The behavioral genetics of colony defense in honeybees: Genetic variability for guarding behavior

Guard honeybees stand at the entrance of colonies and facilitate the exclusion of nonnestmates from the colony. In this study, we examined the hypothesis that genetic variability among individuals in colonies might explain variability in guarding activity. To do this, we cross-fostered honey bees between colonies with high-defensive responses and colonies with low-defensive responses in alarm pheromone tests. Individuals from high-defensive colonies were more likely to guard in their own colonies (controls) than cross-fostered bees from low-defensive colonies. Cross-fostered high-defensive bees also were more likely to guard in low-defense colonies. These results support the hypothesis that interindividual differences in guarding behavior are at least partially under genetic control. A positive correlation between number of guards and response to alarm pheromone demonstrates a link between behaviorally separated components of the overall defensive response.This work was supported by NSF Grant BNS 8605604.     

Determination of Genetically Identical Strains of Four Honeybee Viruses in Bumblebee Positive Samples

In recent years, there has been growing evidence that certain types of honeybee viruses could be transmitted between different pollinators. Within a voluntary monitoring programme, 180 honeybee samples (Apis mellifera carnica) were collected from affected apiaries between 2007 and 2018. Also from August 2017 to August 2018, a total 148 samples of healthy bumblebees (Bombus lapidarius, B. pascuorum, B. terrestris, B. lucorum, B. hortorum, B. sylvarum, B. humilis) were collected at four different locations in Slovenia, and all samples were tested by using RT-PCR methods for six honeybee viruses. Direct sequencing of a total 158 positive samples (acute bee paralysis virus (ABPV n = 33), black queen cell virus (BQCV n = 75), sacbrood bee virus (SBV n = 25) and Lake Sinai virus (LSV n = 25)) was performed from obtained RT-PCR products. The genetic comparison of identified positive samples of bumblebees and detected honeybee field strains of ABPV, BQCV, SBV, and LSV demonstrated 98.74% to 100% nucleotide identity between both species. This study not only provides evidence that honeybees and bumblebees are infected with genetically identical or closely related viral strains of four endemically present honeybee viruses but also detected a high diversity of circulating strains in bumblebees, similar as was observed among honeybees. Important new genetic data for endemic strains circulating in honeybees and bumblebees in Slovenia are presented.     

Risk Assessment for Honeybees from Pesticide-Exposed Pollen

A method for assessing the risk for honeybees from pesticide exposure via pollen is proposed. Four pesticides, selected as markers, were monitored in pollen samples collected in two sampling areas, one located in an intensive agricultural area and the other far from direct pesticide impact. Analytical results were consistent with use patterns of the chemicals and their physico-chemical and persistence properties. For a preliminary estimate of bee exposure via pollen, both by ingestion and by contact, an exposure index was developed, based on physico-chemical properties, persistence and application rates. On the basis of the exposure estimates and acute toxicological data (ingestion and contact LD₅₀), Toxicity Exposure Ratios (TERs) were calculated as indicators of the risk for honeybees due to this particular exposure route. TER values were compared to Hazard Quotient (HQ), calculated as the ratio between application rate and the LC₅₀ value, according to European guidelines, showing a satisfactory agreement. The advantage of the above described procedures is that the environmental fate of the chemicals, and not only application rates, are taken into account. This approach may represent a preliminary tool for a comparative screening of the risk for pollinator insects due to this particular exposure route.     

Rapid dendritic spine stem shortening during one-trial learning: The honeybee's first orientation flight

Our initial study of honeybees using the rapid Golgi method showed that dendritic spines on calycal interneurons had shorter stems due to spine head enlargement in bees with greater cumulative experience. This study sought to determine if spine stem shortening could be induced rapidly the first orientation flight, a one-trial place learning event. Newly emerged bees were reared in a small broodless hive with a virgin queen and allowed to take their first orientation flight at 6 and 8 days of age. Spine profiles of 5 flyers and 5 non-flyers were traced in large scale using light microscopy and a modified camera lucida. Overall spine length and stem length were measured on these tracings using a digitizing tablet. Additional measurements of maximum spine head width, profile area, and perimeter were made using computer image analyses. Examining group differences in spine length as a function of overall spine length, our results revealed a clear association between rapid spine stem shortening and the first orientation flight lasting several minutes. This effect, however, was restricted to only the long spines. Flight-induced stem shortening was accompanied by elongated swelling of the spine head without an appreciable expansion of the spine perimeter.     

Identification of the proteome composition occurring during the course of embryonic development of bees (Apis mellifera)

To investigate the proteome during embryonic development of honeybees, Apis mellifera , proteins were identified by two-dimensional gel electrophoresis, mass spectrometry and protein engine identification tools that were applied to MASCOT and Xproteo search engines. 312, 320, 315 proteins were detected in 24, 48 and 72 h embryos. Thirty-eight highly abundant proteins were identified at the three time points by MS fingerprinting. All 21 proteins could be identified as products of annotated genes of the honeybee. Identified proteins included six proteins related to the metabolism of carbohydrates and energy production, six proteins belonging to the heat shock protein family, three cytoskeletal proteins, four proteins related to the antioxidant system of the embryo and two proteins related to growth regulation of the embryo. Quantitative proteomics was applied to analyze differences in amounts of these proteins during the three above mentioned developmental stages. Our data present an initial molecular picture of honeybee embryos, and will hopefully pave the way for future research on this animal.     

Rapid diagnosis of infectious diseases: The role of giant African pouched rats,dogs and honeybees

Early detection of infection in an individual is an important strategy to prevent transmission and spread of the infection – in the community or among patients admitted in a health care facility. Over the past decade, there have been rapid advances in technology in an attempt to provide rapid, accurate tests for diagnosis of infectious diseases. With this in perspective, the use of animals’ superior olfactory sensitivity to sniff out infectious diseases holds promise.     

Optical Recording of Neuronal Activity in the Insect Central Nervous System: Odorant Coding by the Antennal Lobes of Honeybees

Voltage-sensitive dyes and activity-dependent intrinsic optical signals were used to study the spatio-temporal activity in the antennal lobes of honeybees. The intrinsic signals are somewhat slower than the dye signals but show a 10-fold larger intensity change. These intrinsic signals consist of at least two components—one is wavelength-independent and the other strongly wavelength-dependent, with a maximum at ∼500 nm. Local inhibitory connections within the antennal lobes were examined by recording the activity elicited by an electrical stimulus to the antennal nerve of a slice preparation before and after applying picrotoxin to manipulate GABAergic inhibitory synapses. The inhibition starts with a delay of ∼10 ms after onset of the response and has at least two components. The spatial distribution of the inhibition is extremely inhomogeneous, with areas of small inhibition adjacent to areas of large inhibition. Thus inhibitory interactions in the antennal lobes are not evenly distributed among the glomerular organization. Stimulation of an in vivo preparation with an odour yields a spatially restricted activity. However, the spatial map appears highly dynamic in time because the size of the activated area is a function of the time during and after the stimulus.     

Quantitative Trait Loci Associated with Reversal Learning and Latent Inhibition in Honeybees (Apis mellifera)

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.     

Genotype,Task Specialization,and Nest Environment Influence the Stinging Response Thresholds of Individual Africanized and European Honeybees to Electrical Stimulation

This study was conducted to analyze the stinging response thresholds of individual European and Africanized worker honeybees (Apis mellifera L.) to electrical stimulation. Newly emerged workers were identified, and either were placed into an incubator, into their natal colonies, or cross-fostered in common colonies of European or Africanized ancestry. Nest and guard bees of each type were collected and exposed to an electric stimulus of 0.5 mA, and the time they took to sting a leather substrate was recorded. Africanized bees consistently had significant lower thresholds of defensive response than European bees across all of the environments tested. Guards were faster to sting than nest bees only for the Africanized genotype, suggesting that alleles of African origin have pleiotropic effects on guarding and stinging. This is the first study that shows that single individuals specialized in guarding also may have a lower response threshold for stinging. Environmental effects were also evident. In all cases, bees responded faster to the electrical stimulation after being kept in environments other than their natal nest. Moreover, significant genotype by environment and genotype by task specialization interactions were found. Our results fit a model of division of labor based on differences in response thresholds to stimuli among workers of different genotypes and task groups that result in non-additive effects on colony behavior. Edited by Yong-Kyu Kim.