Nuclear translocation of vitellogenin in the honey bee (Apis mellifera).

Vitellogenin (Vg) is a conserved protein used by nearly all oviparous animals to produce eggs. It is also pleiotropic and performs functions in oxidative stress resistance, immunity, and, in honey bees, behavioral development of the worker caste. It has remained enigmatic how Vg affects multiple traits. Here, we asked whether Vg enters the nucleus and acts via DNA-binding. We used cell fractionation, immunohistology, and cell culture to show that a structural subunit of honey bee Vg translocates into cell nuclei. We then demonstrated Vg-DNA binding theoretically and empirically with prediction software and chromatin immunoprecipitation with sequencing (ChIP-seq), finding binding sites at genes influencing immunity and behavior. Finally, we investigated the immunological and enzymatic conditions affecting Vg cleavage and nuclear translocation and constructed a 3D structural model. Our data are the first to show Vg in the nucleus and suggest a new fundamental regulatory role for this ubiquitous protein. Supplementary information: The online version contains supplementary material available at 10.1007/s13592-022-00914-9.

Context-dependent influence of threat on honey bee social network dynamics and brain gene expression.

Adverse social experience affects social structure by modifying the behavior of individuals, but the relationship between an individual's behavioral state and its response to adversity is poorly understood. We leveraged naturally occurring division of labor in honey bees and studied the biological embedding of environmental threat using laboratory assays and automated behavioral tracking of whole colonies. Guard bees showed low intrinsic levels of sociability compared with foragers and nurse bees, but large increases in sociability following exposure to a threat. Threat experience also modified the expression of caregiving-related genes in a brain region called the mushroom bodies. These results demonstrate that the biological embedding of environmental experience depends on an individual's societal role and, in turn, affects its future sociability.

Butenolide Insecticide Flupyradifurone Affects Honey Bee Worker Antiviral Immunity and Survival.

Honey bees face many environmental stressors, including exposure to pesticides and pathogens. A novel butenolide pesticide, flupyradifurone, was recently introduced to the US and shown to have a bee-friendly toxicity profile. Like the much-scrutinized neonicotinoids that preceded it, flupyradifurone targets the insect nervous system. Some neonicotinoids have been shown to interfere with antiviral immunity, which raised the concern that similar effects may be observed with flupyradifurone. In this study, we investigated how flupyradifurone and a neonicotinoid, clothianidin, affect the ability of honey bee workers to combat an infection of Israeli acute paralysis virus (IAPV). We exposed workers to field-realistic doses of the pesticides either with or without co-exposure with the virus, and then tracked survival and changes in viral titers. We repeated the experiment in the spring and fall to look for any seasonal effects. We found that flupyradifurone caused elevated mortality in the fall, but it did not lead to increased virus-induced mortality. Flupyradifurone also appeared to affect virus clearance, as bees co-exposed to the pesticide and virus tended to have higher viral titers after 48 hours than those exposed to the virus alone. Clothianidin had no effect on viral titers, and it actually appeared to increase resistance to viral infection in spring bees.

Social immunity in honey bees: royal jelly as a vehicle in transferring bacterial pathogen fragments between nestmates.

Social immunity is a suite of behavioral and physiological traits that allow colony members to protect one another from pathogens, and includes the oral transfer of immunological compounds between nestmates. In honey bees, royal jelly is a glandular secretion produced by a subset of workers that is fed to the queen and young larvae, and which contains many antimicrobial compounds. A related form of social immunity, transgenerational immune priming (TGIP), allows queens to transfer pathogen fragments into their developing eggs, where they are recognized by the embryo's immune system and induce higher pathogen resistance in the new offspring. These pathogen fragments are transported by vitellogenin (Vg), an egg-yolk precursor protein that is also used by nurses to synthesize royal jelly. Therefore, royal jelly may serve as a vehicle to transport pathogen fragments from workers to other nestmates. To investigate this, we recently showed that ingested bacteria are transported to nurses' jelly-producing glands, and here, we show that pathogen fragments are incorporated into the royal jelly. Moreover, we show that consuming pathogen cells induces higher levels of an antimicrobial peptide found in royal jelly, defensin-1.

Social immunity in honey bees: royal jelly as a vehicle in transferring bacterial pathogen fragments between nestmates.

Social immunity is a suite of behavioral and physiological traits that allow colony members to protect one another from pathogens, and includes the oral transfer of immunological compounds between nestmates. In honey bees, royal jelly is a glandular secretion produced by a subset of workers that is fed to the queen and young larvae, and which contains many antimicrobial compounds. A related form of social immunity, transgenerational immune priming (TGIP), allows queens to transfer pathogen fragments into their developing eggs, where they are recognized by the embryo's immune system and induce higher pathogen resistance in the new offspring. These pathogen fragments are transported by vitellogenin (Vg), an egg-yolk precursor protein that is also used by nurses to synthesize royal jelly. Therefore, royal jelly may serve as a vehicle to transport pathogen fragments from workers to other nestmates. To investigate this, we recently showed that ingested bacteria are transported to nurses' jelly-producing glands, and here, we show that pathogen fragments are incorporated into the royal jelly. Moreover, we show that consuming pathogen cells induces higher levels of an antimicrobial peptide found in royal jelly, defensin-1.

Pesticide-Virus Interactions in Honey Bees: Challenges and Opportunities for Understanding Drivers of Bee Declines.

Honey bees are key agricultural pollinators, but beekeepers continually suffer high annual colony losses owing to a number of environmental stressors, including inadequate nutrition, pressures from parasites and pathogens, and exposure to a wide variety of pesticides. In this review, we examine how two such stressors, pesticides and viruses, may interact in additive or synergistic ways to affect honey bee health. Despite what appears to be a straightforward comparison, there is a dearth of studies examining this issue likely owing to the complexity of such interactions. Such complexities include the wide array of pesticide chemical classes with different modes of actions, the coupling of many bee viruses with ectoparasitic Varroa mites, and the intricate social structure of honey bee colonies. Together, these issues pose a challenge to researchers examining the effects pesticide-virus interactions at both the individual and colony level.

Honey bee virus causes context-dependent changes in host social behavior.

Anthropogenic changes create evolutionarily novel environments that present opportunities for emerging diseases, potentially changing the balance between host and pathogen. Honey bees provide essential pollination services, but intensification and globalization of honey bee management has coincided with increased pathogen pressure, primarily due to a parasitic mite/virus complex. Here, we investigated how honey bee individual and group phenotypes are altered by a virus of concern, Israeli acute paralysis virus (IAPV). Using automated and manual behavioral monitoring of IAPV-inoculated individuals, we find evidence for pathogen manipulation of worker behavior by IAPV, and reveal that this effect depends on social context; that is, within versus between colony interactions. Experimental inoculation reduced social contacts between honey bee colony members, suggesting an adaptive host social immune response to diminish transmission. Parallel analyses with double-stranded RNA (dsRNA)-immunostimulated bees revealed these behaviors are part of a generalized social immune defensive response. Conversely, inoculated bees presented to groups of bees from other colonies experienced reduced aggression compared with dsRNA-immunostimulated bees, facilitating entry into susceptible colonies. This reduction was associated with a shift in cuticular hydrocarbons, the chemical signatures used by bees to discriminate colony members from intruders. These responses were specific to IAPV infection, suggestive of pathogen manipulation of the host. Emerging bee pathogens may thus shape host phenotypes to increase transmission, a strategy especially well-suited to the unnaturally high colony densities of modern apiculture. These findings demonstrate how anthropogenic changes could affect arms races between human-managed hosts and their pathogens to potentially affect global food security.

Honey bees as bioindicators of changing global agricultural landscapes.

There is a growing need to understand relationships between agricultural intensification and global change. Monitoring solutions, however, often do not include pollinator communities that are of importance to ecosystem integrity. Here, we put forth the honey bee as an economical and broadly available bioindicator that can be used to assess and track changes in the quality of agricultural ecosystems. We detail a variety of simple, low-cost procedures that can be deployed within honey bee hives to gain generalizable information about ecosystem quality at multiple scales, and discuss the potential of the honey bee system in both environmental and ecological bioindication.

The role of Vitellogenin in the transfer of immune elicitors from gut to hypopharyngeal glands in honey bees (Apis mellifera).

Female insects that survive a pathogen attack can produce more pathogen-resistant offspring in a process called trans-generational immune priming. In the honey bee (Apis mellifera), the egg-yolk precursor protein Vitellogenin transports fragments of pathogen cells into the egg, thereby setting the stage for a recruitment of immunological defenses prior to hatching. Honey bees live in complex societies where reproduction and communal tasks are divided between a queen and her sterile female workers. Worker bees metabolize Vitellogenin to synthesize royal jelly, a protein-rich glandular secretion fed to the queen and young larvae. We ask if workers can participate in trans-generational immune priming by transferring pathogen fragments to the queen or larvae via royal jelly. As a first step toward answering this question, we tested whether worker-ingested bacterial fragments can be transported to jelly-producing glands, and what role Vitellogenin plays in this transport. To do this, we fed fluorescently labelled Escherichia coli to workers with experimentally manipulated levels of Vitellogenin. We found that bacterial fragments were transported to the glands of control workers, while they were not detected at the glands of workers subjected to RNA interference-mediated Vitellogenin gene knockdown, suggesting that Vitellogenin plays a role in this transport. Our results provide initial evidence that trans-generational immune priming may operate at a colony-wide level in honey bees.

Differences in group size and the extent of individual participation in group hunting may contribute to differential prey-size use among social spiders.

We have previously shown that the range of prey sizes captured by co-occurring species of group-hunting social spiders correlates positively with their level of sociality. Here, we show that this pattern is probably caused by differences among species in colony size and the extent to which individuals participate in group hunting. We assess levels of participation for each species from the fraction of individuals responding to the struggling prey that partake as attackers and from the extent to which the number of attackers increases with colony size. Of two species that form equally large colonies, the one that captures on average larger prey engaged as attackers a significantly larger fraction of individuals that responded to struggling prey and also increased its number of attackers in larger colonies when presented with large prey items. Surprisingly, a third co-occurring species previously found to capture smaller insects than the other two exhibited the highest levels of participation. This species, however, typically forms small single-family colonies, thereby being limited in the size of insects it can capture. It is thus a combination of colony size and the extent of individual participation (or cooperation) that probably determines patterns of resource use in this community of co-occurring social predators.

A Quantitative Index of Sociality and Its Application to Group-Living Spiders and Other Social Organisms.

Species are often classified in discrete categories, such as solitary, subsocial, social and eusocial based on broad qualitative features of their social systems. Often, however, species fall between categories or species within a category may differ from one another in ways that beg for a quantitative measure of their sociality level. Here, we propose such a quantitative measure in the form of an index that is based on three fundamental features of a social system: (1) the fraction of the life cycle that individuals remain in their social group, (2) the proportion of nests in a population that contain multiple vs. solitary individuals and (3) the proportion of adult members of a group that do not reproduce, but contribute to communal activities. These are measures that should be quantifiable in most social systems, with the first two reflecting the tendencies of individuals to live in groups as a result of philopatry, grouping tendencies and intraspecific tolerance, and the third potentially reflecting the tendencies of individuals to exhibit reproductive altruism. We argue that this index can serve not only as a way of ranking species along a sociality scale, but also as a means of determining how level of sociality correlates with other aspects of the biology of a group of organisms. We illustrate the calculation of this index for the cooperative social spiders and the African mole-rats and use it to analyse how sex ratios and interfemale spacing correlate with level of sociality in spider species in the genus Anelosimus.