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Honey bees are key insect pollinators, providing important economic and ecological value for human beings and ecosystems. This has triggered the development of several monitoring methods for assessing the temporal development of colony size, food storage, brood and pathogens. Nonetheless, most of these methods are based on visual assessments that are observer-dependent and prone to bias. Furthermore, the impact on colony development (invasiveness), as well as accuracy, were rarely considered when implementing new methods. In this study, we present and test a novel accurate and observer-independent method for honey bee colony assessment, capable of being fully standardized. Honey bee colony size is quantified by assessing the weight of adult bees, while brood and provision are assessed by taking photos and conducting image analysis of the combs with the image analysis software Deepbee®. The invasiveness and accuracy of the method were investigated using field data from two experimental apiaries in Portugal, comparing results from test and control colonies. At the end of each field experiment, most of the tested colonies had the same colony size, brood levels and honey production as the control colonies. Nonetheless, continuous weight data indicated some disturbance in tested colonies in the first year of monitoring. The overall accuracy of the image analysis software was improved by training, indicating that it is possible to adapt the software to local conditions. We conclude that the use of this fully quantitative method offers a more accurate alternative to classic visual colony assessments, with negligible impact on colony development. Honey bees are key insect pollinators, providing important economic and ecological valuefor human beings and ecosystems. This has triggered the development of several monitoringmethods for assessing the temporal development of colony size, food storage, broodand pathogens. Nonetheless, most of these methods are based on visual assessments thatare observer-dependent and prone to bias. Furthermore, the impact on colony development(invasiveness), as well as accuracy, were rarely considered when implementing new methods.In this study, we present and test a novel accurate and observer-independent method forhoney bee colony assessment, capable of being fully standardized. Honey bee colony size isquantified by assessing the weight of adult bees, while brood and provision are assessed bytaking photos and conducting image analysis of the combs with the image analysis softwareDeepbeeVR . The invasiveness and accuracy of the method were investigated using field datafrom two experimental apiaries in Portugal, comparing results from test and control colonies.At the end of each field experiment, most of the tested colonies had the same colony size,brood levels and honey production as the control colonies. Nonetheless, continuous weightdata indicated some disturbance in tested colonies in the first year of monitoring. The overallaccuracy of the image analysis software was improved by training, indicating that it ispossible to adapt the software to local conditions. We conclude that the use of this fullyquantitative method offers a more accurate alternative to classic visual colony assessments,with negligible impact on colony development.

Olfaction is a crucial sensory modality underlying foraging, social and mating behaviors in many insects. Since the olfactory system is at the interface between the animal and its environment, it receives strong evolutionary pressures that promote neuronal adaptations and phenotypic variations across species. Hornets are large eusocial predatory wasps with a highly developed olfactory system, critical for foraging and intra-specific communication. In their natural range, hornet species display contrasting ecologies and olfactory-based behaviors, which might match to adaptive shifts in their olfactory system. The first olfactory processing center of the insect brain, the antennal lobe, is made of morphological and functional units called glomeruli. Using fluorescent staining, confocal microscopy and 3D reconstructions, we compared antennal lobe structure, glomerular numbers and volumes in four hornet species (Vespa crabro, Vespa velutina, Vespa mandarinia and Vespa orientalis) with marked differences in nesting site preferences and predatory behaviors. Despite a conserved organization of their antennal lobe compartments, glomeruli numbers varied strongly between species, including in a subsystem thought to process intraspecific cuticular signals. Moreover, specific adaptations involving enlarged glomeruli appeared in two species, V. crabro and V. mandarinia, but not in the others. We discuss the possible function of these adaptations based on species-specific behavioral differences.

Bees and other pollinators play vital roles in biodiversity and food security, and they are a source of income and inspiration. Yet, bees’ biodiversity and abundance are decreasing every day. Their decline, driven by pesticides (1–3), poses serious threats to the environment, ecosystems, and human health. The European pesticide authorization framework states that a pesticide can only be put on the market if it has no harmful effects on human health or animal health and no unacceptable effects on the environment (4). EU ministries met in June to decide what constitutes an “acceptable” effect of a pesticide on bees (5). Despite scientific evidence indicating that the current level of protection is barely adequate, they tentatively agreed to provide even less protection moving forward (5). We call on EU governments to apply the maximum level of protection of pollinators from pesticides (6).

To understand the real causes of disorders and abnormal mortalities of bees (honeybees and wild bees) in the world, the scientific method requires that each category of potential stressors be analyzed in a balanced manner. In this article, I show that the first version of the section dealing with the evaluation of the toxic effects of pesticides on bees, in the IPBES assessment report on pollinators and pollination, revealed an incomplete and biased literature review in many places, especially downplaying the risks that pesticides in general, and neonicotinoids in particular, pose for pollinating insects. Then, according to the rules of IPBES, an independent peer review by external experts of this first version allowed the published report to be more in line with the reality of scientific knowledge, which shows, for example, that sublethal effects of pesticide exposure can impair the ability of bees to provide pollination. However, some other key points remain unchanged in the published version.

Current approaches to risk assessment in bees do not take into account co-exposures from multiple stressors. The European Food Safety Authority (EFSA) is deploying resources and efforts to move towards a holistic risk assessment approach of multiple stressors in bees. This paper describes the general principles of pesticide risk assessment in bees, including recent developments at EFSA dealing with risk assessment of single and multiple pesticide residues and biological hazards. The EFSA Guidance Document on the risk assessment of plant protection products in bees highlights the need for the inclusion of an uncertainty analysis, other routes of exposures and multiple stressors such as chemical mixtures and biological agents. The EFSA risk assessment on the survival, spread and establishment of the small hive beetle, Aethina tumida, an invasive alien species, is provided with potential insights for other bee pests such as the Asian hornet, Vespa velutina. Furthermore, data gaps are identified at each step of the risk assessment, and recommendations are made for future research that could be supported under the framework of Horizon 2020. Finally, the recent work conducted at EFSA is presented, under the overarching MUST-B project ("EU efforts towards the development of a holistic approach for the risk assessment on MUltiple STressors in Bees") comprising a toolbox for harmonised data collection under field conditions and a mechanistic model to assess effects from pesticides and other stressors such as biological agents and beekeeping management practices, at the colony level and in a spatially complex landscape. Future perspectives at EFSA include the development of a data model to collate high quality data to calibrate and validate the model to be used as a regulatory tool. Finally, the evidence collected within the framework of MUST-B will support EFSA's activities on the development of a holistic approach to the risk assessment of multiple stressors in bees. In conclusion, EFSA calls for collaborative action at the EU level to establish a common and open access database to serve multiple purposes and different stakeholders.

International audience; Invasive species are now recognized as a major cause of native biodiversity loss worldwide. In the current deleterious context for pollinators, the invasive yellow-legged hornet, Vespa velutina, represents an additional threat to the domestic honeybee, Apis mellifera, in Europe. Therefore, understanding the impact of this predator on honeybee colonies is of major importance. In the present study, we tried to assess the impact of V. velutina on the honeybee foraging and defence behaviour based on the video monitoring of two hives. Balling behaviour is reported here for the first time under natural conditions in A. mellifera against V. velutina in Europe. Although these results are preliminary and should be carefully considered, we found that the number of hornets impacted honeybee foraging and defence behaviours. More defensive behaviours were notified in the hive, which survives slightly longer. This may suggest that selecting for more defensive colonies may provide an interesting perspective.

In eusocial insect species, the nest represents the fundamental element of the colony. Extranidal activities (foraging, nest maintenance, defence) are fundamental for the development and the survival of the colony. Therefore, they may represent interesting targets to disrupt to limit their expansion in case of pest species such as Vespa velutina, an alien predator of domestic honeybees in Europe. An accurate knowledge of the pattern of activity of this pest’s colonies is therefore required. Due to the highly defensive nature of this hornet, a video monitoring was realized on two colonies during their growth from August to November. Three major behaviours were monitored: nest maintenance, patrolling on the nest and foraging flights. Although of different size and monitored during different years, the two colonies exhibited similar patterns of daily and seasonal variation. This work is a first step in the study of this pest especially in view of control program.

Social learning is taxonomically widespread in the animal kingdom [1], and although it is long thought to be a hallmark of vertebrates, recent studies revealed that it also exists in insects [2-5]. The adaptive functions of social learning are well known, but its underlying mechanisms remain debated [2, 5, 6]. Social insects critically depend on the social transmission of information for successful food search and their colonies' fitness [7] and are tractable models for studying the social cues and cognitive mechanisms involved [2-5]. Besides the well-known dance language allowing them to communicate the location of food sources among nestmates [8], honeybees also learn chemosensory information about these sources both outside and within the hive [9, 10]. In the latter case, they associate the floral scent carried by returning foragers on their body with the nectar provided through mouth-to-mouth trophallaxis, similar to the manner in which foragers directly learn odorant-nectar reward associations at the foraging patch [9-11]. Strikingly, however, neither the dance nor trophallaxis is strictly necessary for foragers recruited within the hive to find the right floral source, and simple body contact between foragers may be sufficient [12]. What is the reinforcing agent in this case? We show here that simple social contact acts as appetitive reinforcement and can be used in associative olfactory learning. We demonstrate that this social reinforcement is mediated by bees' antennal movements and modulated by bees' behavioral development. These results unveil a social learning mechanism that may play a facilitating role in resource exploitation by social groups.

Honey bee populations from the island of Cyprus were analyzed using microsatellite and mitochondrial DNA markers. A total of 268 colonies were sampled in Cyprus, at six different locations-Kyrenia, Katydata, Flassou, Alabra, Troulloi, and Alassa-covering a wide area of the island. Results showed that the Cyprian honey bee Apis mellifera cypria could be distin-guished from other Apis mellifera subspecies based on a „double pattern“ of mitochondrial DNA belonging to the C1 lineage and microsatellite DNA belonging to the O lineage. All populations were homogeneous, except the population from Kyrenia, probably due to the introduction of queens or colonies belonging to the C2, C6, and m7 lineages.Keywords: Apis mellifera cypria, genetic characterization, mitochondrial DNA,microsatellite DNA. 1 CNRS, Laboratoire Evolution, Genomes et Speciation, UPR 9034, CNRS, 91198, Gif-sur-Yvette cedex, France 2 Universite Paris-Sud 11, 91405 Orsay cedex, France 3 Current address: Laboratory of Animal Physiology, School of Biology, Aristo-tle University of Thessaloniki, 54124 Thessaloniki, Greece

A honey bee queen mates with a number of drones a few days after she emerges as an adult. Spermatozoa of different drones are stored in her spermatheca and used for the rest of the queen’s life to fertilize eggs. Sperm usage is thought to be random, so that the patriline distribution within a honey bee colony would remain constant over time. In this study we assigned the progeny of a naturally mated honey bee queen to patrilines using microsatellite markers at the queen’s age of two, three and four years. No significant changes in patriline distribution occurred within each of two foraging seasons, with samples taken one and five months apart, respectively. Overall and pair-wise comparisons between the three analyzed years reached significant levels. Over the three-year period we found a trend for patrilines to become more equally represented with time. It is important to note that this study was performed with a single queen, and thus individual and population variation in sperm usage patterns must be assessed. We discuss long-term changes in patriline composition due to mixing processes in the queen’s spermatheca, following incomplete mixing of different drones’ sperm after mating.

We report the characterization of fifteen microsatellite markers in Vespa velutina, an invasive hornet from Asia that has been unintentionally introduced in France before 2005. It is expanding rapidly, covering one third of the French territory and northern Spain, and causes severe losses to honeybee colonies. An enrichment protocol was used to isolate microsatellite loci, and polymorphism was explored in an invasive population from France and in a population from the native mainland location in China. These markers showed a number of alleles per locus and per population ranging from 1 to 11, and expected heterozygosities ranging from 0.151 to 0.891. These polymorphic markers will be useful to identify the source of the invading population and to discover the invasion pathways.

The black honeybee (Apis mellifera mellifera; M lineage) which is affected by widespread colony losses, colony introgression and population shifts within its natural range of distribution, needs proper protection and management planning. The DraI COI-COII test discriminates between and within honeybee lineages (M, C, A and O). With this test, a global assessment of the diversity found in the M lineage (i.e. description of haplotype frequencies, fragment length and size, nucleotide sequences) is made with colonies newly sampled in France and Belgium (n = 3,261 colonies) and previously described in the literature (n = 3,367). A total of 91 M haplotypes were described including 52 new ones. The haplotypes M4 and M4’ were the most frequent followed by M17, M7, M6, M19, M8, M17’ and by many rare ones. Finally, these data were used to infer hypotheses on the putative origin of the M haplotypic diversity and to determine a protocol for the establishment of protected areas of black honeybee colonies. This protocol is based on the DraI COI-COII test and on case-studies of protected areas of black honeybees from France.

Associative proboscis extension learning differs widely among bees of a colony. This variety of performances is often related to differences in sucrose responsiveness, which determines learning performance. Sucrose responsiveness is partly determined genetically. We studied for the first time effects of paternal genes on associative learning independent of sucrose responsiveness. To do this, we used wild-type workers stemming from five unrelated patrilines. Bees of the patrilines were first tested for sucrose responsiveness. Only bees with equal sucrose responsiveness were analysed for associative olfactory learning, memory and discrimination. The bees of different patrilines did not differ in their acquisition, memory or discrimination of odours when they had similar sucrose responsiveness. But patrilines differed significantly in their sucrose responsiveness. This shows genetic effects on sensory responsiveness but no independent effects on associative learning.

International audience; The yellow-legged hornet, Vespa velutina, was accidentally introduced from Southeast Asia and then invaded France and Korea over the last 10 years. Since its introduction, its predation on honeybee colonies has rapidly become an economic problem in invaded countries. Using mitochondrial cytochrome C oxidase and 22 nuclear microsatellite loci, we showed that native hornet populations were well differentiated and highly diverse. In contrast, introduced populations from France and Korea suffered a genetic bottleneck, which did not prevent their rapid geographic expansion. Analysis of the genetic data indicates that French and Korean populations likely arose from two independent introduction events. The most probable source population is from an area between the Chinese provinces of Zhejiang and Jiangsu. This invasion route is in agreement with knowledge on trade and historical records. By studying colonies of V. velutina, we demonstrated its polyandry, which is very rare among Vespidae. This mating behavior could have favored the success of this Asian hornet in Europe and Korea. Combined, the population and colony results suggest that very few or possibly only one single multi-mated female gave rise to the invasion.

Honeybees have evolved numerous mechanisms for increasing colony-level foraging efficiency, mainly the combined system of scout-recruit division of labour and recruitment communication. A successful forager performs waggle dances on the surface of the comb where it interacts with nectar receivers and dance followers. A forager uses tremble dance when it experiences difficulty finding a receiver bee to unload food upon return to the hive. A bee colony containing numerous subfamilies may increase its efficiency in dance communication if dances are realized by particular groups of specialized individuals or subfamilies rather than by undifferentiated workers. In this study, we determined the subfamily frequencies of waggle and tremble dancers in a colony headed by a naturally mated queen, where the 17 subfamilies can be identified by microsatellite genetic markers. Our results demonstrate that a genetic component is associated with the dance communication in honeybees. More than half of the waggle dances and the tremble dances were performed by workers from only four subfamilies in each case.

Depuis 2009, des variétés de tournesol et de colza, dites « tolérantes aux herbicides » (TH), font leur apparition en France. Destinées à faciliter le désherbage, ces variétés résistent à un herbicide donné ce qui permet a priori d’appliquer l’herbicide sans risque pour la culture. La culture de telles variétés a suscité une contestation sociale se traduisant par l’arrachage de tournesols TH dénoncés comme « OGM cachés ». Dans ce contexte, les ministères chargés de l’Agriculture et de l’Environnement ont demandé à l’Inra et au CNRS de réaliser un bilan des connaissances disponibles sur l’ensemble des impacts de la culture de ces variétés.

International audience; We investigated the prey-predator interactions between the European honeybee, Apis mellifera, and the invasive yellow-legged hornet, Vespa velutina, which first invaded France in 2004 and thereafter spread to neighbouring European countries (Spain, Portugal and Italy). Our goal was to determine how successfully honeybees are able to defend their colonies against their new predator in Europe. Experiments were conducted in the southwest of France-the point of entry of the hornet in Europe-under natural and semi-controlled field conditions. We investigated a total of eight apiaries and 95 colonies subjected to either low or high levels of predation. We analyzed hornet predatory behaviour and collective response of colonies under attack. The results showed that A. mellifera in France exhibit an inefficient and unorganized defence against V. velutina, unlike in other regions of Europe and other areas around the globe where honeybees have co-evolved with their natural Vespa predators.

Although kinship discrimination and nepotism have been extensively investigated in honeybees, Apis mellifera L., the significance of kin discrimination among worker bees is still unclear. One of the behavioural contexts in which nepotistic behaviour of worker bees has been described is the dance communication system: bees were reported to have a strong tendency to dance with their supersisters in colonies consisting of only two subfamilies. Using molecular markers, we reinvestigated this apparent case of nepotism. We found no evidence for subfamily discrimination among dancers and their followers in a colony consisting of only two subfamilies or in a colony consisting of 17 subfamilies.

In honeybees, the cuticular hydrocarbon profiles are partly genetically based and differ between subfamilies, which suggests that they might be used by the workers as labels for subfamily recognition. This ability could potentially form the basis for nepotistic conflicts between subfamilies that would be detrimental to the inclusive fitness of the colony. Here we have compared the subfamily hydrocarbon profiles of 5-day-old workers maintained in isolation with those kept in their parental colony. We demonstrate that the cuticular hydrocarbon profiles tend to be less distant between most subfamilies within the hive compared with those held in isolation. The main consequence of this partial homogenization of the majority of subfamily signatures may result in a reduction of the number of recognizable subfamilies in the colony. Nevertheless, a few subfamilies retain very distinct cuticular hydrocarbon profiles.

SUMMARYCuticular hydrocarbon profiles of drone honey bees (Apis mellifera carnica) of different ages were compared by gas chromatography. Chromatograms of drones from three different colonies were similar and, despite some variations, showed the same patterns as a function of age. We suggest that in addition to behavioural changes, the surface hydrocarbon profiles of the drones might be used as a cue for the worker bees to discriminate between young and old individuals.

WOS: 000319878900011 Most studies that have shown negative sublethal effects of the pesticide imidacloprid on honeybees concern behavioral effects; only a few concern physiological effects. Therefore, we investigated sublethal effects of imidacloprid on the development of the hypopharyngeal glands (HPGs) and respiratory rhythm in honeybees fed under laboratory conditions. We introduced newly emerged honeybees into wooden mesh-sided cages and provided sugar solution and pollen pastry ad libitum. Imidacloprid was administered in the food: 2 mu g/kg in the sugar solution and 3 mu g/kg in the pollen pastry. The acini, the lobes of the HPGs of imidacloprid-treated honeybees, were 14.5 % smaller in diameter in 9-day-old honeybees and 16.3 % smaller in 14-day-old honeybees than in the same-aged untreated honeybees; the difference was significant for both age groups. Imidacloprid also significantly affected the bursting pattern of abdominal ventilation movements (AVM) by causing a 59.4 % increase in the inter-burst interval and a 56.99 % decrease in the mean duration of AVM bursts. At the same time, the quantity of food consumed (sugar solution and pollen pastry) per honeybee per day was the same for both treated and untreated honeybees. EUEuropean Union (EU); Greek Ministry of Agricultural Development and Food; Greek-French bilateral collaboration project [PLATON 09 FR75] This work was supported by a joint project from the EU and the Greek Ministry of Agricultural Development and Food (2008-2010) and by a Greek-French bilateral collaboration project, PLATON 09 FR75. We thank Dimitra Fouka, laboratory technician in Hellenic Institute of Apiculture, for technical assistance. We are also profoundly indebted to Sharilynn Wardrop, for stylistic and linguistic improvements as well to the anonymous reviewers for valuable suggestions.

International audience; Contrary to native predators, which have co-evolved with their prey, alien predators often benefit from native prey naïveté. Vespa velutina, a honeybee predator originating from Eastern China, was introduced into France just before 2004. The present study, based on video recordings of two beehives at an early stage of the invasion process, intends to analyse the alien hornet hunting behaviour on the native prey, Apis mellifera, and to understand the interaction between the activity of the predator and the prey during the day and the season. Chasing hornets spent most of their time hovering facing the hive, to catch flying honeybees returning to the hive. The predation pressure increased during the season confirming previous study based on predator trapping. The number of honeybee captures showed a maximum peak for an intermediate number of V. velutina, unrelated to honeybee activity, suggesting the occurrence of competition between hornets. The number of honeybees caught increased during midday hours while the number of hornets did not vary, suggesting an increase in their efficacy. These results suggest that the impact of V. velutina on honeybees is limited by its own biology and behaviour and did not match the pattern of activity of its prey. Also, it could have been advantageous during the invasion, limiting resource depletion and thus favouring colonisation. This lack of synchronization may also be beneficial for honeybee colonies by giving them an opportunity to increase their activity when the hornets are less effective.

In the polyandrous honey bee, Apis mellifera, workers can potentially increase their inclusive fitness by rearing full-sister queens. If the mother queen dies suddenly, workers feed a few larvae in worker cells with royal jelly and rear them into queens (emergency queen rearing). Using DNA microsatellite markers we determined the patriline of emergency queens reared in two colonies headed by naturally-mated queens before being made queenless. We found that some patrilines were reared more than others in one colony, but not in the other. These differences between colonies suggest that selective rearing is not always present and this might explain the mixed results of previous nepotism studies in the honey bee.

Honey bee (Apis mellifera L.) colonies consist of many patrilines (Estoup et al., 1994) and workers can potentially increase their inclusive fitness during queen replacement if they can favour a full-sister offspring queen (Visscher, 1998). There is evidence of preferential rearing of some patrilines in emergency queen rearing (Tilley and Oldroyd, 1997; Osborne and Oldroyd, 1999; Châline et al., 2003), although the reason for this is unclear. Some patrilines could act nepotistically or a “royalty allele” could make some larvae more attractive to workers (Osborne and Oldroyd, 1999). Early studies on behavioural interactions and larval preferences during queen rearing used colonies with limited numbers of patrilines or cross-fostering of larvae between unrelated colonies with naturally mated queens. They showed weak or no evidence of nepotism (Breed et al., 1984; Noonan, 1986; Page et al., 1989). To avoid the problem of unnaturally low number of patrilines and the use of artificial insemination, we used microsatellites to analyse the kinship of feeding workers and queen larvae during emergency queen rearing, for the first time in a colony with a naturally mated queen. The experiment was carried out in double blind in a queenless observation hive containing individually marked bees. This was done by marking bees emerging from two brood combs of the test colony over 6 consecutive days. Before the experimental colony set-up, the marked bees were returned to their mother colony to ensure a normal ontogeny. Twentyfour hours before the set-up, 50 queen cells were initiated in a queen rearing colony by grafting larvae from the test colony. Thirteen of these were then attached to a brood comb from the test colony and transferred in an observation hive with 2000 of the marked workers. During the experiment, three additional queen cells were built naturally. Each queen cell was observed individually for 1/2 h every day until capping. The total observation time was 2130 min. The bees feeding each queen larvae were recorded, as this seemed the best behaviour for the

The hazard posed to honeybees by systemic insecticides is determined by toxicity tests that are designed to study the effects of insecticides applied on the aerial parts of plants, but are not adapted to systemic substances used as soil or seed treatments. Based on the available data found in the literature, this paper pro- poses modes of honeybees exposure to systemic insecticides by estimating their pollen and nectar consump- tion. Estimates are given for larvae and for the categories of adults which consume the highest amounts of - pollen, the nurse bees, and - nectar, the wax-producing bees, the brood attending bees, the winter bees, and the foraging bees. As a case study, we illustrate these estimates with the example of imidacloprid because its concentrations in sunflower nectar and in sunflower and maize pollens of seed-dressed plants have been precisely determined, and because its levels of lethal, sublethal, acute, and chronic toxicities have been exten- sively investigated. Apis mellifera / systemic insecticide / exposure / imidacloprid / nectar / pollen

In May 2013, the European Commission (EC) adopted a 2‐year restriction on the use of three neonicotinoid pesticides – clothianidin and imidacloprid, both produced by Bayer, and thiametoxam, produced by Syngenta – on crops that attract bees and other pollinators to protect declining populations of pollinating bees. The decision was based on a risk assessment by the European Food Safety Authority (EFSA) [1]. In August 2013, the two companies sued the EC. Syngenta claims the decision was based on a flawed process, on an inaccurate and incomplete EFSA assessment, and was made without the full support of the EU Member States. The EFSA assessments focused on three main routes by which bees could be exposed to the pesticides: from residues in nectar and pollen in the flowers of treated plants; from dust produced when treated seeds are sown or granules are applied; and from residues in guttation fluid produced by treated plants. They found …

International audience; Odorant-binding proteins (OBPs) are small abundant extracellular proteins thought to participate in perireceptor events of odor-pheromone detection by carrying, deactivating, and/or selecting odor stimuli. The honeybee queen pheromone is known to play a crucial role in colony organization, in addition to drone sex attraction. We identified, for the first time in a social insect, a binding protein called antennal-specific protein 1 (ASP1), which binds at least one of the major queen pheromone components. ASP1 was characterized by cDNA cloning, expression in Pichia pastoris, and pheromone binding. In situ hybridization showed that it is specifically expressed in the auxiliary cell layer of the antennal olfactory sensilla. The ASP1 sequence revealed it as a divergent member of the insect OBP family. The recombinant protein presented the exact characteristics of the native protein, as shown by mass spectrometry, and N-terminal sequencing and exclusion-diffusion chromatography showed that recombinant ASP1 is dimeric. ASP1 interacts with queen pheromone major components, opposite to another putative honeybee OBP, called ASP2. ASP1 biosynthetic accumulation, followed by nondenaturing electrophoresis during development, starts at day 1 before emergence, in concomitance with the functional maturation of olfactory neurons. The isobar ASP1b isoform appears simultaneously to ASP1a in workers, but only at approximately 2 weeks after emergence in drones. Comparison of in vivo and heterologous expressions suggests that the difference between ASP1 isoforms might be because of dimerization, which might play a physiological role in relation with mate attraction.

Summary Asian honeybees have been shown to kill hornets by ‘thermo- balling', in which they surround a hornet to form a ball within which the temperature increases to a lethal level. We report here that Cyprian honeybees, Apis mellifera cypria , kill their major enemy, the Oriental hornet, Vespa orientalis , in a different way — by asphyxia- balling, in which the Cyprian honeybees mob the hornet and smother it to death.

55 ref. 6 graph.; International audience; Four distinct hexamerin subunits (referred to as "hexamerins" in the following text) have been identified in the developing honeybee, Apis mellifera, by N-terminal protein sequencing. Hexamerins are abundant in the hemolymph of late larval and early pupal stages, and gradually decline during metamorphosis and adult development. Three hexamerins in the 70 kDa range have been found (Hex70a, Hex70b, Hex70c). In worker and drone, Hex70a is the only hexamerin present in large amount in later adult stages. Hex70b and c exhibit a similar developmental profile, disappearing in the drone just before adult emergence, and in the worker just after. Hex70b or Hex70c are still detectable in the adult queen. Hex80/110 likely exist in at least 3 different subunits, 1 of 110 kDa, and 2 of around 80 kDa, which all share a common N-terminus. They disappear during metamorphosis earlier than Hex70b and c. All these hexamerins have been found also in the antenna, suggesting their utilization in building up of antennal cuticle structures.

The PPR Panel was asked to deliver a scientific opinion on the science behind the development of a risk assessment of plant protection products on bees (Apis mellifera, Bombus spp. and solitary bees). Specific protection goals options were suggested based on the ecosystem services approach. The different routes of exposure were analysed in detail for different categories of bees. The existing test guidelines were evaluated and suggestions for improvement and further research needs were listed. A simple prioritisation tool to assess cumulative effects of single pesticides using mortality data is suggested. Effects from repeated and simultanous exposure and synergism are discussed. Proposals for separate risk assessment schemes, one for honey bees and one for bumble bees and solitary bees, were developed.

Dans le domaine biomedical, qui est l’objet principal de cet article, la litterature montre une correlation directe entre le financement d’une recherche par un industriel et la communication de resultats qui lui sont favorables. Cet effet, appele « le biais de financement », persiste depuis les travaux des annees 1990 jusqu’aux travaux recents. Il peut etre elargi a d’autres domaines tels que l’agro-alimentaire, l’etude des risques environnementaux et sanitaires, etc. Les conflits d’interets peuvent influencer la recherche par des voies inconscientes ou volontaires. Nous analysons les modes d’action possibles des conflits d’interets sur le deroulement de l’activite scientifique, ainsi que l’efficacite des diverses solutions qui ont ete proposees pour gerer ces influences.

Un des parametres les plus importants permettant d'evaluer l'activite d'une colonie d'abeilles est l'evolution, au cours du temps, du nombre des entrees et des sorties d'insectes de leur ruche. Le compteur presente ici est controle par un microprocesseur, et presente plusieurs avantages par rapport aux dispositifs precedemment decrits. Il est bien adapte aux dimensions d'une ruche standard (a 10 cadres), et comporte 32 canaux bidirectionnels. Les detecteurs a infra-rouge presents dans chaque canal ne sont separes que de 0,1 mm, et sont controles par un algorithme sequentiel asynchrone qui augmente la precision du compteur

Nous avons entrepris des etudes multidisciplinaires destinees a elucider les mecanismes neuronaux et comportementaux responsables des capacites d'adaptation de l'abeille aux modifications des conditions environnementales, en nous basant sur l'etude des reponses de cet insecte a des signaux chimiques complexes (fig 1). Des etudes comportementales de la discrimination olfactive ont ete entreprises en utilisant pour stimulus des emissions volatiles florales. L'etude du comportement de butinage, conduite en conditions naturelles et au laboratoire, sur des plantes cultivees telles que le tournesol, associee a l'identification chimique des emissions florales, a montre que:―les preferences inter-genotypes, liees a la qualite du nectar, etaient etablies sur la base d'un nombre limite de constituants discriminatifs au sein des aromes floraux

International audience; Capping of worker Apis mellifera cells is elicited by four fatty acid methyl esters (Methyl palmitate, methyl oleate, methyl linoleate, and methyl linolenate) that are present on the surface of the worker and drone larvae only a few hours before the cell is closed. The amount of the pheromone reaches its maximum value when the cell has just been capped, at 8.5 and at 10.25 days of age, respectively, for worker and drone larvae. There after, the amount of the pheromone decreases to its initial level. These data suggest that the esters also have a role in the capping of the drone cells, the temporal signal allowing the worker bees to recognize the age of the larvae and then to do the appropriate behavior. Two pheromonal components, methyl palmitate and methyl linolenate, and the inactive ethyl palmitate are kairomones attractive to Varroa females. Their secretion by the larvae follows the same pattern of development as the pheromonal signal. The longer and greater kairomonal signal in drone larvae, compared to worker secretion, could explain the preference of Varroa towards drone brood.

The influence of different temperatures (from 26 to 39.5°C) and relative humidities (40 and 70%) on the development of Varma jacobsoni (Oudemans) was studied by placing newly capped and parasitized worker bee broods into thermostatically controlled chambers. In one set of experiments the temperature was kept constant, and in the second set, the parasitized worker broods were placed at a temperature of 40,41, or 42°C for a time varying from 0 to 24 h and then returned to 32.5°C. The optimal temperature for development of the mites was between 32.5 and 33.4°C, which corresponds to the brood temperature of Apis mellifera L. Above 36.5°C, reproduction of varroa females was significantly reduced, and above 38°C, mites began to die without reproduction. Jumps of temperature were unfavorable to the development of the mites. The regulation of brood temperature by bees and occasional temperature peaks may be key factors in resistance of honey bees to varroa mites.

Guards of Cyprian honey bee colonies, Apis mellifera cypria, display a great defensive behaviour against hornets' attacks. The deformed wing virus (DWV) and the kakugo virus (KV) genomes are very similar, but unlike KV, the presence of DWV is not related to honey bees' aggressiveness. This discrepancy is further discussed.