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Varying degrees of Apis mellifera ligustica introgression in protected populations of the black honeybee, Apis mellifera mellifera, in northwest Europe.

The natural distribution of honeybee subspecies in Europe has been significantly affected by human activities during the last century. Non-native subspecies of honeybees have been introduced and propagated, so that native black honeybee (Apis mellifera mellifera) populations lost their identity by gene-flow or went extinct. After previous studies investigated the remaining gene-pools of native honeybees in France and Spain, we here assess the genetic composition of eight northwest European populations of the black honeybee, using both mitochondrial (restriction fragment length polymorphisms of the intergenic transfer RNAleu-COII region) and nuclear (11 microsatellite loci) markers. Both data sets show that A. m. mellifera populations still exist in Norway, Sweden, Denmark, England, Scotland and Ireland, but that they are threatened by gene flow from commercial honeybees. Both Bayesian admixture analysis of the microsatellite data and DraI-RFLP (restriction fragment length polymorphism) analysis of the intergenic region indicated that gene-flow had hardly occurred in some populations, whereas almost 10% introgression was observed in other populations. The most introgressed population was found on the Danish Island of Laeso, which is the last remaining native Danish population of A. m. mellifera and the only one of the eight investigated populations that is protected by law. We discuss how individual admixture analysis can be used to monitor the restoration of honeybee populations that suffer from unwanted hybridization with non-native subspecies.

Animals↗

[Use of a PCR method for controlling pure-breeding of honeybees Apis mellifera mellifera L. in the southern Urals].

A key problem of honeybee (Apis mellifera mellifera) breeding in the Southern Urals is its cross-breeding with the Caucasian honeybee Apis mellifera caucasica. Mitochondrial DNA (mtDNA) in these subspecies differ in the length of a fragment localized between genes CO-I and CO-II, which can be used as a marker. A pair of 20-mer primers for PCR was chosen by means of computer design in order to determine the fragment size in both of the subspecies. The amplified fragment was shown to have a length of 350 bp in A. m. caucasica and 600 bp in A. m. mellifera. The difference in length results from the different ratio between two main elements P and Q, which comprise a major part of this sequence in these subspecies: a copy of P element and two copies of Q element in A. m. mellifera, and a copy of Q element only in A. m. caucasica. This sharply defined distinction allows us to use PCR for differentiating the subspecies, estimating the heterogeneity in the colonies, and rejecting queens in the selection process because of the maternal inheritance of the studied character. The nucleotide sequence of the amplified mtDNA fragment of A. m. mellifera was determined.

Amino Acid Sequence↗

Bacterial diversity in worker adults of Apis mellifera capensis and Apis mellifera scutellata (Insecta: Hymenoptera) assessed using 16S rRNA sequences.

High-fidelity PCR of 16S rRNA sequences was used to identify bacteria associated with worker adults of the honeybee subspecies Apis mellifera capensis and Apis mellifera scutellata. An expected approximately 1.5-kb DNA band, representing almost the entire length of the 16S rRNA gene, was amplified from both subspecies and cloned. Ten unique sequences were obtained: one sequence each clustered with Bifidobacterium (Gram-positive eubacteria), Lactobacillus (Gram-positive eubacteria), and Gluconacetobacter (Gram-negative alpha-proteobacteria); two sequences each clustered with Simonsiella (beta-proteobacteria) and Serratia (gamma-proteobacteria); and three sequences each clustered with Bartonella (alpha-proteobacteria). Although the sequences relating to these six bacterial genera initially were obtained from either A. m. capensis or A. m. scutellata or both, newly designed honeybee-specific 16S rRNA primers subsequently amplified all sequences from all individual workers of both subspecies. Attempts to amplify these sequences from eggs have failed. However, the wsp primers designed to amplify Wolbachia DNA from arthropods, including these bees, consistently produced a 0.6-kb DNA band from individual eggs, indicating that amplifiable bacterial DNA was present. Hence, the 10 bacteria could have been acquired orally from workers or from other substrates. This screening of 16S rRNA sequences from A. m. capensis and A. m. scutellata found sequences related to Lactobacillus and Bifidobacterium which previously had been identified from other honeybee subspecies, as well as sequences related to Bartonella, Gluconacetobacter, Simonsiella/Neisseria, and Serratia, which have not been identified previously from honeybees.

Animals↗

Toxicity of cypermethrin and fenitrothion on the hemolymph carbohydrates, head acetylcholinesterase, and thoracic muscle Na+, K+-ATPase of emerging honeybees (Apis mellifera mellifera. L).

Comparative effects of sublethal doses (0, 0.1, 0.2. 0.4, 0.8, and 1 nmol/bee) of cypermethrin and fenitrothion have been studied on emerging honeybees. The insecticides were injected intrathoracially between the third and the fourth segment. Biochemical effects were determined over a 3-h period. Both cypermethrin and fenitrothion led to a significant hypoglucosemia and hypotrehalosemia 15 min after injection, but cypermethrin seemed more active than fenitrothion at the same doses. A recovery phase appeared for glucosemia and trehalosemia, 60 min after injection. The higher toxicity of cypermethrin than fenitrothion also appeared in this period, where it took a longer time for honeybees to reestablish carbohydrate levels following cypermethrin than fenitrothion injections. The low values of the correlation coefficients (r) for glucose versus trehalose levels led to the supposition that no typical functional interaction between glucose and trehalose could be considered to be involved in this experience. Na+, K+-ATPases activity was significantly inhibited (P< 0.05) by cypermethrin and maximum percentage inhibition was reached (45%) at 1 nmol/bee. The kinetic analysis of honeybee's acetylcholinesterase inhibition by fenitrothion, indicated that this insecticide acts (P< 0.05) on acetylcholinesterase activity. The percentage inhibition exceeded 60% at 0.2 nmol/bee. This result revealed that in general cypermethrin and fenitrothion share common biochemical effects on carbohydrates, although their neurotoxic effects on honeybees might be different.

Acetylcholinesterase↗

[Magnetic parameters of the bees Apis mellifera mellifera (L.) obtained by SQUID-magnetometry].

It was found using SQUID-magnetometry that the dependence of the magnetization of a bee on the intensity of the applied magnetic fields has the form of a hysteresis loop. By using the parameters of the loop, the magnetic phases were identified and their characteristics were determined. It was found that the diamagnetic component is significant and has a susceptibility = -1,3335 x 10(-8) Gs x cm3/Oe. The ferromagnetite phase is represented by a multidomain magnetite with the coercive force of 70-80 Oe and magnetic saturation momnts of 4 x 10(-5) EMU (Gs x cm3) and residual moment of 3.07 x 10(-6) EMU.

Animals↗

The formamidine amitraz as a hyperglycemic alpha-agonist in worker honeybees (Apis mellifera mellifera L.) in vivo.

Intra-abdominal injection of amitraz (0.25 nmol per honeybee, i.e., approx 2.3 nmol/g) to emerging worker bees, in vivo, led to a significant hypertrehalosemia (300-400%) followed by a hyperglucosemia (approximately equal to 600%). Maxima were reached at 0.5 and 2 h, respectively. A strong negative correlation between glucosemia and trehalosemia appeared after injection of pure phentolamine (1 nmol per bee), suggesting stimulation of trehalase activities. Simultaneous administration of the alpha-blocker at greater than or equal to 0.25 nmol per individual suppressed the hyperglycemic response of amitraz. The formamidine pesticide thus likely acts on the honeybee alpha-aminergic system.

Adrenergic alpha-Agonists↗

Analysis of peptides in the brain and corpora cardiaca-corpora allata of the honey bee, Apis mellifera using MALDI-TOF mass spectrometry.

The neuropeptide profiles and diversity of the brain and retrocerebral organs (corpora cardiaca-corpora allata; CC-CA) of adult workers of the honey bee Apis mellifera carnica (dark European strain) were investigated using a combination of HPLC and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) with post-source decay (PSD) and collision-induced dissociation (CID) fragmentation. Using evidence from genomic sources, including BLAST searches of the honey bee genome, comparisons with other species and de novo sequencing by PSD and CID fragmentation, a total of 13 mass ions could be assigned to peptides predicted from the A. mellifera genomic database. Peptides positively identified were A. mellifera tachykinin-related peptides 3 and 4 (APMGFQGMRa; APMGFYGTRa) and leucomyosuppressin (pEDVDHVFLRFa). Peptides tentatively identified were A. mellifera tachykinin-related peptides 2 and 5 (ALMGFQGVRa; ARMGFHGMRa), A. mellifera allatostatins 2, 3 and 4 (GRDYSFGLa; RQYSFGLa; GRQPYSFGLa), A1-SIFamide (AYRKPPFNGSIFa), Q1-leucomyosuppressin (QDVDHVFLRFa) and A. mellifera pyrokinins PK 1, PK 2 and Q1-PK 2 (TSQDITSGMWFGPRLa; pEITQFTPRLa; QITQFTPRLa). Allatostatins, tachykinin-related peptides and A1-SIFamide were not detected in CC-CA extract, which appears to contain predominantly leucomyosuppressin, Q1-leucomyosuppressin, PK 1, PK 2, Q1-PK 2 and some unidentified masses. No ion signal was detected that would correspond to the hypertrehalosaemic peptide (=Manse-AKH), which has been isolated from the Italian race of the honey bee (A. mellifera ligustica), but not from A. mellifera carnica.

Amino Acid Sequence↗

Species-specific structuring of gut bacterial and fungal communities in honey bees Apis cerana and Apis mellifera.

Honey bee gut microbiome studies have primarily emphasized bacteria, leaving fungal communities comparatively overlooked despite their ecological and functional importance. Whole-genome shotgun metagenomics of Apis cerana and Apis mellifera revealed fungal assemblages dominated by Ascomycota, with Basidiomycota and Microsporidia in minor proportions, alongside gut bacterial communities composed mainly of Pseudomonadota, Bacillota, and Actinomycetota. The bacterial diversity was markedly higher in A. mellifera (Shannon&#x2009;=&#x2009;5.90; Simpson&#x2009;=&#x2009;0.98) than in A. cerana (Shannon&#x2009;=&#x2009;4.01; Simpson&#x2009;=&#x2009;0.94; p&#x2009;>&#x2009;0.05), while fungal diversity remained comparable between species (p&#x2009;>&#x2009;0.05). Beta-diversity analyses revealed strong host-specific clustering for both bacterial (PERMANOVA R2&#x2009;=&#x2009;0.7989, p&#x2009;>&#x2009;0.05) and fungal communities (R2&#x2009;=&#x2009;0.7218, p&#x2009;>&#x2009;0.05), indicating distinct microbial organization driven by host species. Bacterial-fungal co-occurrence patterns exhibited host-specific structuring, suggesting differential inter-kingdom community organization between A. cerana and A. mellifera. Linear Discriminant Analysis Effect Size (LEfSe) identified 93 discriminatory fungal taxa (45 enriched in A. cerana, 48 in A. mellifera), highlighting yeast-dominated signatures in A. mellifera and Basidiomycota-affiliated enrichments in A. cerana. KEGG and CAZy profiling revealed host- and kingdom-specific functional differences, with bacterial communities of A. mellifera showing distinct representation of carbohydrate metabolism and nutrient-cycling functions, while fungal communities exhibited a comparatively narrower functional repertoire. Together, these findings provide a high-resolution view of honey bee bacterial and fungal microbiomes, highlighting strong host-driven divergence in taxonomy, function, and cross-kingdom interactions.

Animals↗

Honeybee, Apis mellifera (Hymenoptera: Apidae), leaf damage on Alnus species in Uganda: a blessing or curse in agroforestry?

It is a dictum that Apis mellifera Linnaeus is innocuous in agricultural ecosystems. This study provides the first record of A. mellifera as a significant defoliator of Alnus species. Careful field observations coupled with microscopic examination provided convincing evidence implicating A. mellifera as the cause of leaf perforation on Alnus species in Uganda. Apis mellifera was observed foraging selectively on young Alnus leaves and buds in search of a sticky substance, apparently propolis. In so doing, the bee created wounds that enlarged and caused tattering of Alnus leaves as they matured. Biological surveys indicated that the damage was prevalent and occurred widely, particularly on Alnus acuminata Kunth in Uganda. Incidence of the Apis mellifera damage on Alnus acuminata peaked in the dry season, with up to 90% of leaves emerging per shoot per month damaged, and was lowest in the wet months during peak leaf emergence. Apis mellifera leaf damage was consistently higher on Alnus acuminata than A. nepalensis D. Don., on saplings than mature trees, and on sun exposed than shaded leaves. The activity of honeybees may be detrimental to the productivity of Alnus, yet the substance for which the insect forages on Alnus is a resource with potential economic importance.

Agriculture↗

Ferritin in iron containing granules from the fat body of the honeybees Apis mellifera and Scaptotrigona postica.

It is already known that the behaviour of the honeybee Apis mellifera is influenced by the Earth's magnetic field. Recently it has been proposed that iron-rich granules found inside the fat body cells of this honeybee had small magnetite crystals that were responsible for this behaviour. In the present work, we studied the iron containing granules from queens of two species of honeybees (A. mellifera and Scaptotrigona postica) by electron microscopy methods in order to clarify this point. The granules were found inside rough endoplasmic reticulum cisternae. Energy dispersive X-ray analysis of granules from A. mellifera showed the presence of iron, phosphorus and calcium. The same analysis performed on the granules of S. postica also indicated the presence of these elements along with the additional element magnesium. The granules of A. mellifera were composed of apoferritin-like particles in the periphery while in the core, clusters of organised particles resembling holoferritin were seen. The larger and more mineralised granules of S. postica presented structures resembling ferritin cores in the periphery, and smaller electron dense particles inside the bulk. Electron spectroscopic images of the granules from A. mellifera showed that iron, oxygen and phosphorus were co-localised in the ferritin-like deposits. These results indicate that the iron-rich granules of these honeybees are formed by accumulation of ferritin and its degraded forms together with elements present inside the rough endoplasmic reticulum, such as phosphorus, calcium and magnesium. It is suggested that the high level of phosphate in the milieu would prevent the crystallisation of iron oxides in these structures, making very unlikely their participation in magnetoreception mechanisms. They are most probably involved in iron homeostasis.

Animals↗

Heterogeneity in regional GC content and differential usage of codons and amino acids in GC-poor and GC-rich regions of the genome of Apis mellifera.

The honeybee (Apis mellifera) has a genome with a wide variation in GC content showing 2 clear modal GC values, in some ways reminiscent of an isochore-like structure. To gain insight into causes and consequences of this pattern, we used a comparative approach to study the genome-wide alignment of primarily coding sequence of A. mellifera with Drosophila melanogaster and Anopheles gambiae. The latter 2 species show a higher average GC content than A. mellifera and no indications of bimodality, suggesting that the GC-poor mode is a derived condition in honeybee. In A. mellifera, synonymous sites of genes generally adopt the GC content of the region in which they reside. A large proportion of genes in GC-poor regions have not been assigned to the honeybee assembly because of the low sequence complexity of their genome neighborhood. The synonymous substitution rate between A. mellifera and the other species is very close to saturation, but analyses of nonsynonymous substitutions as well as amino acid substitutions indicate that the GC-poor regions are not evolving faster than the GC-rich regions. We describe the codon usage and amino acid usage and show that they are remarkably heterogeneous within the honeybee genome between the 2 different GC regions. Specifically, the genes located in GC-poor regions show a much larger deviation in both codon usage bias and amino acid usage from the Dipterans than the genes located in the GC-rich regions.

Amino Acids↗

Structural differences in the drone olfactory system of two phylogenetically distant Apis species, A. florea and A. mellifera.

Male insects that are attracted by sex pheromones to find their female mates over long distances have specialized olfactory subsystems. Morphologically, these subsystems are characterized by a large number of receptor neurons sensitive to components of the female's pheromones and hypertrophied glomerular subunits ('macroglomeruli' or 'macroglomerular complexes') in the antennal lobes, in which the axons of the receptor neurons converge. The olfactory subsystems are adapted for an increased sensitivity to perceive minute amounts of pheromones. In Apis mellifera, drones have 18,600 olfactory poreplate sensilla per antenna, each equipped with receptor neurons sensitive to the queen's sex pheromone, and four voluminous macroglomeruli (MG1-MG4) in the antennal lobes. In contrast, we show that drones of the phylogenetically distant species, Apis florea, have only 1,200 poreplate sensilla per antenna and only two macroglomeruli in their antennal lobes. These macroglomeruli are homologous in anatomical position to the two most prominent macroglomeruli in A. mellifera, the MG1 and MG2, but they are much smaller in size. The morphological and anatomical differences described here suggest major modifications in the sex-pheromone processing subsystem of both species: (1) less pheromone sensitivity in A. florea and (2) a more complex sex-pheromone processing and thus a more complex sex-pheromone communication in A. mellifera. Research in honey bee sex-pheromone communication dates back to the 1960s, when Gary (1962) demonstrated that in Apis mellifera the queen's mandibular gland secretion and especially its main component, 9-ODA (9-keto-2(E)-decenoic acid), is highly attractive to drones on their nuptial flight. Later, cross-species attraction experiments showed that other honey bee species, Apis florea, A. cerana, and A. dorsata probably also use the queen's mandibular gland secretion as a mating attractant (Butler et al. 1967; Sanasi et al. 1971). Besides its function in mating behavior, the queen's mandibular gland secretion is the main pheromone regulating queen-worker interactions (Free 1987). In this context, several studies have demonstrated the behavioral significance of single components (Slessor et al. 1988) and differences in the composition of the secretion between Apis species (Plettner et al. 1996, 1997; Keeling et al. 2000). Regarding the interspecific differences in the queen's signal, the question arises whether this variation is reflected in the olfactory system of drones and workers of the various species.

Animals↗

Insights into social insects from the genome of the honeybee Apis mellifera.

Here we report the genome sequence of the honeybee Apis mellifera, a key model for social behaviour and essential to global ecology through pollination. Compared with other sequenced insect genomes, the A. mellifera genome has high A+T and CpG contents, lacks major transposon families, evolves more slowly, and is more similar to vertebrates for circadian rhythm, RNA interference and DNA methylation genes, among others. Furthermore, A. mellifera has fewer genes for innate immunity, detoxification enzymes, cuticle-forming proteins and gustatory receptors, more genes for odorant receptors, and novel genes for nectar and pollen utilization, consistent with its ecology and social organization. Compared to Drosophila, genes in early developmental pathways differ in Apis, whereas similarities exist for functions that differ markedly, such as sex determination, brain function and behaviour. Population genetics suggests a novel African origin for the species A. mellifera and insights into whether Africanized bees spread throughout the New World via hybridization or displacement.

Animals↗

The flight physiology of reproductives of Africanized, European, and hybrid honeybees (Apis mellifera).

Neotropical African honeybees (Apis mellifera scutellata), in the process of spreading throughout tropical and subtropical regions of the Americas, hybridize with and mostly replace European honeybees (primarily Apis mellifera mellifera and Apis mellifera ligustica). To help understand this process, we studied the effect of lineage (African, European, or hybrid) on the flight physiology of honeybee reproductives. Flight metabolic rates were higher in queens and drones of African lineage than in European or hybrid bees, as has been previously found for foraging workers. These differences were associated with higher thorax/body mass ratios and higher thorax-specific metabolic rates in African lineage bees. Queens were reared in common colonies, so these metabolic and morphological differences are likely to be genetic in origin. African drones had higher wing beat frequencies and thorax temperatures than European or hybrid bees. Hybrids were intermediate for many parameters, but hybrid queen mass-specific flight metabolic rates were low relative to Africans and were nonlinearly affected by the proportion of African lineage, consistent with some negative heterosis for this trait.

Analysis of Variance↗

Geographical overlap of two mitochondrial genomes in Spanish honeybees (Apis mellifera iberica).

Restriction enzyme cleavage maps of mitochondrial DNA from the Spanish honeybee, Apis mellifera iberica (Hymenoptera: Apidae), were compared with those from the European subspecies A. m. mellifera, A. m. ligustica, and A. m. carnica, and the African subspecies A. m. intermissa and A. m. scutellata. The mitochondrial DNA (mtDNA) of the two African subspecies can be distinguished by restriction fragment polymorphisms revealed by Hinf I digests. Two distinct mtDNA types were found among Spanish honeybees: a west European mellifera-like type, which predominates in the north of Spain, and an African intermissa-like type, which predominates in the south. Spain appears to be a region of contact and hybridization between the two subspecies A. m. intermissa and A. m. mellifera, which respectively represent African and west European honeybee lineages. This natural boundary between European and African honeybee populations in the Old World may provide a model for predicting the eventual outcome of the colonization of North America by introduced African honeybees.

Africa↗

Bombus impatiens (Hymenoptera: Apidae): an alternative to Apis mellifera (Hymenoptera: Apidae) for lowbush blueberry pollination.

The pollination effectiveness of the commercially reared bumble bee Bombus impatiens Cresson, was compared in field studies to the honey bee, Apis mellifera L., for lowbush blueberry, Vaccinium angustifolium Ait. A preliminary study indicated that B. impatiens had potential as an alternative pollinator. In a 3-yr study, percentage fruit set, percentage harvested berries, berry weight, and seeds per berry were compared in blueberry fields stocked at 7.5 A. mellifera hives per hectare to 5, 7.5, or 10 B. impatiens colonies per hectare. Percentage of harvested berries (yield) was significantly higher in fields stocked with B. impatiens at 10 colonies per hectare. No other parameters measuring pollinator effectiveness were significantly different at 5, 7.5, or 10 colonies per hectare. Flower handling time was significantly faster for B. impatiens and it more frequently collected blueberry pollen. All parameters of pollinator effectiveness were similar for B. impatiens, A. mellifera, and native wild bees in a follow-up study. Overall, B. impatiens was a suitable alternative to A. mellifera.

Animals↗

Esterase isozymes of Apis mellifera: substrate and inhibition characteristics, developmental ontogeny, and electrophoretic variability.

Starch gel electrophoresis utilizing different types of substrates and inhibitors made it possible to detect several esterases in crude extracts of Apis mellifera. Our results suggest that there are six Apis mellifera esterase isozymes (esterases 1-6) that differ not only in electrophoretic mobility but also in substrate specificity and inhibition properties. Some of the esterase isozymes are controlled by more than one allele. The frequency of these genetic variants was analyzed in four populations of Apis mellifera from several localities. Esterases 1, 2, and 4 do not exhibit developmental changes, but the electrophoretic profile of esterases 3, 4, and 6 varies during ontogenetic development.

Alleles↗