Germline ATM variants predispose to melanoma: a joint analysis across the GenoMEL and MelaNostrum consortia.

PURPOSE: Ataxia-Telangiectasia Mutated (ATM) has been implicated in the risk of several cancers, but establishing a causal relationship is often challenging. Although ATM single-nucleotide polymorphisms have been linked to melanoma, few functional alleles have been identified. Therefore, ATM impact on melanoma predisposition is unclear. METHODS: From 22 American, Australian, and European sites, we collected 2,104 familial, multiple primary (MPM), and sporadic melanoma cases who underwent ATM genotyping via panel, exome, or genome sequencing, and compared the allele frequency (AF) of selected ATM variants classified as loss-of-function (LOF) and variants of uncertain significance (VUS) between this cohort and the gnomAD non-Finnish European (NFE) data set. RESULTS: LOF variants were more represented in our study cohort than in gnomAD NFE, both in all (AF = 0.005 and 0.002, OR = 2.6, 95% CI = 1.56-4.11, p < 0.01), and familial + MPM cases (AF = 0.0054 and 0.002, OR = 2.97, p < 0.01). Similarly, VUS were enriched in all (AF = 0.046 and 0.033, OR = 1.41, 95% CI = 1.6-5.09, p < 0.01) and familial + MPM cases (AF = 0.053 and 0.033, OR = 1.63, p < 0.01). In a case-control comparison of two centers that provided 1,446 controls, LOF and VUS were enriched in familial + MPM cases (p = 0.027, p = 0.018). CONCLUSION: This study, describing the largest multicenter melanoma cohort investigated for ATM germline variants, supports the role of ATM as a melanoma predisposition gene, with LOF variants suggesting a moderate-risk.

Economic Dependence and Vulnerability of United States Agricultural Sector on Insect-Mediated Pollination Service.

Deficits in insect-mediated pollination service undermine ecosystem biodiversity and function, human nutrition, and economic welfare. Global pollinator supply continues to decline, while production of pollination-dependent crops increases. Using publicly available price and production data and existing pollination field studies, we quantify economic dependence of United States crops on insect-mediated pollination service at the county level and update existing coefficients of insect dependence of sample crops when possible. Economic value dependent on pollination service totals 34.0 billion USD in 2012. Twenty percent of US counties produce 80% of total economic value attributable to insect pollinators. We compile county-level data and consider the spatial relationship between economic value dependent on insect-mediated pollination, region-specific forage suitability, and crop-specific agricultural areas within US landscapes. We identify vulnerable, highly dependent areas where habitat for wild pollinators has been reduced. These results can help inform future efforts to conserve and bolster managed and wild pollinator populations to ensure sustainable production of key agricultural crops.

Macronutrient ratios in pollen shape bumble bee (Bombus impatiens) foraging strategies and floral preferences.

To fuel their activities and rear their offspring, foraging bees must obtain a sufficient quality and quantity of nutritional resources from a diverse plant community. Pollen is the primary source of proteins and lipids for bees, and the concentrations of these nutrients in pollen can vary widely among host-plant species. Therefore we hypothesized that foraging decisions of bumble bees are driven by both the protein and lipid content of pollen. By successively reducing environmental and floral cues, we analyzed pollen-foraging preferences of Bombus impatiens in (i) host-plant species, (ii) pollen isolated from these host-plant species, and (iii) nutritionally modified single-source pollen diets encompassing a range of protein and lipid concentrations. In our semifield experiments, B impatiens foragers exponentially increased their foraging rates of pollen from plant species with high protein:lipid (P:L) ratios; the most preferred plant species had the highest ratio (∼4.6:1). These preferences were confirmed in cage studies where, in pairwise comparisons in the absence of other floral cues, B impatiens workers still preferred pollen with higher P:L ratios. Finally, when presented with nutritionally modified pollen, workers were most attracted to pollen with P:L ratios of 5:1 and 10:1, but increasing the protein or lipid concentration (while leaving ratios intact) reduced attraction. Thus, macronutritional ratios appear to be a primary factor driving bee pollen-foraging behavior and may explain observed patterns of host-plant visitation across the landscape. The nutritional quality of pollen resources should be taken into consideration when designing conservation habitats supporting bee populations.

Effective Heisenberg Model and Exchange Interaction for Strongly Correlated Systems.

We consider the extended Hubbard model and introduce a corresponding Heisenberg-like problem written in terms of spin operators. The derived formalism is reminiscent of Anderson's idea of the effective exchange interaction and takes into account nonlocal correlation effects. The results for the exchange interaction and spin susceptibility in the magnetic phase are expressed in terms of single-particle quantities. This fact not only can be used for realistic calculations of multiband systems but also allows us to reconsider a general description of many-body effects in the most interesting physical regimes, where the physical properties of the system are dominated by collective (bosonic) fluctuations. In the strongly spin-polarized limit, when the local magnetic moment is well defined, the exchange interaction reduces to a standard expression of the density functional theory that has been successfully used in practical calculations of magnetic properties of real materials.

Maternal methyl donor and cofactor supplementation in late pregnancy increases ß-cell numbers at 16 days of life in growth-restricted twin lambs.

Restricted growth before birth (IUGR) increases adult risk of Type 2 diabetes by impairing insulin sensitivity and secretion. Altered fetal one-carbon metabolism is implicated in developmental programming of adult health and disease by IUGR. Therefore, we evaluated effects of maternal dietary supplementation with methyl donors and cofactors (MMDS), designed to increase fetal supply, on insulin action in the spontaneously IUGR twin lamb. In vivo glucose-stimulated insulin secretion and insulin sensitivity were measured at days 12-14 in singleton controls (CON, n = 7 lambs from 7 ewes), twins (IUGR, n = 8 lambs from 8 ewes), and twins from ewes that received MMDS (2 g rumen-protected methionine, 300 mg folic acid, 1.2 g sulfur, 0.7 mg cobalt) daily from 120 days after mating (~0.8 of term) until delivery (IUGR+MMDS, n = 8 lambs from 4 ewes). Body composition and pancreas morphometry were assessed in lambs at day 16 IUGR reduced size at birth and increased neonatal fractional growth rate. MMDS normalized long bone lengths but not other body dimensions of IUGR lambs at birth. IUGR did not impair glucose control or insulin action at days 12-14, compared with controls. MMDS increased metabolic clearance rate of insulin and increased ß-cell numerical density and tended to improve insulin sensitivity, compared with untreated IUGR lambs. This demonstrates that effects of late-pregnancy methyl donor supplementation persist until at least the third week of life. Whether these effects of MMDS persist beyond early postnatal life and improve metabolic outcomes after IUGR in adults and the underlying mechanisms remain to be determined.

Genome-wide analysis of signatures of selection in populations of African honey bees (Apis mellifera) using new web-based tools.

BACKGROUND: With the development of inexpensive, high-throughput sequencing technologies, it has become feasible to examine questions related to population genetics and molecular evolution of non-model species in their ecological contexts on a genome-wide scale. Here, we employed a newly developed suite of integrated, web-based programs to examine population dynamics and signatures of selection across the genome using several well-established tests, including F ST, pN/pS, and McDonald-Kreitman. We applied these techniques to study populations of honey bees (Apis mellifera) in East Africa. In Kenya, there are several described A. mellifera subspecies, which are thought to be localized to distinct ecological regions. RESULTS: We performed whole genome sequencing of 11 worker honey bees from apiaries distributed throughout Kenya and identified 3.6 million putative single-nucleotide polymorphisms. The dense coverage allowed us to apply several computational procedures to study population structure and the evolutionary relationships among the populations, and to detect signs of adaptive evolution across the genome. While there is considerable gene flow among the sampled populations, there are clear distinctions between populations from the northern desert region and those from the temperate, savannah region. We identified several genes showing population genetic patterns consistent with positive selection within African bee populations, and between these populations and European A. mellifera or Asian Apis florea. CONCLUSIONS: These results lay the groundwork for future studies of adaptive ecological evolution in honey bees, and demonstrate the use of new, freely available web-based tools and workflows ( http://usegalaxy.org/r/kenyanbee ) that can be applied to any model system with genomic information.

Neurophysiological mechanisms underlying sex- and maturation-related variation in pheromone responses in honey bees (Apis mellifera).

In the honey bee (Apis mellifera), social organization is primarily mediated by pheromones. Queen-produced 9-oxo-2-decenoic acid (9-ODA) functions as both a social and sex pheromone, eliciting attraction in both female workers and male drones, but also affecting other critical aspects of worker physiology and behavior. These effects are also maturation related, as younger workers and sexually mature drones are most receptive to 9-ODA. While changes in the peripheral nervous system drive sex-related differences in sensitivity to 9-ODA, the mechanisms driving maturation-related shifts in receptivity to 9-ODA remain unknown. Here, we investigate the hypothesis that changes at the peripheral nervous system may be mediating plastic responses to 9-ODA by characterizing expression levels of AmOR11 (the olfactory receptor tuned to 9-ODA) and electrophysiological responses to 9-ODA. We find that receptor expression correlates significantly with behavioral receptivity to 9-ODA, with nurses and sexually mature drones exhibiting higher levels of expression than foragers and immature drones, respectively. Electrophysiological responses to 9-ODA were not found to correlate with behavioral receptivity or receptor expression, however. Thus, while receptor expression at the periphery exhibits a level of plasticity that correlates with behavior, the mechanisms driving maturation-dependent responsiveness to 9-ODA appear to function primarily in the central nervous system.

Testing the plasticity of insulin secretion and ß-cell function in vivo: responses to chronic hyperglycaemia in the sheep.

Plasticity of insulin secretion is essential to maintain the action of insulin during insulin resistance and to prevent diabetes. Investigation of the plasticity of insulin secretion and its regulation is challenging, and the objective of this study was to develop a novel large-animal-based model. The effect of chronic moderate hyperglycaemia on the plasticity of insulin secretion, ß-cell mass and function was determined in sheep. Adolescent sheep (120 days old) were infused with 25% glucose for 16 days to increase blood glucose by 50% (n = 10), and control animals (n = 9) were infused with saline. Glucose- and arginine-stimulated insulin secretion, insulin sensitivity and glucose effectiveness were measured in vivo before and during treatment (days 10-14), and ß-cell mass was measured at the end of treatment. Hyperglycaemia increased blood glucose (+53%) and plasma insulin (+403%; each P < 0.003) and did not alter whole-body insulin sensitivity. Hyperglycaemia increased glucose-stimulated insulin secretion (particularly second phase; five-fold) and arginine-stimulated insulin secretion (particularly first phase; four-fold). Hyperglycaemia reduced ß-cell mass (∼50%, P = 0.038) and increased glucose- and arginine-stimulated insulin secretion relative to ß-cell mass five-fold (P = 0.060) and 20-fold (P = 0.007), respectively. Chronic hyperglycaemia therefore induces marked adaptation and upregulation of glucose- and arginine-stimulated insulin secretion by enhancing ß-cell function rather than increasing ß-cell mass in the sheep, consistent with long-term adaptations seen in humans. This marked plasticity of insulin secretion in response to moderate hyperglycaemia provides a novel model for the investigation of factors affecting its capacity and underlying determinants.

Soil moisture affects plant-pollinator interactions in an annual flowering plant.

Many environmental factors impact plant and pollinator communities. However, variation in soil moisture and how it mediates the plant-pollinator interactions has yet to be elucidated. We hypothesized that long-term variation in soil moisture can exert a strong selective pressure on the floral and vegetative traits of plants, leading to changes in pollinator visitation. We demonstrated that there are three phenotypic populations of Gentiana aristata in our study alpine region in the Qinghai-Tibetan Plateau that vary in floral colour and other traits. Pink (dry habitat) and blue (intermediate habitat) flower populations are visited primarily by bumblebees, and white (wet habitat) flower populations are visited by flies. These patterns of visitation are driven by vegetative and floral traits and are constant when non-endemic plants are placed in the intermediate habitats. Additionally, the floral communities in different habitats vary, with more insect-pollinated forbs in the dry and intermediate habitats versus the wet habitats. Through a common garden and reciprocal transplant experiment, we demonstrated that plant growth traits, pollinator attractiveness and seed production are highest when the plant population is raised in its endemic habitat. This suggests that these plant populations have evolved to pollinator communities associated with habitat differences. This article is part of the theme issue 'Natural processes influencing pollinator health: from chemistry to landscapes'.

Impacts of soil nutrition on floral traits, pollinator attraction, and fitness in cucumbers (Cucumis sativus L.).

Annual plants allocate soil nutrients to floral display and pollinator rewards to ensure pollination success in a single season. Nitrogen and phosphorus are critical soil nutrients whose levels are altered by intensive land use that may affect plants' fitness via pollinator attractiveness through floral display and rewards. In a controlled greenhouse study, we studied in cucumbers (Cucumis sativus) how changes in soil nitrogen and phosphorus influence floral traits, including nectar and pollen reward composition. We evaluated how these traits affect bumble bee (Bombus impatiens, an important cucumber pollinator) visitation and ultimately fruit yield. While increasing nitrogen and phosphorus increased growth and floral display, excess nitrogen created an asymptotic or negative effect, which was mitigated by increasing phosphorus. Male floral traits exhibited higher plasticity in responses to changes in soil nutrients than female flowers. At 4:1 nitrogen:phosphorus ratios, male flowers presented increased nectar volume and pollen number resulting in increased bumble bee visitation. Interestingly, other pollinator rewards remained consistent across all soil treatments: male and female nectar sugar composition, female nectar volume, and pollen protein and lipid concentrations. Therefore, although cucumber pollination success was buffered in conditions of nutrient stress, highly skewed nitrogen:phosphorus soil ratios reduced plant fitness via reduced numbers of flowers and reward quantity, pollinator attraction, and ultimately yield.

Thrice out of Asia and the adaptive radiation of the western honey bee.

The origin of the western honey bee Apis mellifera has been intensely debated. Addressing this knowledge gap is essential for understanding the evolution and genetics of one of the world's most important pollinators. By analyzing 251 genomes from 18 native subspecies, we found support for an Asian origin of honey bees with at least three expansions leading to African and European lineages. The adaptive radiation of honey bees involved selection on a few genomic "hotspots." We found 145 genes with independent signatures of selection across all bee lineages, and these genes were highly associated with worker traits. Our results indicate that a core set of genes associated with worker and colony traits facilitated the adaptive radiation of honey bees across their vast distribution.

The transcription factor Krüppel homolog 1 is linked to hormone mediated social organization in bees.

BACKGROUND: Regulation of worker behavior by dominant queens or workers is a hallmark of insect societies, but the underlying molecular mechanisms and their evolutionary conservation are not well understood. Honey bee and bumble bee colonies consist of a single reproductive queen and facultatively sterile workers. The queens' influences on the workers are mediated largely via inhibition of juvenile hormone titers, which affect division of labor in honey bees and worker reproduction in bumble bees. Studies in honey bees identified a transcription factor, Krüppel-homolog 1 (Kr-h1), whose expression in worker brains is significantly downregulated in the presence of a queen or queen pheromone and higher in forager bees, making this gene an ideal candidate for examining the evolutionary conservation of socially regulated pathways in Hymenoptera. RESULTS: In contrast to honey bees, bumble bees foragers do not have higher Kr-h1 levels relative to nurses: in one of three colonies levels were similar in nurses and foragers, and in two colonies levels were higher in nurses. Similarly to honey bees, brain Kr-h1 levels were significantly downregulated in the presence versus absence of a queen. Furthermore, in small queenless groups, Kr-h1 levels were downregulated in subordinate workers with undeveloped ovaries relative to dominant individuals with active ovaries. Brain Kr-h1 levels were upregulated by juvenile hormone treatment relative to a vehicle control. Finally, phylogenetic analysis indicates that KR-H1 orthologs are presence across insect orders. Though this protein is highly conserved between honey bees and bumble bees, there are significant differences between orthologs of insects from different orders. CONCLUSIONS: Our results suggest that Kr-h1 is associated with juvenile hormone mediated regulation of reproduction in bumble bees. The expression of this transcription factor is inhibited by the queen and associated with endocrine mediated regulation of social organization in two species of bees. Thus, KR-H1 may transcriptionally regulate a conserved genetic module that is part of a pathway that has been co-opted to function in social behavior, and adjusts the behavior of workers to their social environmental context.

A screening-level assessment of the pollinator-attractiveness of ornamental nursery stock using a honey bee foraging assay.

In urban and suburban landscapes characterized by extensive designed greenspaces, the support of pollinator communities hinges significantly on floral resources provided by ornamental plants. The attractiveness of ornamental plants to pollinators, however, cannot be presumed, and some studies suggest that a majority of ornamental plant varieties receive little or no pollinator visitation. Here, we harness the sampling power of the western honey bee, a generalist pollinator whose diet breadth overlaps substantially with that of other pollinators, to survey the utilization of ornamental plants grown at three commercial nurseries in Connecticut, USA. Using a combination of DNA metabarcoding and microscopy, we identify, to genus-level, pollen samples from honey bee colonies placed within each nursery, and we compare our results with nursery plant inventories to identify the subset of cultivated genera that were visited during pollen foraging. Samples were collected weekly from May to September, encompassing the majority of the growing season. Our findings show that some plant genera known to be cultivated as ornamentals in our system, particularly ornamental trees and shrubs (e.g. Hydrangea, Rosa, Spiraea, Syringa, Viburnum), functioned as major pollen sources, but the majority of plants inventoried at our nurseries provided little or no pollen to honey bees. These results are in agreement with a growing body of literature highlighting the special importance of woody plants as resources for flower-visiting insects. We encourage further exploration of the genera highlighted in our data as potential components of pollinator-friendly ornamental greenspace.

Pollen Protein: Lipid Macronutrient Ratios May Guide Broad Patterns of Bee Species Floral Preferences.

Pollinator nutritional ecology provides insights into plant-pollinator interactions, coevolution, and the restoration of declining pollinator populations. Bees obtain their protein and lipid nutrient intake from pollen, which is essential for larval growth and development as well as adult health and reproduction. Our previous research revealed that pollen protein to lipid ratios (P:L) shape bumble bee foraging preferences among pollen host-plant species, and these preferred ratios link to bumble bee colony health and fitness. Yet, we are still in the early stages of integrating data on P:L ratios across plant and bee species. Here, using a standard laboratory protocol, we present over 80 plant species' protein and lipid concentrations and P:L values, and we evaluate the P:L ratios of pollen collected by three bee species. We discuss the general phylogenetic, phenotypic, behavioral, and ecological trends observed in these P:L ratios that may drive plant-pollinator interactions; we also present future research questions to further strengthen the field of pollination nutritional ecology. This dataset provides a foundation for researchers studying the nutritional drivers of plant-pollinator interactions as well as for stakeholders developing planting schemes to best support pollinators.

Studies of granulosa cell maturation in dominant and subordinate bovine follicles: novel extracellular matrix focimatrix is co-ordinately regulated with cholesterol side-chain cleavage CYP11A1.

During growth of antral ovarian follicles granulosa cells first become associated with a novel type of extracellular matrix, focimatrix, and at larger sizes follicles become either subordinate or dominant. To examine this, bovine subordinate (9.0+/-S.E.M. 0.4 mm; n=16), partially dominant (12.0+/-0.6 mm; n=18) and fully dominant (15.0+/-0.4 mm; n=14) follicles were examined by real time RT-PCR analyses of granulosa cells and by immunohistochemistry of focimatrix. Changes in the expression of FSH receptor, LH receptor, cholesterol side-chain cleavage (CYP11A1), 3beta-hydroxysteroid dehydrogenase, aromatase (CYP19A1) and inhibin-alpha and beta-B were observed as expected for follicle sizes examined. After adjusting for size differences, only CYP11A1 was significantly different between the groups, and elevated in dominant follicles. Also after adjusting for differences in size there were no significant differences in expression of focimatrix components collagen type IV alpha-1 (COL4A1), laminin beta-2, nidogen 1 (NID1), and perlecan (HSPG2) or the volume density of NID1 and -2 and HSPG2. The volume density of focimatrix components in laminin 111 was significantly elevated in dominant follicles. Adjusting for analysis of more than one follicle per animal and for multiple correlations, CYP11A1 mRNA levels were highly correlated with the focimatrix genes COL4A1, NID1 and -2 and HSPG2. Thus, focimatrix may potentially regulate CYP11A1 expression, and the regulation of both could be important in follicular dominance.

A candidate pheromone receptor and two odorant receptors of the hawkmoth Manduca sexta.

In this study, we cloned and characterized three Manduca sexta odorant receptors (ORs). One receptor is a putative pheromone receptor expressed exclusively in a cell associated with male-specific type-I trichoid sensilla. We describe the results of real-time PCR (RT-PCR) and quantitative real-time PCR (qRT-PCR) experiments that show MsextaOR1 is expressed only in male antennae. In situ hybridization labels a single cell associated with type-1 trichoid sensilla, which houses two neurons that have been previously determined to respond to the major components of the pheromone blend. The second receptor, MsextaOR2, was discovered using degenerate primers designed to conserved motifs of a unique group ORs that share as much as 88% identity. Comparison of RT-PCR, qRT-PCR, and in situ hybridization results with those of ORs in the Drosophila melanogaster Or83b subfamily shows a strong sequence and expression pattern similarity. The third receptor, MsextaOR3, was found by 5'-end sequencing of a normalized and subtracted cDNA library from male M. sexta antennae. RT-PCR and qRT-PCR show that this receptor is expressed only in male and female antennae. These are the first ORs, including a putative pheromone receptor, to be described from M. sexta.

Impaired beta-cell function and inadequate compensatory increases in beta-cell mass after intrauterine growth restriction in sheep.

Poor growth before birth increases the risk of non-insulin-dependent diabetes mellitus (NIDDM) and impairs insulin secretion relative to sensitivity. We investigated the effects of intrauterine growth restriction in sheep on insulin secretion, beta-cell mass, and function from before birth to young adulthood and its molecular basis. Pancreas was collected from control and placentally restricted sheep as fetuses (d 143 gestation), lambs (aged 42 d), and young adults (aged 556 d), following independent measures of in vivo insulin secretion and sensitivity. beta-Cells and islets were counted after immunohistochemical staining for insulin. In lambs, gene expression was measured by RT-PCR and expressed relative to 18S. beta-Cell mass correlated positively with fetal weight but negatively with birth weight in adult males. Glucose-stimulated insulin disposition and beta-cell function correlated negatively with fetal weight but positively with birth weight in adult males. Placental restriction increased pancreatic expression of IGF-II and IGF-I but decreased that of voltage-gated calcium channel, alpha1D subunit (CACNA1D) in lambs. In male lambs, pancreatic IGF-II and insulin receptor expression correlated strongly and positively with beta-cell mass and CACNA1D expression with glucose-stimulated insulin disposition. Restricted growth before birth in the sheep does not impair insulin secretion, relative to sensitivity, before birth or in young offspring. IGF-II and insulin receptor are implicated as key molecular regulators of beta-cell mass compensation, whereas impaired expression of the voltage-gated calcium channel may underlie impaired beta-cell function after intrauterine growth restriction. With aging, the insulin secretory capacity of the beta-cell is impaired in males, and their increases in beta-cell mass are inadequate to maintain adequate insulin secretion relative to sensitivity.

How do oral and maxillofacial surgeons manage concussion?

Craniofacial trauma results in distracting injuries that are easy to see, and as oral and maxillofacial surgeons (OMFS) we gravitate towards injuries that can be seen and are treatable surgically. However, we do tend not to involve ourselves (and may potentially overlook) injuries that are not obvious either visually or radiographically, and concussion is one such. We reviewed the records of 500 consecutive patients who presented with facial fractures at the Queen Elizabeth Hospital, Birmingham, to identify whether patients had been screened for concussion, and how they had been managed. Of the 500 cases 186 (37%) had concussion, and 174 (35%) had a more severe traumatic brain injury. The maxillofacial team documented loss of consciousness in 314 (63%) and pupillary reactions in 215 (43%). Ninety-three (19%) were referred for a neurosurgical opinion, although most of these were patients who presented with a Glasgow coma scale (GCS) of ≤13. Only 37 patients (7%) were referred to the traumatic brain injury clinic. Recent reports have indicated that 15% of all patients diagnosed with concussion have symptoms that persist for longer than two weeks. These can have far-reaching effects on recovery, and have an appreciable effect on the psychosocial aspects of the patients' lives. As we have found, over one third of patients with craniofacial trauma are concussed. We think, therefore, that all patients who have been referred to OMFS with craniofacial trauma should be screened for concussion on admission, and at the OMFS follow up clinic. In addition, there should be an agreement between consultants that such patients should be referred to the traumatic brain injury clinic for follow up.

Consistent pollen nutritional intake drives bumble bee (Bombus impatiens) colony growth and reproduction across different habitats.

Foraging behavior is a critical adaptation by insects to obtain appropriate nutrients from the environment for development and fitness. Bumble bees (Bombus spp.) form annual colonies which must rapidly increase their worker populations to support rearing reproductive individuals before the end of the season. Therefore, colony growth and reproduction should be dependent on the quality and quantity of pollen resources in the surrounding landscape. Our previous research found that B. impatiens foraging preferences to different plant species were shaped by pollen protein:lipid nutritional ratios (P:L), with foragers preferring pollen species with a ~5:1 P:L ratio. In this study, we placed B. impatiens colonies in three different habitats (forest, forest edge, and valley) to determine whether pollen nutritional quality collected by the colonies differed between areas that may differ in resource abundance and diversity. We found that habitat did not influence the collected pollen nutritional quality, with colonies in all three habitats collecting pollen averaging a 4:1 P:L ratio. Furthermore, there was no difference in the nutritional quality of the pollen collected by colonies that successfully reared reproductives and those that did not. We found however, that "nutritional intake," calculated as the colony-level intake rate of nutrient quantities (protein, lipid, and sugar), was strongly related to colony growth and reproductive output. Therefore, we conclude that B. impatiens colony performance is a function of the abundance of nutritionally appropriate floral resources in the surrounding landscape. Because we did not comprehensively evaluate the nutrition provided by the plant communities in each habitat, it remains to be determined how B. impatiens polylectic foraging strategies helps them select among the available pollen nutritional landscape in a variety of plant communities to obtain a balance of key macronutrients.

Investigating the viral ecology of global bee communities with high-throughput metagenomics.

Bee viral ecology is a fascinating emerging area of research: viruses exert a range of effects on their hosts, exacerbate impacts of other environmental stressors, and, importantly, are readily shared across multiple bee species in a community. However, our understanding of bee viral communities is limited, as it is primarily derived from studies of North American and European Apis mellifera populations. Here, we examined viruses in populations of A. mellifera and 11 other bee species from 9 countries, across 4 continents and Oceania. We developed a novel pipeline to rapidly and inexpensively screen for bee viruses. This pipeline includes purification of encapsulated RNA/DNA viruses, sequence-independent amplification, high throughput sequencing, integrated assembly of contigs, and filtering to identify contigs specifically corresponding to viral sequences. We identified sequences for (+)ssRNA, (-)ssRNA, dsRNA, and ssDNA viruses. Overall, we found 127 contigs corresponding to novel viruses (i.e. previously not observed in bees), with 27 represented by >0.1% of the reads in a given sample, and 7 contained an RdRp or replicase sequence which could be used for robust phylogenetic analysis. This study provides a sequence-independent pipeline for viral metagenomics analysis, and greatly expands our understanding of the diversity of viruses found in bee communities.