bee species (Tetragona clavipes) of 24 sampled in 145 colonies (0.69%) did we detect the presence of more than one allele for glycerol-3-phosphate dehydrogenase (EC 1.1.1.8), an enzyme that is involved in flight. In 34 colonies containing 9 wasp species, 5 colonies of only 2 species (Polybia paulista and P. sericea) showed variation in larval G-3-PDH (14.7%). The small amount of variation observed for the G-3-PDH-1 locus in the bee and wasp species analyzed in the present study agrees with that reported for the G-3-PDH system in other insects.

Alleles↗

Bee venom inhibits superoxide production by human neutrophils.

Investigation of the antiinflammatory properties of bee venom demonstrates that it inhibits production of superoxide anion by human neutrophils in a potent, selective, nontoxic, dose-dependent fashion, both pre- and poststimulation by particulate and soluble activators of the neutrophil oxidative metabolism burst. The effect is not due to receptor competition, superoxide dismutase, and/or catalase activity, scavenging, or indicator media effects. These findings may explain the antiinflammatory effects of whole bee venom in experimental systems, its widespread use in folk medicine, and lead to the development of potent, new antiinflammatory substances for therapeutic use in man.

Bee Venoms↗

Retrograde labelling of photoreceptors in different regions of the compound eyes of bees and ants.

The geometry of retinal receptor arrays and the projection patterns of photoreceptor axons are unravelled in the compound eyes of bees and ants by backfilling large populations of photoreceptors with horseradish peroxidase or Lucifer yellow. Such retrograde labelling techniques are applied in the insect retina for the first time. They brilliantly label populations of specific receptor types within more than 100 ommatidia. Such preparations are obtained in three distinctly different parts of the eye: (1) the part of the retina that is positioned at the uppermost dorsal margin of the eye and specialized for the detection of polarized skylight; (2) the remainder of the dorsal retina; and (3) the ventral retina. As shown by the large populations of labelled photoreceptors, the retinal receptor arrays differ strikingly between different parts of the eye. Furthermore, there exist similarities as well as marked differences between bees and ants. Former hypotheses concerning the location of photoreceptor terminals in the first and second visual neuropil have all been based on a few individual Golgi-labelled photoreceptors. The results presented in this paper, based on retrograde mass impregnations of selected types of photoreceptors, confirm some of the former hypotheses and reject others. The new findings are important for understanding how insects analyse polarized skylight.

Animals↗

Human fatalities caused by wasp and bee stings in Sweden.

Bee and wasp stings represent an uncommon cause of death. In a retrospective study of all fatalities due to venomous animals in Sweden during a 10-year-period, we found 19 fatalities due to wasps, 1 due to bees but none due to snake bite. This gives an annual incidence of 0.2 per million inhabitants. Most of the deceased were older than 50 years and had an underlying cardiovascular disease while previous severe reactions after insect stings were uncommon. The number of stings was not a factor of importance. Symptoms and death occurred within minutes after the sting. The autopsy findings were in most cases non-specific. Although uncommon, the possibility of a fatal insect sting should be considered in unwitnessed deaths occurring outdoors in summertime.

Adult↗

Selenium in pollen gathered by bees foraging on fly ash-grown plants.

Pollen collected from the rear legs of honey bees found foraging on asters voluntarily growing on a fly ash dump in Endwell, New York, was found to contain a high concentration of selenium. Pollen from bees foraging on asters growing on field soil contained no detectable selenium.

Animals↗

Clinal variation and selection of MDH allozymes in honey bee populations.

Latitudinal clines of malate dehydrogenase-1 (MDH-1) allozymes occur within honey bee populations on three continents: Europe, North America and South America. The North and South American populations are introduced and demonstrate that Mdh allelic clines were established within the last 150 years. The frequency of the 'medium' electrophoretic allele increases in frequency with increasing latitude while the 'fast' allele decreases with latitude on all the three continents. The clines are best explained by the average daily high temperature for July on all continents. These parallel clines provide evidence for selection on Mdh alleles in honey bees.

Alleles↗

Utilization and metabolism of dietary sterols in the honey bee and the yellow fever mosquito.

The honey bee, Apis mellifera, does not convert C28 and C29 phytosterols to cholesterol as found in most previous studies of phytophagous or omnivorous insects, but instead the workers and queens selectively transfer 24-methylenecholesterol, sitosterol and isofucosterol from their endogenous sterol pools to the brood larvae regardless of the sterol in the worker diet. Administering radiolabeled sterols by feeding and injection has made it possible to trace this selective transfer through a second generation of the honey bee. In further comparative sterol metabolism studies, the yellow fever mosquito, Aedes aegypti, was shown to be capable of dealkylating and converting a radiolabeled C29 dietary sterol ([14C]sitosterol) to cholesterol. Metabolic studies with several radiolabeled dietary sterols and an inhibitor of steroid metabolism in the yellow fever mosquito further verified this capability.

Aedes↗

A non-policing honey bee colony (Apis mellifera capensis).

In the Cape honey bee Apis mellifera capensis, workers lay female eggs without mating by thelytokous parthenogenesis. As a result, workers are as related to worker-laid eggs as they are to queen-laid eggs and therefore worker policing is expected to be lower, or even absent. This was tested by transferring worker- and queen-laid eggs into three queenright A. m. capensis discriminator colonies and monitoring their removal. Our results show that worker policing is variable in A. m. capensis and that in one colony worker-laid eggs were not removed. This is the first report of a non-policing queenright honey bee colony. DNA microsatellite and morphometric analysis suggests that the racial composition of the three discriminator colonies was different. The variation in policing rates could be explained by differences in degrees of hybridisation between A. m. capensis and A. m. scutellata, although a larger survey is needed to confirm this.

Animals↗

Effects of cross-feeding anarchistic and wild type honey bees: anarchistic workers are not queen-like.

Unlike normal (wild type) honey bee ( Apis mellifera) colonies, 'anarchistic' colonies are characterised by workers that activate their ovaries in the presence of the queen and brood and by the ability of their workers to lay eggs that evade worker policing. In the Cape honey bee ( A. m. capensis), female larvae can manipulate non- capensis nurse workers such that they receive more larval food and develop into worker-queen intermediates or intercastes. We speculated that, in anarchistic colonies, larvae might produce signals that result in excessive feeding of female larvae. Excessively fed female larvae may then develop into reproductively active workers. In this study we cross-fostered anarchistic and wild type brood and investigated the effect of cross-fostering on the amount of food fed to larvae and on the morphology of the resulting workers. We show that anarchistic larvae do not manipulate wild type nurse workers into feeding them more, nor do anarchistic workers develop into worker-queen intermediates. On the contrary, anarchistic larvae are fed less than wild type larvae and anarchistic workers seem to be poor nurses in that they feed larvae less, irrespective of brood genotype.

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↗