The impacts of invertebrate RNA virus population dynamics on virulence and infection outcomes are poorly understood. Deformed wing virus (DWV), the main viral pathogen of honey bees, negatively impacts bee health, which can lead to colony death. Despite previous reports on the reduction of DWV diversity following the arrival of the parasitic mite Varroa destructor, the key DWV vector, we found high genetic diversity of DWV in infested United States honey bee colonies. Phylogenetic analysis showed that divergent US DWV genotypes are of monophyletic origin and were likely generated as a result of diversification after a genetic bottleneck. To investigate the population dynamics of this divergent DWV, we designed a series of novel infectious cDNA clones corresponding to coexisting DWV genotypes, thereby devising a reverse-genetics system for an invertebrate RNA virus quasispecies. Equal replication rates were observed for all clone-derived DWV variants in single infections. Surprisingly, individual clones replicated to the same high levels as their mixtures and even the parental highly diverse natural DWV population, suggesting that complementation between genotypes was not required to replicate to high levels. Mixed clone–derived infections showed a lack of strong competitive exclusion, suggesting that the DWV genotypes were adapted to coexist. Mutational and recombination events were observed across clone progeny, providing new insights into the forces that drive and constrain virus diversification. Accordingly, our results suggest that Varroa influences DWV dynamics by causing an initial selective sweep, which is followed by virus diversification fueled by negative frequency-dependent selection for new genotypes. We suggest that this selection might reflect the ability of rare lineages to evade host defenses, specifically antiviral RNA interference (RNAi). In support of this hypothesis, we show that RNAi induced against one DWV strain is less effective against an alternate strain from the same population.

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Dynamic evolution in the key honey bee pathogen deformed wing virus: Novel insights into virulence and competition using reverse genetics

Land-use change and habitat loss have profoundly disturbed the resource availability for many organisms in farmlands, including bees. To counteract the resulting decline of bees and to maintain their pollination service to crops, bee pollinator-friendly schemes have been developed. We assessed the most established bee pollinator-friendly schemes which mainly aim at enhancing the availability of floral food resources and, or, nesting sites. We found that the availability of non-floral resources was typically overlooked in these schemes, although more than 30% of bee species worldwide depend on non-floral resources. In this opinion paper, we call for more attention on the role of non-floral resources in such conservation schemes. In fact, at least two of the most species rich Apoidea families, the Apidae and Megachilidae, need non-floral resources for nest building, defence, protection and health. For example, resin is known to improve health and resistance to pests and pathogens in stingless bees and the western honey bee Apis mellifera. Beyond social bees, many solitary bee species, in particularly within the Euglossini, Centridini and Megachilidae, also use resin, leaf pieces, trichome secretions and other materials for the construction and protection of brood cells and the nest. Besides protection, non-floral resources can also provide alternative food resources for bees, e.g. sugary secretions from other insects. Surprisingly little is known on this insect–insect interaction although some studies suggest that honey production by beekeepers can largely depend on this alternative food resource. The apparent knowledge gap on the role of non-floral resources for bees can thus directly concern stakeholders. Interestingly, many non-floral resources are provided by woody vegetation, but most of the bee pollinator-friendly schemes are currently orientated towards enhancing flower availability in field margins and flower strips. We therefore suggest more attention in the protection of trees and hedgerows in conservation, restoration and management schemes to best supporting bee health by providing combined access to nesting, floral and non-floral foraging resources.

https://link.springer.com/content/pdf/10.1007/s10841-019-00206-1.pdf

Beyond flowers: including non-floral resources in bee conservation schemes

The western honey bee remains the most important pollinator for agricultural crops. Disease and stressors threaten honey bee populations and productivity during winter- and summertime, creating costs for beekeepers and negative impacts on agriculture. To combat diseases and improve overall bee health, researchers are constantly developing honey bee medicines using the tools of microbiology, molecular biology and chemistry. Below, we present a manifesto alongside standardized protocols that outline the development and a systematic approach to test natural products as ‘bee medicines’. These will be accomplished in both artificial rearing conditions and in colonies situated in the field. Output will be scored by gene expression data of host immunity, bee survivorship, reduction in pathogen titers, and more subjective merits of the compound in question. Natural products, some of which are already encountered by bees in the form of plant resins and nectar compounds, provide promising low-cost candidates for safe prophylaxis or treatment of bee diseases.

Natural Product Medicines for Honey Bees: Perspective and Protocols.

Mushrooms, the fruiting bodies of fungi, are known for a long time in different cultures around the world to possess medicinal properties and are used to treat various human diseases. Mushrooms that are parts of traditional medicine in Asia had been extensively studied and this has led to identification of their bioactive ingredients. North America, while home to one of the world's largest and diverse ecological systems, has not subjected its natural resources especially its diverse array of mushroom species for bioprospecting purposes: Are mushrooms native to North America a good source for drug discovery? In this review, we compile all the published studies up to September 2020 on the bioprospecting of North American mushrooms. Out of the 79 species that have been investigated for medicinal properties, 48 species (60%) have bioactivities that have not been previously reported. For a mere 16 selected species, 17 new bioactive compounds (10 small molecules, six polysaccharides and one protein) have already been isolated. The results from our literature search suggest that mushrooms native to North America are indeed a good source for drug discovery.

Medicinal Properties and Bioactive Compounds from Wild Mushrooms Native to North America.

We use the term social-medication to describe the deliberate consumption or use of plant compounds by social insects that are detrimental to a pathogen or parasite at the colony level, result in increased inclusive fitness to the colony, and have potential costs either at the individual or colony level in the absence of parasite infection. These criteria for social-medication differ from those for self-medication in that inclusive fitness costs and benefits are distinguished from individual costs and benefits. The consumption of pollen and nectar may be considered a form of social immunity if they help fight infection, resulting in a demonstrated increase in colony health and survival. However, the dietary use of pollen and nectar per se is likely not a form of social-medication unless there is a detriment or cost to their consumption in the absence of parasite infection, such as when they contain phytochemicals that are toxic at certain doses. We provide examples among social bees (bumblebees, stingless bees and honey bees) in which the consumption or use of plant compounds have a demonstrated role in parasite defense and health of the colony. We indicate where more work is needed to distinguish between prophylactic and therapeutic effects of these compounds, and whether the effects are observed at the individual or colony level.

Social-medication in bees: the line between individual and social regulation

Waves of highly infectious viruses sweeping through global honey bee populations have contributed to recent declines in honey bee health Bees have been observed foraging on mushroom mycelium, suggesting that they may be deriving medicinal or nutritional value from fungi Fungi are known to produce a wide array of chemicals with antimicrobial activity, including compounds active against bacteria, other fungi, or viruses We tested extracts from the mycelium of multiple polypore fungal species known to have antiviral properties Extracts from amadou (Fomes) and reishi (Ganoderma) fungi reduced the levels of honey bee deformed wing virus (DWV) and Lake Sinai virus (LSV) in a dose-dependent manner In field trials, colonies fed Ganoderma resinaceum extract exhibited a 79-fold reduction in DWV and a 45,000-fold reduction in LSV compared to control colonies These findings indicate honey bees may gain health benefits from fungi and their antimicrobial compounds

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Extracts of Polypore Mushroom Mycelia Reduce Viruses in Honey Bees

Entomopathogenic fungi show great promise as pesticides in terms of their relatively high target specificity, low non-target toxicity, and low residual effects in agricultural fields and the environment. However, they also frequently have characteristics that limit their use, especially concerning tolerances to temperature, ultraviolet radiation, or other abiotic factors. The devastating ectoparasite of honey bees, Varroa destructor, is susceptible to entomopathogenic fungi, but the relatively warm temperatures inside honey bee hives have prevented these fungi from becoming effective control measures. Using a combination of traditional selection and directed evolution techniques developed for this system, new strains of Metarhizium brunneum were created that survived, germinated, and grew better at bee hive temperatures (35 °C). Field tests with full-sized honey bee colonies confirmed that the new strain JH1078 is more virulent against Varroa mites and controls the pest comparable to current treatments. These results indicate that entomopathogenic fungi are evolutionarily labile and capable of playing a larger role in modern pest management practices.

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David Sumerlin

Papers

Extracts of Polypore Mushroom Mycelia Reduce Viruses in Honey Bees

Directed evolution of Metarhizium fungus improves its biocontrol efficacy against Varroa mites in honey bee colonies.