Keith S. Delaplane

Agrochemical exposure is a major contributor to ecological declines worldwide, including the loss of crucial pollinator species. In addition to direct toxicity, field-relevant doses of pesticides can increase species’ vulnerabilities to other stressors, including parasites. Experimental field demonstrations of potential interactive effects of pesticides and additional stressors are rare, as are tests of mechanisms via which pollinators tolerate pesticides. Here, we controlled honey bee colony exposure to field-relevant concentrations of 2 neonicotinoid insecticides (clothianidin and thiamethoxam) in pollen and simultaneously manipulated intracolony genetic heterogeneity. We showed that exposure increased rates of Varroa destructor (Anderson and Trueman) parasitism and that while increased genetic heterogeneity overall improved survivability, it did not reduce the negative effect size of neonicotinoid exposure. This study is, to our knowledge, the first experimental field demonstration of how neonicotinoid exposure can increase V. destructor populations in honey bees and also demonstrates that colony genetic diversity cannot mitigate the effects of neonicotinoid pesticides.

Honey bee parasites remain a critical challenge to management and conservation. Because managed honey bees are maintained in colonies kept in apiaries across landscapes, the study of honey bee parasites allows the investigation of spatial principles in parasite ecology and evolution. We used a controlled field experiment to study the relationship between population growth rate and virulence (colony survival) of the parasite Varroa destructor (Anderson and Trueman). We used a nested design of 10 patches (apiaries) of 14 colonies to examine the spatial scale at which Varroa population growth matters for colony survival. We tracked Varroa population size and colony survival across a full year and found that Varroa populations that grow faster in their host colonies during the spring and summer led to larger Varroa populations across the whole apiary (patch) and higher rates of neighboring colony loss. Crucially, this increased colony loss risk manifested at the patch scale, with mortality risk being related to spatial adjacency to colonies with fast-growing Varroa strains rather than with Varroa growth rate in the colony itself. Thus, within-colony population growth predicts whole-apiary virulence, demonstrating the need to consider multiple scales when investigating parasite growth-virulence relationships.

Polyandry in social Hymenoptera is associated with reduced within-colony relatedness and increased colony-level ecologic fitness. One explanation for this sees increasing within-nest genetic diversity as a mechanism for improving group task efficiency and colony competitiveness. A queen on her mating flight captures nearly 90% of her breeding population’s allele potential by her tenth effective mating (m_e ~ 10 males). Under this population allele capture (PAC) model, colony fitness gains track mating number in an asymptotic manner, leveling out after about the tenth mating. A supporting theory we call the genotype composition (GC) model sees genetic novelty at mating levels higher than the m_e ~ 10 asymptote, the hyperpolyandry zone, resulting from unique genotype compositions whose number are potentially infinite. Colony fitness gains under the GC model will track mating number in a linear manner. We set up field colonies with Apis mellifera queens each instrumentally mated with 1, 2, 4, 8, 16, or 32 males, creating a polyandry gradient bracketing the qualitative divide of m_e ~ 10, measured tokens of colony level fitness, and collected observation hive data. Our results lead us to conclude that (1) ancestral colony traits fundamental to eusociality (cooperative brood care) respond to mating level changes at or below m_e ~ 10 in a manner consistent with the PAC model, whereas (2) more derived specialized colony phenotypes (resistance to the non-native parasite Varroa destructor) continue improving with increasing m_e in a manner consistent with the GC model. By either model, (3) the mechanism for increasing colony fitness is an increase in worker task specialisms and task efficiency. Significance statement: Polyandry is a female’s practice of mating with many males, storing their sperm, and using it to produce genetically diverse offspring. In complex social bees, a queen captures nearly 90% of her breeding population’s diversity potential by her tenth mating; however, queens in nature routinely mate with many more than ten males. We tested two models that, together, explain how social bee colonies ecologically benefit from queen mating numbers ranging from 2 to potential infinity. A population allele capture (PAC) model focuses on colony fitness gains at mating numbers at or below 10, and we provide evidence that it was at these polyandry levels that significant gains were made in an ancestral eusocial trait, cooperative brood care. A genotype composition (GC) model focuses on colony fitness gains at higher mating numbers, and we believe these gains are centered around more recently evolved ecologic specialisms such as parasite resistance.

A significant amount of researcher and practitioner effort has focused on developing new chemical controls for the parasitic Varroa destructor mite in beekeeping. One outcome of that has been the development and testing of “glycerol–oxalic acid” mixtures to place in colonies for extended periods of time, an off-label use of the otherwise legal miticide oxalic acid. The majority of circulated work on this approach was led by practitioners and published in nonacademic journals, highlighting a lack of effective partnership between practitioners and scientists and a possible failure of the extension mandate in beekeeping in the United States. Here, we summarize the practitioner-led studies we could locate and partner with a commercial beekeeper in the Southeast of the United States to test the “shop towel–oxalic acid–glycerol” delivery system developed by those practitioners. Our study, using 129 commercial colonies between honey flows in 2017 split into 4 treatment groups, showed no effectiveness in reducing Varroa parasitism in colonies exposed to oxalic acid–glycerol shop towels. We highlight the discrepancy between our results and those circulated by practitioners, at least for the Southeast, and the failure of extension to support practitioners engaged in research.

The nest-scavenging beetle Aethina tumida remains a persistent problem for beekeepers in parts of the Southeast United States, where warm wet soils allow beetle populations to grow rapidly and overwhelm colonies, especially during the summer dearth. Furthermore, small hive beetle infestation prevents beekeepers from easily provisioning colonies with additional pollen or protein feed (patties), preventing holistic management of honey bee health via improved nutrition, and reducing the economic potential of package and nucleus colony rearing in the Southeast. Here, we demonstrate using both in vitro laboratory trials and a small in vivo field trial that the differential specificity of anthranilic diamide insecticides (specifically, chlorantraniliprole) between bees and beetles allows for the control and prevention of small hive beetle infestation in honey bee colonies even when feeding with large patties. Honey bees show orders of magnitude higher tolerance to chlorantraniliprole compared to small hive beetles, opening new avenues for improving bee health including during spring splits and throughout the summer.

The ectoparasitic mite, Varroa destructor (Anderson and Trueman), is the leading cause of western honey bee colony, Apis mellifera (L.), mortality in the United States. Due to mounting evidence of resistance to certain approved miticides, beekeepers are struggling to keep their colonies alive. To date, there are varied but limited approved options for V. destructor control. Vaporized oxalic acid (OA) has proven to be an effective treatment against the dispersal phase of V. destructor but has its limitations since the vapor cannot penetrate the protective wax cap of honey bee pupal cells where V. destructor reproduces. In the Southeastern United States, honey bee colonies often maintain brood throughout the year, limiting the usefulness of OA. Prior studies have shown that even repeated applications of OA while brood is present are ineffective at decreasing mite populations. In the summer of 2021, we studied whether incorporating a forced brood break while vaporizing with OA would be an effective treatment against V. destructor. Ninety experimental colonies were divided into 2 blocks, one with a brood break and the other with no brood break. Within the blocks, each colony was randomly assigned 1 of 3 treatments: no OA, 2 g OA, or 3 g OA. The combination of vaporizing with OA and a forced brood break increased mite mortality by 5× and reduced mite populations significantly. These results give beekeepers in mild climates an additional integrated pest management method for controlling V. destructor during the summer season.

Six studies were done on feeding and foraging behaviors of three species of subterranean termites, Coptotermes formosanus Shiraki, Reticulitermes flavipes (Kollar), and R. virginicus (Banks) (Isoptera: Rhinotermitidae). When presented with four equal wood blocks, C. formosanus did not forage randomly but concentrated on a few preferred blocks. When R. flavipes and C. formosanus were each placed in foraging arenas with linear, successive wood blocks, C. formosanus consumed a greater percentage dry mass of a wood block before moving to the next block (10.3%) than did R. flavipes (4.2%). Groups of C. formosanus were offered wood blocks that differed in initial moisture content. Wood feeding rate, number of workers, and number of soldiers were highest in the high moisture treatment. Large changes in wood moisture occurred and were affected by the presence of termites. Groups of termites from five colonies of C. formosanus were presented with wood blocks that had been previously damaged: (1) by nestmates, (2) by conspecifics from another colony, (3) by R. virginicus, and (4) no damage. Coptotermes formosanus preferred wood previously damaged by conspecifics, regardless of colony origin, over wood damaged by R. virginicus or undamaged wood. Additionally, they preferred wood damaged by R. virginicus over undamaged wood. Wood surface area (mm$\sp2$) exposed per unit feeding was higher for C. formosanus and R. flavipes than for R. virginicus. Wood surface area was sometimes reduced, rather than increased, as a result of feeding by R. virginicus. Groups of C. formosanus were dyed with 0%, 0.5% or 1% concentrations of the dye, Sudan Red 7B. Dyed termites had lower numbers of symbiotic protozoans, lower feeding rates, and lower survivorship than did non-dyed termites. These studies suggest that toxic baits for remedial control of termites should be placed at areas of strong foraging since termites are predisposed to stay at rewarding sites. Baits should be highly moist. Areas of structures with previous termite damage should be carefully monitored for reinfestation. These species may differ in their roles as wood decomposers. the popular termite marker, Sudan Red 7B, is not totally innocuous to C. formosanus.

The American beekeeping industry continually experiences colony mortality with annual losses as high as 43%. A leading cause of this is the exotic, ectoparasitic mite, Varroa destructor Anderson & Trueman (Mesostigmata: Varroidae). Integrated Pest Management (IPM) options are used to keep mite populations from reaching lethal levels, however, due to resistance and/or the lack of suitable treatment options, novel controls for reducing mites are warranted. Oxalic acid for controlling V. destructor has become a popular treatment regimen among commercial and backyard beekeepers. Applying vaporized oxalic acid inside a honey bee hive is a legal application method in the U.S., and results in the death of exposed mites. However, if mites are in the reproductive stage and therefore under the protective wax capping, oxalic acid is ineffective. One popular method of applying oxalic is vaporizing multiple times over several weeks to try and circumvent the problem of mites hiding in brood cells. By comparing against control colonies, we tested oxalic acid vaporization in colonies treated with seven applications separated by 5 d (35 d total). We tested in apiaries in Georgia and Alabama during 2019 and 2020, totaling 99 colonies. We found that adult honey bees Linnaeus (Hymenoptera: Apidae), and developing brood experienced no adverse impacts from the oxalic vaporization regime. However, we did not find evidence that frequent periodic application of oxalic during brood-rearing periods is capable of bringing V. destructor populations below treatment thresholds.

Honey bees (Apis mellifera L. Hymeoptera: Apidae) use hydrogen peroxide (synthesized by excreted glucose oxidase) as an important component of social immunity. However, both tolerance of hydrogen peroxide and the production of glucose oxidase in honey is costly. Hydrogen peroxide may also be encountered by honey bees at high concentrations in nectar while foraging, however despite its presence both in their foraged and stored foods, it is unclear if and how bees monitor concentrations of, and their behavioral responses to, hydrogen peroxide. The costs of glucose oxidase production and the presence of hydrogen peroxide in both nectar and honey suggest hypotheses that honey bees preferentially forage on hydrogen peroxide supplemented feed syrups at certain concentrations, and avoid feed syrups supplemented with hydrogen peroxide at concentrations above some tolerance threshold. We test these hypotheses and find that, counter to expectation, honey bees avoid glucose solutions supplemented with field-relevant hydrogen peroxide concentrations and either avoid or don’t differentiate supplemented sucrose solutions when given choice assays. This is despite honey bees showing high tolerance for hydrogen peroxide in feed solutions, with no elevated mortality until concentrations of hydrogen peroxide exceed 1% (v/v) in solution, with survival apparent even at concentrations up to 10%. The behavioral interaction of honey bees with hydrogen peroxide during both within-colony synthesis in honey and when foraging on nectar therefore likely relies on interactions with other indicator molecules, and maybe constrained evolutionarily in its plasticity, representing a constitutive immune mechanism.