Comparing the Appetitive Learning Performance of Six European Honeybee Subspecies in a Common Apiary.

The Western honeybee (Apis mellifera L.) is one of the most widespread insects with numerous subspecies in its native range. How far adaptation to local habitats has affected the cognitive skills of the different subspecies is an intriguing question that we investigate in this study. Naturally mated queens of the following five subspecies from different parts of Europe were transferred to Southern Germany: A. m. iberiensis from Portugal, A. m. mellifera from Belgium, A. m. macedonica from Greece, A. m. ligustica from Italy, and A. m. ruttneri from Malta. We also included the local subspecies A. m. carnica in our study. New colonies were built up in a common apiary where the respective queens were introduced. Worker offspring from the different subspecies were compared in classical olfactory learning performance using the proboscis extension response. Prior to conditioning, we measured individual sucrose responsiveness to investigate whether possible differences in learning performances were due to differential responsiveness to the sugar water reward. Most subspecies did not differ in their appetitive learning performance. However, foragers of the Iberian honeybee, A. m. iberiensis, performed significantly more poorly, despite having a similar sucrose responsiveness. We discuss possible causes for the poor performance of the Iberian honeybees, which may have been shaped by adaptation to the local habitat.

Authoritative subspecies diagnosis tool for European honey bees based on ancestry informative SNPs.

BACKGROUND: With numerous endemic subspecies representing four of its five evolutionary lineages, Europe holds a large fraction of Apis mellifera genetic diversity. This diversity and the natural distribution range have been altered by anthropogenic factors. The conservation of this natural heritage relies on the availability of accurate tools for subspecies diagnosis. Based on pool-sequence data from 2145 worker bees representing 22 populations sampled across Europe, we employed two highly discriminative approaches (PCA and FST) to select the most informative SNPs for ancestry inference. RESULTS: Using a supervised machine learning (ML) approach and a set of 3896 genotyped individuals, we could show that the 4094 selected single nucleotide polymorphisms (SNPs) provide an accurate prediction of ancestry inference in European honey bees. The best ML model was Linear Support Vector Classifier (Linear SVC) which correctly assigned most individuals to one of the 14 subspecies or different genetic origins with a mean accuracy of 96.2% ± 0.8 SD. A total of 3.8% of test individuals were misclassified, most probably due to limited differentiation between the subspecies caused by close geographical proximity, or human interference of genetic integrity of reference subspecies, or a combination thereof. CONCLUSIONS: The diagnostic tool presented here will contribute to a sustainable conservation and support breeding activities in order to preserve the genetic heritage of European honey bees.

Geographical Distribution and Selection of European Honey Bees Resistant to Varroa destructor.

Developing resistance to the varroa mite in honey bees is a major goal for apicultural science and practice, the development of selection strategies and the availability of resistant stock. Here we present an extended literature review and survey of resistant populations and selection programs in the EU and elsewhere, including expert interviews. We illustrate the practical experiences of scientists, beekeepers, and breeders in search of resistant bees. We describe numerous resistant populations surviving without acaricide treatments, most of which developed under natural infestation pressure. Their common characteristics: reduced brood development; limited mite population growth; and low mite reproduction, may cause conflict with the interests of commercial beekeeping. Since environmental factors affect varroa mite resistance, particular honey bee strains must be evaluated under different local conditions and colony management. The resistance traits of grooming, hygienic behavior and mite reproduction, together with simple testing of mite population development and colony survival, are significant in recent selection programs. Advanced breeding techniques and genetic and physiological selection tools will be essential in the future. Despite huge demand, there is no well-established market for resistant stock in Europe. Moreover, reliable experience or experimental evidence regarding the resistance of stocks under different environmental and management conditions is still lacking.

Evaluation of Suppressed Mite Reproduction (SMR) Reveals Potential for Varroa Resistance in European Honey Bees (Apis mellifera L.).

In the fight against the Varroa destructor mite, selective breeding of honey bee (Apis mellifera L.) populations that are resistant to the parasitic mite stands as a sustainable solution. Selection initiatives indicate that using the suppressed mite reproduction (SMR) trait as a selection criterion is a suitable tool to breed such resistant bee populations. We conducted a large European experiment to evaluate the SMR trait in different populations of honey bees spread over 13 different countries, and representing different honey bee genotypes with their local mite parasites. The first goal was to standardize and validate the SMR evaluation method, and then to compare the SMR trait between the different populations. Simulation results indicate that it is necessary to examine at least 35 single-infested cells to reliably estimate the SMR score of any given colony. Several colonies from our dataset display high SMR scores indicating that this trait is present within the European honey bee populations. The trait is highly variable between colonies and some countries, but no major differences could be identified between countries for a given genotype, or between genotypes in different countries. This study shows the potential to increase selective breeding efforts of V. destructor resistant populations.

A long-term field study on the effects of dietary exposure of clothianidin to varroosis-weakened honey bee colonies.

Clothianidin is a commonly used systemic insecticide in seed treatments. Residues of clothianidin can occur in nectar and pollen as a result of within-plant-translocation. Foraging bees can collect contaminated nectar or pollen. Concerns have been brought forward that exposure to pesticide residues might affect colonies especially if they are weakened by varroosis. However, there are few scientific studies investigating such multiple-stressor scenarios in the context of the entire colony. To close this gapa field trial with 24 colonies was set up. The study design comprised four groups of six colonies each fed with uncontaminated sugar syrup ('C0'), or syrup spiked with 10 µg L-1 clothianidin ('C10'), 50 µg L-1 clothianidin ('C50') or 200 µg L-1 clothianidin ('C200'). C10 represented a residue concentration that may exceptionally occur and therefore a worst-case scenario, the higher dietary concentrations exceed and do not reflect fieldrealistic levels. A substantial load of 8 mites of Varroa destructor per ten gram bees in autumn was adjusted. The colonies were followed up for 328 days. The amount of brood and the strength of each colony were regularly assessed. Colony health, bee mortality, overwintering success, hive weights, and levels of in-hive residues were determined. Varroosis turned out to be the significant key factor for the endpoint colony strength. Clothianidin did not have a statistically significant impact on C0, C10 and C50 colonies. No statistical evidence was found for an interaction between varroosis andexposure to clothianidin.

Quantitative patterns of vertical transmission of deformed wing virus in honey bees.

Deformed wing virus (DWV) is an important pathogen in a broad range of insects, including honey bees. Concordant with the spread of Varroa, DWV is present in the majority of honey bee colonies and can result in either low-level infections with asymptomatic bees that nonetheless exhibit increased colony loss under stress, or high-level infections with acute effects on bee health and viability. DWV can be transmitted vertically or horizontally and evidence suggests that horizontal transmission via Varroa is associated with acute symptomatic infections. Vertical transmission also occurs and is presumably important for the maintenance of DWV in honey bee populations. To further our understanding the vertical transmission of DWV through queens, we performed three experiments: we studied the quantitative effectiveness of vertical transmission, surveyed the prevalence of successful egg infection under commercial conditions, and distinguished among three possible mechanisms of transmission. We find that queen-infection level predicts the DWV titers in their eggs, although the transmission is not very efficient. Our quantitative assessment of DWV demonstrates that eggs in 1/3 of the colonies are infected with DWV and highly infected eggs are rare in newly-installed spring colonies. Additionally, our results indicate that DWV transmission occurs predominantly by virus adhering to the surface of eggs (transovum) rather than intracellularly. Our combined results suggest that the queens' DWV vectoring capacity in practice is not as high as its theoretical potential. Thus, DWV transmission by honey bee queens is part of the DWV epidemic with relevant practical implications, which should be further studied.

Performance of honey bee colonies under a long-lasting dietary exposure to sublethal concentrations of the neonicotinoid insecticide thiacloprid.

BACKGROUND: Substantial honey bee colony losses have occurred periodically in the last decades. The drivers for these losses are not fully understood. The influence of pests and pathogens are beyond dispute, but in addition, chronic exposure to sublethal concentrations of pesticides has been suggested to affect the performance of honey bee colonies. This study aims to elucidate the potential effects of a chronic exposure to sublethal concentrations (one realistic worst-case concentration) of the neonicotinoid thiacloprid to honey bee colonies in a three year replicated colony feeding study. RESULTS: Thiacloprid did not significantly affect the colony strength. No differences between treatment and control were observed for the mortality of bees, the infestation with the parasitic mite Varroa destructor and the infection levels of viruses. No colony losses occurred during the overwintering seasons. Furthermore, thiacloprid did not influence the constitutive expression of the immunity-related hymenoptaecin gene. However, upregulation of hymenoptaecin expression as a response to bacterial challenge was less pronounced in exposed bees than in control bees. CONCLUSION: Under field conditions, bee colonies are not adversely affected by a long-lasting exposure to sublethal concentrations of thiacloprid. No indications were found that field-realistic and higher doses exerted a biologically significant effect on colony performance. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Deformed wing virus can be transmitted during natural mating in honey bees and infect the queens.

Deformed wing virus is an important contributor to honey bee colony losses. Frequently queen failure is reported as a cause for colony loss. Here we examine whether sexual transmission during multiple matings of queens is a possible way of virus infection in queens. In an environment with high prevalence of deformed wing virus, queens (n = 30) were trapped upon their return from natural mating flights. The last drone's endophallus (n = 29), if present, was removed from the mated queens for deformed wing virus quantification, leading to the detection of high-level infection in 3 endophalli. After oviposition, viral quantification revealed that seven of the 30 queens had high-level deformed wing virus infections, in all tissues, including the semen stored in the spermathecae. Two groups of either unmated queens (n = 8) with induced egg laying, or queens (n = 12) mated in isolation with drones showing comparatively low deformed wing virus infections served as control. None of the control queens exhibited high-level viral infections. Our results demonstrate that deformed wing virus infected drones are competitive to mate and able to transmit the virus along with semen, which occasionally leads to queen infections. Virus transmission to queens during mating may be common and can contribute noticeably to queen failure.

Effects of genotype, environment, and their interactions on honey bee health in Europe.

There are several reports of honey bee populations in Europe which survive without treatment for Varroa. However, when evaluated outside their native area, higher survival and resistance traits were not observed in colonies of a survivor population. Varroa infestation is strongly influenced by environmental factors, probably affecting threshold levels on a European scale. In a Europe-wide experiment colonies of local origin survived significantly longer than colonies of non-local origin, clearly indicating the presence of genotype-environment interactions. Transmission by Varroa selects for virulent strains of DWV, but it is currently unknown how these may interact with different genotypes of bees. The distribution of Nosema ceranae is significantly affected by environment, but there is at least one Nosema-resistant population.

Honey bee surveillance: a tool for understanding and improving honey bee health.

Honey bee surveillance systems are increasingly used to characterize honey bee health and disease burdens of bees in different regions and/or over time. In addition to quantifying disease prevalence, surveillance systems can identify risk factors associated with colony morbidity and mortality. Surveillance systems are often observational, and prove particularly useful when searching for risk factors in real world complex systems. We review recent examples of surveillance systems with particular emphasis on how these efforts have helped increase our understanding of honey bee health.

Hinf-I digestion of cytochrome oxidase I region is not a diagnostic test for A. m. lamarckii

Restriction fragment length polymorphism of whole mitochondrial DNA or PCR amplified mtDNA regions are known to be useful in discriminating among honey bee lineages and also some individual subspecies. In this study, PCR-amplified fragments of cytochrome oxidase I (CO-I) and cytochrome B (Cyt B) of honey bees sampled from different countries (Cyprus, Turkey, Ethiopia, Syria and Egypt) were digested with Hinf I and Bgl II restriction enzymes, respectively. Eastern Europe and Mediterranean honey bee subspecies were separated by the Cyt B/Bgl II analysis, although Hinf I digestion of the CO-I region yielded much finer resolution within different honey bee lineages. Here we report that CO-I/Hinf-I is a discriminative test for the mitochondrial "O" lineage, rather than a diagnostic site for A. m. lamarckii.

Recent Mediterranean fruit fly (Diptera: Tephritidae) infestations in Florida--a genetic perspective.

Microsatellite and mitochondrial DNA (mtDNA) variability data were used to study infestations of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) in Florida in 1997 and 1998. A total of 132 flies collected in monitoring traps or as larvae removed from fruit were examined at three polymorphic mtDNA restriction sites and two microsatellite loci. All of the flies sampled in Florida in 1997 displayed the mitochondrial AAB haplotype and represent a novel introduction of Mediterranean fruit flies into the state. All flies collected in central Florida in 1998 also displayed the AAB haplotype. Microsatellite analysis of these specimens from 1998 detected only alleles that were present in 1997. These results strongly indicate that the 1998 Florida outbreaks were derived from the Florida populations from the previous year. According to our analyses, the Mediterranean region is the most likely source for the 1997 Florida infestation. Flies from a small outbreak in Miami Springs, Dade County, FL, early in 1998 had a different mtDNA haplotype, characterized by the AAC restriction pattern. Microsatellites of these specimens showed significant differences in their allelic distribution from AAB flies, indicating an origin from a separate source population. South America is the most likely source for the Miami Springs flies.