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Abstract We investigate the emergence, mutation profile, and dissemination of SARS-CoV-2 lineage B.1.214.2, first identified in Belgium in January 2021. This variant, featuring a 3-amino acid insertion in the spike protein similar to the Omicron variant, was speculated to enhance transmissibility or immune evasion. Initially detected in international travelers, it substantially transmitted in Central Africa, Belgium, Switzerland, and France, peaking in April 2021. Our travel-aware phylogeographic analysis, incorporating travel history, estimated the origin to the Republic of the Congo, with primary European entry through France and Belgium, and multiple smaller introductions during the epidemic. We correlate its spread with human travel patterns and air passenger data. Further, upon reviewing national reports of SARS-CoV-2 outbreaks in Belgian nursing homes, we found this strain caused moderately severe outcomes (8.7% case fatality ratio). A distinct nasopharyngeal immune response was observed in elderly patients, characterized by 80% unique signatures, higher B- and T-cell activation, increased type I IFN signaling, and reduced NK, Th17, and complement system activation, compared to similar outbreaks. This unique immune response may explain the variant's epidemiological behavior and underscores the need for nasal vaccine strategies against emerging variants.

Abstract Understanding the complex interactions between external and internal factors that influence pollinator foraging behaviour is essential to understand ecosystem functioning, design agricultural practices or develop effective conservation strategies. However, it remains challenging to collect large and reliable data sets with reasonable personnel and workload. In this study, we present a wireless and cost‐effective robotic flower equipped with internet of things (IoT) technology that automatically offers nectar to visiting insects while monitoring visitation time and duration. The robotic flower is easy to manipulate and settings such as nectar refill rates can be remotely altered, making it ideal for field settings. The system transmits data completely wirelessly and autonomously, is mobile and easy to clean. The prototype settings allow for approximately 2 weeks of uninterrupted data collection for each battery charge. As a proof‐of‐concept application, a foraging preference dual choice experiment with bumblebees was performed. On average, more than 7000 flower visits per colony were registered daily with a set‐up consisting of 16 robotic flowers. The data show a gradual preference shift away from the pre‐trained low concentration, confirming the hypothesis of favouring sugar water with higher concentration. The robotic flower provides accurate and reliable data on insect behaviour, significantly reducing the price and/or labour costs. Although primarily designed for (bumble)bees, the system could be easily adapted for other flower‐visiting insects. The robotic flower is user‐friendly and can be easily adapted to address a wide range of research questions in pollination ecology, conservation biology, biocontrol and ecotoxicology, and allows for detailed studies on how nectar traits, flower colour and shape or pollutants affect foraging behaviour.

Summary: Volatile aroma compounds are important chemical cues for insects. Behavioral responses to specific odors differ strongly between insect species, and the exact causative molecules are often unknown. Beer is frequently used in insect traps because it combines hundreds of plant and microbial aromas that attract many insects. Here, we analyzed responses of the pest fruit fly Drosophila suzukii and benign Drosophila melanogaster to beers with different chemical compositions. Using extensive chemical and behavioral assays, we identified ecologically relevant chemicals that influence drosophilid behavior and that induce different odor-evoked activity patterns in the antennal lobe of the two species obtained by functional imaging. Specific mixes of compounds increased the species-specificity and sex-specificity of lures in both laboratory and greenhouse settings. Together, our study shows how examining insect responses to highly complex natural mixtures of aroma compounds provides insight into insect-specific behavioral responses and also opens avenues for improved pest control.

Abstract Background When formulating and evaluating COVID-19 vaccination strategies, an emphasis has been placed on preventing severe disease that overburdens healthcare systems and leads to mortality. However, more conventional outcomes such as quality-adjusted life years (QALYs) and inequality indicators are warranted as additional information for policymakers. Methods We adopted a mathematical transmission model to describe the infectious disease dynamics of SARS-COV-2, including disease mortality and morbidity, and to evaluate (non)pharmaceutical interventions. Therefore, we considered temporal immunity levels, together with the distinct transmissibility of variants of concern (VOCs) and their corresponding vaccine effectiveness. We included both general and age-specific characteristics related to SARS-CoV-2 vaccination. Our scenario study is informed by data from Belgium, focusing on the period from August 2021 until February 2022, when vaccination for children aged 5-11 years was initially not yet licensed and first booster doses were administered to adults. More specifically, we investigated the potential impact of an earlier vaccination programme for children and increased or reduced historical adult booster dose uptake. Results Through simulations, we demonstrate that increasing vaccine uptake in children aged 5-11 years in August–September 2021 could have led to reduced disease incidence and ICU occupancy, which was an essential indicator for implementing non-pharmaceutical interventions and maintaining healthcare system functionality. However, an enhanced booster dose regimen for adults from November 2021 onward could have resulted in more substantial cumulative QALY gains, particularly through the prevention of elevated levels of infection and disease incidence associated with the emergence of Omicron VOC. In both scenarios, the need for non-pharmaceutical interventions could have decreased, potentially boosting economic activity and mental well-being. Conclusions When calculating the impact of measures to mitigate disease spread in terms of life years lost due to COVID-19 mortality, we highlight the impact of COVID-19 on the health-related quality of life of survivors. Our study underscores that disease-related morbidity could constitute a significant part of the overall health burden. Our quantitative findings depend on the specific setup of the interventions under review, which is open to debate or should be contextualised within future situations.

Abstract The perception and appreciation of food flavor depends on many interacting chemical compounds and external factors, and therefore proves challenging to understand and predict. Here, we combine extensive chemical and sensory analyses of 250 different beers to train machine learning models that allow predicting flavor and consumer appreciation. For each beer, we measure over 200 chemical properties, perform quantitative descriptive sensory analysis with a trained tasting panel and map data from over 180,000 consumer reviews to train 10 different machine learning models. The best-performing algorithm, Gradient Boosting, yields models that significantly outperform predictions based on conventional statistics and accurately predict complex food features and consumer appreciation from chemical profiles. Model dissection allows identifying specific and unexpected compounds as drivers of beer flavor and appreciation. Adding these compounds results in variants of commercial alcoholic and non-alcoholic beers with improved consumer appreciation. Together, our study reveals how big data and machine learning uncover complex links between food chemistry, flavor and consumer perception, and lays the foundation to develop novel, tailored foods with superior flavors.

Abstract Social insects owe their widespread success to their ability to efficiently coordinate behaviour to carry out complex tasks. Several leaf-cutter ant species employ an advanced type of division of labour known as task partitioning, where the task of retrieving leaves is distributed between workers that cut and drop and those that collect the fallen leaves. It is not entirely clear how such highly coordinated behaviour can evolve, as it would seem to require the simultaneous mutations of multiple traits during the same generation. Here, we use an agent-based simulation model to show how task partitioning in leaf-cutter ants can gradually evolve by exploiting stigmergy (indirect coordination through the environment) through gravity (leaves falling from the treetop on the ground forming a cache). Our simple model allows independent variation in two core behavioural dimensions: the tendency to drop leaves and the tendency to pick up dropped leaves. Task partitioning readily evolves even under these minimal assumptions through adaptation to an arboreal environment where traveling up and down the tree is costly. Additionally, we analyse ant movement dynamics to demonstrate how the ants achieve efficient task allocation through task switching and negative feedback control.

Abstract Cooperative behaviour can evolve through conditional strategies that direct cooperation towards interaction partners who have themselves been cooperative in the past. Such strategies are common in human cooperation, but they can be vulnerable to manipulation: individuals may try to exaggerate their past cooperation to elicit reciprocal contributions or improve their reputation for future gains. Little is known about the prevalence and the ramifications of misrepresentation in human cooperation, neither in general nor about its cultural facets (self-sacrifice for the group is valued differently across cultures). Here, we present a large-scale interactive decision making experiment (N = 870), performed in China and the USA, in which individuals had repeated cooperative interactions in groups. Our results show that (1) most individuals from both cultures overstate their contributions to the group if given the opportunity, (2) misrepresentation of cooperation is detrimental to cooperation in future interactions, and (3) the possibility to build up a personal reputation amplifies the effects of misrepresentation on cooperation in China, but not in the USA. Our results suggest that misrepresentation of cooperation is likely to be an important factor in (the evolution of) human social behaviour, with, depending on culture, diverging impacts on cooperation outcomes.

Juvenile hormone (JH) affects multiple physiological traits in insects. In social insects, besides development, JH has been demonstrated to influence caste determination and the production of chemical compounds. In social wasps, JH triggers behavioral maturation, gonadotropic effects, and hydrocarbon modulation. Here, we investigated whether JH displays the same function in fertility and fertility cue production in females of the German wasp Vespula germanica, previously shown in the related species Vespula vulgaris. By experimentally treating workers with JH-analog, an anti-JH, and acetone solvent control, we tested whether JH modulates the cuticular chemical expression (CHCs), the Dufour’s gland chemical composition, and hence the compounds found over the egg’s surface. Additionally, we explored whether JH has a gonadotropic effect on workers. Workers treated with the JH-analog acquired a chemical profile that was intermediate between the queen and other treated workers. Interestingly, the same pattern was also seen in the Dufour’s glands and eggs, although more subtle. Furthermore, workers treated with the JH-analog were more fertile when compared to the controls, supporting the fact that JH acts as a gonadotropic hormone. Our results indicate a similar function of JH in societies of related wasp species V. germanica and V. vulgaris.

Microbes are constantly confronted with changes and challenges in their environment. A proper response to these environmental cues is needed for optimal cellular functioning and fitness. Interestingly, past exposure to environmental cues can accelerate or boost the response when this condition returns, even in daughter cells that have not directly encountered the initial cue. Moreover, this behavior is mostly epigenetic and often goes hand in hand with strong heterogeneity in the strength and speed of the response between isogenic cells of the same population, which might function as a bet-hedging strategy. In this review, we discuss examples of history-dependent behavior (HDB) or “memory,” with a specific focus on HDB in fluctuating environments. In most examples discussed, the lag time before the response to an environmental change is used as an experimentally measurable proxy for HDB. We highlight different mechanisms already implicated in HDB, and by using HDB in fluctuating carbon conditions as a case study, we showcase how the metabolic state of a cell can be a key determining factor for HDB. Finally, we consider possible evolutionary causes and consequences of such HDB.

Wasps (Vespidae) are important organisms to understand the evolution of social behaviour. Wasps show different levels of sociality, which includes solitary to highly eusocial organisms. In social insect species, queens and workers differ in physiology and morphology. The Neotropical swarm-founding wasps (Epiponini) show a variety of caste syndromes. In this clade, the caste-flexibility is a unique characteristic, in which workers can become queens and swarm to start a new nest. The investigation of the caste system comparing several Epiponini species show a clear-cut morphological distinction between queens and workers, with a morphological continuum between queens and workers. However, whether cuticular hydrocarbons (CHCs) are used as cues for caste recognition in swarm-founding wasps is still unknown. We studied whether CHCs may display caste-linked differences in eleven species of Epiponini wasps and if CHCs differences would follow morphological patterns. Our results suggest that queens and workers of Epiponini wasps are chemically different from each other at two levels, qualitatively and quantitatively, or merely quantitatively. This variation seems to exist regardless of their morphological traits and may be useful to help us understanding how chemical communication evolved differently in these species.

The buff-tailed bumblebee Bombus terrestris presents two distinct colony phenotypes in which some colonies already start producing males very early in the season, while others switch to producing sexuals much later in the season, and specialize mainly in the production of virgin queens. Despite having been extensively investigated in the past, the mechanisms underlying these two distinct phenotypes remain poorly understood. Here we use a combination of behavioral and genetic data to demonstrate that early switchers are in fact diploid male producing colonies, while late switchers produced normal, haploid males. Hence, at a proximate level, early switching to male production was caused by queens mating with males that by chance carried the same sex allele. These patterns were not the result of strong inbreeding within the study population, as the average inbreeding coefficient was very low and in fact slightly negative (−0.11). Moreover, an effect of inbreeding was further excluded because hybrids produced by crossing two distinct outbred populations also produced diploid males in early switching colonies. Finally, we found that diploid males only emerged after a first cohort of workers was produced, while we would have expected diploid males to be produced simultaneously with workers in a 50:50 ratio. We demonstrate that adult diploid males had a distinct cuticular hydrocarbon profile and propose that queens likely cull diploid males during early colony development to reduce the cost of diploid male production.

In the highly eusocial wasp, Vespula vulgaris, queens produce honest signals to alert their subordinate workers of their fertility status, and therefore they are reproductively suppressed and help in the colony. The honesty of the queen signals is likely maintained due to hormonal regulation, which affects fertility and fertility cue expression. Here, we tested if hormonal pleiotropy could support the hypothesis that juvenile hormone controls fertility and fertility signaling in workers. In addition, we aimed to check oocyte size as a proxy of fertility. To do that, we treated V. vulgaris workers with synthetic versions of juvenile hormone (JH) analogue and a JH inhibitor, methoprene and precocene, respectively. We dissected the treated females to check ovary activation and analyzed their chemical profile. Our results showed that juvenile hormone has an influence on the abundance of fertility linked compounds produced by workers, and it also showed to increase oocyte size in workers. Our results corroborate the hypothesis that juvenile hormone controls fertility and fertility signaling in workers, whereby workers are unable to reproduce without alerting other colony members of their fertility. This provides supports the hypothesis that hormonal pleiotropy contributes to keeping the queen fertility signals honest.

Humans are known to use social heuristics to make intuitive decisions on whether to cooperate. Here, the authors show with evolutionary simulations that social heuristics can be an adaptive solution to uncertainties about the consequences of cooperation and can greatly increase cooperation levels.

Abstract In insect societies, both queens and workers produce chemicals that reliably signal caste membership and reproductive status. The mechanisms that help to maintain the honesty of such queen and fertility signals, however, remain poorly studied. Here we test if queen signal honesty could be based on the shared endocrine control of queen fertility and the production of specific signals. In support of this “hormonal pleiotropy” hypothesis, we find that in the common wasp, application of methoprene (a juveline hormone analogue) caused workers to acquire a queen-like cuticular hydrocarbon profile, resulting in the overproduction of known queen pheromones as well as some compounds typically linked to worker fertility. By contrast, administration of precocene-I (a JH inhibitor) had a tendency to have the opposite effect. Furthermore, a clear gonadotropic effect of JH in queens was suggested by the fact that circulating levels of JH were ca. 2 orders of magnitude higher in queens than those in workers and virgin, non-egg-laying queens, even if methoprene or precocene treatment did not affect the ovary development of workers. Overall, these results suggest that queen signal honesty in this system is maintained by queen fertility and queen signal production being under shared endocrine control.

In several highly eusocial insect species with morphologically distinct castes, queen-characteristic cuticular hydrocarbons (CHCs) have been shown to act as queen signals that suppress the reproduction of nestmate workers. However, it is not known whether such queen pheromones might also play a role in regulating reproductive division of labor in primitively eusocial insects that lack morphologically defined castes. Here, we experimentally tested whether a blend of CHCs which occurred in higher concentrations in the dominant breeding female acts as a queen pheromone, and inhibits reproduction by subordinate females in the primitively eusocial paper wasp Polistes satan. In contrast to earlier findings with highly eusocial species, our results show that although specific compounds were enhanced in dominant, reproductively active females, a blend of five of these compounds did not inhibit development of the ovaries of female nestmates. Instead, the dominant female had to be physically present to prevent subordinate females from reproducing. Our results are in line with earlier evidence suggesting that fertility-linked compounds in primitively eusocial wasps act only as cues and help to regulate reproduction when used in conjunction with aggressive dominance behavior, meaning the physical presence of the dominant female. Alternatively, our results support the hypothesis that queen pheromones in highly eusocial insects were co-opted from fertility cues that were already present in primitively eusocial ancestors, even when initially, such compounds were merely produced as by-products of ovarian activation, without actually serving to signal reproductive status.

The spotted wing drosophila, Drosophila suzukii, is an invasive pest species from Southeast Asia that was recently introduced in Europe and North America. As this fruit fly lays its eggs in ripening soft-skinned fruit, it causes great damage to a variety of crops, including cherries, blueberries, blackberries, raspberries, grapes, plums and strawberries. Consequently, there is a great demand for an effective and species-specific lure, which requires the development of successful attractants. Until now, there is no lure available that is species-specific and can detect the presence of D. suzukii before infestation. As blackberry (Rubus fruticosus) is one of the preferred host crops of D. suzukii, the volatile compounds of R. fruticosus berries are here identified and quantified using multiple headspace SPME (solid phase micro extraction) GC-MS (gas chromatography–mass spectrometry). Subsequently, the attractivity of 33 of the identified compounds was tested with a two-choice laboratory bioassay. Acetaldehyde, hexyl acetate, linalool, myrtenol, L-limonene and camphene came out as significantly attractive to D. suzukii. The first four attractive compounds induced the strongest effect and therefore provided the best prospects to be implemented in a potential lure. These findings could contribute towards the development of more effective attractants for monitoring and mass trapping D. suzukii.

Sphecophaga vesparum often parasitizes nests of vespid wasps such as Vespula vulgaris and Vespula germanica. Inside the colonies, the ectoparasitic larvae feed on the immature forms of the wasps. There are two adult forms of S. vesparum. The large, winged adults emerge from either rigid yellow cocoons or the orange cocoons used for overwintering. The small, brachypterous females emerge from soft, white cocoons. The species is facultative deuterotokous, producing mostly parthenogenic females and infrequently producing males. Here, we describe the production of chemical compounds related to the different developmental forms of the parasitoid S. vesparum (larvae, pupae and adults). We also compare the chemical profiles of the parasitoid wasp adults to those of their two main host species, Vespula vulgaris and Vespula germanica. The results show differences in hydrocarbon composition of larvae, pupae and adults of S. vesparum. Our results also suggest a partial mimicry of each of the two host species, mostly relating to linear alkanes present in both parasitoids and the host vespid wasp species. This matching is likely due to the recycling of the prey’s hydrocarbons, as has been found in other species of parasitoids.

ABSTRACT When faced with environmental changes, microbes often enter a temporary growth arrest during which they reprogram the expression of specific genes to adapt to the new conditions. A prime example of such a lag phase occurs when microbes need to switch from glucose to other, less-preferred carbon sources. Despite its industrial relevance, the genetic network that determines the duration of the lag phase has not been studied in much detail. Here, we performed a genome-wide Bar-Seq screen to identify genetic determinants of the Saccharomyces cerevisiae glucose-to-galactose lag phase. The results show that genes involved in respiration, and specifically those encoding complexes III and IV of the electron transport chain, are needed for efficient growth resumption after the lag phase. Anaerobic growth experiments confirmed the importance of respiratory energy conversion in determining the lag phase duration. Moreover, overexpression of the central regulator of respiration, HAP4, leads to significantly shorter lag phases. Together, these results suggest that the glucose-induced repression of respiration, known as the Crabtree effect, is a major determinant of microbial fitness in fluctuating carbon environments. IMPORTANCE The lag phase is arguably one of the prime characteristics of microbial growth. Longer lag phases result in lower competitive fitness in variable environments, and the duration of the lag phase is also important in many industrial processes where long lag phases lead to sluggish, less efficient fermentations. Despite the immense importance of the lag phase, surprisingly little is known about the exact molecular processes that determine its duration. Our study uses the molecular toolbox of S. cerevisiae combined with detailed growth experiments to reveal how the transition from fermentative to respirative metabolism is a key bottleneck for cells to overcome the lag phase. Together, our findings not only yield insight into the key molecular processes and genes that influence lag duration but also open routes to increase the efficiency of industrial fermentations and offer an experimental framework to study other types of lag behavior.

In a recent study, Amsalem, Orlova & Grozinger (2015) performed experiments with Bombus impatiens bumblebees to test the hypothesis that saturated cuticular hydrocarbons are evolutionarily conserved signals used to regulate reproductive division of labor in many Hymenopteran social insects. They concluded that the cuticular hydrocarbon pentacosane (C25), previously identified as a queen pheromone in a congeneric bumblebee, does not affect worker reproduction in B. impatiens. Here we discuss some shortcomings of Amsalem et al.’s study that make its conclusions unreliable. In particular, several confounding effects may have affected the results of both experimental manipulations in the study. Additionally, the study’s low sample sizes (mean n per treatment = 13.6, range: 4–23) give it low power, not 96–99% power as claimed, such that its conclusions may be false negatives. Inappropriate statistical tests were also used, and our reanalysis found that C25 substantially reduced and delayed worker egg laying in B. impatiens. We review the evidence that cuticular hydrocarbons act as queen pheromones, and offer some recommendations for future queen pheromone experiments.

Yeast cells produce various volatile metabolites that are key contributors to the pleasing fruity and flowery aroma of fermented beverages. Several of these fruity metabolites, including isoamyl acetate and ethyl acetate, are produced by a dedicated enzyme, the alcohol acetyl transferase Atf1. However, despite much research, the physiological role of acetate ester formation in yeast remains unknown. Using a combination of molecular biology, neurobiology, and behavioral tests, we demonstrate that deletion of ATF1 alters the olfactory response in the antennal lobe of fruit flies that feed on yeast cells. The flies are much less attracted to the mutant yeast cells, and this in turn results in reduced dispersal of the mutant yeast cells by the flies. Together, our results uncover the molecular details of an intriguing aroma-based communication and mutualism between microbes and their insect vectors. Similar mechanisms may exist in other microbes, including microbes on flowering plants and pathogens.

Division of labor is ubiquitous in biological systems, as evidenced by various forms of complex task specialization observed in both animal societies and multicellular organisms. Although clearly adaptive, the way in which division of labor first evolved remains enigmatic, as it requires the simultaneous co-occurrence of several complex traits to achieve the required degree of coordination. Recently, evolutionary swarm robotics has emerged as an excellent test bed to study the evolution of coordinated group-level behavior. Here we use this framework for the first time to study the evolutionary origin of behavioral task specialization among groups of identical robots. The scenario we study involves an advanced form of division of labor, common in insect societies and known as "task partitioning", whereby two sets of tasks have to be carried out in sequence by different individuals. Our results show that task partitioning is favored whenever the environment has features that, when exploited, reduce switching costs and increase the net efficiency of the group, and that an optimal mix of task specialists is achieved most readily when the behavioral repertoires aimed at carrying out the different subtasks are available as pre-adapted building blocks. Nevertheless, we also show for the first time that self-organized task specialization could be evolved entirely from scratch, starting only from basic, low-level behavioral primitives, using a nature-inspired evolutionary method known as Grammatical Evolution. Remarkably, division of labor was achieved merely by selecting on overall group performance, and without providing any prior information on how the global object retrieval task was best divided into smaller subtasks. We discuss the potential of our method for engineering adaptively behaving robot swarms and interpret our results in relation to the likely path that nature took to evolve complex sociality and task specialization.

When invasive species move to new environments they typically experience population bottlenecks that limit the probability that pathogens and parasites are also moved. The invasive species may thus be released from biotic interactions that can be a major source of density-dependent mortality, referred to as enemy release. We examined for evidence of enemy release in populations of the common wasp (Vespula vulgaris), which attains high densities and represents a major threat to biodiversity in its invaded range. Mass spectrometry proteomic methods were used to compare the microbial communities in wasp populations in the native (Belgium and England) and invaded range (Argentina and New Zealand). We found no evidence of enemy release, as the number of microbial taxa was similar in both the introduced and native range. However, some evidence of distinctiveness in the microbial communities was observed between countries. The pathogens observed were similar to a variety of taxa observed in honey bees. These taxa included Nosema, Paenibacillus, and Yersina spp. Genomic methods confirmed a diversity of Nosema spp., Actinobacteria, and the Deformed wing and Kashmir bee viruses. We also analysed published records of bacteria, viruses, nematodes and fungi from both V. vulgaris and the related invader V. germanica. Thirty-three different microorganism taxa have been associated with wasps including Kashmir bee virus and entomophagous fungi such as Aspergillus flavus. There was no evidence that the presence or absence of these microorganisms was dependent on region of wasp samples (i.e. their native or invaded range). Given the similarity of the wasp pathogen fauna to that from honey bees, the lack of enemy release in wasp populations is probably related to spill-over or spill-back from bees and other social insects. Social insects appear to form a reservoir of generalist parasites and pathogens, which makes the management of wasp and bee disease difficult.

The amount of the trail pheromone substance, 3-ethyl-2,5-dimethylpyrazine, of the leaf-cutting ant Atta sexdens sexdens (Hymenoptera: Formicidae) increases with increasing size of the ant from 0 to 35 ng per individual. The compounds 2,5-dimethylpyrazine and trimethylpyrazine are also consistently present. Trails of venom gland extracts are followed with unequal fidelity by different castes. Minor workers follow best, mediums least well. Workers walk sinuously on narrow trails and less sinuously on wider trails up to 9 mm. Trails wider than 9 mm are not followed. The optimum concentration of pheromone on synthetic trails lies between 15 and 150 pg cm-1. Given a choice of concentrations at a trail branch, workers always choose the more concentrated. Workers showed slight preference for a trail made with venom gland secretion from their own, over that from an alien colony, but there is no preference for a trail that contains Dufour gland secretion additionally.

It is easy to see why stingless bees (Meliponini) were chosen for this special issue, entitled Stingless bees: Integrating basic biology and conservation. Interest in these bees is increasing each day, with their role as ecosystem services providers in the pollination of wild and cultivated plants, and their fascinating biology and social behavior. Yet, despite having been kept by the Native Americans for hundreds of years for their honey, they remain much understudied compared to their better known relatives, the bumblebees and honeybees. This is in part due to their massive biodiversity, with known species now numbering over 500 – all with a tropical distribution. The diversity and rich social behavior of the stingless bees has often been compared to the knowledge of the other group of eusocial bees, including the bumblebees (Bombini) and honeybees (Apini). Yet, the development of new scientific techniques and research methods, and progress in deciphering their phylogeny, now allows for an updated interpretation of the evolution, biology and conservation of Meliponini bees.

Since the last decade, unusually high honey bee colony losses have been reported mainly in North-America and Europe. Here, we report on a comprehensive bee pathogen screening in Belgium covering 363 bee colonies that were screened for 18 known disease-causing pathogens and correlate their incidence in summer with subsequent winter mortality. Our analyses demonstrate that, in addition to Varroa destructor, the presence of the trypanosomatid parasite Crithidia mellificae and the microsporidian parasite Nosema ceranae in summer are also predictive markers of winter mortality, with a negative synergy being observed between the two in terms of their effects on colony mortality. Furthermore, we document the first occurrence of a parasitizing phorid fly in Europe, identify a new fourth strain of Lake Sinai Virus (LSV), and confirm the presence of other little reported pathogens such as Apicystis bombi, Aphid Lethal Paralysis Virus (ALPV), Spiroplasma apis, Spiroplasma melliferum and Varroa destructor Macula-like Virus (VdMLV). Finally, we provide evidence that ALPV and VdMLV replicate in honey bees and show that viruses of the LSV complex and Black Queen Cell Virus tend to non-randomly co-occur together. We also noticed a significant correlation between the number of pathogen species and colony losses. Overall, our results contribute significantly to our understanding of honey bee diseases and the likely causes of their current decline in Europe.

The long-term decline of managed honeybee hives in the world has drawn significant attention to the scientific community and bee-keeping industry. A high pathogen load is believed to play a crucial role in this phenomenon, with the bee viruses being key players. Most of the currently characterized honeybee viruses (around twenty) are positive stranded RNA viruses. Techniques based on RNA signatures are widely used to determine the viral load in honeybee colonies. High throughput screening for viral loads necessitates the development of a multiplex polymerase chain reaction approach in which different viruses can be targeted simultaneously. A new multiparameter assay, called "BeeDoctor", was developed based on multiplex-ligation probe dependent amplification (MLPA) technology. This assay detects 10 honeybee viruses in one reaction. "BeeDoctor" is also able to screen selectively for either the positive strand of the targeted RNA bee viruses or the negative strand, which is indicative for active viral replication. Due to its sensitivity and specificity, the MLPA assay is a useful tool for rapid diagnosis, pathogen characterization, and epidemiology of viruses in honeybee populations. "BeeDoctor" was used for screening 363 samples from apiaries located throughout Flanders; the northern half of Belgium. Using the "BeeDoctor", virus infections were detected in almost eighty percent of the colonies, with deformed wing virus by far the most frequently detected virus and multiple virus infections were found in 26 percent of the colonies.

The eusocial societies of honeybees, where the queen is the only fertile female among tens of thousands sterile worker bees, have intrigued scientists for centuries. The proximate factors, which cause the inhibition of worker bee ovaries, remain largely unknown; as are the factors which cause the activation of worker ovaries upon the loss of queen and brood in the colony. In an attempt to reveal key players in the regulatory network, we made a proteomic comparison of hemolymph profiles of workers with completely activated ovaries vs. rudimentary ovaries. An unexpected finding of this study is the correlation between age matched worker sterility and the enrichment of Picorna-like virus proteins. Fertile workers, on the other hand, show the upregulation of potential components of the immune system. It remains to be investigated whether viral infections contribute to worker sterility directly or are the result of a weaker immune system of sterile workers.