Gene expression in bumble bee larvae differs qualitatively between high and low concentration imidacloprid exposure levels.

Neonicotinoid pesticides negatively impact bumble bee health, even at sublethal concentrations. Responses to the neonicotinoid imidacloprid have been studied largely at individual adult and colony levels, focusing mostly on behavioral and physiological effects. Data from developing larvae, whose health is critical for colony success, are deficient, particularly at the molecular level where transcriptomes can reveal disruption of fundamental biological pathways. We investigated gene expression of Bombus impatiens larvae exposed through food provisions to two field-realistic imidacloprid concentrations (0.7 and 7.0 ppb). We hypothesized both concentrations would alter gene expression, but the higher concentration would have greater qualitative and quantitative effects. We found 678 genes differentially expressed under both imidacloprid exposures relative to controls, including mitochondrial activity, development, and DNA replication genes. However, more genes were differentially expressed with higher imidacloprid exposure; uniquely differentially expressed genes included starvation response and cuticle genes. The former may partially result from reduced pollen use, monitored to verify food provision use and provide additional context to results. A smaller differentially expressed set only in lower concentration larvae, included neural development and cell growth genes. Our findings show varying molecular consequences under different field-realistic neonicotinoid concentrations, and that even low concentrations may affect fundamental biological processes.

Global Trends in Bumble Bee Health.

Bumble bees (Bombus) are unusually important pollinators, with approximately 260 wild species native to all biogeographic regions except sub-Saharan Africa, Australia, and New Zealand. As they are vitally important in natural ecosystems and to agricultural food production globally, the increase in reports of declining distribution and abundance over the past decade has led to an explosion of interest in bumble bee population decline. We summarize data on the threat status of wild bumble bee species across biogeographic regions, underscoring regions lacking assessment data. Focusing on data-rich studies, we also synthesize recent research on potential causes of population declines. There is evidence that habitat loss, changing climate, pathogen transmission, invasion of nonnative species, and pesticides, operating individually and in combination, negatively impact bumble bee health, and that effects may depend on species and locality. We distinguish between correlational and causal results, underscoring the importance of expanding experimental research beyond the study of two commercially available species to identify causal factors affecting the diversity of wild species.

Test of the invasive pathogen hypothesis of bumble bee decline in North America.

Emergent fungal diseases are critical factors in global biodiversity declines. The fungal pathogenNosema bombiwas recently found to be widespread in declining species of North American bumble bees (Bombus), with circumstantial evidence suggesting an exotic introduction from Europe. This interpretation has been hampered by a lack of knowledge of global genetic variation, geographic origin, and changing prevalence patterns ofN. bombiin declining North American populations. Thus, the temporal and spatial emergence ofN. bombiand its potential role in bumble bee decline remain speculative. We analyzeNosemaprevalence and genetic variation in the United States and Europe from 1980, before an alleged introduction in the early 1990s, to 2011, extractingNosemaDNA fromBombusnatural history collection specimens from across this time period.Nosema bombiprevalence increased significantly from low detectable frequency in the 1980s to significantly higher frequency in the mid- to late-1990s, corresponding to a period of reported massive infectious outbreak ofN. bombiin commercial bumble bee rearing stocks in North America. Despite the increased frequency, we find no conclusive evidence of an exoticN. bombiorigin based on genetic analysis of globalNosemapopulations; the widespreadNosemastrain found currently in declining United States bumble bees was present in the United States before commercial colony trade. Notably, the USN. bombiis not detectably different from that found predominantly throughout Western Europe, with both regions characterized by low genetic diversity compared with high levels of diversity found in Asia, where commercial bee breeding activities are low or nonexistent.

Phylogenomics Controlling for Base Compositional Bias Reveals a Single Origin of Eusociality in Corbiculate Bees.

As increasingly large molecular data sets are collected for phylogenomics, the conflicting phylogenetic signal among gene trees poses challenges to resolve some difficult nodes of the Tree of Life. Among these nodes, the phylogenetic position of the honey bees (Apini) within the corbiculate bee group remains controversial, despite its considerable importance for understanding the emergence and maintenance of eusociality. Here, we show that this controversy stems in part from pervasive phylogenetic conflicts among GC-rich gene trees. GC-rich genes typically have a high nucleotidic heterogeneity among species, which can induce topological conflicts among gene trees. When retaining only the most GC-homogeneous genes or using a nonhomogeneous model of sequence evolution, our analyses reveal a monophyletic group of the three lineages with a eusocial lifestyle (honey bees, bumble bees, and stingless bees). These phylogenetic relationships strongly suggest a single origin of eusociality in the corbiculate bees, with no reversal to solitary living in this group. To accurately reconstruct other important evolutionary steps across the Tree of Life, we suggest removing GC-rich and GC-heterogeneous genes from large phylogenomic data sets. Interpreted as a consequence of genome-wide variations in recombination rates, this GC effect can affect all taxa featuring GC-biased gene conversion, which is common in eukaryotes.

Deep sequencing and ecological characterization of gut microbial communities of diverse bumble bee species.

Gut bacterial communities of bumble bees are correlated with defense against pathogens. Further understanding this host-microbe association is vitally important as bumble bees are currently experiencing global population declines, potentially due in part to emergent diseases. In this study, we used pyrosequencing and community fingerprinting (ARISA) to characterize the gut microbial communities of nine bumble species from across the Bombus phylogeny. Overall, we delimited 74 bacterial taxa (operational taxonomic units or OTUs) belonging to Betaproteobacteria, Gammaproteobacteria, Bacilli, Actinobacteria, Flavobacteria and Alphaproteobacteria. Each bacterial community was taxonomically simple, containing an average of 1.9 common (relative abundance per sample > 5%) bacterial OTUs. The most abundant and prevalent (occurring in 92% of the samples) bacterial OTU, based on 16S rRNA sequences, closely matched that of the previously described Betaproteobacteria species Snodgrassella alvi. Bacteria that were first described in bee-related external environments dominated a number of gut bacterial communities, suggesting that they are not strictly dependent on the internal gut environment. The ARISA data showed a correlation between bacterial community structures and the geographic locations where the bees were sampled, suggesting that at least a subset of the bacterial species may be transmitted environmentally. Using light and fluorescent microscopy, we demonstrated that the gut bacteria form a biofilm on the internal epithelial surface of the ileum, corroborating results obtained from Apis mellifera.

USBombus, a database of contemporary survey data for North American Bumble Bees (Hymenoptera, Apidae, Bombus) distributed in the United States.

BACKGROUND: Bumble bees (Hymenoptera: Apidae, Bombus) are pollinators of wild and economically important flowering plants. However, at least four bumble bee species have declined significantly in population abundance and geographic range relative to historic estimates, and one species is possibly extinct. While a wealth of historic data is now available for many of the North American species found to be in decline in online databases, systematic survey data of stable species is still not publically available. The availability of contemporary survey data is critically important for the future monitoring of wild bumble bee populations. Without such data, the ability to ascertain the conservation status of bumble bees in the United States will remain challenging. NEW INFORMATION: This paper describes USBombus, a large database that represents the outcomes of one of the largest standardized surveys of bumble bee pollinators (Hymenoptera, Apidae, Bombus) globally. The motivation to collect live bumble bees across the United States was to examine the decline and conservation status of Bombus affinis, B. occidentalis, B. pensylvanicus, and B. terricola. Prior to our national survey of bumble bees in the United States from 2007 to 2010, there have only been regional accounts of bumble bee abundance and richness. In addition to surveying declining bumble bees, we also collected and documented a diversity of co-occuring bumble bees. However we have not yet completely reported their distribution and diversity onto a public online platform. Now, for the first time, we report the geographic distribution of bumble bees reported to be in decline (Cameron et al. 2011), as well as bumble bees that appeared to be stable on a large geographic scale in the United States (not in decline). In this database we report a total of 17,930 adult occurrence records across 397 locations and 39 species of Bombus detected in our national survey. We summarize their abundance and distribution across the United States and association to different ecoregions. The geospatial coverage of the dataset extends across 41 of the 50 US states, and from 0 to 3500 m a.s.l. Authors and respective field crews spent a total of 512 hours surveying bumble bees from 2007 to 2010. The dataset was developed using SQL server 2008 r2. For each specimen, the following information is generally provided: species, name, sex, caste, temporal and geospatial details, Cartesian coordinates, data collector(s), and when available, host plants. This database has already proven useful for a variety of studies on bumble bee ecology and conservation. However it is not publicly available. Considering the value of pollinators in agriculture and wild ecosystems, this large database of bumble bees will likely prove useful for investigations of the effects of anthropogenic activities on pollinator community composition and conservation status.

Strepsiptera, phylogenomics and the long branch attraction problem.

Insect phylogeny has recently been the focus of renewed interest as advances in sequencing techniques make it possible to rapidly generate large amounts of genomic or transcriptomic data for a species of interest. However, large numbers of markers are not sufficient to guarantee accurate phylogenetic reconstruction, and the choice of the model of sequence evolution as well as adequate taxonomic sampling are as important for phylogenomic studies as they are for single-gene phylogenies. Recently, the sequence of the genome of a strepsipteran has been published and used to place Strepsiptera as sister group to Coleoptera. However, this conclusion relied on a data set that did not include representatives of Neuropterida or of coleopteran lineages formerly proposed to be related to Strepsiptera. Furthermore, it did not use models that are robust against the long branch attraction artifact. Here we have sequenced the transcriptomes of seven key species to complete a data set comprising 36 species to study the higher level phylogeny of insects, with a particular focus on Neuropteroidea (Coleoptera, Strepsiptera, Neuropterida), especially on coleopteran taxa considered as potential close relatives of Strepsiptera. Using models robust against the long branch attraction artifact we find a highly resolved phylogeny that confirms the position of Strepsiptera as a sister group to Coleoptera, rather than as an internal clade of Coleoptera, and sheds new light onto the phylogeny of Neuropteroidea.

Geographical patterns of genetic divergence in the widespread Mesoamerican bumble bee Bombus ephippiatus (Hymenoptera: Apidae).

Bumble bees (Bombus Latreille) are an important group of social insects, well recognized throughout northern temperate regions as important pollinators of wild and agricultural plants. Little is known about the biology of this group in southern portions of the Americas, especially in Mesoamerica, a region of geological and ecological complexity from Mexico through Central America. One ubiquitous Mesoamerican species, Bombus ephippiatus, is enigmatic. Like many other Bombus, this species is homogeneous in body structure yet exhibits striking intraspecific color pattern polymorphism across its range, leading to uncertainty about its genealogical boundaries. It has been grouped taxonomically with B. wilmattae, a species narrowly restricted to southern Mexico and northern Guatamala. Furthermore, the relationships between these two taxa and a third species, B. impatiens, found only in America north of Mexico, have been controversial. Our phylogenetic analysis of DNA sequences from mitochondrial COI and nuclear PEPCK and CAD resolves the phylogeny of these three taxa as (B. impatiens, (B. ephippiatus, B. wilmattae)). Additional data from eight nuclear microsatellite markers reveal complex patterns of genetic divergence and isolation among populations of B. ephippiatus across its extensive geographic range, providing evidence for multiple independent evolutionary lineages. These lineages correspond not only to geographic and habitat variation across their range, but also to distinct color pattern groups present in the species. Knowledge of the phylogeny and genetic divergence of the B. ephippiatus group will provide a framework for understanding evolutionary and ecological origins of color pattern polymorphism in bumble bees, as well as providing insight into geographical factors enhancing speciation in Mesoamerica.

Interspecific geographic distribution and variation of the pathogens Nosema bombi and Crithidia species in United States bumble bee populations.

Several bumble bee (Bombus) species in North America have undergone range reductions and rapid declines in relative abundance. Pathogens have been suggested as causal factors, however, baseline data on pathogen distributions in a large number of bumble bee species have not been available to test this hypothesis. In a nationwide survey of the US, nearly 10,000 specimens of 36 bumble bee species collected at 284 sites were evaluated for the presence and prevalence of two known Bombus pathogens, the microsporidium Nosema bombi and trypanosomes in the genus Crithidia. Prevalence of Crithidia was ≤10% for all host species examined but was recorded from 21% of surveyed sites. Crithidia was isolated from 15 of the 36 Bombus species screened, and were most commonly recovered from Bombus bifarius, Bombus bimaculatus, Bombus impatiens and Bombus mixtus. Nosema bombi was isolated from 22 of the 36 US Bombus species collected. Only one species with more than 50 sampled bees, Bombus appositus, was free of the pathogen; whereas, prevalence was highest in Bombus occidentalis and Bombus pensylvanicus, two species that are reportedly undergoing population declines in North America. A variant of a tetranucleotide repeat in the internal transcribed spacer (ITS) of the N. bombi rRNA gene, thus far not reported from European isolates, was isolated from ten US Bombus hosts, appearing in varying ratios in different host species. Given the genetic similarity of the rRNA gene of N. bombi sampled in Europe and North America to date, the presence of a unique isolate in US bumble could reveal one or more native North American strains and indicate that N. bombi is enzootic across the Holarctic Region, exhibiting some genetic isolation.

Patterns of range-wide genetic variation in six North American bumble bee (Apidae: Bombus) species.

The increasing evidence for population declines in bumble bee (Bombus) species worldwide has accelerated research efforts to explain losses in these important pollinators. In North America, a number of once widespread Bombus species have suffered serious reductions in range and abundance, although other species remain healthy. To examine whether declining and stable species exhibit different levels of genetic diversity or population fragmentation, we used microsatellite markers to genotype populations sampled across the geographic distributions of two declining (Bombus occidentalis and Bombus pensylvanicus) and four stable (Bombus bifarius; Bombus vosnesenskii; Bombus impatiens and Bombus bimaculatus) Bombus species. Populations of declining species generally have reduced levels of genetic diversity throughout their range compared to codistributed stable species. Genetic diversity can be affected by overall range size and degree of isolation of local populations, potentially confounding comparisons among species in some cases. We find no evidence for consistent differences in gene flow among stable and declining species, with all species exhibiting weak genetic differentiation over large distances (e.g. >1000 km). Populations on islands and at high elevations experience relatively strong genetic drift, suggesting that some conditions lead to genetic isolation in otherwise weakly differentiated species. B. occidentalis and B. bifarius exhibit stronger genetic differentiation than the other species, indicating greater phylogeographic structure consistent with their broader geographic distributions across topographically complex regions of western North America. Screening genetic diversity in North American Bombus should prove useful for identifying species that warrant monitoring, and developing management strategies that promote high levels of gene flow will be a key component in efforts to maintain healthy populations.

Survey of bumble bee (Bombus) pathogens and parasites in Illinois and selected areas of northern California and southern Oregon.

Pathogens have been implicated as potential factors in the recent decline of some North American bumble bee (Bombus) species, but little information has been reported about the natural enemy complex of bumble bees in the United States. We targeted bumble bee populations in a state-wide survey in Illinois and several sites in California and Oregon where declines have been reported to determine presence and prevalence of natural enemies. Based on our observations, most parasites and pathogens appear to be widespread generalists among bumble bee species, but susceptibility to some natural enemies appeared to vary.

Genes involved in convergent evolution of eusociality in bees.

Eusociality has arisen independently at least 11 times in insects. Despite this convergence, there are striking differences among eusocial lifestyles, ranging from species living in small colonies with overt conflict over reproduction to species in which colonies contain hundreds of thousands of highly specialized sterile workers produced by one or a few queens. Although the evolution of eusociality has been intensively studied, the genetic changes involved in the evolution of eusociality are relatively unknown. We examined patterns of molecular evolution across three independent origins of eusociality by sequencing transcriptomes of nine socially diverse bee species and combining these data with genome sequence from the honey bee Apis mellifera to generate orthologous sequence alignments for 3,647 genes. We found a shared set of 212 genes with a molecular signature of accelerated evolution across all eusocial lineages studied, as well as unique sets of 173 and 218 genes with a signature of accelerated evolution specific to either highly or primitively eusocial lineages, respectively. These results demonstrate that convergent evolution can involve a mosaic pattern of molecular changes in both shared and lineage-specific sets of genes. Genes involved in signal transduction, gland development, and carbohydrate metabolism are among the most prominent rapidly evolving genes in eusocial lineages. These findings provide a starting point for linking specific genetic changes to the evolution of eusociality.

Patterns of widespread decline in North American bumble bees.

Bumble bees (Bombus) are vitally important pollinators of wild plants and agricultural crops worldwide. Fragmentary observations, however, have suggested population declines in several North American species. Despite rising concern over these observations in the United States, highlighted in a recent National Academy of Sciences report, a national assessment of the geographic scope and possible causal factors of bumble bee decline is lacking. Here, we report results of a 3-y interdisciplinary study of changing distributions, population genetic structure, and levels of pathogen infection in bumble bee populations across the United States. We compare current and historical distributions of eight species, compiling a database of >73,000 museum records for comparison with data from intensive nationwide surveys of >16,000 specimens. We show that the relative abundances of four species have declined by up to 96% and that their surveyed geographic ranges have contracted by 23-87%, some within the last 20 y. We also show that declining populations have significantly higher infection levels of the microsporidian pathogen Nosema bombi and lower genetic diversity compared with co-occurring populations of the stable (nondeclining) species. Higher pathogen prevalence and reduced genetic diversity are, thus, realistic predictors of these alarming patterns of decline in North America, although cause and effect remain uncertain.

Comparative genetic analyses of historical and contemporary collections highlight contrasting demographic histories for the bumble bees Bombus pensylvanicus and B. impatiens in Illinois.

Direct comparison of genetic patterns between museum specimens and contemporary collections can be a powerful approach for detecting recent demographic changes. Using microsatellite markers, we examined historical and contemporary genetic variation from an apparently declining bumble bee species, Bombus pensylvanicus, and from a stable species, Bombus impatiens, in central Illinois. For each species, we genotyped specimens from the Illinois Natural History Survey collected from three populations between 1969-1972 and from a resurvey of the same areas conducted in 2008. Population structure in B. pensylvanicus increased markedly over the last four decades (from theta(ST) = 0.001 to 0.027) while no structure was detected in B. impatiens for either time period (theta(ST) = -0.006 to -0.003). Changes in genetic diversity were not significant for either species, although small reductions were observed for B. pensylvanicus in all three populations. Coalescent simulations incorporating both contemporary and historical samples suggest that this small change is not surprising for recent population declines, as large reductions in genetic diversity were only apparent under the most severe bottleneck scenarios. These results demonstrate how comparisons of genetic patterns between temporal periods and species can help elucidate potential threats to population health and suggest several strategies that might be useful in the conservation of B. pensylvanicus in the Midwestern USA.

Multigene phylogeny reveals eusociality evolved twice in vespid wasps.

Eusocial wasps of the family Vespidae are thought to have derived their social behavior from a common ancestor that had a rudimentary caste-containing social system. In support of this behavioral scenario, the leading phylogenetic hypothesis of Vespidae places the eusocial wasps (subfamilies Stenogastrinae, Polistinae, and Vespinae) as a derived monophyletic clade, thus implying a single origin of eusocial behavior. This perspective has shaped the investigation and interpretation of vespid social evolution for more than two decades. Here we report a phylogeny of Vespidae based on data from four nuclear gene fragments (18S and 28S ribosomal DNA, abdominal-A and RNA polymerase II) and representatives from all six extant subfamilies. In contrast to the current phylogenetic perspective, our results indicate two independent origins of vespid eusociality, once in the clade Polistinae+Vespinae and once in the Stenogastrinae. The stenogastrines appear as an early diverging clade distantly related to the vespines and polistines and thus evolved their distinctive form of social behavior from a different ancestor than that of Polistinae+Vespinae. These results support earlier views based on life history and behavior and have important implications for interpreting transitional stages in vespid social evolution.

Hot bumble bees at good food: thoracic temperature of feeding Bombus wilmattae foragers is tuned to sugar concentration.

The ability of bees to generate metabolic heat plays an important role in their ability to forage and pollinate because they must achieve a minimum temperature to activate their flight muscles. In honey bees and stingless bees, the thoracic temperature of feeding foragers is correlated with the caloric value of sucrose solution provided at feeders outside the nest. We provide the first detailed data showing that this phenomenon also occurs in the closely related bumble bee and thus may be homologous in all social bees of the Apidae. Using infrared thermography, we measured T(th) for Bombus wilmattae foragers (mass 0.17+/-0.11 g, length 15.0+/-1.5 mm) from six wild colonies, foraging on a range of sucrose concentrations (0.5-2.5 mol l(-1), 16-65% by mass) in foraging arenas. For all colonies, we measured significant increases in DeltaT(th) (P<0.0001) with increasing sucrose concentration, with significant differences (P<0.0001) between colonies due to different linear regression slopes (0.28-2.4) and y-intercepts (2.7-5.5). We suggest that this modulation of pitching T(th) to sucrose concentration is a general phenomenon in all social bees and may be a widespread adaptation facilitating rapid food collection in flying Hymenoptera.