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Bumble bees are declining across the globe. The causes of this decline have been attributed to a variety of stressors, including pesticides. Fungicides are a type of pesticide that has been understudied in the context of bumble bee health. As a result, fungicides are often applied to flowering plants without consideration of pollinator exposure. Recent work demonstrates that fungicides have sublethal effects in bumble bees, but little is known about how much fungicide it takes to cause these sublethal effects. To address this gap in the literature, we fed microcolonies of the common eastern bumble bee (Bombus impatiens CressonHymenoptera: ApidaeHymenoptera: ApidaeHymenoptera: ApidaeHymenoptera: Apidae) pollen contaminated with a range of fungicide concentrations. We chose these concentrations based on the range of fungicide concentrations in pollen and nectar that were reported in the literature. Results revealed that later-stage pupae and newly emerged males are potentially sensitive to fungicide exposure, showing smaller size and reduced fat reserves at intermediate levels of contamination. Compared to the control, intermediated levels of fungicide-contaminated pollen led to increased pupal mortality and delayed male emergence. Contrary to expectations, higher fungicide levels did not exhibit a linear relationship with negative impacts, suggesting nuanced effects. Because body size and emergence timing are important aspects of bumble bee reproductive behavior, results have implications for mating success, potentially disrupting colony development. [ABSTRACT FROM AUTHOR]

Bumble bees are the primary commercial pollinators for greenhouse‐grown crops and managing them requires insight into the Bombus life cycle. However, some aspects of the life cycle are not fully understood. Once mated, adult Bombus gynes can be stored for a short term at a constant low temperature (CLT). However, survival rates for some species are low during CLT, particularly when storage periods exceed a month. To address this, we tested the hypothesis that Bombus gynes stored in a fluctuating thermal regime (FTR) have increased survival compared to gynes stored in CLT. Specifically, the FTR had a baseline temperature of 2°C, with a daily fluctuation to 20°C relative to the CLT of 2°C with no warming pulse. We tested the effects of FTR on gyne survival and post‐cold storage lipid content in three species used for commercial pollination in North America: B. vosnesenskii, B. huntii and B. impatiens. We further assessed how additional factors influence cold storage survival, including gyne body weight, body size and colony age. Survival responses varied among species. Bombus vosnesenskii and B. huntii had a lower survival rate in FTR compared to CLT, but B. impatiens gynes had a significantly higher survival rate in FTR. We did not observe significant effects of FTR on lipid stores for any species, but survival increased with an increasing initial weight prior to cold storage. Moreover, gynes produced later in the colony life cycle had lower body weight and decreased survival compared to early emerging gynes. Our study provides insight into how cold storage conditions interact with colony and gyne characteristics to influence captive overwintering survival. Our findings have economic applications for improving commercial rearing and efforts for conservation rearing of at‐risk Bombus species. [ABSTRACT FROM AUTHOR]

Bumble bees are integral pollinators of native and cultivated plant communities, but species are undergoing significant changes in range and abundance on a global scale. Climate change and land cover alteration are key drivers in pollinator declines; however, limited research has evaluated the cumulative effects of these factors on bumble bee assemblages. This study tests bumble bee assemblage (calculated as richness and abundance) responses to climate and land use by modeling species‐specific habitat requirements, and assemblage‐level responses across geographic regions. We integrated species richness, abundance, and distribution data for 18 bumble bee species with site‐specific bioclimatic, landscape composition, and landscape configuration data to evaluate the effects of multiple environmental stressors on bumble bee assemblages throughout 433 agricultural fields in Florida, Indiana, Kansas, Kentucky, Maryland, South Carolina, Utah, Virginia, and West Virginia from 2018 to 2020. Distinct east versus west groupings emerged when evaluating species‐specific habitat associations, prompting a detailed evaluation of bumble bee assemblages by geographic region. Maximum temperature of warmest month and precipitation of driest month had a positive impact on bumble bee assemblages in the Corn Belt/Appalachian/northeast, southeast, and northern plains regions, but a negative impact on the mountain region. Further, forest land cover surrounding agricultural fields was highlighted as supporting more rich and abundant bumble bee assemblages. Overall, climate and land use combine to drive bumble bee assemblages, but how those processes operate is idiosyncratic and spatially contingent across regions. From these findings, we suggested regionally specific management practices to best support rich and abundant bumble bee assemblages in agroecosystems. Results from this study contribute to a better understanding of climate and landscape factors affecting bumble bees and their habitats throughout the United States. [ABSTRACT FROM AUTHOR]

Declines in bumble bee species range and abundances are documented across multiple continents and have prompted the need for research to aid species recovery and conservation. The rusty patched bumble bee (Bombus affinis) is the first federally listed bumble bee species in North America. We conducted a range-wide population genetics study of B. affinis from across all extant conservation units to inform conservation efforts. To understand the species' vulnerability and help establish recovery targets, we examined population structure, patterns of genetic diversity, and population differentiation. Additionally, we conducted a site-level analysis of colony abundance to inform prioritizing areas for conservation, translocation, and other recovery actions. We find substantial evidence of population structuring along an east-to-west gradient. Putative populations show evidence of isolation by distance, high inbreeding coefficients, and a range-wide male diploidy rate of ~15%. Our results suggest the Appalachians represent a genetically distinct cluster with high levels of private alleles and substantial differentiation from the rest of the extant range. Site-level analyses suggest low colony abundance estimates for B. affinis compared to similar datasets of stable, co-occurring species. These results lend genetic support to trends from observational studies, suggesting that B. affinis has undergone a recent decline and exhibit substantial spatial structure. The low colony abundances observed here suggest caution in overinterpreting the stability of populations even where B. affinis is reliably detected interannually. These results help delineate informed management units, provide context for the potential risks of translocation programs, and help set clear recovery targets for this and other threatened bumble bee species. [ABSTRACT FROM AUTHOR]

Bombus vosnesenskii Radowszkowski, 1862 is one of three bumble bee species commercially available for pollination services in North America; however, little is documented about B. vosnesenskii colony life cycle or the establishment of ex situ rearing, mating, and overwintering practices. In this study, we documented nest success, colony size, and gyne production; recorded the duration of mating events; assessed overwintering survival of mated gynes; and evaluated second-generation nest success for colonies established from low- and high-elevation wild-caught B. vosnesenskii gynes. Of the 125 gynes installed, 62.4% produced brood cells (nest initiation) and 43.2% had at least 1 worker eclose (nest establishment). High-elevation B. vosnesenskii gynes had significantly higher nest initiation and establishment success than low-elevation gynes. However, low-elevation colonies were significantly larger with queens producing more gynes on average. Mating was recorded for 200 low-elevation and 37 high-elevation gynes, resulting in a mean duration of 62 and 51 min, respectively. Mated gynes were then placed into cold storage for 54 days to simulate overwintering, which resulted in 59.1% of low-elevation gynes surviving and 91.9% of high-elevation gynes surviving. For second-generation low-elevation gynes, 26.4% initiated nesting and 14.3% established nesting. Second-generation high-elevation gynes did not initiate nesting despite CO2 narcosis treatments. Overall, these results increase our understanding of B. vosnesenskii nesting, mating, and overwintering biology from 2 elevations. Furthermore, this study provides information on successful husbandry practices that can be used by researchers and conservationists to address knowledge gaps and enhance the captive rearing of bumble bees. [ABSTRACT FROM AUTHOR]

Pollen is an essential component of bee diets, and rearing bumble bees (Bombus spp.) for commercial use necessitates feeding pollen in mass quantities. This pollen is collected from honey bee (Apis mellifera L.) colonies because neither an artificial diet nor an economical, large-scale pollen collection process from flowers is available. The provenance of honey bee-collected pollen is often unknown, and in some cases has crossed international borders. Both deformed wing virus (DWV) and the fungal pathogen Ascosphaera apis (Claussen) Olive & Spiltoir (cause of chalkbrood disease); occur in honey bee-collected pollen, and infections have been observed in bumble bees. We used these pathogens as general surrogates for viruses and spore-forming fungal diseases to test the efficacy of 3 sterilization methods, and assessed whether treatment altered pollen quality for the bumble bee. Using honey bee-collected pollen spiked with known doses of DWV and A. apis , we compared gamma irradiation (GI), ozone fumigation (OZ), and ethylene oxide fumigation (EO) against an untreated positive control and a negative control. Following sterilization treatments, we tested A. apis spore viability, detected viral presence with PCR, and tested palatability to the bumble bee Bombus impatiens Cresson. We also measured bacterial growth from pollens treated with EO and GI. GI and EO outperformed OZ treatment in pathogen suppression. EO had the highest sterilizing properties under commercial conditions and retained palatability and supported bee development better than other treatments. These results suggest that EO sterilization reduces pathogen risks while retaining pollen quality as a food source for rearing bumble bees. [ABSTRACT FROM AUTHOR]

Of the 265 known bumble bee (Bombus) species, knowledge of colony lifecycle is derived from relatively few species. As interest in Bombus commercialization and conservation grows, it is becoming increasingly important to understand colony growth dynamics across a variety of species since variation exists in nest success, colony growth, and reproductive output. In this study, we reported successful nest initiation and establishment rates of colonies and generated a timeline of colony development for 15 western North American Bombus species, which were captively reared from wild-caught gynes from 2009 to 2019. Additionally, we assessed variation in colony size among 5 western North American Bombus species from 2015 to 2018. Nest initiation and establishment rates varied greatly among species, ranging from 5–76.1% and 0–54.6%, respectively. Bombus griseocollis had the highest rates of nest success across the 11-yr period, followed by B. occidentalis , B. vosnesenskii , and B. huntii. Furthermore, days to nest initiation and days to nest establishment varied among species, ranging from 8.4 to 27.7 days and 32.7 to 47 days. Colony size also differed significantly among species with B. huntii and B. vosnesenskii producing more worker/drone cells than B. griseocollis , B. occidentalis , and B. vancouverensis. Additionally, gyne production differed significantly among species with B. huntii colonies producing more gynes than B. vosnesenskii. Results from this study increase knowledge of systematic nesting biology for numerous western North American Bombus species under captive rearing conditions, which can further improve rearing techniques available to conservationists and researchers. [ABSTRACT FROM AUTHOR]

Understanding historical range shifts and population size variation provides an important context for interpreting contemporary genetic diversity. Methods to predict changes in species distributions and model changes in effective population size (Ne) using whole genomes make it feasible to examine how temporal dynamics influence diversity across populations. We investigate Ne variation and climate‐associated range shifts to examine the origins of a previously observed latitudinal heterozygosity gradient in the bumble bee Bombus vancouverensis Cresson (Hymenoptera: Apidae: Bombus Latreille) in western North America. We analyze whole genomes from a latitude‐elevation cline using sequentially Markovian coalescent models of Ne through time to test whether relatively low diversity in southern high‐elevation populations is a result of long‐term differences in Ne. We use Maxent models of the species range over the last 130,000 years to evaluate range shifts and stability. Ne fluctuates with climate across populations, but more genetically diverse northern populations have maintained greater Ne over the late Pleistocene and experienced larger expansions with climatically favorable time periods. Northern populations also experienced larger bottlenecks during the last glacial period, which matched the loss of range area near these sites; however, bottlenecks were not sufficient to erode diversity maintained during periods of large Ne. A genome sampled from an island population indicated a severe postglacial bottleneck, indicating that large recent postglacial declines are detectable if they have occurred. Genetic diversity was not related to niche stability or glacial‐period bottleneck size. Instead, spatial expansions and increased connectivity during favorable climates likely maintain diversity in the north while restriction to high elevations maintains relatively low diversity despite greater stability in southern regions. Results suggest genetic diversity gradients reflect long‐term differences in Ne dynamics and also emphasize the unique effects of isolation on insular habitats for bumble bees. Patterns are discussed in the context of conservation under climate change. [ABSTRACT FROM AUTHOR]

The acute decline in global biodiversity includes not only the loss of rare species, but also the rapid collapse of common species across many different taxa. The loss of pollinating insects is of particular concern because of the ecological and economic values these species provide. The western bumble bee (Bombus occidentalis) was once common in western North America, but this species has become increasingly rare through much of its range. To understand potential mechanisms driving these declines, we used Bayesian occupancy models to investigate the effects of climate and land cover from 1998 to 2020, pesticide use from 2008 to 2014, and projected expected occupancy under three future scenarios. Using 14,457 surveys across 2.8 million km2 in the western United States, we found strong negative relationships between increasing temperature and drought on occupancy and identified neonicotinoids as the pesticides of greatest negative influence across our study region. The mean predicted occupancy declined by 57% from 1998 to 2020, ranging from 15 to 83% declines across 16 ecoregions. Even under the most optimistic scenario, we found continued declines in nearly half of the ecoregions by the 2050s and mean declines of 93% under the most severe scenario across all ecoregions. This assessment underscores the tenuous future of B. occidentalis and demonstrates the scale of stressors likely contributing to rapid loss of related pollinator species throughout the globe. Scaled-up, international species-monitoring schemes and improved integration of data from formal surveys and community science will substantively improve the understanding of stressors and bumble bee population trends. [ABSTRACT FROM AUTHOR]

Comprehensive decisions on the management of commercially produced bees, depend largely on associated knowledge of genetic diversity. In this study, we present novel microsatellite markers to support the breeding, management, and conservation of the blue orchard bee, Osmia lignaria Say (Hymenoptera: Megachilidae). Native to North America, O. lignaria has been trapped from wildlands and propagated on-crop and used to pollinate certain fruit, nut, and berry crops. Harnessing the O. lignaria genome assembly, we identified 59,632 candidate microsatellite loci in silico, of which 22 were tested using molecular techniques. Of the 22 loci, 12 loci were in Hardy-Weinberg equilibrium (HWE), demonstrated no linkage disequilibrium (LD), and achieved low genotyping error in two Intermountain North American wild populations in Idaho and Utah, USA. We found no difference in population genetic diversity between the two populations, but there was evidence for low but significant population differentiation. Also, to determine if these markers amplify in other Osmia , we assessed 23 species across the clades apicata , bicornis , emarginata , and ribifloris. Nine loci amplified in three species/subspecies of apicata , 22 loci amplified in 11 species/subspecies of bicornis , 11 loci amplified in seven species/subspecies of emarginata , and 22 loci amplified in two species/subspecies of ribifloris. Further testing is necessary to determine the capacity of these microsatellite loci to characterize genetic diversity and structure under the assumption of HWE and LD for species beyond O. lignaria. These markers will inform the conservation and commercial use of trapped and managed O. lignaria and other Osmia species for both agricultural and nonagricultural systems. [ABSTRACT FROM AUTHOR]

Commercialized bumble bees (Bombus) are primary pollinators of several crops within open field and greenhouse settings. However, the common eastern bumble bee (Bombus impatiens Cresson, 1863) is the only species widely available for purchase in North America. As an eastern species, concerns have been expressed over their transportation outside of their native range. Therefore, there is a need to identify regionally appropriate candidates for commercial crop pollination services, especially in the western U.S.A. In this study, we evaluated the commercialization potential of brown-belted bumble bees (Bombus griseocollis De Geer, 1773), a broadly distributed species throughout the U.S.A. by assessing nest initiation and establishment rates of colonies produced from wild-caught gynes, creating a timeline of colony development, and identifying lab-reared workers' critical thermal maxima (CTMax) and lethal temperature (ecological death). From 2019 to 2021, 70.6% of the wild-caught B. griseocollis gynes produced brood in a laboratory setting. Of these successfully initiated nests, 74.8% successfully established a nest (produced a worker), providing guidance for future rearing efforts. Additionally, lab-reared workers produced from wild-caught B. griseocollis gynes had an average CTMax of 43.5°C and an average lethal temperature of 46.4°C, suggesting B. griseocollis can withstand temperatures well above those commonly found in open field and greenhouse settings. Overall, B. griseocollis should continue to be evaluated for commercial purposes throughout the U.S.A. [ABSTRACT FROM AUTHOR]

In June of 2013 an application of dinotefuran on an ornamental planting of European linden trees (Tilia cordata Mill. [Malvales: Malvalceae]) in a shopping mall parking lot in Wilsonville, Oregon provoked the largest documented pesticide kill of bumble bees in North America. Based on geographic information systems and population genetic analysis, we estimate that between 45,830 and 107,470 bumble bees originating from between 289 and 596 colonies were killed during this event. Dinotefuran is a neonicotinoid that is highly effective in exterminating and/or harming target pest insects and non-target beneficial insects. Analysis to detect the concentration of pesticides in flowers that received foliar application revealed that the minimum reported dinotefuran concentration of a sampled T. cordata flower was 7.4 ppm, or in excess of 737% above the LC50 of the beneficial pollinator, the honey bee (Apis mellifera Linnaeus, 1758 [Hymenoptera: Apidae]). Furthermore, sampled Vosnesensky bumble bees (Bombus vosnesenskii Radoskowski, 1862 [Hymenoptera: Apidae]) were found to have an average dinotefuran concentration of 0.92 ppm at the time of death, which exceeds the maximum LC50 of A. mellifera (0.884 ppm). Our study underscores the lethal impact of the neonicotinoid pesticide dinotefuran on pollinating insect populations in a suburban environment. To our knowledge, the documentation and impact of pesticide kills on wild populations of beneficial insects has not been widely reported in the scientific literature. It is likely that the vast majority of mass pesticide kills of beneficial insects across other environments go unnoticed and unreported. [ABSTRACT FROM AUTHOR]

Pollinators play pivotal roles in maintaining agricultural and natural plant communities, yet some bee populations are declining. Loss of pollinator habitats as a result of agricultural intensification and urbanization have reduced bee abundance and diversity. Additionally, climate change has affected bee distributions and led to disruption of plant-pollinator synchrony, impacting ecosystem processes. However, how these factors concurrently influence bee assemblages is poorly understood. Therefore, we evaluated bumble bee (Bombus) assemblages in relation to the proportion of agricultural, semi-natural, and urban land cover and interannual variation in temperature, precipitation, and relative humidity in Utah agroecosystems using Bombus captured as bycatch in pest monitoring traps from 2014 to 2018. Bombus assemblage composition was highest in agricultural sites with increased agricultural land cover in the surrounding area, low temperatures, and high relative humidity during the growing season; and lowest in sites with increased urban land cover, high temperatures, and low relative humidity. Functional dispersion did not differ among these groups indicating a range of tongue lengths, body sizes, hair lengths, and hair types were present within all agricultural sites. Further, high beta-diversity, as indicated by unique Bombus assemblages among sites, suggests that all agricultural sites in this study have potential conservation value for maintaining Bombus communities. Therefore, it is important that diverse habitats for pollinators are maintained through targeted management techniques. Additionally, our collection of Bombus from mid-May to mid-September identified phenological overlap within Bombus assemblages, which helps ensure pollination services are provided even if a particular species is lost due to environmental disturbances. However, while there is overlap in functional traits and phenology, considerations should be given due to widespread pollinator declines. Overall, evaluating landscape and climate variables together may yield more realistic results and better inform effective management and land-use planning strategies to prevent ecological homogenization and to foster future resiliency of Bombus populations. • The combined effects of land cover and climate impact Bombus in agroecosystems. • Bombus composition was highest with crop land, low temperatures and high humidity. • Bombus composition was lowest with urban land, high temperatures and low humidity. • Phenological overlap among species provides functional redundancy. • High Bombus turnover suggests all surveyed sites have conservation value. [ABSTRACT FROM AUTHOR]