Western Honey Bee Apis Mellifera Research Articles

Analyzing Gut Microbial Community in Varroa destructor-Infested Western Honeybee (Apis mellifera).

The western honeybee Apis mellifera L., a vital crop pollinator and producer of honey and royal jelly, faces numerous threats including diseases, chemicals, and mite infestations, causing widespread concern. While extensive research has explored the link between gut microbiota and their hosts. However, the impact of Varroa destructor infestation remains understudied. In this study, we employed massive parallel amplicon sequencing assays to examine the diversity and structure of gut microbial communities in adult bee groups, comparing healthy (NG) and Varroa-infested (VG) samples. Additionally, we analyzed Varroa-infested hives to assess the whole body of larvae. Our results indicated a notable prevalence of the genus Bombella in larvae and the genera Gillamella, unidentified Lactobacillaceae, and Snodgrassella in adult bees. However, no statistically significant difference was observed between NG and VG. Furthermore, our PICRUSt analysis demonstrated distinct KEGG classification patterns between larval and adult bee groups, with larvae displaying a higher abundance of genes involved in cofactor and vitamin production. Notably, despite the complex nature of the honeybee bacterial community, methanogens were found to be present in low abundance in the honeybee microbiota.

Locomotion behavioural responses of Varroa destructor exposed to western honey bee (Apis mellifera) semiochemicals

Locomotion behavioural responses of Varroa destructor exposed to western honey bee (Apis mellifera) semiochemicals

Age-related response to mite parasitization and viral infection in the honey bee suggests a trade-off between growth and immunity.

Host age at parasites' exposure is often neglected in studies on host-parasite interactions despite the important implications for epidemiology. Here we compared the impact of the parasitic mite Varroa destructor, and the associated pathogenic virus DWV on different life stages of their host, the western honey bee Apis mellifera. The pre-imaginal stages of the honey bee proved to be more susceptible to mite parasitization and viral infection than adults. The higher viral load in mite-infested bees and DWV genotype do not appear to be the drivers of the observed difference which, instead, seems to be related to the immune-competence of the host. These results support the existence of a trade-off between immunity and growth, making the pupa, which is involved in the highly energy-demanding process of metamorphosis, more susceptible to parasites and pathogens. This may have important implications for the evolution of the parasite's virulence and in turn for honey bee health. Our results highlight the important role of host's age and life stage at exposure in epidemiological modelling. Furthermore, our study could unravel new aspects of the complex honey bee-Varroa relationship to be addressed for a sustainable management of this parasite.

Expansion microscopy in honeybee brains for high-resolution neuroanatomical analyses in social insects

The diffraction limit of light microscopy poses a problem that is frequently faced in structural analyses of social insect brains. With the introduction of expansion microscopy (ExM), a tool became available to overcome this limitation by isotropic physical expansion of preserved specimens. Our analyses focus on synaptic microcircuits (microglomeruli, MG) in the mushroom body (MB) of social insects, high-order brain centers for sensory integration, learning, and memory. MG undergo significant structural reorganizations with age, sensory experience, and during long-term memory formation. However, the changes in subcellular architecture involved in this plasticity have only partially been accessed yet. Using the western honeybee Apis mellifera as an experimental model, we established ExM for the first time in a social insect species and applied it to investigate plasticity in synaptic microcircuits within MG of the MB calyces. Using combinations of antibody staining and neuronal tracing, we demonstrate that this technique enables quantitative and qualitative analyses of structural neuronal plasticity at high resolution in a social insect brain.

Honey bees (Apis mellifera) preselected for Varroa sensitive hygiene discriminate between live and dead Varroa destructor and inanimate objects

Varroa destructor is one of the main causes of colony losses of the western honey bee (Apis mellifera). Many efforts exist to breed honey bees resistant to V. destructor. Varroa sensitive hygiene (VSH) is a commonly selected behavioural trait; VSH workers remove the pupae of mite infested brood cells with high efficiency, interrupting the reproduction of the mite. The cues and triggers for this behaviour are not yet fully understood. To determine what elicits this removal behaviour, we examined preselected VSH workers´ responses to four different groups of objects inserted into freshly capped cells: live mites, dead mites, odour reduced mites, and glass beads. These were also compared to control cells that were opened and closed without inserting any object. The pupae in cells containing inorganic objects (glass beads) were removed at similar rates to the control, demonstrating that an object alone does not trigger a removal response. Dead and odour reduced mites were removed at a higher frequency than control cells, but less frequently than live mites. Workers sometimes removed items resting near the top of the cell without removing the pupa. Our results demonstrate that although mite odour from dead mites triggers removal behaviour, the pupa of cells containing live mites were removed more frequently, suggesting that other cues (i.e. odour from feeding wound) or signals (i.e. pupal movement to signal distress) are important. Future research should focus on elucidating these other cues or signals from the brood and mites, as mite presence alone seems to be insufficient.

The presence of identical deformed wing virus sequence variants in co-occurring Apis species in Northern Thailand may represent a potential epidemiological threat to native honey bees of Southeast Asia

Widespread native honey bee species in South and East Asia (Apis cerana, Apis dorsata and Apis florea) and the imported western honey bee (Apis mellifera) share habitats and potentially also share pathogens. Chief among the threats facing A. mellifera in Europe and North America is deformed wing virus (DWV), including its two principal genotypes: A and B (DWV-A and DWV-B respectively). Though DWV-A has been recorded in Asia’s native Apis species, it is not known if DWV-B, or both DWV-A and DWV-B, are currently widespread in Asia and, if so, whether viral transmission is primarily intraspecific or interspecific. This study aims to fill these knowledge gaps by (i) determining the DWV genotype in four co-occurring Apis host species using qPCR and (ii) inferring viral transmission between them using nucleotide sequences of DWV from Apis host species collected at three independent localities in Northern Thailand. We found DWV-A and -B in all four Apis species, the exotic A. mellifera and the native A. cerana, A. dorsata and A. florea. That DWV-A sequences were identical across Apis species at the same locality, with a similar pattern for DWV-B sequences, suggests that DWV’s epidemiology is largely driven by ongoing interspecific transmission (spillover) of DWV across co-occurring native and exotic Apis species. Both genotypes of DWV represent a serious threat to Asia’s exotic and native honey bee species.

Bombella pluederhausensis sp. nov., Bombella pollinis sp. nov., Bombella saccharophila sp. nov. and Bombella dulcis sp. nov., four Bombella species isolated from the environment of the western honey bee Apis mellifera.

Four strains of members of the genus Bombella were isolated from samples associated with the western honey bee Apis mellifera, which could not be assigned to a species with a validly published name. Strains TMW 2.2543T, TMW 2.2556T, TMW 2.2558T and TMW 2.2559T exhibit in silico DNA-DNA hybridisation (isDDH) and orthologous average nucleotide identity (orthoANI) values below species delineation thresholds compared with all described species of the genus Bombella and with each other. TMW 2.2556T and TMW 2.2558T form their own clade within the genus. The major respiratory quinone of all strains was Q-10. The composition of cellular fatty acids was diverse between strains. All strains stained Gram-negative, were rod-shaped, strictly aerobic, pellicle-forming, catalase-positive, oxidase-negative, mesophilic and grew over a wide pH range; they were halosensitive but glucose-tolerant. Unlike the other studied strains, TMW 2.2558T was non-motile. Phylogenetic, chemotaxonomic and physiological analyses revealed a clear distinction between all the strains and species with validly published names. All the data support the proposition of four novel species within the genus Bombella, namely Bombella pluederhausensis sp. nov., Bombella pollinis sp. nov., Bombella saccharophila sp. nov. and Bombella dulcis sp. nov., with the respective type strains Bombella pluederhausensis sp. nov. TMW 2.2543T (= DSM 114872T, = LMG 32791T), Bombella pollinis sp. nov. TMW 2.2556T (= DSM 114874T, = LMG 32792T), Bombella saccharophila sp. nov. TMW 2.2558T (= DSM 114875T, = LMG 32793T) and Bombella dulcis sp. nov. TMW 2.2559T (= DSM 114877T, = LMG 32794T). Moreover, three genomes available in the NCBI database that have not yet been described as species with validly published names could be assigned to the proposed species. Bombella sp. ESL0378 and Bombella sp. ESL0385 to Bombella pollinis sp. nov. and Bombella sp. AS1 to Bombella saccharophila sp. nov.

Report of amoebic disease in a colony of Western honey bees (Apis mellifera).

The amoeba Malpighamoeba mellificae is the etiologic agent of amoebic (amoeba) disease of Western honey bees (Apis mellifera). M. mellificae damages the Malpighian tubules, which is believed to weaken and kill the host bee. Here, the authors describe the detection of this organism in a honey bee colony in the Yukon Territory, Canada. The Malpighian tubules of 14% (7/50) of the adult worker bees were discolored dark brown. Fifteen bees screened using conventional polymerase chain reaction for the 18S gene of M. mellificae were positive for the pathogen. Histologically, the lumens of Malpighian tubules were packed with amoebae, causing dilation of the tubules and attenuation and loss of the tubular epithelium. This phylogenetic analysis places M. mellificae in a new clade, a sister group to the Entamoebidae. This work provides a foundation for further investigation into the distribution, prevalence, and pathology associated with M. mellificae infection.

Host brood traits, independent of adult behaviours, reduce Varroa destructor mite reproduction in resistant honeybee populations

The ectoparasitic mite Varroa destructor is an invasive species of Western honey bees (Apis mellifera) and the largest pathogenic threat to their health world-wide. Its successful invasion and expansion is related to its ability to exploit the worker brood for reproduction, which results in an exponential population growth rate in the new host. With invasion of the mite, wild honeybee populations have been nearly eradicated from Europe and North America, and the survival of managed honeybee populations relies on mite population control treatments. However, there are a few documented honeybee populations surviving extended periods without control treatments due to adapted host traits that directly impact Varroa mite fitness. The aim of this study was to investigate if Varroa mite reproductive success was affected by traits of adult bee behaviours or by traits of the worker brood, in three mite-resistant honey bee populations from Sweden, France and Norway. The mite’s reproductive success was measured and compared in broods that were either exposed to, or excluded from, adult bee access. Mite-resistant bee populations were also compared with a local mite-susceptible population, as a control group. Our results show that mite reproductive success rates and mite fecundity in the three mite-resistant populations were significantly different from the control population, with the French and Swedish populations having significantly lower reproductive rates than the Norwegian population. When comparing mite reproduction in exposed or excluded brood treatments, no differences were observed, regardless of population. This result clearly demonstrates that Varroa mite reproductive success can be suppressed by traits of the brood, independent of adult worker bees.

Novel eDNA approaches to monitor Western honey bee (Apis mellifera L.) microbial and arthropod communities

AbstractPests and pathogens are a continuous threat to the health of Western honey bee Apis mellifera L. Monitoring honey bee colonies for arthropod pests, disease‐causing bacteria and fungi, and early detection of new invasions is essential to maintain the pollination services provided by honey bees. We investigated the feasibility of using eDNA metabarcoding to detect pests and pathogens in bee hives and across their foraging environment. We sampled 13 sources for eDNA within and outside hives from our test apiary to determine where the most informative eDNA could be obtained, with most sources sampled thrice. This resulted in 61 samples, 20 negative controls, and three positive controls. Furthermore, we compared two eDNA collection techniques—wiping surfaces with moistened forensic swabs and using a spray/wash technique that aggregated surface eDNA into a container, before collecting the eDNA on a filter. We used DNA metabarcoding with universal primer sets to target arthropod, bacterial, and fungal communities. Our results showed that most sources yielded sufficient eDNA and that results of the swab and spray/wash methods were similar when they could be applied to the same surface. We detected DNA from honey bee bacterial symbionts, mycotoxin‐producing fungi and Brachymyrmex sp. rover ants. Common pests and pathogens of concern to honey bees (i.e., [small hive beetle (Aethina tumida), Varroa destructor, and Melissococcus plutonius]) were detected. This matched our visual observations of clinical signs of these pests and pathogens in the hives we tested. DNA from some species was source specific, which has implications for using eDNA as a monitoring tool. Collectively, our data show that eDNA metabarcoding can accurately detect DNA from arthropods and microbes honey bees contact and can be used as a comprehensive molecular predictor tool for colony health surveys.

이중가닥 RNA를 이용한 재래꿀벌 낭충봉아부패병 바이러스 질병 치료제의 과거와 현재

Apiculture has been taking an important role for agriculture to provide the pollinators. In Korea, the major pollinators for agricultural and horticultural production are the Western honeybee ( Apis mellifera) and the Asian honeybee ( Apis cerana). Until late 2000s, the Asian honeybee has been responsible for 20% of the Korean apiculture, however, the majority of the Asian honeybee population was decimated by the sacbrood virus (SBV). In the meantime, molecular technologies have been developed to diagnose the SBV infected honeybees and to suppress the SBV by the RNA interference techniques. Recently, the industrialization of double-stranded RNA (dsRNA) as SBV inhibitor is at hand by virtue of the technological progress. This review article is to summarize the numerous efforts of the last decade to overcome the threats of SBV outbreak to protect Asian honeybees in Korea.

Seasonal differences in Varroa destructor population growth in western honey bee (Apis mellifera) colonies

Varroa destructor is a major threat for apiculture worldwide. A successful approach to control this parasite must include the application of effective treatments at the correct time. To understand the effect that treatment timing has on Varroa populations at different seasons, we conducted an experiment using a dataset comprising two separate field trials over multiple years, both trials containing four apiary sites composed of 20 honey bee colonies across an area representative of north central Florida environments. Before the start of the season, colonies were treated with two acaricides simultaneously to bring the Varroa populations to ∼0.25 mites/100 bees. Following treatment, we monitored the mite populations monthly via alcohol washes. Our results show that the temporal efficacy of Varroa treatments varies across seasons. We observed that it takes about 4–5 months after treatment in winter and spring for mite populations to return to the standard economical threshold (3 mites/100 bees). Nevertheless, there is a steeper increase in mite populations (<3 months to exceed the economic threshold) after treating colonies in summer and fall. The level of infestation that leads to colony collapse and the rate of colony decline also varied by season. To our knowledge, this is the first study evaluating seasonal effects on Varroa population growth and the first model of Varroa population growth in Florida, USA. Our results serve as a foundation for Varroa treatment models, aiding beekeepers in the future as a part of a holistic approach to control this devastating honey bee parasite.

Foraging distances, habitat preferences and seasonal colony performance of honeybees in Central European forest landscapes

Abstract Unmanaged forest is considered as native habitat of the Western honeybee Apis mellifera and is assumed to provide important pollen and nectar sources. However, resource supply might be spatially and temporally restricted in managed forests and landscape‐scale studies in European forest regions are currently lacking. Capitalizing on a unique communication behaviour, the waggle dance, we investigated honeybee foraging in a deciduous forest region in southern Germany. We analysed 2022 waggle dances performed by bees of 12 colonies placed in the centre of landscapes with varying degree of forest cover (50%–99% at a 2‐km radius) from March to August 2019, thereby identifying foraging distances and habitat preferences over almost an entire foraging season. By connecting dance information with colony weight recordings, we assessed the role of landscape composition and the contribution of different habitat types to colony weight gain. Foraging distances generally increased with the amount of forest in the surrounding landscape pointing to inferior food resources in the forest. Yet, this effect strongly depended on the season and was more pronounced for pollen than for nectar foraging. Even though colonies in forest‐dominated landscapes had to fly further, colony weight was not significantly affected by forest cover. Compared to expectations based on the proportions of different habitats, colonies foraged more frequently in grassland and cropland than in deciduous and coniferous forests, with late summer being an especially difficult period for pollen foraging in forests. During a phase of colony weight gain in early summer, the use of forests for nectar/honeydew foraging was close to the expectation, highlighting forests as an important complementary source of carbohydrates during short periods of the year. Policy implication. The ecological and economic value of managed forest as habitat for honeybees could be significantly increased by the continuous provision of floral resources, especially for pollen foraging. Therefore, we recommend diversifying forest stands with insect‐pollinated trees, permitting secondary successions in forest gaps, and at larger scales, creating forest landscapes with high habitat diversity. This will not only help native, wild‐living honeybees and beekeepers' colonies residing in forest areas but will also foster social bees in general.

Intraspecific Variability in Proteomic Profiles and Biological Activities of the Honey Bee Hemolymph.

Pollinator declines have raised major concerns for the maintenance of biodiversity and food security, calling for a better understanding of environmental factors that affect their health. Here we used hemolymph analysis to monitor the health status of Western honey bees Apis mellifera. We evaluated the intraspecific proteomic variations and key biological activities of the hemolymph of bees collected from four Egyptian localities characterized by different food diversities and abundances. Overall, the lowest protein concentrations and the weakest biological activities (cytotoxicity, antimicrobial and antioxidant properties) were recorded in the hemolymph of bees artificially fed sucrose solution and no pollen. By contrast, the highest protein concentrations and biological activities were recorded in bees that had the opportunity to feed on various natural resources. While future studies should expand comparisons to honey bee populations exposed to more different diets and localities, our results suggest hemolymph samples can be used as reliable indicators of bee nutrition.

A robotic honeycomb for interaction with a honeybee colony.

Robotic technologies have shown the capability to interact with living organisms and even to form integrated mixed societies composed of living and artificial agents. Biocompatible robots, incorporating sensing and actuation capable of generating and responding to relevant stimuli, can be a tool to study collective behaviors previously unattainable with traditional techniques. To investigate collective behaviors of the western honeybee (Apis mellifera), we designed a robotic system capable of observing and modulating the bee cluster using an array of thermal sensors and actuators. We initially integrated the system into a beehive populated with about 4000 bees for several months. The robotic system was able to observe the colony by continuously collecting spatiotemporal thermal profiles of the winter cluster. Furthermore, we found that our robotic device reliably modulated the superorganism's response to dynamic thermal stimulation, influencing its spatiotemporal reorganization. In addition, after identifying the thermal collapse of a colony, we used the robotic system in a "life-support" mode via its thermal actuators. Ultimately, we demonstrated a robotic device capable of autonomous closed-loop interaction with a cluster comprising thousands of individual bees. Such biohybrid societies open the door to investigation of collective behaviors that necessitate observing and interacting with the animals within a complete social context, as well as for potential applications in augmenting the survivability of these pollinators crucial to our ecosystems and our food supply.

Significant, but not biologically relevant: Nosema ceranae infections and winter losses of honey bee colonies

The Western honey bee Apis mellifera, which provides about 90% of commercial pollination, is under threat from diverse abiotic and biotic factors. The ectoparasitic mite Varroa destructor vectoring deformed wing virus (DWV) has been identified as the main biotic contributor to honey bee colony losses worldwide, while the role of the microsporidium Nosema ceranae is still controversially discussed. In an attempt to solve this controversy, we statistically analyzed a unique data set on honey bee colony health collected from a cohort of honey bee colonies over 15 years and comprising more than 3000 data sets on mite infestation levels, Nosema spp. infections, and winter losses. Multivariate statistical analysis confirms that V. destructor is the major cause of colony winter losses. Although N. ceranae infections are also statistically significantly correlated with colony losses, determination of the effect size reveals that N. ceranae infections are of no or low biological relevance.

Density of predating Asian hornets at hives disturbs the 3D flight performance of honey bees and decreases predation success.

Automated 3D image-based tracking systems are new and promising devices to investigate the foraging behavior of flying animals with great accuracy and precision. 3D analyses can provide accurate assessments of flight performance in regard to speed, curvature, and hovering. However, there have been few applications of this technology in ecology, particularly for insects. We used this technology to analyze the behavioral interactions between the Western honey bee Apis mellifera and its invasive predator the Asian hornet, Vespa velutina nigrithorax. We investigated whether predation success could be affected by flight speed, flight curvature, and hovering of the Asian hornet and honey bees in front of one beehive. We recorded a total of 603,259 flight trajectories and 5175 predator-prey flight interactions leading to 126 successful predation events, representing 2.4% predation success. Flight speeds of hornets in front of hive entrances were much lower than that of their bee prey; in contrast to hovering capacity, while curvature range overlapped between the two species. There were large differences in speed, curvature, and hovering between the exit and entrance flights of honey bees. Interestingly, we found hornet density affected flight performance of both honey bees and hornets. Higher hornet density led to a decrease in the speed of honey bees leaving the hive, and an increase in the speed of honey bees entering the hive, together with more curved flight trajectories. These effects suggest some predator avoidance behavior by the bees. Higher honey bee flight curvature resulted in lower hornet predation success. Results showed an increase in predation success when hornet number increased up to 8 individuals, above which predation success decreased, likely due to competition among predators. Although based on a single colony, this study reveals interesting outcomes derived from the use of automated 3D tracking to derive accurate measures of individual behavior and behavioral interactions among flying species.

Respiration levels of honey bee castes as a possible indicator in Varroa mite attraction

It has been demonstrated that fatty acids esters play an important role in the caste selection Varroa mites. However, there may be another mechanism of attraction in the preferred host location. We hypothesize that it may be due to differential respiration rates between larvae, but no investigations have been reported. This research was developed to determine if there is a measurable level and significant difference in honey bee caste respiration that may be exploited by Varroa mites in choice selection. In this study, we measured the respiration of all stages of the Western honey bee Apis mellifera, L. castes to determine the rate of continuous respiration. An assay was conducted using a portable photosynthesis system for gas exchange and chlorophyll fluorescence measurement (Li-Core LI-6400XT) to measure the respiration rate. In this study, a comparative analysis resulted in respiration levels that indicated a significant difference in the CO2 levels of adult worker (777.0 ± 0.002 μg CO2 g insect-1 min-1) and drone (375.5 µg CO2) honey bees. The comparison of worker (5.59 ± 0.002 μg CO2 g insect-1 min-1) and drone (7.20 ± 0.002 μg CO2 g insect-1 min-1) pupae showed no significant difference in the respiration levels; however, there was a noticeable difference in respiration levels of the worker (6.33 ± 0.002 μg CO2 g insect-1 min-1) and drone (10.07 ± 0.002 μg CO2 g insect-1 min-1) larvae. Future research will utilize the knowledge obtained to examine the mite’s attraction to the honey bee castes in host location. This information is a crucial step for the advancement of research studies and the development of management protocols for Varroa.

Decline in wild bee species richness associated with honey bee (Apis mellifera L.) abundance in an urban ecosystem.

The spatial heterogeneity of urban landscapes, relatively low agrochemical use, and species-rich floral communities often support a surprising diversity of wild pollinators in cities. However, the management of Western honey bees (Apis mellifera L.) in urban areas may represent a new threat to wild bee communities. Urban beekeeping is commonly perceived as an environmentally friendly practice or a way to combat pollinator declines, when high-density beekeeping operations may actually have a negative influence on native and wild bee populations through floral resource competition and pathogen transmission. On the Island of Montréal, Canada there has been a particularly large increase in beekeeping across the city. Over the years following a large bee diversity survey ending in 2013, there was an influx of almost three thousand honey bee colonies to the city. In this study, we examined the wild bee communities and floral resources across a gradient of honey bee abundances in urban greenspaces in 2020, and compared the bee communities at the same sites before and after the large influx of honey bees. Overall, we found a negative relationship between urban beekeeping, pollen availability, and wild bee species richness. We also found that honey bee abundance had the strongest negative effect on small (inter-tegular span <2.25 mm) wild bee species richness. Small bee species may be at higher risk in areas with abundant honey bee populations as their limited foraging range may reduce their access to floral resources in times of increased competition. Further research on the influence of urban beekeeping on native and wild pollinators, coupled with evidence-based beekeeping regulations, is essential to ensure cities contain sufficient resources to support wild bee diversity alongside managed honey bees.

Western honey bee (Apis mellifera L.) attraction to commercial pollen substitutes and wildflower pollen in vitro

AbstractMany beekeepers feed their western honey bee (Apis mellifera) colonies artificial pollen substitutes to provide colonies with adequate nutrition during times of limited pollen quantity or quality. We provided caged worker bees commercially available pollen substitutes (AP23, MegaBee, UltraBee) and wildflower pollen in a choice‐test to determine their relative attraction to/preference for the diets. We measured diet consumption by honey bees and observed honey bee behaviour to evaluate bee preferences for certain diets. The bees interacted with and consumed more wildflower pollen than they did any of the commercially available pollen substitutes. Our data suggest that bees have a strong preference for wildflower pollen over commercially available pollen substitutes.