Per Kryger

Varroa-Virus Interaction in Collapsing Honey Bee Colonies

Varroa mites and viruses are the currently the high-profile suspects in collapsing bee colonies. Therefore, seasonal variation in varroa load and viruses (Acute-Kashmir-Israeli complex (AKI) and Deformed Wing Virus (DWV)) were monitored in a year-long study. We investigated the viral titres in honey bees and varroa mites from 23 colonies (15 apiaries) under three treatment conditions: Organic acids (11 colonies), pyrethroid (9 colonies) and untreated (3 colonies). Approximately 200 bees were sampled every month from April 2011 to October 2011, and April 2012. The 200 bees were split to 10 subsamples of 20 bees and analysed separately, which allows us to determine the prevalence of virus-infected bees. The treatment efficacy was often low for both treatments. In colonies where varroa treatment reduced the mite load, colonies overwintered successfully, allowing the mites and viruses to be carried over with the bees into the next season. In general, AKI and DWV titres did not show any notable response to the treatment and steadily increased over the season from April to October. In the untreated control group, titres increased most dramatically. Viral copies were correlated to number of varroa mites. Most colonies that collapsed over the winter had significantly higher AKI and DWV titres in October compared to survivors. Only treated colonies survived the winter. We discuss our results in relation to the varroa-virus model developed by Stephen Martin.

High degree of polyandry in Apis dorsata queens detected by DNA microsatellite variability

Workers of six colonies of the giant honeybee Apis dorsata from Sabah, Malaysia (five colonies) and Java (one colony) were genotyped using single locus DNA fingerprinting. The colonies from Sabah nested in colony aggregations of 5 and 28 nests respectively on two trees. Three DNA microsatellite loci (A14, A76, A88) with a total of 27 alleles provided sufficient genetic variability to classify the workers into distinct sub-families revealing the degree of polyandry of the queens. Queens mated on average with 30.17 ± 5.98 drones with a range from 19 to 53. The average effective number of matings per queen was 25.56 ± 11.63. In the total sample of 192 workers, 22 individuals were found that were not offspring of the colonys queen. Three of these were potentially drifted offspring workers from genotyped queens of colonies nesting on the same tree.

The acoustic near field of a dancing honeybee

SummaryThe acoustic near field close to honeybees performing the wagging dance was investigated with pairs of small, matched microphones placed in various positions around the dancing bees. The dance ‘sounds’ are produced by the wings, which act as an asymmetrical dipole emitter. Close to the abdomen, the ‘sound’ pressures in the air spaces above and below the plane of the wings are totally out of phase. A zone of very intense acoustical short-circuiting exists close to the edges of the wings, where pressure gradients of about 1 Pa/mm are observed in the dorso-ventral direction (perpendicular to the plane of the wings). The pressure gradients drive air movements with velocity amplitudes up to about 1 m/s. The pressure gradients are much smaller in directions radially away from the bee and decrease rapidly with increasing distance from the wings. The ‘sound’ pressure detected by a stationary probe at one side of the bee is strongly modulated at 12–13 Hz as a result of the bees side-to-side wagging. Surprisingly little ‘sound’ is found near the dancers head. The positions of the follower bees reflect the properties of the acoustic field: The follower bees place their antennae in the zone of maximum acoustical short-circuiting where the air particle movements are most intense. These observations suggest 1) how follower bees can avoid mixing up the messages carried by the dance ‘sounds’ when two or more bees are dancing only a few cm apart and 2) how the followers might extract information about a dancers spatial orientation from the acoustic near field she produces. The observations also provide clues regarding the nature of the putative ‘sound’ receivers.

Whole-genome scan in thelytokous-laying workers of the Cape honeybee (Apis mellifera capensis): central fusion, reduced recombination rates and centromere mapping using half-tetrad analysis.

While workers of almost all subspecies of honeybee are able to lay only haploid male eggs, Apis mellifera capensis workers are able to produce diploid female eggs by thelytokous parthenogenesis. Cytological analyses have shown that during parthenogenesis, egg diploidy is restored by fusion of the two central meiotic products. This peculiarity of the Cape bee preserves two products of a single meiosis in the daughters and can be used to map centromere positions using half-tetrad analysis. In this study, we use the thelytokous progenies of A. m. capensis workers and a sample of individuals from a naturally occurring A. m. capensis thelytokous clone to map centromere position for most of the linkage groups of the honeybee. We also show that the recombination rate is reduced by >10-fold during the meiosis of A. m. capensis workers. This reduction is restricted to thelytokous parthenogenesis of capensis workers and is not observed in the meiosis of queen within the same subspecies or in arrhenotokous workers of another subspecies. The reduced rate of recombination seems to be associated with negative crossover interference. These results are discussed in relation to evolution of thelytokous parthenogenesis and maintenance of heterozygosity and female sex after thelytoky.

Multiple nuptial flights, sperm transfer and the evolution of extreme polyandry in honeybee queens

The honeybee, Apis mellifera, has an extremely polyandrous mating system, which often involves multiple nuptial flights by its queens. To understand the evolution of extreme polyandry, we investigated the cost of multiple nuptial flights in relation to potential benefits. We analysed, with eight DNA microsatellite loci, the paternity of worker offspring of naturally mated queens. Queens that were restricted to one nuptial flight, but may have taken additional nuptial flights if allowed to do so, had significantly fewer matings than queens that started oviposition after a single nuptial flight. Furthermore, the number of sperm stored in a spermatheca increased significantly with the number of matings. We suggest that queens adjust their nuptial flight frequencies according to their mating success in their previous nuptial flights. The number of copulations seems to serve as a signal for the initiation of oviposition. In the light of these findings, we reconsider and discuss the significance of the sperm limitation hypothesis for the evolution of extreme polyandry in A. mellifera.

Standard methods for characterising subspecies and ecotypes of Apis mellifera

Summary The natural diversity of honey bees in Europe is eroding fast. A multitude of reasons lead to a loss of both genetic diversity and specific adaptations to local conditions. To preserve locally adapted bees through breeding efforts and to maintain regional strains in conservation areas, these valuable populations need to be identified. In this paper, we give an overview of methods that are currently available and used for recognition of honey bee subspecies and ecotypes, or that can be utilised to verify the genetic origin of colonies for breeding purposes. Beyond summarising details of morphometric, allozyme and DNA methods currently in use, we report recommendations with regard to strategies for sampling, and suggest methods for statistical data analysis. In particular, we emphasise the importance of reference data and consistency of methods between laboratories to yield comparable results.

The size of wild honeybee populations ( Apis mellifera ) and its implications for the conservation of honeybees

The density of wild honeybee colonies (Apis mellifera) in the African dry highland savannahs was estimated in three Nature Reserves in Gauteng, South Africa (Ezemvelo, Leeuwfontein, Suikerbosrand) based on the genotypes of drones which were caught at drone congregation areas. Densities were estimated to range between 12.4 and 17.6 colonies per square kilometer. In addition colony densities were estimated in two German National parks (Müritz and Hochharz) and a commercial mating apiary. The density of colonies was significantly lower at the German sampling sites with estimates of 2.4–3.2 colonies per square kilometer, which closely matches the nation-wide density of colonies kept by beekeepers. This shows that the densities of colonies observed in wild populations under the harsh conditions of the African dry savannahs exceeds that of Germany by far, in spite of intensive beekeeping. The intensity of apiculture in Europe is therefore unlikely to compensate for the loss of habitats suitable for wild honeybees due to agriculture, forestry and other cultivation of land.

Conserving diversity and vitality for honey bee breeding.

Summary Beekeepers in Europe, North America and other parts of the world have repeatedly been afflicted by elevated and sometimes unexplained colony losses. Multiple factors have been considered in connection with increased winter losses. In addition to national programmes investigating possible causes for increased honey bee mortality, scientists collaborate at an international level on different aspects of bee health within the COLOSS network. Within this network, Working Group 4 explores aspects of genetic diversity in relation to the vitality and health of honey bee populations. In this paper, we briefly review the genetic diversity of honey bees in Europe, discuss the effects of beekeeping and selective breeding on honey bee populations under the aspect of genetic diversity and bee health, and review the current status of EU legislation with respect to protection of native bee populations. We introduce and discuss recent approaches in honey bee selective breeding to improve disease resistance by introducing traits related to colony vitality. Finally, we present the aims of WG4 within the COLOSS network and briefly introduce our experimental approach.

Survival and immune response of drones of a Nosemosis tolerant honey bee strain towards N. ceranae infections.

Honey bee colonies (Apis mellifera) have been selected for low level of Nosema in Denmark over decades and Nosema is now rarely found in bee colonies from these breeding lines. We compared the immune response of a selected and an unselected honey bee lineage, taking advantage of the haploid males to study its potential impact on the tolerance toward Nosema ceranae, a novel introduced microsporidian pathogen. After artificial infections of the N. ceranae spores, the lineage selected for Nosema tolerance showed a higher N. ceranae spore load, a lower mortality and an up-regulated immune response. The differences in the response of the innate immune system between the selected and unselected lineage were strongest at day six post infection. In particular genes of the Toll pathway were up-regulated in the selected strain, probably is the main immune pathway involved in N. ceranae infection response. After decades of selective breeding for Nosema tolerance in the Danish strain, it appears these bees are tolerant to N. ceranae infections.

Standard methods for rearing and selection of Apis mellifera queens

Summary Here we cover a wide range of methods currently in use and recommended in modern queen rearing, selection and breeding. The recommendations are meant to equally serve as standards for both scientific and practical beekeeping purposes. The basic conditions and different management techniques for queen rearing are described, including recommendations for suitable technical equipment. As the success of breeding programmes strongly depends on the selective mating of queens, a subchapter is dedicated to the management and quality control of mating stations. Recommendations for the handling and quality control of queens complete the queen rearing section. The improvement of colony traits usually depends on a comparative testing of colonies. Standardized recommendations for the organization of performance tests and the measurement of the most common selection characters are presented. Statistical methods and data preconditions for the estimation of breeding values which integrate pedigree and performance data from as many colonies as possible are described as the most efficient selection method for large populations. Alternative breeding programmes for small populations or certain scientific questions are briefly mentioned, including also an overview of the young and fast developing field of molecular selection tools. Because the subject of queen rearing and selection is too large to be covered within this paper, plenty of references are given to facilitate comprehensive studies.