Buff-tailed Bumblebee Bombus Terrestris Research Articles

Effects of diesel exhaust particles on the health and survival of the buff-tailed bumblebee Bombus terrestris after acute and chronic oral exposure

The diesel-powered transportation sector is a major producer of environmental pollution in the form of micro- and nanoscale diesel exhaust particles (DEP). Pollinators, such as wild bees, may inhale DEP or ingest it orally through plant nectar. However, if these insects are adversely affected by DEP is largely unknown. To investigate potential health threats of DEP to pollinators, we exposed individuals of Bombus terrestris to different concentrations of DEP. We analysed the polycyclic aromatic hydrocarbons (PAH) content of DEP since these are known to elicit adverse effects on invertebrates. We investigated the dose-dependent effects of those well-characterized DEP on survival and fat body content, as a proxy for the insects’ health condition, in acute and chronic oral exposure experiments. Acute oral exposure to DEP showed no dose-dependent effects on survival or fat body content of B. terrestris. However, we could show dose-dependent effects after chronic oral exposure with high doses of DEP where significantly increased mortality was observed. Further, there was no dose-dependent effect of DEP on the fat body content after exposure. Our results give insights into how the accumulation of high concentrations of DEP e.g., near heavily trafficked sites, can influence insect pollinators’ health and survival.

Efficient visual learning by bumble bees in virtual-reality conditions: Size does not matter.

Recent developments allowed establishing virtual-reality (VR) setups to study multiple aspects of visual learning in honey bees under controlled experimental conditions. Here, we adopted a VR environment to investigate the visual learning in the buff-tailed bumble bee Bombus terrestris. Based on responses to appetitive and aversive reinforcements used for conditioning, we show that bumble bees had the proper appetitive motivation to engage in the VR experiments and that they learned efficiently elemental color discriminations. In doing so, they reduced the latency to make a choice, increased the proportion of direct paths toward the virtual stimuli and walked faster toward them. Performance in a short-term retention test showed that bumble bees chose and fixated longer on the correct stimulus in the absence of reinforcement. Body size and weight, although variable across individuals, did not affect cognitive performances and had a mild impact on motor performances. Overall, we show that bumble bees are suitable experimental subjects for experiments on visual learning under VR conditions, which opens important perspectives for invasive studies on the neural and molecular bases of such learning given the robustness of these insects and the accessibility of their brain.

Taking advantage of transparency: A proof-of-principle for the analysis of the uptake of labeled microplastic particles by organisms of different functional feeding guilds using an adapted CUBIC protocol

The analysis of the ingestion of microplastics (MP) by biota is frequently performed through invasive procedures such as chemical digestion protocols or by histological analysis of thin sections. Different, promising approaches for the observation of ingested MP particles pose so called tissue clearing methods. They are currently applied to organs, tissue samples, or whole organisms, rendering the sample transparent and enable to look inside an otherwise opaque environment. To date, there is a lack of methods to detect labeled MP inside an opaque organism's digestive tract without interfering with the sample's integrity. Therefore, our goal was to adapt the CUBIC tissue clearing protocol (Clear, Unobstructed Brain/Body Imaging Cocktails and Computational Analysis) for aquatic and terrestrial organisms of various functional feeding groups for the analysis of the uptake of fluorescent labeled microplastic (MP) particles. We included the buff-tailed bumblebee Bombus terrestris, the compost worm Eisenia fetida, the woodlouse Porcellio scaber, the freshwater shrimp Gammarus roeselii, and the quagga mussel Dreissena bugensis in the analysis. The adapted CUBIC method has led to transparency in all normally opaque organisms. It further offers a simple way of locating fluorescent labeled MP inside the digestive system of the different organisms while leaving them intact.

Dominance of Fructose-Associated Fructobacillus in the Gut Microbiome of Bumblebees (Bombus terrestris) Inhabiting Natural Forest Meadows.

Simple SummaryA vast array of microorganisms colonize invertebrates and vertebrates. Most of these microbes reside in the digestive tract, where they constitute the intestinal (gut) microbiome. Some microbes are commensal, coexisting with their host without causing harm, while others can be mutualistic or pathogenic. Mutualistic microorganisms perform many health-related functions such as promoting digestion and acquisition of nutrients; hormone regulation; maintenance and control of the immune system; regulation of homeostasis and stress physiology of the body; insecticide resistance; production of certain vitamins; and providing protection against pathogenic microorganisms, parasites, and diseases. Bee-specific bacterial genera such as Lactobacillus, Snodgrassella, and Gilliamella dominate the gut communities of many bumblebees. This study confirmed Lactobacillus, Snodgrassella, and Gilliamella as dominant gut bacteria of the buff-tailed bumblebee Bombus terrestris in the agricultural landscape. However, we show that the guts of B. terrestris from natural forest habitats can be dominated by fructose-associated Fructobacillus spp. Our findings may have important implications for understanding the ecological role of bumblebees and the reasons for the decline of key pollinators.Bumblebees are key pollinators in agricultural landscapes. However, little is known about how gut microbial communities respond to anthropogenic changes. We used commercially produced colonies of buff-tailed bumblebees (Bombus terrestris) placed in three habitats. Whole guts (midgut, hindgut, and rectum) of B. terrestris specimens were dissected from the body and analyzed using 16S phylogenetic community analysis. We observed significantly different bacterial community composition between the agricultural landscapes (apple orchards and oilseed rape (Brassica napus) fields) and forest meadows, whereas differences in gut communities between the orchards and oilseed rape fields were nonsignificant. Bee-specific bacterial genera such as Lactobacillus, Snodgrassella, and Gilliamella dominated gut communities of B. terrestris specimens. In contrast, the guts of B. terrestris from forest meadows were dominated by fructose-associated Fructobacillus spp. Bacterial communities of workers were the most diverse. At the same time, those of males and young queens were less diverse, possibly reflecting greater exposure to the colony’s inner environment compared to the environment outside the colony, as well as bumblebee age. Our results suggest that habitat quality, exposure to environmental microbes, nectar quality and accessibility, and land use significantly affect gut bacterial composition in B. terrestris.

A micro-CT-based standard brain atlas of the bumblebee

In recent years, bumblebees have become a prominent insect model organism for a variety of biological disciplines, particularly to investigate learning behaviors as well as visual performance. Understanding these behaviors and their underlying neurobiological principles requires a clear understanding of brain anatomy. Furthermore, to be able to compare neuronal branching patterns across individuals, a common framework is required, which has led to the development of 3D standard brain atlases in most of the neurobiological insect model species. Yet, no bumblebee 3D standard brain atlas has been generated. Here we present a brain atlas for the buff-tailed bumblebee Bombus terrestris using micro-computed tomography (micro-CT) scans as a source for the raw data sets, rather than traditional confocal microscopy, to produce the first ever micro-CT-based insect brain atlas. We illustrate the advantages of the micro-CT technique, namely, identical native resolution in the three cardinal planes and 3D structure being better preserved. Our Bombus terrestris brain atlas consists of 30 neuropils reconstructed from ten individual worker bees, with micro-CT allowing us to segment neuropils completely intact, including the lamina, which is a tissue structure often damaged when dissecting for immunolabeling. Our brain atlas can serve as a platform to facilitate future neuroscience studies in bumblebees and illustrates the advantages of micro-CT for specific applications in insect neuroanatomy.

Gene expression and epigenetics reveal species-specific mechanisms acting upon common molecular pathways in the evolution of task division in bees

A striking feature of advanced insect societies is the existence of workers that forgo reproduction. Two broad types of workers exist in eusocial bees: nurses who care for their young siblings and the queen, and foragers who guard the nest and forage for food. Comparisons between these two worker subcastes have been performed in honeybees, but data from other bees are scarce. To understand whether similar molecular mechanisms are involved in nurse-forager differences across distinct species, we compared gene expression and DNA methylation profiles between nurses and foragers of the buff-tailed bumblebee Bombus terrestris and the stingless bee Tetragonisca angustula. These datasets were then compared to previous findings from honeybees. Our analyses revealed that although the expression pattern of genes is often species-specific, many of the biological processes and molecular pathways involved are common. Moreover, the correlation between gene expression and DNA methylation was dependent on the nucleotide context, and non-CG methylation appeared to be a relevant factor in the behavioral changes of the workers. In summary, task specialization in worker bees is characterized by a plastic and mosaic molecular pattern, with species-specific mechanisms acting upon broad common pathways across species.

Evidence for and against deformed wing virus spillover from honey bees to bumble bees: a reverse genetic analysis

Deformed wing virus (DWV) is a persistent pathogen of European honey bees and the major contributor to overwintering colony losses. The prevalence of DWV in honey bees has led to significant concerns about spillover of the virus to other pollinating species. Bumble bees are both a major group of wild and commercially-reared pollinators. Several studies have reported pathogen spillover of DWV from honey bees to bumble bees, but evidence of a sustained viral infection characterized by virus replication and accumulation has yet to be demonstrated. Here we investigate the infectivity and transmission of DWV in bumble bees using the buff-tailed bumble bee Bombus terrestris as a model. We apply a reverse genetics approach combined with controlled laboratory conditions to detect and monitor DWV infection. A novel reverse genetics system for three representative DWV variants, including the two master variants of DWV—type A and B—was used. Our results directly confirm DWV replication in bumble bees but also demonstrate striking resistance to infection by certain transmission routes. Bumble bees may support DWV replication but it is not clear how infection could occur under natural environmental conditions.

Effects of chemical and biological plant protection products on R&D colonies of the buff-tailed bumblebee Bombus terrestris: (2.5 Part 1)

Bumblebees (Bombus terrestris) are exposed daily to Plant Protection Products through their foraging and feeding activities. Through all possible means of contact with pesticides, consumption through sugarwater is the most severe. In the present study, lethal and sublethal effects of the consumption of sugarwater solutions with the pesticides Sivanto WG (flupyradifuron), Exalt SC (spinetoram) and Oikos EC (azadirachtin) were studied using a sequential dilution testing scheme of 1/1 and 1/10 of the maximum field recommended concentration (MFRC). For the weekly assessment, parameters such as the survival of the mother queen, of workers and drones, the formation of gynes, and the weight and volume of the colonies were recorded. Moreover, by the end of the colony’s life, the total number of formed workers/drones, the number of newborn gynes and queen brood were also recorded. The IOBC side-effect classes for laboratory trials were applied in order for the results to be categorized and conclusions made. Both tested concentrations of Sivanto WG (flupyradifuron) were slightly harmful for queen, worker and drone populations, Exalt SC (spinetoram) was harmful at 1/1 dilution but only slightly harmful at the 1/10 dilution, and both concentrations of Oikos EC (azadirachtin) were slightly harmful for workers and drones but toxic for queens at both dilutions.

Effects of Bacillus thuringiensis subsp. aizawai GC91 (Agree WG) on R&D colonies of the Buff-Tailed Bumblebee Bombus terrestris (2.5 Part 2)

Bacillus thuringiensis subsp. aizawai, a widely used biological plant protection product, was tested on buff-tailed bumblebee Bombus terrestris, using an updated laboratory method on full standardized R&D colonies. The maximum field recommended concentration (MFRC) was applied through topical, oral pollen and oral sugar water treatment. Parameters such as survival of the mother queen and workers, formation of gynes, weight and volume of the colonies were recorded during the study, while the total number of formed workers/drones, the number of newborn gynes and queen brood were taken also at the end of the colonies’ life. For the evaluation of the results the data were calculated and categorized according to the IOBC side-effect classes, used for laboratory trials. According to the results, no toxic effect was recorded for all parameters taken from the bumblebee colonies when they were exposed to B. thuringiensis aizawai GC91.

Mating precedes selective immune priming which is maintained throughout bumblebee queen diapause

BackgroundUnderstanding the mechanisms by which organisms adapt to unfavourable conditions is a fundamental question in ecology and evolutionary biology. One such mechanism is diapause, a period of dormancy typically found in nematodes, fish, crustaceans and insects. This state is a key life-history event characterised by arrested development, suppressed metabolism and increased stress tolerance and allows an organism to avoid prolonged periods of harsh and inhospitable environmental conditions. For some species, diapause is preceded by mating which can have a profound effect on female behaviour, physiology and key biological processes, including immunity. However, our understanding of how mating impacts long-term immunity and whether these effects persist throughout diapause is currently limited. To address this, we explored molecular changes in the haemolymph of the ecologically important pollinator, the buff-tailed bumblebee Bombus terrestris. B. terrestris queens mate prior to entering diapause, a non-feeding period of arrested development that can last 6–9 months. Using mass-spectrometry-based proteomics, we quantified changes in the pre-diapause queen haemolymph after mating, as well as the subsequent protein expression of mated queens during and post-diapause.ResultsOur analysis identified distinct proteome profiles associated with diapause preparation, maintenance and termination. More specifically, mating pre-diapause was followed by an increase in the abundance of antimicrobial peptides, key effectors of the immune system. Furthermore, we identified the elevated abundance of these proteins to be maintained throughout diapause. This finding was in contrast to the general reduction observed in immune proteins during diapause suggestive of selective immune priming and expression during diapause. Diapause also affected the expression of proteins involved in cuticular maintenance, olfaction, as well as proteins of unknown function, which may have roles in diapause regulation.ConclusionsOur results provide clear molecular evidence for the consequences and benefits of mating at the immune level as it precedes the selective increased abundance of antimicrobial peptides that are sustained throughout diapause. In addition, our results provide novel insights into the molecular mechanisms by which bumblebees prepare for, survive, and recover from diapause, insights that may have implications for our general understanding of these processes in other insect groups.

The effects of the neonicotinoid imidacloprid on gene expression and DNA methylation in the buff-tailed bumblebee Bombus terrestris.

Neonicotinoids are effective insecticides used on many important arable and horticultural crops. They are nicotinic acetylcholine receptor agonists which disrupt the function of insect neurons and cause paralysis and death. In addition to direct mortality, there are numerous sublethal effects of low doses of neonicotinoids on bees. We hypothesize that some of these large array of effects could be a consequence of epigenetic changes in bees induced by neonicotinoids. We compared whole methylome (BS-seq) and RNA-seq libraries of the brains of buff-tailed bumblebee Bombus terrestris workers exposed to field-realistic doses of the neonicotinoid imidacloprid to libraries from control workers. We found numerous genes which show differential expression between neonicotinoid-treated bees and control bees, but no differentially methylated cytosines in any context. We found CpG methylation to be focused mainly in exons and associated with highly expressed genes. We discuss the implications of our results for future legislation.

Identification and functional characterisation of a novel N-cyanoamidine neonicotinoid metabolising cytochrome P450, CYP9Q6, from the buff-tailed bumblebee Bombus terrestris

Recent work has shown that two bumblebee (Bombus terrestris) cytochrome P450s of the CYP9Q subfamily, CYP9Q4 and CYP9Q5, are important biochemical determinants of sensitivity to neonicotinoid insecticides. Here, we report the characterisation of a third P450 gene CYP9Q6, previously mis-annotated in the genome of B. terrestris, encoding an enzyme that metabolises the N-cyanoamidine neonicotinoids thiacloprid and acetamiprid with high efficiency. The genomic location and complete ORF of CYP9Q6 was corroborated by PCR and its metabolic activity characterised in vitro by expression in an insect cell line. CYP9Q6 metabolises both thiacloprid and acetamiprid more rapidly than the previously reported CYP9Q4 and CYP9Q5. We further demonstrate a direct, in vivo correlation between the expression of the CYP9Q6 enzyme in transgenic Drosophila melanogaster and an increased tolerance to thiacloprid and acetamiprid. We conclude that CYP9Q6 is an efficient metaboliser of N-cyanoamidine neonicotinoids and likely plays a key role in the high tolerance of B. terrestris to these insecticides.

Alternative splicing associated with phenotypic plasticity in the bumble bee Bombus terrestris.

Phenotypic plasticity is when one genome can produce more than one phenotype. The caste system found in many social insects is an important example of plasticity. Several studies have examined gene expression in social insect developmental and caste differences. Changes in gene expression, however, are not the only source of phenotypic plasticity. Here, we investigate the role of alternative splicing in the buff-tailed bumble bee Bombus terrestris. We found that 5,458 genes in B.terrestris (40%) express more than one isoform. Larvae have the lowest level of splicing events, followed by adults and then pupae. We found that when an isoform is expressed in a given caste in the larval stage, it tends to be expressed in all castes at the larval stage. The same is true at the pupal stage. However, we see more complicated interactions between the adult castes with reproductive females showing different isoform expression compared to nonreproductive females and male adults showing the most distinct patterns. We found 455 isoform switching genes, that is genes, where one developmental stage, sex or caste uses a specific isoform and another type uses a different isoform. Among genes displaying isoform switching are some involved in the ecdysteriod pathway, an important system in insect behaviour.

Molecular cloning and characterization of the SIFamide precursor and receptor in a hymenopteran insect, Bombus terrestris

SIFamides (SIFa) are a family of neuropeptides that are highly conserved among arthropods. In insects, this peptide is mainly expressed in four medial interneurons in the pars intercerebralis and affects sexual behavior, sleep regulation and pupal mortality. Furthermore, an influence on the hatching rate has been observed. The first SIFa receptor (SIFR) was pharmacologically characterized in Drosophila melanogaster and is homologous to the vertebrate gonadotropin-inhibitory hormone (GnIH) receptor (NPFFR). In this study, we pharmacologically characterized the SIFR of the buff-tailed bumblebee Bombus terrestris. We demonstrated an intracellular increase in calcium ions and cyclic AMP (cAMP) upon ligand binding with an EC50 value in the picomolar and nanomolar range, respectively. In addition, we studied the agonistic properties of a range of related and modified peptides. By means of quantitative real time PCR (qPCR), we examined the relative transcript levels of Bomte-SIFa and Bomte-SIFR in a variety of tissues.

Interspecific sensitivity of bees towards dimethoate and implications for environmental risk assessment.

Wild and domesticated bee species are exposed to a variety of pesticides which may drive pollinator decline. Due to wild bee sensitivity data shortage, it is unclear if the honey bee Apis mellifera is a suitable surrogate species in the current EU risk assessment scheme. Furthermore, the underlying causes for sensitivity differences in bees are not established. We assessed the acute toxicity (median lethal dose, LD50) of dimethoate towards multiple bee species, generated a species sensitivity distribution and derived a hazardous dose (HD5). Furthermore, we performed a regression analysis with body weight and dimethoate toxicity. HD5 lower 95% confidence limit was equal to honey bee mean LD50 when applying a safety factor of 10. Body weight proved to be a predictor of interspecific bee sensitivity but did not explain the pattern completely. Using acute toxicity values from honey bees and a safety factor of 10 seems to cover the interspecific sensitivity range of bees in the case of dimethoate. Acute endpoints of proposed additional test species, the buff-tailed bumblebee Bombus terrestris and the red mason bee Osmia bicornis, do not improve the risk assessment for the entire group. However, this might not apply to other insecticides such as neonicotinoids.

How landscape, pollen intake and pollen quality affect colony growth in Bombus terrestris

Abundance and diversity of bumblebees have been declining over the past decades. To successfully conserve bumblebee populations, we need to understand how landscape characteristics affect the quantity and quality of floral resources collected by colonies and subsequently colony performance. We therefore investigated how amount and composition of pollen collected by buff-tailed bumblebee Bombus terrestris colonies was affected by the surrounding landscape (i.e. the proportion of forest, urban, semi-natural habitats) and how they were related to colony growth. Thirty B. terrestris colonies were placed at grassland sites differing in surrounding landscape. Colonies were established in spring when availability of flowering plants was highest, and their weight gain was monitored for 1 month. We additionally recorded the quantity and compared plant taxonomic composition and nutritional quality (i.e. amino acid composition) of pollen stored. Bumblebee colonies varied little in the pollen spectra collected despite differences in surrounding landscape composition. They collected on average 80 % of pollen from woody plants, with 34 % belonging to the genus Acer. Early colony growth positively correlated with total amount of woody pollen and protein collected and decreased with increasing proportions of semi-natural habitats and total amino acid concentrations. Our results suggest that woody plant species represent highly important pollen sources for the generalist forager B. terrestris early in the season. We further show that colony growth of B. terrestris is predominantly affected by the quantity, not quality, of forage, indicating that several abundant plant species flowering throughout the bumblebees’ foraging season may cover the colonies’ nutritional needs.

Reproductive workers show queenlike gene expression in an intermediately eusocial insect, the buff‐tailed bumble bee Bombus terrestris

Bumble bees represent a taxon with an intermediate level of eusociality within Hymenoptera. The clear division of reproduction between a single founding queen and the largely sterile workers is characteristic for highly eusocial species, whereas the morphological similarity between the bumble bee queen and the workers is typical for more primitively eusocial hymenopterans. Also, unlike other highly eusocial hymenopterans, division of labour among worker subcastes is plastic and not predetermined by morphology or age. We conducted a differential expression analysis based on RNA-seq data from 11 combinations of developmental stage and caste to investigate how a single genome can produce the distinct castes of queens, workers and males in the buff-tailed bumble bee Bombus terrestris. Based on expression patterns, we found males to be the most distinct of all adult castes (2411 transcripts differentially expressed compared to nonreproductive workers). However, only relatively few transcripts were differentially expressed between males and workers during development (larvae: 71 and pupae: 162). This indicates the need for more distinct expression patterns to control behaviour and physiology in adults compared to those required to create different morphologies. Among female castes, reproductive workers and their nonreproductive sisters displayed differential expression in over ten times more transcripts compared to the differential expression found between reproductive workers and their mother queen. This suggests a strong shift towards a more queenlike behaviour and physiology when a worker becomes fertile. This contrasts with eusocial species where reproductive workers are more similar to nonreproductive workers than the queen.

Nutrient balancing of the adult worker bumblebee (Bombus terrestris) depends on the dietary source of essential amino acids.

ABSTRACTAnimals carefully regulate the amount of protein that they consume. The quantity of individual essential amino acids (EAAs) obtained from dietary protein depends on the protein source, but how the proportion of EAAs in the diet affects nutrient balancing has rarely been studied. Recent research using the Geometric Framework for Nutrition has revealed that forager honeybees who receive much of their dietary EAAs from floral nectar and not from solid protein have relatively low requirements for dietary EAAs. Here, we examined the nutritional requirements for protein and carbohydrates of foragers of the buff-tailed bumblebee Bombus terrestris. By using protein (sodium caseinate) or an equimolar mixture of the 10 EAAs, we found that the intake target (nutritional optimum) of adult workers depended on the source and proportion of dietary EAAs. When bees consumed caseinate-containing diets in a range of ratios between 1:250 and 1:25 (protein to carbohydrate), they achieved an intake target (IT) of 1:149 (w/w). In contrast to those fed protein, bees fed the EAA diets had an IT more biased towards carbohydrates (1:560 w/w) but also had a greater risk of death than those fed caseinate. We also tested how the dietary source of EAAs affected free AAs in bee haemolymph. Bees fed diets near their IT had similar haemolymph AA profiles, whereas bees fed diets high in caseinate had elevated levels of leucine, threonine, valine and alanine in the haemolymph. We found that like honeybees, bumblebee workers prioritize carbohydrate intake and have a relatively low requirement for protein. The dietary source of EAAs influenced both the ratio of protein/EAA to carbohydrate and the overall amount of carbohydrate eaten. Our data support the idea that EAAs and carbohydrates in haemolymph are important determinants of nutritional state in insects.

Electrostatic sensitivity of hairs in the buff-tailed bumblebee Bombus terrestris

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ZoologyBumblebees: Behaviour, Ecology, and Conservation. Second Edition. By DaveGoulson. Oxford and New York: Oxford University Press. $120.00 (hardcover); $55.00 (paper). xi 317 p. 16 pl.; ill.; index. ISBN: 9780199553068 (hc); 9780199553075 (pb). 2010.

Previous articleNext article No AccessNew Biological BooksZoology Bumblebees: Behaviour, Ecology, and Conservation. Second Edition. By DaveGoulson. Oxford and New York: Oxford University Press. $120.00 (hardcover); $55.00 (paper). xi 317 p. 16 pl.; ill.; index. ISBN: 9780199553068 (hc); 9780199553075 (pb). 2010.JamesC.NiehJamesC.NiehSection of Ecology, Behavior & Evolution, Division of Biological Sciences, University of California, San Diego, La Jolla, California Search for more articles by this author Section of Ecology, Behavior & Evolution, Division of Biological Sciences, University of California, San Diego, La Jolla, CaliforniaPDFPDF PLUSFull Text Add to favoritesDownload CitationTrack CitationsPermissionsReprints Share onFacebookTwitterLinkedInRedditEmail SectionsMoreDetailsFiguresReferencesCited by The Quarterly Review of Biology Volume 85, Number 3September 2010 Published in association with Stony Brook University Article DOIhttps://doi.org/10.1086/655083 Views: 5Total views on this site Citations: 1Citations are reported from Crossref Copyright 2010 by The University of Chicago Press. All rights reserved.PDF download Crossref reports the following articles citing this article:Viviana Di Pietro, Helena Mendes Ferreira, Annette Van Oystaeyen, Felix Wäckers, Tom Wenseleers, Ricardo Caliari Oliveira Distinct Colony Types Caused by Diploid Male Production in the Buff-Tailed Bumblebee Bombus terrestris, Frontiers in Ecology and Evolution 10 (Apr 2022).https://doi.org/10.3389/fevo.2022.844251