The effects of protein supplementation, fumagillin treatment, and colony management on the productivity and long-term survival of honey bee (Apis mellifera) colonies.

In this study, we intensively measured the longitudinal productivity and survival of 362 commercially managed honey bee colonies in Canada, over a two-year period. A full factorial experimental design was used, whereby two treatments were repeated across apiaries situated in three distinct geographic regions: Northern Alberta, Southern Alberta and Prince Edward Island, each having unique bee management strategies. In the protein supplemented treatment, colonies were continuously provided a commercial protein supplement containing 25% w/w pollen, in addition to any feed normally provided by beekeepers in that region. In the fumagillin treatment, colonies were treated with the label dose of Fumagilin-B® each year during the fall. Neither treatment provided consistent benefits across all sites and dates. Fumagillin was associated with a large increase in honey production only at the Northern Alberta site, while protein supplementation produced an early season increase in brood production only at the Southern Alberta site. The protein supplement provided no long-lasting benefit at any site and was also associated with an increased risk of death and decreased colony size later in the study. Differences in colony survival and productivity among regions, and among colonies within beekeeping operations, were far larger than the effects of either treatment, suggesting that returns from extra feed supplements and fumagillin were highly contextually dependent. We conclude that use of fumagillin is safe and sometimes beneficial, but that beekeepers should only consider excess protein supplementation when natural forage is limiting.

Systematic review of residual toxicity studies of pesticides to bees and veracity of guidance on pesticide labels.

Residues of pesticides on crops can result in mortality to foraging bees. Pesticide applicators in the U.S. encounter a statement on pesticide labels, which coarsely indicate which products dissipate over the course of an evening. There is reason to suspect that these statements may not align with residual toxicity data, given previous findings. Without a complete database of residual toxicity estimates; however, it is not possible to determine whether the residual toxicity components of statements on pesticide labels similarly diverge from published studies. We compiled 50 studies on residual toxicity trials with formulated pesticides and calculated the residual time to 25% mortality (RT25) of each assay for three different bee species (Apis mellifera, Nomia melanderi, and Megachile rotundata). Our findings were compared to a U.S. Environmental Protection Agency (EPA) published database of RT25 values. Of the RT25 values that we could compare, we found that over 90% of the values support a similar conclusion to the EPA. Next, we compared our values and the EPA's values to the statements on 155 EPA registered pesticide product labels. Of these labels, a little less than a third presented their residual toxicity in a manner inconsistent with their calculated RT25 and current EPA labeling guidelines. Moreover, over a third of labels contained an active ingredient which was neither listed under the EPA's RT25 database nor had a published study to estimate this value. We provide the first evidence that many pesticide labels may convey residual toxicity information to applicators that is not correct and could lead to bees being exposed to toxic residues on plants.

Honey bee (Apis mellifera) colony strength and its effects on pollination and yield in highbush blueberries (Vaccinium corymbosum).

Many pollination studies with honey bees have examined the effect of colony density on crop yield and yet overlook the effect of variation in the population size of these colonies. High colony density in northern highbush blueberry has been met with concerns from beekeepers who feel higher densities will intensify outbreaks of European foulbrood (EFB, Melissococcus plutonius, Truper and dé Clari), a honey bee brood disease. The purpose of this study was to confirm the prevalence of EFB in colonies pollinating blueberries and to determine whether field-level variation in the population of adult workers in colonies explained variation in blueberry fruit set and/or yield. We addressed these objectives over the course of two production seasons at 13 commercial blueberry fields in Oregon, USA, stocked with identical densities of 10 colonies/ha. We confirmed that all colonies had negligible symptoms of EFB at the start of blueberry pollination, but 53% of colonies in 2019 and 41% in 2020 had symptoms immediately following the pollination season. We also validated a method for rapidly assessing adult honey bee colony populations, namely by counting the rate of foragers returning to colonies, and it was found to be strongly correlated to true internal adult bee population independent of year and ambient temperature at the time of evaluation. Using returning forager counts, we determined there was considerable variation in the average population of colonies at each field, ranging from an estimated 10,300 to 30,700 adult worker bees per colony. While average colony strength did not predict variation in fruit set, it was related to variation in yield, independent of year. Our linear model of flight count (as a proxy for colony strength) predicts estimated yield increases of up to 25,000 kg/ha of blueberries could be achieved by colonies stronger than the recommended six frame minimum, suggesting that higher pollination benefits could be achieved without increasing hive density if stronger colonies are promoted.

Partial shading by solar panels delays bloom, increases floral abundance during the late-season for pollinators in a dryland, agrivoltaic ecosystem.

Habitat for pollinators is declining worldwide, threatening the health of both wild and agricultural ecosystems. Photovoltaic solar energy installation is booming, frequently near agricultural lands, where the land underneath ground-mounted photovoltaic panels is traditionally unused. Some solar developers and agriculturalists in the United States are filling the solar understory with habitat for pollinating insects in efforts to maximize land-use efficiency in agricultural lands. However, the impact of the solar panel canopy on the understory pollinator-plant community is unknown. Here we investigated the effects of solar arrays on plant composition, bloom timing and foraging behavior of pollinators from June to September (after peak bloom) in full shade plots and partial shade plots under solar panels as well as in full sun plots (controls) outside of the solar panels. We found that floral abundance increased and bloom timing was delayed in the partial shade plots, which has the potential to benefit late-season foragers in water-limited ecosystems. Pollinator abundance, diversity, and richness were similar in full sun and partial shade plots, both greater than in full shade. Pollinator-flower visitation rates did not differ among treatments at this scale. This demonstrates that pollinators will use habitat under solar arrays, despite variations in community structure across shade gradients. We anticipate that these findings will inform local farmers and solar developers who manage solar understories, as well as agriculture and pollinator health advocates as they seek land for pollinator habitat restoration in target areas.

Field rates of Sivanto™ (flupyradifurone) and Transform® (sulfoxaflor) increase oxidative stress and induce apoptosis in honey bees (Apis mellifera L.).

Pesticide exposures can have detrimental impacts on bee pollinators, ranging from immediate mortality to sub-lethal impacts. Flupyradifurone is the active ingredient in Sivanto™ and sulfoxaflor is the active ingredient in Transform®. They are both relatively new insecticides developed with an intent to reduce negative effects on bees, when applied to bee-attractive crops. With the growing concern regarding pollinator health and pollinator declines, it is important to have a better understanding of any potential negative impacts, especially sub-lethal, of these pesticides on bees. This study reports novel findings regarding physiological stress experienced by bees exposed to field application rates of these two insecticides via a Potter Tower sprayer. Two contact exposure experiments were conducted-a shorter 6-hour study and a longer 10-day study. Honey bee mortality, sugar syrup and water consumption, and physiological responses (oxidative stress and apoptotic protein assays) were assessed in bees exposed to Sivanto™ and Transform®, and compared to bees in control group. For the longer, 10-day contact exposure experiment, only the Sivanto™ group was compared to the control group, as high mortality recorded in the sulfoxaflor treatment group during the shorter contact exposure experiment, made the latter group unfeasible to test in the longer 10-days experiment. In both the studies, sugar syrup and water consumptions were significantly different between treatment groups and controls. The highest mortality was observed in Transform® exposed bees, followed by the Sivanto™ exposed bees. Estimates of reactive oxygen/nitrogen species indicated significantly elevated oxidative stress in both pesticide treatment groups, when compared to controls. Caspase-3 protein assays, an indicator of onset of apoptosis, was also significantly higher in the pesticide treatment groups. These differences were largely driven by post exposure duration, indicating sub-lethal impacts. Further, our findings also emphasize the need to revisit contact exposure impacts of Sivanto™, given the sub-lethal impacts and mortality observed in our long-term (10-day) contact exposure experiment.

Labels of insecticides to which Oregon honey bee (Apis mellifera L.) hives could be exposed do not align with federal recommendations in their communication of acute and residual toxicity to honey bees.

BACKGROUND: Teaching pesticide applicators how to determine a product's toxicity to honey bees (Apis mellifera L.) by understanding the label is a way to reduce honey bee exposure to pesticides. Applicators are currently taught how to interpret labels that follow the United States Environmental Protection Agency's (USEPA's) recommendations for how this toxicity should be communicated. To effectively educate applicators, it must be determined if labels follow USEPA recommendations in how they communicate toxicity to honey bees, and if they do not, what patterns exist in these deviations from the recommendations (hereafter, deviations). RESULTS: As a case study, we analyzed the USEPA master labels of insecticides used in 16 situations where there is concern over Oregon honey bee colony exposure to pesticides. Of the 232 labels analyzed, 31.5% deviated from USEPA recommendations in at least one way. Labels of both commercial and garden products deviated from USEPA recommendations. Products with labels that deviated from USEPA recommendations were found across application situation and active ingredient subgroup. The percentage of products per application situation and per chemical subgroup whose label deviated from USEPA recommendations varied. The most common deviation represented a direct contradiction between the statement used and the toxicity class of the product's active ingredient. CONCLUSION: Applicators will be underprepared to interpret the pesticide labels they see in the field if they are exclusively taught how to interpret labels that follow USEPA recommendations. Applicators should be taught how to interpret a product label that uses non-USEPA-recommended statements to communicate toxicity to honey bees. © 2019 Society of Chemical Industry.

Pollinators enhance crop yield and shorten the growing season by modulating plant functional characteristics: A comparison of 23 canola varieties.

Insect pollination of flowers should change the within-season allocation of resources in plants. But the nature of this life-history response, particularly regarding allocation to roots, photosynthetic structures, and flowers, is empirically unresolved. This study uses a greenhouse experiment to investigate the effect of insect pollination on the reproductive output of 23 varieties of a globally important crop-canola (Brassica napus). Overall, insect pollination modified the functional characteristics (flower timing & effort, plant size & shape, seed packaging, root biomass) of canola, increasing seed production and quality, and pollinator dependence. Reproductive output and pollinator dependence were defined by strong trait trade-offs, which ranged from more pollinator-dependent plants favouring early reproductive effort, to less pollinator-dependent plants favouring a prolonged phenology with smaller plant size and lower seed quality. Seed production decreased with pollinator dependence in the absence of pollinators. The agricultural preference for hybrid varieties will increase seed production compared to open-pollinated varieties, but, even so, pollinators typically enhance seed production of both types. Our study elucidates how insect pollination alters the character and function of a globally important crop, supporting optimization of yield via intensification of insect pollination, and highlights the beneficial effects of insect pollination early in the season.

Pesticides and pollinators: A socioecological synthesis.

The relationship between pesticides and pollinators, while attracting no shortage of attention from scientists, regulators, and the public, has proven resistant to scientific synthesis and fractious in matters of policy and public opinion. This is in part because the issue has been approached in a compartmentalized and intradisciplinary way, such that evaluations of organismal pesticide effects remain largely disjoint from their upstream drivers and downstream consequences. Here, we present a socioecological framework designed to synthesize the pesticide-pollinator system and inform future scholarship and action. Our framework consists of three interlocking domains-pesticide use, pesticide exposure, and pesticide effects-each consisting of causally linked patterns, processes, and states. We elaborate each of these domains and their linkages, reviewing relevant literature and providing empirical case studies. We then propose guidelines for future pesticide-pollinator scholarship and action agenda aimed at strengthening knowledge in neglected domains and integrating knowledge across domains to provide decision support for stakeholders and policymakers. Specifically, we emphasize (1) stakeholder engagement, (2) mechanistic study of pesticide exposure, (3) understanding the propagation of pesticide effects across levels of organization, and (4) full-cost accounting of the externalities of pesticide use and regulation. Addressing these items will require transdisciplinary collaborations within and beyond the scientific community, including the expertise of farmers, agrochemical developers, and policymakers in an extended peer community.

Integrative Genomics Reveals the Genetics and Evolution of the Honey Bee's Social Immune System.

Social organisms combat pathogens through individual innate immune responses or through social immunity-behaviors among individuals that limit pathogen transmission within groups. Although we have a relatively detailed understanding of the genetics and evolution of the innate immune system of animals, we know little about social immunity. Addressing this knowledge gap is crucial for understanding how life-history traits influence immunity, and identifying if trade-offs exist between innate and social immunity. Hygienic behavior in the Western honey bee, Apis mellifera, provides an excellent model for investigating the genetics and evolution of social immunity in animals. This heritable, colony-level behavior is performed by nurse bees when they detect and remove infected or dead brood from the colony. We sequenced 125 haploid genomes from two artificially selected highly hygienic populations and a baseline unselected population. Genomic contrasts allowed us to identify a minimum of 73 genes tentatively associated with hygienic behavior. Many genes were within previously discovered QTLs associated with hygienic behavior and were predictive of hygienic behavior within the unselected population. These genes were often involved in neuronal development and sensory perception in solitary insects. We found that genes associated with hygienic behavior have evidence of positive selection within honey bees (Apis), supporting the hypothesis that social immunity contributes to fitness. Our results indicate that genes influencing developmental neurobiology and behavior in solitary insects may have been co-opted to give rise to a novel and adaptive social immune phenotype in honey bees.

Peptide biomarkers used for the selective breeding of a complex polygenic trait in honey bees.

We present a novel way to select for highly polygenic traits. For millennia, humans have used observable phenotypes to selectively breed stronger or more productive livestock and crops. Selection on genotype, using single-nucleotide polymorphisms (SNPs) and genome profiling, is also now applied broadly in livestock breeding programs; however, selection on protein/peptide or mRNA expression markers has not yet been proven useful. Here we demonstrate the utility of protein markers to select for disease-resistant hygienic behavior in the European honey bee (Apis mellifera L.). Robust, mechanistically-linked protein expression markers, by integrating cis- and trans- effects from many genomic loci, may overcome limitations of genomic markers to allow for selection. After three generations of selection, the resulting marker-selected stock outperformed an unselected benchmark stock in terms of hygienic behavior, and had improved survival when challenged with a bacterial disease or a parasitic mite, similar to bees selected using a phenotype-based assessment for this trait. This is the first demonstration of the efficacy of protein markers for industrial selective breeding in any agricultural species, plant or animal.

Viruses of managed alfalfa leafcutting bees (Megachille rotundata Fabricus) and honey bees (Apis mellifera L.) in Western Canada: Incidence, impacts, and prospects of cross-species viral transmission.

We examined whether alfalfa leafcutting bees (ALCB, Megachille rotundata) experienced a higher incidence of seven viruses commonly found honey bees (Apis mellifera) when placed alongside honey bees for hybrid canola seed pollination. Although two viruses - sacbrood virus (SBV) and deformed wing virus (DWV) - were detected in ALCB adults, their presence appeared independent of whether honey bees were present in the same field or not. A further survey of viruses among ALCB adults in three different alfalfa seed growing regions in Western Canada confirmed the ubiquity of sacbrood virus (SBV) as well as the infrequent presence of acute bee paralysis virus (ABPV), both of which had not been previously reported on ALCB. Moreover, SBV and ABPV were detected in the cocoon stage and only in one region. Co-infection among pools of ALCB adults with both of these viruses was more closely correlated with decreasing levels of cocoon viability than infection levels in cocoons themselves. This research suggests ongoing viral transmission between honey bees and ALCB in the same fields is likely low but that co-infection with these viruses may lower ALCB productivity.

A Bio-Economic Case Study of Canadian Honey Bee (Hymenoptera: Apidae) Colonies: Marker-Assisted Selection (MAS) in Queen Breeding Affects Beekeeper Profits.

Over the past decade in North America and Europe, winter losses of honey bee (Hymenoptera: Apidae) colonies have increased dramatically. Scientific consensus attributes these losses to multifactorial causes including altered parasite and pathogen profiles, lack of proper nutrition due to agricultural monocultures, exposure to pesticides, management, and weather. One method to reduce colony loss and increase productivity is through selective breeding of queens to produce disease-, pathogen-, and mite-resistant stock. Historically, the only method for identifying desirable traits in honey bees to improve breeding was through observation of bee behavior. A team of Canadian scientists have recently identified markers in bee antennae that correspond to behavioral traits in bees and can be tested for in a laboratory. These scientists have demonstrated that this marker-assisted selection (MAS) can be used to produce hygienic, pathogen-resistant honey bee colonies. Based on this research, we present a beekeeping case study where a beekeeper's profit function is used to evaluate the economic impact of adopting colonies selected for hygienic behavior using MAS into an apiary. Our results show a net profit gain from an MAS colony of between 2% and 5% when Varroa mites are effectively treated. In the case of ineffective treatment, MAS generates a net profit benefit of between 9% and 96% depending on the Varroa load. When a Varroa mite population has developed some treatment resistance, we show that MAS colonies generate a net profit gain of between 8% and 112% depending on the Varroa load and degree of treatment resistance.

A search for protein biomarkers links olfactory signal transduction to social immunity.

BACKGROUND: The Western honey bee (Apis mellifera L.) is a critical component of human agriculture through its pollination activities. For years, beekeepers have controlled deadly pathogens such as Paenibacillus larvae, Nosema spp. and Varroa destructor with antibiotics and pesticides but widespread chemical resistance is appearing and most beekeepers would prefer to eliminate or reduce the use of in-hive chemicals. While such treatments are likely to still be needed, an alternate management strategy is to identify and select bees with heritable traits that allow them to resist mites and diseases. Breeding such bees is difficult as the tests involved to identify disease-resistance are complicated, time-consuming, expensive and can misidentify desirable genotypes. Additionally, we do not yet fully understand the mechanisms behind social immunity. Here we have set out to discover the molecular mechanism behind hygienic behavior (HB), a trait known to confer disease resistance in bees. RESULTS: After confirming that HB could be selectively bred for, we correlated measurements of this behavior with protein expression over a period of three years, at two geographically distinct sites, using several hundred bee colonies. By correlating the expression patterns of individual proteins with HB scores, we identified seven putative biomarkers of HB that survived stringent control for multiple hypothesis testing. Intriguingly, these proteins were all involved in semiochemical sensing (odorant binding proteins), nerve signal transmission or signal decay, indicative of the series of events required to respond to an olfactory signal from dead or diseased larvae. We then used recombinant versions of two odorant-binding proteins to identify the classes of ligands that these proteins might be helping bees detect. CONCLUSIONS: Our data suggest that neurosensory detection of odors emitted by dead or diseased larvae is the likely mechanism behind a complex and important social immunity behavior that allows bees to co-exist with pathogens.

Genome sequencing and comparative genomics of honey bee microsporidia, Nosema apis reveal novel insights into host-parasite interactions.

BACKGROUND: The microsporidia parasite Nosema contributes to the steep global decline of honey bees that are critical pollinators of food crops. There are two species of Nosema that have been found to infect honey bees, Nosema apis and N. ceranae. Genome sequencing of N. apis and comparative genome analysis with N. ceranae, a fully sequenced microsporidia species, reveal novel insights into host-parasite interactions underlying the parasite infections. RESULTS: We applied the whole-genome shotgun sequencing approach to sequence and assemble the genome of N. apis which has an estimated size of 8.5 Mbp. We predicted 2,771 protein- coding genes and predicted the function of each putative protein using the Gene Ontology. The comparative genomic analysis led to identification of 1,356 orthologs that are conserved between the two Nosema species and genes that are unique characteristics of the individual species, thereby providing a list of virulence factors and new genetic tools for studying host-parasite interactions. We also identified a highly abundant motif in the upstream promoter regions of N. apis genes. This motif is also conserved in N. ceranae and other microsporidia species and likely plays a role in gene regulation across the microsporidia. CONCLUSIONS: The availability of the N. apis genome sequence is a significant addition to the rapidly expanding body of microsprodian genomic data which has been improving our understanding of eukaryotic genome diversity and evolution in a broad sense. The predicted virulent genes and transcriptional regulatory elements are potential targets for innovative therapeutics to break down the life cycle of the parasite.

Correlation of proteome-wide changes with social immunity behaviors provides insight into resistance to the parasitic mite, Varroa destructor, in the honey bee (Apis mellifera).

BACKGROUND: Disease is a major factor driving the evolution of many organisms. In honey bees, selection for social behavioral responses is the primary adaptive process facilitating disease resistance. One such process, hygienic behavior, enables bees to resist multiple diseases, including the damaging parasitic mite Varroa destructor. The genetic elements and biochemical factors that drive the expression of these adaptations are currently unknown. Proteomics provides a tool to identify proteins that control behavioral processes, and these proteins can be used as biomarkers to aid identification of disease tolerant colonies. RESULTS: We sampled a large cohort of commercial queen lineages, recording overall mite infestation, hygiene, and the specific hygienic response to V. destructor. We performed proteome-wide correlation analyses in larval integument and adult antennae, identifying several proteins highly predictive of behavior and reduced hive infestation. In the larva, response to wounding was identified as a key adaptive process leading to reduced infestation, and chitin biosynthesis and immune responses appear to represent important disease resistant adaptations. The speed of hygienic behavior may be underpinned by changes in the antenna proteome, and chemosensory and neurological processes could also provide specificity for detection of V. destructor in antennae. CONCLUSIONS: Our results provide, for the first time, some insight into how complex behavioural adaptations manifest in the proteome of honey bees. The most important biochemical correlations provide clues as to the underlying molecular mechanisms of social and innate immunity of honey bees. Such changes are indicative of potential divergence in processes controlling the hive-worker maturation.

Ecological adaptation of diverse honey bee (Apis mellifera) populations.

BACKGROUND: Honey bees are complex eusocial insects that provide a critical contribution to human agricultural food production. Their natural migration has selected for traits that increase fitness within geographical areas, but in parallel their domestication has selected for traits that enhance productivity and survival under local conditions. Elucidating the biochemical mechanisms of these local adaptive processes is a key goal of evolutionary biology. Proteomics provides tools unique among the major 'omics disciplines for identifying the mechanisms employed by an organism in adapting to environmental challenges. RESULTS: Through proteome profiling of adult honey bee midgut from geographically dispersed, domesticated populations combined with multiple parallel statistical treatments, the data presented here suggest some of the major cellular processes involved in adapting to different climates. These findings provide insight into the molecular underpinnings that may confer an advantage to honey bee populations. Significantly, the major energy-producing pathways of the mitochondria, the organelle most closely involved in heat production, were consistently higher in bees that had adapted to colder climates. In opposition, up-regulation of protein metabolism capacity, from biosynthesis to degradation, had been selected for in bees from warmer climates. CONCLUSIONS: Overall, our results present a proteomic interpretation of expression polymorphisms between honey bee ecotypes and provide insight into molecular aspects of local adaptation or selection with consequences for honey bee management and breeding. The implications of our findings extend beyond apiculture as they underscore the need to consider the interdependence of animal populations and their agro-ecological context.

The innate immune and systemic response in honey bees to a bacterial pathogen, Paenibacillus larvae.

BACKGROUND: There is a major paradox in our understanding of honey bee immunity: the high population density in a bee colony implies a high rate of disease transmission among individuals, yet bees are predicted to express only two-thirds as many immunity genes as solitary insects, e.g., mosquito or fruit fly. This suggests that the immune response in bees is subdued in favor of social immunity, yet some specific immune factors are up-regulated in response to infection. To explore the response to infection more broadly, we employ mass spectrometry-based proteomics in a quantitative analysis of honey bee larvae infected with the bacterium Paenibacillus larvae. Newly-eclosed bee larvae, in the second stage of their life cycle, are susceptible to this infection, but become progressively more resistant with age. We used this host-pathogen system to probe not only the role of the immune system in responding to a highly evolved infection, but also what other mechanisms might be employed in response to infection. RESULTS: Using quantitative proteomics, we compared the hemolymph (insect blood) of five-day old healthy and infected honey bee larvae and found a strong up-regulation of some metabolic enzymes and chaperones, while royal jelly (food) and energy storage proteins were down-regulated. We also observed increased levels of the immune factors prophenoloxidase (proPO), lysozyme and the antimicrobial peptide hymenoptaecin. Furthermore, mass spectrometry evidence suggests that healthy larvae have significant levels of catalytically inactive proPO in the hemolymph that is proteolytically activated upon infection. Phenoloxidase (PO) enzyme activity was undetectable in one or two-day-old larvae and increased dramatically thereafter, paralleling very closely the age-related ability of larvae to resist infection. CONCLUSION: We propose a model for the host response to infection where energy stores and metabolic enzymes are regulated in concert with direct defensive measures, such as the massive enhancement of PO activity.

Evaluation of the shaking technique for the economic management of American foulbrood disease of honey bees (Hymenoptera: Apidae).

Shaking is a nonantibiotic management technique for the bacterial disease American foulbrood (AFB) (Paenibacillus larvae sensu Genersch et al.), in which infected nesting comb is destroyed and the adult honey bees, Apis mellifera L. (Hymenoptera: Apidae), are transferred onto uncontaminated nesting material. We hypothesized that colonies shaken onto frames of uninfected drawn comb would have similar reductions in AFB symptoms and bacterial spore loads than those shaken onto frames of foundation, but they would attain higher levels of production. We observed that colonies shaken onto drawn comb, or a combination of foundation and drawn comb, exhibited light transitory AFB infections, whereas colonies shaken onto frames containing only foundation failed to exhibit clinical symptoms. Furthermore, concentrations of P. larvae spores in honey and adult worker bees sampled from colonies shaken onto all comb and foundation treatments declined over time and were undetectable in adult bee samples 3 mo after shaking. In contrast, colonies that were reestablished on the original infected comb remained heavily infected resulting in consistently high levels of spores, and eventually, their death. In a subsequent experiment, production of colonies shaken onto foundation was compared with that of colonies established from package (bulk) bees or that of overwintered colonies. Economic analysis proved shaking to be 24% more profitable than using package bees. These results suggest that shaking bees onto frames of foundation in the spring is a feasible option for managing AFB in commercial beekeeping operations where antibiotic use is undesirable or prohibited.