SummaryHoney bee queens are highly polyandrous and mate in flight. Instrumental insemination is an essential tool that provides complete control of honey bee mating for research and breeding purposes. The technique requires specialized equipment to anesthetize and immobilize the queen and to collect and deliver semen from the drones. Semen is harvested from mature drones by hand eversion of the endophallus and collected into a syringe. The queen is placed in a chamber and anesthetized during the procedure of insertion of semen into the oviducts. Queens are introduced into colonies and their performance can equal to that of naturally mated queens, given proper technique and care. Métodos estándar para la inseminación artificial de reinas de Apis mellifera ResumenLas reinas de las abejas melíferas presentan un elevado grado de poliandria y se aparean durante el vuelo. La inseminación artificial es una herramienta esencial que proporciona un control completo del apareamiento de las abejas con fines de investigación y de cría. La técnica requiere de un equipo especializado para anestesiar e inmovilizar a la reina y para colectar y administrar el semen de los zánganos. El semen se obtiene de zánganos maduros por eversión manual del endofalo y se recoge en una jeringa. La reina se coloca en una cámara y se mantiene anestesiada durante el proceso de inserción del semen en los oviductos. Las reinas se introducen en las colonias y su rendimiento puede ser igual al de las reinas que se aparearon de forma natural, si se realiza la técnica y se da la atención adecuada.

-Instrumental insemination, a reliable method to control honey bee mating, is an essential tool for research and stock improvement. A review of studies compare colony performance of instrumentally inseminated queens, IIQs, and naturally mated queens, NMQs. Factors affecting queen performance are also reviewed. The collective results of the data demonstrate that the different methodologies used, in the treatment of queens, has a significant affect on performance rather than the insemination procedure. Beekeeping practices can optimize or inhibit performance. The competitive performance of IIQs is demonstrated when queens are given proper care. The advantage of selection and a known semen dosage can result in higher performance levels of IIQs.Apis mellifera / queen honey bees / instrumental insemination / colony performance

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Our primary objective was to identify techniques to transform the genome of the honey bee (Apis mellifera) with foreign DNA constructs. The strategy we adopted was to linearize foreign DNA and introduce it with sperm during the instrumental insemination of virgin queen honey bees. We analysed extracts from larvae within the same cohort and isolated the predicted fragment by means of PCR amplification of genomic DNA. Larvae that carried the construct also expressed the introduced DNA. We propagated several transgenic lines for up to three generations, which demonstrates its heritability. Once carried by a queen, the construct can be detected in that queen's larvae over several months. However, there was no evidence of integration of the construct, at least as determined by genomic Southern analysis. Nevertheless, this demonstrates the general viability of the technique for introduction of DNA, and it should be augmented by further use of transposable elements that enhance integration.

Proboscis extension conditioning of honeybee workers was used to study the processing of odorants when bees were conditioned to binary mixtures. Responses to a set of pure floral odors and pheromones after conditioning have already been described. When bees are conditioned to certain mixtures of odorants, the response to both components is equal to that when they are tested alone. However, mixtures of an aliphatic aldehyde and an alcohol elicit asymmetric response patterns; that is, the response to the aldehyde is much stronger than that to the alcohol. A bee's response to the alcohol after it had been trained in an aldehyde background is significantly lower than when the bee is trained to respond to the same alcohol in the background of another odorant. Such response patterns are not necessarily caused by a behavioral decrement resulting from a compound-unique perceptual effect produced by the mixture. Furthermore, studies of blocking show that behavioral acquisition in response to one component can be hindered or blocked by pretraining with the other component. These results suggest that honeybees can perceive the individual components of some binary mixtures. The similarities in neural processing in olfactory systems of vertebrates and invertebrates mean that such studies could elucidate behavioral mechanisms of olfaction in a wide phylogenetic spectrum of animals.

A study was conducted to identify quantitative trait loci (QTLs) that affect learning in honeybees. Two F1 supersister queens were produced from a cross between two established lines that had been selected for differences in the speed at which they reverse a learned discrimination between odors. Different families of haploid drones from two of these F1 queens were evaluated for two kinds of learning performance--reversal learning and latent inhibition--which previously showed correlated selection responses. Random amplified polymorphic DNA markers were scored from recombinant, haploid drone progeny that showed extreme manifestations of learning performance. Composite interval mapping procedures identified two QTLs for reversal learning (lrn2 and lrn3: LOD, 2.45 and 2.75, respectively) and one major QTL for latent inhibition (lrn1: LOD, 6.15). The QTL for latent inhibition did not map to either of the linkage groups that were associated with reversal learning. Identification of specific genes responsible for these kinds of QTL associations will open up new windows for better understanding of genes involved in learning and memory.