New Insights into Honey Bee (Apis mellifera) Pheromone Communication. Is the Queen Mandibular Pheromone Alone in Colony Regulation?

Background: In social insects, the queen is essential to the functioning and homeostasis of the colony. This influencehas been demonstrated to be mediated through pheromone communication. However, the only social insect forwhich any queen pheromone has been identified is the honey bee (Apis mellifera) with its well-known queenmandibular pheromone (QMP). Although pleiotropic effects on colony regulation are accredited to the QMP, thispheromone does not trigger the full behavioral and physiological response observed in the presence of the queen,suggesting the presence of additional compounds. We tested the hypothesis of a pheromone redundancy in honeybee queens by comparing the influence of queens with and without mandibular glands on worker behavior andphysiology.Results: Demandibulated queens had no detectable (E)-9-oxodec-2-enoic acid (9-ODA), the major compound in QMP,yet they controlled worker behavior (cell construction and queen retinue) and physiology (ovary inhibition) asefficiently as intact queens.Conclusions: We demonstrated that the queen uses other pheromones as powerful as QMP to control the colony. Itfollows that queens appear to have multiple active compounds with similar functions in the colony (pheromoneredundancy). Our findings support two hypotheses in the biology of social insects: (1) that multiple semiochemicalswith synonymous meaning exist in the honey bee, (2) that this extensive semiochemical vocabulary exists because itconfers an evolutionary advantage to the colony

honeybee workers exhibit conserved molecular responses to diverse pathogens

Background Organisms typically face infection by diverse pathogens, and hosts are thought to have developed specific responses to each type of pathogen they encounter. The advent of transcriptomics now makes it possible to test this hypothesis and compare host gene expression responses to multiple pathogens at a genome-wide scale. Here, we performed a meta-analysis of multiple published and new transcriptomes using a newly developed bioinformatics approach that filters genes based on their expression profile across datasets. Thereby, we identified common and unique molecular responses of a model host species, the honey bee (Apis mellifera), to its major pathogens and parasites: the Microsporidia Nosema apis and Nosema ceranae, RNA viruses, and the ectoparasitic mite Varroa destructor, which transmits viruses. Results We identified a common suite of genes and conserved molecular pathways that respond to all investigated pathogens, a result that suggests a commonality in response mechanisms to diverse pathogens. We found that genes differentially expressed after infection exhibit a higher evolutionary rate than non- differentially expressed genes. Using our new bioinformatics approach, we unveiled additional pathogen-specific responses of honey bees; we found that apoptosis appeared to be an important response following microsporidian infection, while genes from the immune signalling pathways, Toll and Imd, were differentially expressed after Varroa/virus infection. Finally, we applied our bioinformatics approach and generated a gene co-expression network to identify highly connected (hub) genes that may represent important mediators and regulators of anti-pathogen responses. Conclusions Our meta-analysis generated a comprehensive overview of the host metabolic and other biological processes that mediate interactions between insects and their pathogens. We identified key host genes and pathways that respond to phylogenetically diverse pathogens, representing an important source for future functional studies as well as offering new routes to identify or generate pathogen resilient honey bee stocks. The statistical and bioinformatics approaches that were developed for this study are broadly applicable to synthesize information across transcriptomic datasets. These approaches will likely have utility in addressing a variety of biological questions

Honey bee survival mechanisms against the parasite Varroa destructor: a systematic review of phenotypic and genomic research efforts

The ectoparasitic mite Varroa destructor is the most significant pathological threat to the western honey bee, Apis mellifera, leading to the death of most colonies if left untreated. An alternative approach to chemical treatments is to selectively enhance heritable honey bee traits of resistance or tolerance to the mite through breeding programs, or select for naturally surviving untreated colonies. We conducted a literature review of all studies documenting traits of A. mellifera populations either selectively bred or naturally selected for resistance and tolerance to mite parasitism. This allowed us to conduct an analysis of the diversity, distribution and importance of the traits in different honey bee populations that can survive V. destructor globally. In a second analysis, we investigated the genetic bases of these different phenotypes by comparing 'omics studies (genomics, transcriptomics, and proteomics) of A. mellifera resistance and tolerance to the parasite. Altogether, this review provides a detailed overview of the current state of the research projects and breeding efforts against the most devastating parasite of A. mellifera. By highlighting the most promising traits of Varroa-surviving bees and our current knowledge on their genetic bases, this work will help direct future research efforts and selection programs to control this pest. Additionally, by comparing the diverse populations of honey bees that exhibit those traits, this review highlights the consequences of anthropogenic and natural selection in the interactions between hosts and parasites. (C) 2020 The Authors. Published by Elsevier Ltd on behalf of Australian Society for Parasitology

Pan-european assessment, monitoring, and mitigation of stressors on the health of bees

As part of the activities foreseen by the POSHBEE project, we performed experiments to verify the toxicokinetics of three pesticides (the insecticide sulfoxaflor, the fungicide azoxystrobin and the herbicide glyphosate) in the bodies of social bees (Apis mellifera and Bombus terrestris) and solitary bees (Osmia bicornis). For each species all castes and sexes were studied. Based on the results from dose-response assays, sublethal doses were used to treat individuals, which were exposed to the chosen pesticides orally and by contact. Bees were then sampled at multiple time points post- exposure, to capture the breakdown of the active ingredients in the bodies of the organisms. Results of the chemical analyses on bee samples were used for the evaluation of the dynamics of the oral and contact acute exposure in the three species. Overall, the active ingredients sulfoxaflor, azoxystrobin and glyphosate degrade in all species, sex or caste. Nevertheless, there were some exceptions: in honey bee workers, glyphosate administered topically and azoxystrobin administered via the oral route seemed to remain stable in the bees’ bodies even 10 days after exposure (11% and 13% of degradation rate, respectively). We also observed a low degradation of sulfoxaflor following topical exposure in bumble bee queens (26% of degradation rate) and in Osmia bee females (22% of degradation rate). In bumble bees exposed topically to glyphosate degradation was lower than 50% for all sexes and castes. These results deserve further attention by researchers to understand the destiny of these molecules in the bee body and their effects therein.Prepared under contract from the European Commission; Grant agreement No. 773921; EU Horizon 2020 Research and Innovation action.Prepared under contract from the European Commission; Grant agreement No. 773921; EU Horizon 2020 Research and Innovation actio

Pan-european assessment, monitoring, and mitigation of stressors on the health of bees

Within the PoshBee Project, we have worked out precise protocols for testing the toxicological endpoints and metabolisation rate of agrochemicals on and in bees. Three agrochemical classes (insecticides, fungicides and herbicides) were tested on three bee groups (honey bees, bumble bees and solitary mason bees). The specific protocols, different for each combination of the pesticide class and bee group, were designed based on the existing toxicity testing methods and modified according to the specificities of the different bee groups, their castes and the different exotoxicological features of the compounds. Sulfoxaflor, Azoxystrobin, and Glyphosate were used respectively as examples of the three pesticide classes. Apis mellifera, Bombus terrestris and Osmia bicornis were chosen as the examples of the three bee groups. The protocols for honey bees and bumble bees were designed based on the official existing guidelines edited by OECD. Nevertheless, given that the OECD guidelines provide recommendations to fit with most agrochemicals, many of the parameters are defined with low precision in order to be flexible and adapt to different categories of molecules. While OECD has no guidelines for testing solitary bees, we used information from the unpublished ring test protocols for Osmia bicornis by the ICPPR non- Apis working group.Prepared under contract from the European Commission; Grant agreement No. 773921; EU Horizon 2020 Research and Innovation action.Prepared under contract from the European Commission; Grant agreement No. 773921; EU Horizon 2020 Research and Innovation action

Pan-european assessment, monitoring, and mitigation of stressors on the health of bees

Within the PoshBee Project we have tested three bee species – honey bees Apis mellifera, bumble bees Bombus terrestris and solitary bees Osmia bicornis – for their sensitivity to pesticides and analysed the clearance of pesticides from bees. For each species, all castes and sexes were studied. We synthesised the mortality data (LD50 or results of limit tests) with the toxicokinetic patterns and analysed this against the background of inter- and intraspecific variation in life-histories of the tested bees. The clearance of sulfoxaflor is relatively similar across all bee species tested and in females after contact treatment it tends to be retained. The toxicity increases over time independently of the clearance from the body. The clearance of azoxystrobin was rapid in Osmia and bumble bees, as well as in honey bee queens, but in honey bee workers there was very little clearance. Similar to sulfoxaflor the toxicity increased over time, although the residues were detected at very low levels. Glyphosate tended to be retained in bumble bees after contact treatment but cleared rapidly after oral treatment. For Osmia bees only in males after contact treatment was the glyphosate almost lost. The toxicity of a pesticide is dependent on the exact dosage, but also the exposure route and time, as well as the speed of detoxification and clearance from a body. The assessment for the hazard that a less toxic pesticide might pose, can be largely dependent on the exposure route. The effects of pesticide toxicity can increase even after the molecules have been cleared out of the body.Prepared under contract from the European Commission; Grant agreement No. 773921; EU Horizon 2020 Research and Innovation action.Prepared under contract from the European Commission; Grant agreement No. 773921; EU Horizon 2020 Research and Innovation action

Pan-european assessment, monitoring, and mitigation of stressors on the health of bees

Inter-individual differences in pesticide sensitivity may trigger variability in the risk posed by pesticides. Therefore, to better inform pesticide risk assessment for bees, we studied the variability of responses to several pesticides based on endogenous (developmental stage, genetic background, caste) and exogenous factors (pesticide co-exposure). We mainly investigated the toxicity of the insecticide sulfoxaflor, the fungicide azoxystrobin and the herbicide glyphosate. We first used LD50 tests to determine the acute oral and contact toxicity of these pesticides across the different bee species, developmental stages (larva vs adult in honey bees), castes (honey bee and bumble bee workers, queens and drones), and genetic backgrounds (honey bee subspecies). We then considered the risks posed by chronic and sublethal exposures to pesticides by implementing behavioural and reproductive endpoints in the screening of pesticide toxicity. Data showed that azoxystrobin and glyphosate under the test conditions were mildly toxic to bees. However, a large variability in bee sensitivity to sulfoxaflor was found, especially across species and individuals of different castes or sex. This variability is therefore important to consider for increasing the safety margin of the risk posed by insecticides in bees. Several effects induced by sublethal concentrations or doses of pesticides are also described, such as the occurrence of a Non-Monotonic Dose-Response (NMDR) and delayed effects in honey bees, impairment of reproductive performances in bumble bees, and a decreased longevity of Osmia adult females (although no effects were found on larval development). Finally, an interaction between pesticides was found when exposure was by contact, but not under oral exposure. In conclusion, the range of effects described here provides very useful insights for better understanding the toxicity of pesticides and therefore the risks they might pose to bees.Prepared under contract from the European Commission; Grant agreement No. 773921; EU Horizon 2020 Research and Innovation action.Prepared under contract from the European Commission; Grant agreement No. 773921; EU Horizon 2020 Research and Innovation action

Large expert-curated database for benchmarking document similarity detection in biomedical literature search

Document recommendation systems for locating relevant literature have mostly relied on methods developed a decade ago. This is largely due to the lack of a large offline gold-standard benchmark of relevant documents that cover a variety of research fields such that newly developed literature search techniques can be compared, improved and translated into practice. To overcome this bottleneck, we have established the RElevant LIterature SearcH consortium consisting of more than 1500 scientists from 84 countries, who have collectively annotated the relevance of over 180 000 PubMed-listed articles with regard to their respective seed (input) article/s. The majority of annotations were contributed by highly experienced, original authors of the seed articles. The collected data cover 76% of all unique PubMed Medical Subject Headings descriptors. No systematic biases were observed across different experience levels, research fields or time spent on annotations. More importantly, annotations of the same document pairs contributed by different scientists were highly concordant. We further show that the three representative baseline methods used to generate recommended articles for evaluation (Okapi Best Matching 25, Term Frequency-Inverse Document Frequency and PubMed Related Articles) had similar overall performances. Additionally, we found that these methods each tend to produce distinct collections of recommended articles, suggesting that a hybrid method may be required to completely capture all relevant articles. The established database server located at https://relishdb.ict.griffith.edu.au is freely available for the downloading of annotation data and the blind testing of new methods. We expect that this benchmark will be useful for stimulating the development of new powerful techniques for title and title/abstract-based search engines for relevant articles in biomedical research.Peer reviewe

Delayed effects of a single dose of a neurotoxic pesticide (sulfoxaflor) on honeybee foraging activity

International audiencePesticide risk-assessment guidelines for honeybees (Apis mellifera) generally require determining the acute toxicity of a chemical over the short-term through fix-duration tests. However, potential long-lasting or delayed effects resulting from an acute exposure (e.g. a single dose) are often overlooked, although the modification of a developmental process may have life-long consequences. To investigate this question, we exposed young honeybee workers to a single sublethal field-realistic dose of a neurotoxic pesticide, sulfoxaflor, at one of two amounts (16 or 60 ng), at the moment when they initiated orientation flights (preceding foraging activity). We then tracked in the field their flight activity and lifespan with automated life-long monitoring devices. Both amounts of sulfoxaflor administered reduced the total number of flights but did not affect bee survival and flight duration. When looking at the time series of flight activity, effects were not immediate but delayed until foraging activity with a decrease in the daily number of foraging flights and consequently in their total number (24 and 33% less for the 16 and 60 ng doses, respectively). The results of our study therefore blur the general assumption in honeybee toxicology that acute exposure results in immediate and rapid effects and call for long-term recording and/or time-to-effect measurements, even upon exposure to a single dose of pesticide

Transcriptome profiling of the honeybee parasite Varroa destructor provides new biological insights into the mite adult life cycle

Abstract Background The parasite Varroa destructor represents a significant threat to honeybee colonies. Indeed, development of Varroa infestation within colonies, if left untreated, often leads to the death of the colony. Although its impact on bees has been extensively studied, less is known about its biology and the functional processes governing its adult life cycle and adaptation to its host. We therefore developed a full life cycle transcriptomic catalogue in adult Varroa females and included pairwise comparisons with males, artificially-reared and non-reproducing females (10 life cycle stages and conditions in total). Results Extensive remodeling of the Varroa transcriptome was observed, with an upregulation of energetic and chitin metabolic processes during the initial and final phases of the life cycle (e.g. phoretic and post-oviposition stages), whereas during reproductive stages in brood cells genes showing functions related to transcriptional regulation were overexpressed. Several neurotransmitter and neuropeptide receptors involved in behavioural regulation, as well as active compounds of salivary glands, were also expressed at a higher level outside the reproductive stages. No difference was detected between artificially-reared phoretic females and their counterparts in colonies, or between females who failed to reproduce and females who successfully reproduced, indicating that phoretic individuals can be reared outside host colonies without impacting their physiology and that mechanisms underlying reproductive failure occur before oogenesis. Conclusions We discuss how these new findings reveal the remarkable adaptation of Varroa to its host biology and notably to the switch from living on adults to reproducing in sealed brood cells. By spanning the entire adult life cycle, our work captures the dynamic changes in the parasite gene expression and serves as a unique resource for deciphering Varroa biology and identifying new targets for mite control