Standard survey methods for estimating colony losses and explanatory risk factors in Apis mellifera

This chapter addresses survey methodology and questionnaire design for the collection of data pertaining to estimation of honey bee colony loss rates and identification of risk factors for colony loss. Sources of error in surveys are described. Advantages and disadvantages of different random and non-random sampling strategies and different modes of data collection are presented to enable the researcher to make an informed choice. We discuss survey and questionnaire methodology in some detail, for the purpose of raising awareness of issues to be considered during the survey design stage in order to minimise error and bias in the results. Aspects of survey design are illustrated using surveys in Scotland. Part of a standardized questionnaire is given as a further example, developed by the COLOSS working group for Monitoring and Diagnosis. Approaches to data analysis are described, focussing on estimation of loss rates. Dutch monitoring data from 2012 were used for an example of a statistical analysis with the public domain R software. We demonstrate the estimation of the overall proportion of losses and corresponding confidence interval using a quasi-binomial model to account for extra-binomial variation. We also illustrate generalized linear model fitting when incorporating a single risk factor, and derivation of relevant confidence intervals

Phylogeography and population genetics of honey bees (Apis mellifera) from Turkey based on COI-COII sequence data

A study that involved DNA sequencing of COI-COII intergenic region of the mitochondrial DNA genome of Apis mellifera honey bees from Turkey was conducted to determine the population genetics and phylogeographic structure of this species from seven distinct areas of Turkey. From the 132 honey bees subjected to DNA sequencing, a total of 12 mitotypes of A. mellifera "C" lineage were observed, of which only one mitotype, C 13, had been reported previously. The most common mitotype, C12, accounted for 47% of the Apis mellifera "C" lineage samples and was found in 13 of the 22 sampled locations. This mitotype was also the basal ancestral mitotype based on TCS spanning tree analysis. The greatest amount of genetic diversity was observed in Bursa, where 4 mitotypes of the A. mellifera "C" lineage were unique to this location. Wright's F-statistics revealed that Artvin and Bursa were the most genetically distinct locations relative to the other sampled locations. Applying a molecular clock, Turkish A. mellifera "C" lineage mitotypes have been diverging for approximately 10,000 to 16,500 yr. based on phylogenetic analysis. In addition, two A. m. syriaca samples were observed from Hatay, Turkey. Phylogenetic analysis which included other A. mellifera subspecies confirms the subspecies relationships of A. mellifera "C" lineage, and A. m. syriaca. this study corroborates other studies that show Turkey to be a reservoir of genetically distinct populations of A. mellifera "C" lineage, which can be useful for developing genetic conservation strategies for A. mellifera

Proboscis conditioning experiments with honeybees, Apis mellifera caucasica, with butyric acid and DEET mixture as conditioned and unconditioned stimuli

Three experiments are described investigating whether olfactory repellents DEET and butyric acid can support the classical conditioning of proboscis extension in the honeybee, Apis mellifera caucasica (Hymenoptera: Apidae). In the first experiment DEET and butyric acid readily led to standard acquisition and extinction effects, which are comparable to the use of cinnamon as a conditioned stimulus. These results demonstrate that the odor of DEET or butyric acid is not intrinsically repellent to honey bees. In a second experiment, with DEET and butyric acid mixed with sucrose as an unconditioned stimulus, proboscis conditioning was not established. After several trials, few animals responded to the unconditioned stimulus. These results demonstrate that these chemicals are gustatory repellents when in direct contact. In the last experiment a conditioned suppression paradigm was used. Exposing animals to butyric acid or DEET when the proboscis was extended by direct sucrose stimulation or by learning revealed that retraction of the proboscis was similar to another novel odor, lavender, and in all cases greatest when the animal was not permitted to feed. These results again demonstrate that DEET or butyric acid are not olfactory repellents, and in addition, conditioned suppression is influenced by feeding state of the bee.Peer reviewedPsychologyZoolog

HBeeID: a molecular tool that identifies honey bee subspecies from different geographic populations

Background: Honey bees are the principal commercial pollinators. Along with other arthropods, they are increasingly under threat from anthropogenic factors such as the incursion of invasive honey bee subspecies, pathogens and parasites. Better tools are needed to identify bee subspecies. Genomic data for economic and ecologically important organisms is increasing, but in its basic form its practical application to address ecological problems is limited. Results: We introduce HBeeID a means to identify honey bees. The tool utilizes a knowledge-based network and diagnostic SNPs identified by discriminant analysis of principle components and hierarchical agglomerative clustering. Tests of HBeeID showed that it identifies African, Americas-Africanized, Asian, and European honey bees with a high degree of certainty even when samples lack the full 272 SNPs of HBeeID. Its prediction capacity decreases with highly admixed samples. Conclusion: HBeeID is a high-resolution genomic, SNP based tool, that can be used to identify honey bees and screen species that are invasive. Its flexible design allows for future improvements via sample data additions from other localities

Morphometric and electrophoretic variation in different honeybee (Apis mellifera L.) populations

Honeybee colonies from Turkey, Naxcivan (Azerbaijan) and Austria were studied for morphometric and electrophoretic variation and for comparison among different subspecies. Out of six enzyme systems, four were found to be polymorphic with 14 allozymes. Discriminant function analysis of 10 morphometric variables resulted in a total of four groups. Cluster analysis of electrophoretic data showed a similar type of groupings. Among the four polymorphic loci, Mdh-1 showed the highest and Est-3 the lowest differentiation among populations. © TÜBİTAK

Map position of phosphoglucomutase (Pgm) locus on autosome IV of house fly (Diptera: Muscidae)

We determined the map position of phosphoglucomutase (Pgm) locus on autosome IV of housefly, Musca domestica L. (Diptera: Muscidae), by using three and five point linkage test crosses. Test strains had visible mutant markers (car, bwb, cyw, and aabys). We analyzed 1,738 offspring in total from two groups of single-pair matings by electrophoresis. Here, we report the linkage of Pgm locus to autosome IV loci curly wing (cyw) and yellow eyes (ye) with recombination frequency of 16.9 and 1.1%, respectively. We combined the distances calculated from this study and the previously published data. An updated linkage map of the M. domestica L. Autosome IV was drawn based on combined data in terms of real map units obtained from the mapping function. © 2006 Entomological Society of America

Mitochondrial DNA variation in honey bee (apis mellifera l.) populations from Turkey

We have studied mitochondrial (mt) DNA variation in 334 honey bee colonies from 7 different geographic regions of Turkey. We have evaluated Dra I restriction profiles of the CO-I CO-II intergenic region, Hinf-I and Taq-I restriction profiles of the CO-I gene and EcoR-I restriction profiles of the whole mtDNA. We obtained three different mtDNA patterns by EcoR-I digestion. The pattern typical for A. m. carnica/A. m. ligustica predominated throughout Turkey (327 colonies, 97.9%). We observed the pattern common in African subspecies only in Hatay province (6 colonies, 1.8%) and a previously unreported pattern in one colony from Balıkesir province (0.3%). Dra-I restriction analysis of the CO-I CO-II intergenic region yielded seven haplotypes. Haplotype 1 (TrDra-1) was the most common one found in Turkey, whereas haplotype 2 (TrDra-2) was widely distributed in Eastern Anatolia. Based on mitochondrial ND2 sequences taken from two samples collected in each region, bees from Hatay clustered with A. m. lamarckii and A. m. meda (morphological A and O lineages), while bees from central Anatolia clustered within the C morphological lineage group. © 2006 International Bee Research Association.North Atlantic Treaty Organization: CRG.309726The authors would like to thank to all who assisted to this project in various ways. L. Garnery provided insight and guidance in methodology pertaining to the Dra-I test. This project was partially supported by a grant (VHAG -1077) from TUBITAK to MK, by an Integrated Ph.D. Program project grant from TUBITAK to AK, and by a NATO collaborative linkage grant (CRG.309726) to AK, LG, and WS. We also thank to anonymous reviewers for their comments on an earlier version of the manuscript