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Simple Summary: Working in lowbush blueberry fields, we studied how to provide a holistic measure of pesticide exposure to bees. Several different taxa of bees can be involved in blueberry pollination, including honey bees, bumble bees (which are native), and other native bees, such as mason, mining, and leafcutter bees. Using chemical analysis, we compared and contrasted samples from different sources to (1) measure pesticide exposure and (2) predict exposure risk in light of established levels of concern established by the U.S. Environmental Protection Agency. We trapped honey bee pollen as foragers entered the hive and, within the three years of the study, found considerable similarity of these samples to treated flowers; the measures were the number and concentration of pesticide residues, as well as the predicted level of risk. We found differences among two or three of these measures when comparing trapped honey bee pollen and trapped mason bee pollen, as well as between trapped honey bee pollen and whole bodies of captured honey bees. In these comparisons, the number of residues detected in honey bee pollen was typically greater, as was the estimated risk. The residues found when comparing whole body analysis of honey bee workers, bumble bee queens, and other native bees showed similarity in total residue concentrations and risk, but there was a higher number of different residues detected on honey bees. This research complements another study (Part I) focused on honey bee pollen trapping in lowbush blueberry and cranberry. Both studies provide a toxicological baseline for pesticide residue exposure to pollinators in the lowbush blueberry agroecosystem. In lowbush blueberry fields, we conducted residue analysis comparing flowers, trapped pollen (honey bee and Osmia spp.), and collected bees (honey bee workers, bumble bee queens, and non-Bombus spp. wild native bees). The study was conducted from 2012 to 2014. The number of pesticide residues, total concentrations, and risk to honey bees (Risk Quotient) on flowers were not significantly different from those determined for trapped honey bee pollen (except in one study year when residues detected in flower samples were significantly lower than residue numbers detected in trapped pollen). The compositions of residues were similar on flowers and trapped pollen. The number of residues detected in honey bee pollen was significantly greater than the number detected in Osmia spp. pollen, while the total concentration of residue was not different between the two types of pollen. The risk to honey bees was higher in trapped honey bee pollen than in trapped Osmia spp. pollen. The analysis of honey bee workers, native bumble bee queens, and native solitary bees showed that although more pesticide residues were detected on honey bee workers, there were no differences among the bee taxa in total residue concentrations or risk (as estimated in terms of risk to honey bees). [ABSTRACT FROM AUTHOR]

Simple Summary: Bumblebee species are crucial pollinators for both crops and natural ecosystems. Managing diversity and securing the services they provide requires sufficient data to manage these systems effectively. In this study, seventeen environmental factors were selected to understand their contribution to shaping species assemblages and to provide baseline information for understanding drivers of community assembly. The study focused on Gansu Province because it has some of the highest richness of bumblebees within China, and China represents a major data gap for bee distribution data. Net primary productivity followed by water vapor pressure were top-ranked drivers of species distributions. The results enhance our understanding of the importance of environmental factors in shaping bumblebee community assemblages. Understanding the influence of factors responsible for shaping community assemblage is crucial for biodiversity management and conservation. Gansu is one of the richest regions for bumblebee species in the world. We explored the distribution data of 52 bumblebee species collected in Gansu and its surroundings between 2002 and 2022, predicting habitat suitability based on 17 environmental variables using MaxEnt. The factors influencing community assemblage were assessed using canonical correspondence analysis. Net primary productivity, water vapor pressure, temperature seasonality, annual precipitation, and precipitation seasonality were some of the most influential drivers of species distributions. Based on Ward's agglomerative cluster analysis, four biogeographic zones are described: the Southern humid zone, the Western Qilian snow mountain zone, the Eastern Loess plateau zone, and the Western dry mountain zone. In the clusters of grid cells based on beta diversity values, the Southern humid zone comprised 42.5% of the grid cells, followed by the Eastern Loess plateau zone (32.5%), the Western dry mountain zone (20%), and the Western Qilian snow mountain zone (5%). Almost all the environmental factors showed a significant contribution to the assemblages of bumblebees of different groups. Our findings highlight the need for better data to understand species biogeography and diversity patterns, and they provide key baseline data for refining conservation strategies. [ABSTRACT FROM AUTHOR]

Simple Summary: Bumblebees are well-suited pollinators of economic crops. This study evaluated the foraging behavior and pollination effects of Bombus terrestris on cherry tomatoes and pears in northern China. B. terrestris pollination can improve cultivation efficiency, increase yield, and produce more economic benefits, which indicates that it has great potential in pollination applications for cherry tomatoes and pears in northern China. Bumblebees (Bombus terrestris) have strong environmental adaptability and high pollen transfer efficiency, making them well-suited pollinators of economic crops. However, bumblebee pollination is still not widely applied in northern China due to the lack of data on foraging behavior and pollination effects. We conducted a three-year experiment involving cherry tomatoes (Solanum lycopersicum L.) and pears (Pyrus spp.) treated with bumblebee pollination to evaluate the foraging behavior and pollination effects on these two crops. Results showed that B. terrestris had enhanced foraging activities as daytime temperatures rose from 18 °C to 26 °C, as indicated by the increased number of bees leaving the hive and returning bees carrying pollen in greenhouses in winter. There were two peaks in the foraging activity of bumblebees in pear orchards in early spring, which was closely related to the temperature change in the daytime. Undoubtedly, cherry tomatoes treated with B. terrestris had higher fruit setting rate, weight, seed number, and fruit yields compared to those with hormone 2,4-dichlorophenoxyacetic acid treatments, as well as a lower rate of deformed fruits. B. terrestris pollination can significantly increase the fruit setting rate and fruit yield of pears, compared with open pollination, and can fully achieve the effect of hand pollination. B. terrestris pollination can improve cultivation efficiency, increase yield, and produce more economic benefits. Moreover, it can also contribute to reducing hormone residues and ensure the safety of agricultural products. We recommend its application to cherry tomatoes in greenhouses in winter and potential application to pears in orchards in early spring in northern China. However, the risk to local bumblebee species of introducing commercially available bumblebees into orchards should be considered and evaluated in future research. This study provides both empirical support and a theoretical basis for the selection of bumblebees as pollinators in the production of economically important crops and the improvement of crop cultivation management in northern China. [ABSTRACT FROM AUTHOR]

Simple Summary: Our study investigated the dynamics of pathogen communities within Chinese bumblebee populations, a key issue for understanding emerging infectious diseases and their impacts on insect biodiversity loss. By employing computational modeling on extensive pathogen data from bumblebees, we uncover that the host species variation significantly influences pathogen assembly and occurrences, compared to environmental factors like climate and location. Moreover, our results reveal significant pathogen–pathogen interactions, with similar pathogens exhibiting facilitatory relationships and distinct pathogen types showing strong negative associations, possibly due to immune response interactions and competitive dynamics within hosts. Our findings highlight the role of host–pathogen coevolution and other ecological interactions in shaping pathogen communities. The implications of this work are substantial for bumblebee conservation, improving our understanding of pathogen dynamics that could inform strategies to stop declines in bumblebee populations. Bumblebees have been considered one of the most important pollinators on the planet. However, recent reports of bumblebee decline have raised concern about a significant threat to ecosystem stability. Infectious diseases caused by multiple pathogen infections have been increasingly recognized as an important mechanism behind this decline worldwide. Understanding the determining factors that influence the assembly and composition of pathogen communities among bumblebees can provide important implications for predicting infectious disease dynamics and making effective conservation policies. Here, we study the relative importance of biotic interactions versus interspecific host resistance in shaping the pathogen community composition of bumblebees in China. We first conducted a comprehensive survey of 13 pathogens from 22 bumblebee species across China. We then applied joint species distribution modeling to assess the determinants of pathogen community composition and examine the presence and strength of pathogen–pathogen associations. We found that host species explained most of the variations in pathogen occurrences and composition, suggesting that host specificity was the most important variable in predicting pathogen occurrences and community composition in bumblebees. Moreover, we detected both positive and negative associations among pathogens, indicating the role of competition and facilitation among pathogens in determining pathogen community assembly. Our research demonstrates the power of a pluralistic framework integrating field survey of bumblebee pathogens with community ecology frameworks to understand the underlying mechanisms of pathogen community assembly. [ABSTRACT FROM AUTHOR]

Simple Summary: Several lab studies have shown that pesticides are causing sublethal effects on bees; however, little is known about their impact in actual field conditions. Continuous surveillance of the bees in the field will help monitor their pesticide exposure and better understand their impact on bee health. This study investigated the potential use of a new optical sensor, coupled with machine learning, to automatically identify flying insects, as an approach to monitoring the sublethal effects of pesticides on insects. To do so, bumblebees, Bombus terrestris, were exposed to field-realistic doses of a neonicotinoid pesticide, and their flight activity was recorded by the sensor. Through the data acquired, the investigation focused on whether the machine learning algorithm could distinguish the flight events that were created by the healthy bumblebees and the bumblebees that had been exposed to sublethal doses of the pesticide. The results showed that the algorithm could differentiate flight events, indicating the promise of the sensor as a continuous monitoring tool for bee health in the field. Among insects, bees are important pollinators, providing many vital ecosystem services. The recent pollinator decline is threatening both their diversity and abundance. One of the main drivers of this decline is the extensive use of pesticides. Neonicotinoids, one of the most popular groups of pesticides, can be toxic to bees. In fact, numerous studies have found that neonicotinoids can cause sublethal effects, which can impair the biology, physiology, and colony survival of the bees. Yet, there are still knowledge gaps, and more research is needed to better understand the interaction between neonicotinoids and bees, especially in the field. A new optical sensor, which can automatically identify flying insects using machine learning, has been created to continuously monitor insect activity in the field. This study investigated the potential use of this sensor as a tool for monitoring the sublethal effects of pesticides on bumblebees. Bombus terrestris workers were orally exposed to field-realistic doses of imidacloprid. Two types of exposures were tested: acute and chronic. The flight activity of pesticide-exposed and non-exposed bumblebees was recorded, and the events of the insect flights recorded by the sensor were used in two ways: to extract the values of the wingbeat frequency and to train machine learning models. The results showed that the trained model was able to recognize differences between the events created by pesticide-exposed bumblebees and the control bumblebees. This study demonstrates the possibility of the optical sensor for use as a tool to monitor bees that have been exposed to sublethal doses of pesticides. The optical sensor can provide data that could be helpful in managing and, ideally, mitigating the decline of pollinators from one of their most major threats, pesticides. [ABSTRACT FROM AUTHOR]

Simple Summary: Honey bees are famous for their capacity to precisely time their visits to flowers to maximize food reward, but it is not known whether similar "time-memory" exists in other bees that forage over shorter distances from their nests. Here, we tested whether bumble bees that live in smaller colonies can associate reward with time of day and color. We trained bumble bee workers to feed on yellow or blue artificial flowers during either the morning or evening, respectively. During the test day, we presented non-rewarding flowers and recorded the behavior of the foraging bees. We found that trained bees preferred yellow flowers during the time corresponding to the morning training time and blue flowers during the time corresponding to the evening training time. These observations show that bumble bees can associate time of day with a specific color and reward, suggesting that time-memory is not limited to species such as honey bees that forage over long distances and time periods. Circadian clocks regulate ecologically important complex behaviors in honey bees, but it is not clear whether similar capacities exist in other species of bees. One key behavior influenced by circadian clocks is time-memory, which enables foraging bees to precisely time flower visitation to periods of maximal pollen or nectar availability and reduces the costs of visiting a non-rewarding flower patch. Bumble bees live in smaller societies and typically forage over shorter distances than honey bees, and it is therefore not clear whether they can similarly associate reward with time of day. We trained individually marked bumble bee (Bombus terrestris) workers to forage for sugar syrup in a flight cage with yellow or blue feeders rewarding either during the morning or evening. After training for over two weeks, we recorded all visitations to colored feeders filled with only water. We performed two experiments, each with a different colony. We found that bees tended to show higher foraging activity during the morning and evening training sessions compared to other times during the day. During the test day, the trained bees were more likely to visit the rewarding rather than the non-rewarding colored feeders at the same time of day during the test sessions, indicating that they associated time of day and color with the sugar syrup reward. These observations lend credence to the hypothesis that bumble bees have efficient time-memory, indicating that this complex behavior is not limited to honey bees that evolved sophisticated social foraging behaviors over large distances. [ABSTRACT FROM AUTHOR]

Simple Summary: Insects can perceive and use polarized light in the sky when navigating by using a special region in their eyes called the dorsal rim area (DRA). The size and characteristics of the DRA can differ among insect species but, until now, there has been no effective method to investigate and compare these differences. We used a combination of 2D photography and X-ray imaging to determine the area of the DRA and demonstrated this method in the buff-tailed bumblebee. We showed that the DRA could be reliably obtained using 2D photographs and we found that the bumblebee body size affected the DRA. Using X-ray imaging, we also showed that the crystalline cones, which are light-focusing structures, were smaller in the DRA compared to the rest of the bumblebee eye. Our method was time-efficient, non-destructive, and could be used to determine photoreceptor characteristics at an individual level. This method will allow for more in-depth studies of the DRA in insects. Polarized light-based navigation in insects is facilitated by a polarization-sensitive part of the eye, the dorsal rim area (DRA). Existing methods to study the anatomy of the DRA are destructive and time-consuming. We presented a novel method for DRA localization, dissection, and measurement using 3D volumetric images from X-ray micro-computed tomography in combination with 2D photographs. Applying the method on size-polymorphic buff-tailed bumblebees, Bombus terrestris, we found that the DRA was easily obtainable from photographs of the dorsal eye region. Allometric analysis of the DRA in relation to body size in B. terrestris showed that it increased with the body size but not at the same rate. By localizing the DRA of individual bumblebees, we could also perform individual-level descriptions and inter-individual comparisons between the ommatidial structures (lens, crystalline cones, rhabdoms) of three different eye regions (DRA, non-DRA, proximate to DRA). One feature distinct to the bumblebee DRA was the smaller dimension of the crystalline cones in comparison to other regions of the eye. Using our novel methodology, we provide the first individual-level description of DRA ommatidial features and a comparison of how the DRA varies with body size in bumblebees. [ABSTRACT FROM AUTHOR]

Simple Summary: Research on nutritional requirements in combination with improving bee protection has recently increased because of the continuous global decline in the abundance and diversity of bees. The decline is currently attributed to several stressors, including climate change, diminishing forage resources, and pesticide use, among other factors. Industrial hemp (Cannabis sativa L.) is a new crop that is grown over a wide geographic area in the United States, providing economic and nutritional benefits to humans. However, the contribution of hemp floral resources (pollen) to bee nutrition is not well understood. We investigated the chemical composition of pollen from four industrial hemp varieties (Canda, CFX-2, Henola, and Joey) and documented the abundance and diversity of bees on the crop using two sampling methods. Results showed differences in composition among the four hemp varieties. Overall, the Joey variety was the most preferred by bees, despite expressing lower protein, amino acid, and saturated and monosaturated fatty acid content. Based on our findings, we concluded that industrial hemp pollen provides some nutritional benefits to bees. However, it is important to understand that multiple sources of pollen are needed for sustained bee survival. Apart from its economic value, industrial hemp (Cannabis sativa L.) is a prolific pollen producer, serving as a food source for bees. However, little is known regarding the extent to which varietal differences in hemp pollen chemistry influences bee preference. Here, we report the chemical profile of pollen from four hemp varieties (Canda, CFX-2, Henola, and Joey) and bee abundance and diversity, using direct visual counts and pan traps. The number and type of bees on each variety was recorded and the chemical composition (proximate analysis and mineral, amino, and fatty acid profiles) of pollen from each hemp variety was determined. During the entire sampling period, three bee types (bumble bees, honey bees, and sweat bees) were recorded, with a combined total of 1826. Among these, sweat bees and bumble bees were the most prevalent and were highest on the Joey variety. The four varieties expressed protein content ranging from 6.05% to 6.89% and the highest in Henola. Seventeen amino acids were expressed in all varieties, with leucine recording the highest content ranging from 4.00 mg/g in Canda to 4.54 mg/g in Henola. In general, Henola expressed high protein, amino acid, and saturated and monosaturated fatty acid contents and recorded significantly fewer bees compared with Joey, which had a low content of these components and a high content of polyunsaturated fatty acids. Our findings suggest that, while industrial hemp offers abundant and accessible pollen that would promote bee health and sustainability of their ecosystem services, the nutritional quality may not be adequate for bee growth and development as an exclusive pollen source. [ABSTRACT FROM AUTHOR]

Simple Summary: Microsporidia are intracellular parasites that have detrimental effects on their various invertebrate and vertebrate hosts. The microsporidia pathogenic to honeybees and bumblebees belong to the genus Vairimorpha (previously Nosema). Among these, V. bombi is the most widespread species worldwide and it infects a variety of bumblebee species. In this study, we investigated the infection of V. bombi among Japanese native bumblebees and the alien species, Bombus terrestris, established in Japan, using PCR and microscopy. We discovered that V. bombi infected the subgenus Bombus s. str. species/subspecies with a low prevalence in areas both with and without the presence of B. terrestris, suggesting that V. bombi may be native to Japan. In contrast, a newly discovered microsporidian parasite that is phylogenetically distinct from V. bombi was found to infect the subgenus Diversobombus species/subspecies with a high prevalence. Hence, if the new microsporidian species does not negatively affect the host and is vertically transmitted from queen to offspring, it may coexist with its host. In addition, this may suggest that V. bombi and the new microsporidium have different life history strategies. Microsporidia are spore-forming intracellular parasites of various invertebrates and vertebrates. Vairimorpha bombi negatively affects the fitness of bumblebees and its prevalence correlates with declining bumblebee populations. The invasive alien species Bombus terrestris colonized Japan and possibly introduced new parasites. To assess the infection prevalence of V. bombi in Japanese bumblebees and B. terrestris, we investigated V. bombi infections using PCR and microscopy. The prevalence of sporulating V. bombi infections in three Bombus s. str. species/subspecies was low, whereas that of non/low-sporulating Vairimorpha sp. infections in three Diversobombus species/subspecies was high. Invasive B. terrestris showed low prevalence of non/low-sporulating V. bombi infections and shared the same V. bombi haplotype with B. hypocrita found in Hokkaido, where B. terrestris is present, and in Honshu, where B. terrestris is absent. Although V. bombi may have been introduced with B. terrestris colonies imported from Europe, it seems to be originally distributed in Japan. Furthermore, a new Vairimorpha sp. was found in Japanese bumblebee species. V. bombi and Vairimorpha sp. showed different organ and host specificities in bumblebees. There are no reports on the specific effects of other Vairimorpha spp. on bumblebees; further studies are needed to clarify the individual characteristics of Vairimorpha spp. [ABSTRACT FROM AUTHOR]

Simple Summary: Bumble bees live through two phases during their life cycle: single queens and colonies. Queens enter diapause after mating as a survival strategy to avoid harsh environmental conditions. The energy requirements of queens during diapause depend mainly on the nutrient reserves at the prediapause stage. Temperature acclimation can improve the queen's survival during diapause. Therefore, it is important to understand the effects of temperature acclimation on the prediapause nutritional reserves and energy consumption of bumble bee queens. In this study, we evaluated the effects of three acclimation temperatures and three acclimation times on the free water, protein, lipid, and total sugar reserves of bumble bee queens during prediapause and the energy consumption of queens during the 3-month diapause period. Overall, lower temperature acclimation increased queens' lipid reserves during prediapause and reduced the consumption of protein, lipids, and total sugars during diapause. Therefore, low-temperature acclimation could be more beneficial for bumble bee queens to improve cold resistance and increase reserves of major nutrient lipids during diapause. A queen's diapause is a key period of the bumble bee life cycle that enables them to survive under unfavorable conditions. During diapause, queens fast, and nutritional reserves depend on the accumulation of nutrients during the prediapause period. Temperature is one of the most important factors affecting queens' nutrient accumulation during prediapause and nutrient consumption during diapause. Here, we used a 6-day-old mated queen of the bumble bee Bombus terrestris to evaluate the effect of temperature (10, 15, and 25 °C) and time (3, 6, and 9 days) on free water, protein, lipids, and total sugars during prediapause and at the end of 3 months of diapause. Stepwise regression analysis revealed that total sugars, free water, and lipids were much more affected by temperature than protein (p < 0.05). Lower temperature acclimation significantly increased (p < 0.05) free water and lipid accumulation by queens during prediapause. In contrast, higher temperature acclimation significantly increased (p < 0.05) protein and total sugar accumulation by queens during prediapause. The effect of temperature acclimation on the queen survival rate was not significantly different (p > 0.05) after 3 months of diapause. Moreover, lower temperature acclimation reduced protein, lipid, and total sugar consumption by queens during diapause. In conclusion, low-temperature acclimation increases queens' lipid accumulation during prediapause and reduces the nutritional consumption of queens during diapause. Low-temperature acclimation during prediapause could benefit queens by improving cold resistance and increasing reserves of major nutrient lipids during diapause. [ABSTRACT FROM AUTHOR]

Simple Summary: Bumblebees, as important pollinating insects, play a vital role in maintaining natural and agricultural ecosystems. Its antennae with sensilla can guide in selecting a nectariferous source, locating nest sites, and communicating between individuals. To explore how bumblebees detect and receive chemical signals from nectariferous plant and foraging behavior, the morphology of antennae with sensilla, including antennal length and type, distribution, and number of antennal sensilla were compared in four bumblebee species, Bombus atiapes, Bombus breviceps, Bombus flavescens, and Bombus terrestris through scanning electron microscopy. The characteristics and differences in the total antennal length, and length of scape, pedicel, and flagellum were recorded among three castes and four species. Furthermore, 13 major types of antennal sensilla in total were observed, including trichodea sensilla (TS A-E), placodea sensilla (PS A-B), basiconica sensilla (BaS), coeloconica sensilla (COS A-B), chaetic sensilla (CS A-B), and Böhm sensilla (BS), of which chaetic sensilla B (CS B), only observed in females of B. atripes, was firstly reported in Apidae. The characteristics and differences in the distributions and number of sensilla were also found among three castes and four species. Therefore, this study may help to understand the effects of antennae with sensilla in the coevolution of species and the external environment. Bumblebees, as pollinators, play an important role in maintaining natural and agricultural ecosystems. Antennae with sensilla of bumblebees as social insects have essential effects in foraging, nest searching, courting, and mating, and are different in species and sexes. Previous studies on the morphology of antennae and sensilla in bumblebees have been limited to a few species and a single caste. To better understand how bumblebees detect and receive the chemical signal from nectariferous plants and foraging behavior, the morphology of antennae with sensilla, including the antennal length, and type, distribution, and number of antennal sensilla in four species, Bombus atripes, Bombus breviceps, Bombus flavescens, and Bombus terrestris was compared by scanning electron microscopy (SEM) herein. The total antennal length of queens are the longest and workers are the shortest in three castes, and in four species the longest of the total antennal length among three castes all are in B. flavescens, which is significantly longer than other species (p < 0.05) and the length of the scape in queens and workers are both longer than males, significantly different in queens (p < 0.05), and not significantly different in workers (p > 0.05), and the length of flagellums in females are not always shorter than males, of which the length of flagellms in queens of B. flavescens are significantly longer than males (p < 0.05), and the length of pedicel and all flagellomeres varies among species and castes. A total of 13 major types of sensilla in total were observed, including trichodea sensilla (TS A-E), placodea sensilla (PS A-B), basiconica sensilla (BaS), coeloconica sensilla (COS A-B), chaetic sensilla (CS A-B), and Böhm sensilla (BS), of which chaetic sensilla B (CS B), only observed in females of B. atripes, was firstly reported in Apidae. Moreover, the number of all sensilla was the most in males, the least was in workers, the number of sensilla varies within castes and species. Furthermore, the morphological characteristics of antennae and the potential functions of sensilla are discussed. [ABSTRACT FROM AUTHOR]

Simple Summary: Bombus breviceps is an important native species of bumblebee in southern China. As a pollinator, it has the potential for domestication and commercial rearing. In this study, in order to identify the nutritional requirements for the egg-laying, hatching, and colony foundation stages, we selected three common local types of pollen, including camellia pollen, oilseed rape pollen, and apricot pollen, as well as mixtures of them, to feed B. breviceps queens to evaluate the reproductive performance, and feasibility of these pollens as a diet and the nutritional requirements. The results showed that the pollen with a higher essential amino acid content had advantages in laying time, the number of eggs laid, larval ejection, and the time of first worker emergence, while the pollen with a higher crude protein content had advantages in the time the colony took to reach ten workers. These results may guide the selection of feed in B. breviceps artificial feeding and help to explore the nutritional requirements in oviposition and the colony stage from the perspective of conventional pollen. Pollen is an important source of nutrition for bumblebees to survive, reproduce, and raise their offspring. To explore the nutritional requirements for the egg laying and hatching of queenright Bombus breviceps colonies, camellia pollen, oilseed rape pollen, apricot pollen, and mixtures of two or three types of pollen in equal proportions were used to feed the queens in this study. The results showed that the camellia pollen with a higher essential amino acid content was superior to the pollen with a lower essential amino acid content in the initial egg-laying time (p < 0.05), egg number (p < 0.05), larval ejection (p < 0.01), time of first worker emergence (p < 0.05), and the average weight of workers in the first batch (p < 0.01). It took less time for colonies under the camellia pollen and camellia–oilseed rape–apricot pollen mix treatments, both with a higher crude protein content, to reach ten workers in the colony (p < 0.01). On the contrary, the queens fed apricot pollen never laid an egg, and larvae fed oilseed rape pollen were all ejected—both pollens with a lower essential amino acid content. The results emphasize that the diet should be rationally allocated to meet the nutritional needs of local bumblebees at various stages when guiding them to lay eggs, hatch, and develop a colony. [ABSTRACT FROM AUTHOR]

Simple Summary: Why do plants have showy flowers? Clearly, many plant species must attract pollinators and have floral adaptations for this. However, some flowers attract insects without showy petals. This suggests that a key function of showy visual signals is to attract naïve, first-time pollinator visitors. This is similar to how a restaurant with a large sign and showy visual signals may be especially important in gaining first-time visits when competing with other establishments or plants. Customers or pollinators will know if the first visit is rewarding and use this to decide whether to revisit. First, they must visit. Most flowers and restaurants would benefit from more visitors. Restaurants usually have empty tables, and many flowers are pollen limited in their reproduction. Here, we manipulated the ray petals of inflorescences of two garden flowers to test the hypothesis that the role of showy visual signals is to attract naïve visitors. On their first inflorescence visit to both species, naïve honey bees and bumble bees were more likely to visit intact inflorescences, than those with ray petals removed. However, by the tenth consecutive inflorescence, bees showed no preference. A positive correlation was observed between the visitation of inflorescences with no petals and the inflorescence number on both study plants, for both bees. These results strongly suggest that a key function of showy petals is to attract naïve, first-time visitors. Flower evolution includes a range of questions concerning the function of showy morphological features such as petals. Despite extensive research on the role of petals in attracting pollinators, there has been little experimental testing of their importance in attracting naïve versus experienced flower-visitors. In an exploratory field study, we manipulated the ray petals of inflorescences of two garden flowers, Rudbeckia hirta and Helenium autumnale, to test the hypothesis that these showy structures primarily function to attract first-time, naïve, visitors. On their first inflorescence visit to both species, naïve honey bees and bumble bees were more likely to visit intact inflorescences, than those with ray petals removed. However, by the tenth consecutive inflorescence on the same visit to the flower patch, test insects showed no preference. A positive correlation was observed between the visitation of inflorescences with zero petals and inflorescence number on both study plants, for both bees. These results suggest that a key function of showy petals is to attract naïve, first-time visitors. Similar to how a restaurant attracts diners with a large sign, showy signals may be vital to enticing first-time visitors when competing with other establishments or plants for customers or pollinators. We hope the findings of this exploratory study will stimulate further work in this area. [ABSTRACT FROM AUTHOR]

Simple Summary: Determining the adaptive mechanisms by which bumblebees adapt to high altitudes can help us to better understand their distribution, providing a basis for the future protection and utilization of bumblebee resources. For this study, the adaptive mechanisms of two dominant bumblebee species in the northeastern Qinghai-Tibet Plateau—Bombus kashmirensis and B. waltoni—were studied through transcriptomics methods. For each species, enrichment analysis of the differentially expressed genes and gene set enrichment analysis were carried out between samples collected at different altitudes (4000 m, 4500 m, and 5000 m). The results indicate that these bumblebees tend to up-regulate energy metabolism-related genes when facing extremely high-altitude environments. Of the enriched pathways up-regulated in higher altitudes, the pentose and glucuronate interconversions pathway presented the most severe up-regulation in multiple comparisons of different altitudes for B. kashmirensis, as well as the AMPK signaling pathway, which was found to be up-regulated in both species. Notably, limited by the extreme high altitudes in this study, oxidative phosphorylation was found to be down-regulated with increasing altitude, which is uncommon in studies on bumblebee adaptation to high altitudes. Determining the adaptive mechanisms by which bumblebees adapt to high altitudes can help us to better understand their distribution, providing a basis for the future protection and utilization of bumblebee resources. For this study, the adaptive mechanisms of two dominant bumblebee species in the northeastern Qinghai-Tibet Plateau—Bombus kashmirensis and B. waltoni—were studied through transcriptomics methods. For each species, enrichment analysis of the differentially expressed genes and gene set enrichment analysis were carried out between samples collected at different altitudes (4000 m, 4500 m, and 5000 m). The results indicate that these bumblebees tend to up-regulate energy metabolism-related genes when facing extremely high-altitude environments. Of the enriched pathways up-regulated in higher altitudes, the pentose and glucuronate interconversions pathway presented the most severe up-regulation in multiple comparisons of different altitudes for B. kashmirensis, as well as the AMPK signaling pathway, which was found to be up-regulated in both species. Notably, limited by the extreme hypoxic conditions in this study, oxidative phosphorylation was found to be down-regulated with increasing altitude, which is uncommon in studies on bumblebee adaptation to high altitudes. [ABSTRACT FROM AUTHOR]

Simple Summary: In recent decades, there has been a significant global decline in pollinators. In addition to the honey bee, it is bumblebees that contribute significantly to the pollination of many wild and farm plants. Many factors, such as habitat loss, climate change, pesticides and pathogens, are contributing to the decline of bumblebee populations. The main parasites of bumblebees include Crithidia bombi, Apicystis bombi and Nosema bombi. In our study, we aimed to obtain the first knowledge of their occurrence in the Czech Republic in the two most abundant bumblebee species (buff-tailed bumblebee Bombus terrestris and red-tailed bumblebee Bombus lapidarius). More than half of the captured buff-tailed bumblebee individuals were infected by C. bombi, less than a quarter of N. bombi individuals and the least by A. bombi. Red-tailed bumblebee individuals were infected less frequently. Surprisingly, more individuals were infected with all three parasite species than with only the combination of N. bombi and A. bombi. Parasite infection is also influenced by the environment. More individuals were infected with C. bombi in urban and forested landscapes than in grasslands and agricultural landscapes. In turn, a higher incidence of N. bombi was found around greenhouses where commercially produced bumblebees were used. Bumblebees are significant pollinators for both wild plants and economically important crops. Due to the worldwide decrease in pollinators, it is crucial to monitor the prevalence and distribution of bumblebee pathogens. Field-caught bumblebee workers and males were examined for the presence of three pathogens during the summer months of the years 2015–2020 (Bombus terrestris/lucorum) and 2015–2017 (Bombus lapidarius). The greatest prevalence was in the case of Crithidia bombi, where significantly more workers (57%) of B. terrestris/lucorum were infected than males (41%). Infection was also confirmed in 37% of B. lapidarius workers. The average prevalence was very similar in the case of Nosema bombi in workers (25%) and males (22%) of B. terrestris/lucorum. In the case of B. lapidarius, 17% of the workers were infected. The lowest number of infected individuals was for Apicystis bombi and the prevalence of infection was significantly higher for males (22%) than workers (8%) of B. terrestris/lucorum. Only 3% of workers and 4% of males of B. terrestris/lucorum were simultaneously infected with three types of pathogens, but no worker was infected with only a combination of N. bombi and A. bombi. The greatest prevalence of C. bombi was found in urban or woodland areas. [ABSTRACT FROM AUTHOR]

The findings of this study revealed that honey bees and bumble bees had different EAG responses to tomato flower volatiles, and the two bee species showed different behavioral choices to the same compound. It has been widely known that bumble bees are effective bee pollinators of tomato as they are capable of buzz pollination to vibrate tomato anthers in order to release pollen grains, and only one visit is sufficient for full tomato pollination [[43]]. Keywords: Apis mellifera; Bombus terrestris; EAG; behavior; tomato flower volatiles EN Apis mellifera Bombus terrestris EAG behavior tomato flower volatiles 987 12 11/17/22 20221101 NES 221101 1. Bees play an important role in the pollination of tomatoes, yet the relationship between foraging choices and olfactory cues is unclear because of the difference in foraging behavior between honey bees and bumble bees on tomato flowers. [Extracted from the article]

Simple Summary: Bumble bees have annual colonies that produce new queens at the end of summer. They mate and accumulate substantial fat reserves before overwintering underground prior to emerging in the spring. Because body size influences how much fat a queen can accumulate in the fall and how quickly she burns through those store in the winter, we asked if queen body condition influences the timing of spring emergence. We measured mass and body condition (indicative of remaining fat reserves) of bumble bee queens in the spring. Large queens and those in better body condition emerged throughout the spring, whereas lighter queens and those with poorer body condition emerged earlier, suggesting that depleted fat reserves could trigger early emergence of bumble bee queens. For insects, the timing of many life history events (phenology) depends on temperature cues. Body size is a critical mediator of insect responses to temperature, so may also influence phenology. The determinants of spring emergence of bumble bee queens are not well understood, but body size is likely important for several reasons. In fall, queens accumulate energy stores to fuel overwinter survival. Accumulation of fat stores prior to and depletion of fat stores during overwintering are likely size-dependent: larger queens can accumulate more lipids and have lower mass-specific metabolic rates. Therefore, larger queens and queens in relatively better condition may have delayed depletion of energy stores, allowing for later spring emergence. To test whether timing of spring emergence is associated with body size and condition, we captured 295 Bombus huntii queens in Laramie, WY, during the 2020 and 2021 growing seasons, weighed them, and measured intertegular width (a size metric unaffected by variation in feeding and hydration state). Early emerging queens were smaller than later emerging queens across years. Mass relative to intertegular width increased as the season progressed suggesting, as predicted, that body condition influences the timing of spring emergence for these crucial pollinators. [ABSTRACT FROM AUTHOR]

Assuming that some wild bee populations are resource limited [[56]], the possible competition with early arriving bumble bees (as well as honey bees) may imply that some native bees need to work harder and invest more time and energy in order to collect the proteins (i.e., pollen) needed for provisioning their offspring. Bumble bees visited both pollen- and nectar-rewarding flowers earlier than the wild bees, and the pollen-rewarding flowers earlier than the carpenter bees, but with no significant difference compared to the honey bee for both type of flowers (Table 4). Bumble bees (and to lesser extent honey bees) arrive to pollen-rewarding flowers earlier in the morning and have the opportunity to deplete pollen rewards before the later arrival of the local wild bees ( I Xylocopa i and other wild bee taxa). [Extracted from the article]

We conducted a bee survey in neonicotinoid-treated commercial potato fields using bowl and vane traps in the 2016 growing season. Traps were placed outside the fields, at the field edges, and 10 and 30 m into the fields. We collected 756 bees representing 58 species, with Lasioglossum spp. comprising 73% of all captured bees. We found seven Bombus spp., of which B. impatiens was the only known visitor of potato flowers in our region. The majority of the bees (68%) were collected at the field edges and in the field margins. Blue vane traps caught almost four-times as many bees and collected 30% more species compared to bowl traps. Bee communities did not differ across trap locations but they were different among trap types. We tested B. impatiens visitation to neonicotinoid treated and untreated potato flowers in field enclosures. The amount of time bees spent at flowers and the duration of visits were not significantly different between the two treatments. Our results demonstrate that a diverse assemblage of bees is associated with an agroecosystem dominated by potatoes despite the apparent lack of pollinator resources provided by the crop. We found no difference in B. impatiens foraging behavior on neonicotinoid-treated compared to untreated plants. [ABSTRACT FROM AUTHOR]

Bees provide vital pollination services to the majority of flowering plants in both natural and agricultural systems. Unfortunately, both native and managed bee populations are experiencing declines, threatening the persistence of these plants and crops. Agricultural chemicals are one possible culprit contributing to bee declines. Even fungicides, generally considered safe for bees, have been shown to disrupt honey bee development and impair bumble bee behavior. Little is known, however, how fungicides may affect bumble bee colony growth. We conducted a controlled cage study to determine the effects of fungicide exposure on colonies of a native bumble bee species (Bombus impatiens). Colonies of B. impatiens were exposed to flowers treated with field-relevant levels of the fungicide chlorothalonil over the course of one month. Colony success was assessed by the number and biomass of larvae, pupae, and adult bumble bees. Bumble bee colonies exposed to fungicide produced fewer workers, lower total bee biomass, and had lighter mother queens than control colonies. Our results suggest that fungicides negatively affect the colony success of a native bumble bee species and that the use of fungicides during bloom has the potential to severely impact the success of native bumble bee populations foraging in agroecosystems. [ABSTRACT FROM AUTHOR]

Simple Summary: A 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. [ABSTRACT FROM AUTHOR]

Simple Summary: Plants in semi-natural areas provide food resources for pollinators that visit pollinator-dependent crop species, such as Oilseed Rape (OSR). Here, we study the patterns of pollinator visitation on OSR and its co-flowering plants in adjacent semi-natural areas. We find that OSR is visited by pollinators that are abundant in the community and that these pollinators also visit co-flowering plant species in semi-natural areas. OSR primarily influences the pollination of plant species which have similar floral traits (i.e., other disc flowers). Plant species that attract a high abundances of bumblebees, wild bees, flies, and beetles influence the pollination of OSR the most. Our results suggest that plant species in semi-natural areas that support the high abundances of common pollinators which are generalized in their visitation are most important to the pollination of OSR, and that such plant species do not necessarily have similar floral traits to OSR. Mass-flowering crops, such as Oilseed Rape (OSR), provide resources for pollinators and benefit from pollination services. Studies that observe the community of interactions between plants and pollinators are critical to understanding the resource needs of pollinators. We observed pollinators on OSR and wild plants in adjacent semi-natural areas in Sachsen-Anhalt, Germany to quantify (1) the co-flowering plants that share pollinators with OSR, (2) the identity and functional traits of plants and pollinators in the network module of OSR, and (3) the identity of the plants and pollinators that act as network connectors and hubs. We found that four common plants share a high percentage of their pollinators with OSR. OSR and these plants all attract abundant pollinators in the community, and the patterns of sharing were not more than would be expected by chance sampling. OSR acts as a module hub, and primarily influences the other plants in its module that have similar functional traits. However, the plants that most influence the pollination of OSR have different functional traits and are part of different modules. Our study demonstrates that supporting the pollination of OSR requires the presence of semi-natural areas with plants that can support a high abundances of generalist pollinators. [ABSTRACT FROM AUTHOR]

Simple Summary: In bumblebee colonies, the authority of male reproduction is not entirely controlled by the queen. Workers who lose mating ability could also produce haploidy males. The conflict over the reproduction of males in bumblebee colonies becomes more evident compared to the higher eusocial honeybees. However, the reproductive characteristics of worker-born males in bumblebees remain unclear. Here, we evaluated the body size and reproductive ability of males produced by worker and queen in Bombus terrestris. Smaller size and slighter individuals' weight but equally high sperm viability was found in worker-born males compared to that of queen-born males. Moreover, worker-born males performed as excellently as queen-born males in copulation and colony development of the mated queens, such as queen egg laying, colony foundation, and colony size. This study contributes the new knowledge of the reproductive biology of eusocial bees. Queen-worker conflict over the reproduction of males exists in the majority of haplodiplioidy hymenpteran species such as bees, wasps, and ants, whose workers lose mating ability but can produce haploid males in colony. Bumblebee is one of the representatives of primitively eusocial insects with plastic division labor and belongs to monandrous and facultative low polyandry species that have reproductive totipotent workers, which are capable of competing with mother queen to produce haploid males in the queenright colony compared to higher eusocial species, e.g., honeybees. So, bumblebees should be a better material to study worker reproduction, but the reproductive characteristics of worker-born males (WMs) remain unclear. Here, we choose the best-studied bumblebee Bombus terrestris to evaluate the morphological characteristics and reproductive ability of WMs from the queenless micro-colonies. The sexually matured WMs showed smaller in forewing length and weight, relatively less sperm counts but equally high sperm viability in comparison with the queen-born males (QMs) of the queenright colony. Despite with smaller size, the WMs are able to successfully mate with the virgin queens in competition with the QMs under laboratory conditions, which is quite different from the honeybees reported. In addition, there was no difference in the colony development, including the traits such as egg-laying rate, colony establishment rate, and populations of offspring, between the WM- and the QM-mated queens. Our study highlights the equivalent reproductive ability of worker-born males compared to that of queens, which might exhibit a positive application or special use of bumblebee rearing, especially for species whose males are not enough for copulation. Further, our finding contributes new evidence to the kin selection theory and suggests worker reproduction might relate to the evolution of sociality in bees. [ABSTRACT FROM AUTHOR]

Simple Summary: The DNA of mitochondria can be transferred into the nucleus and form nuclear mitochondrial DNAs (NUMTs). In this study, we identified and characterized NUMTs in genus-wide bumblebee species. The number of NUMTs in bumblebee is much lower than those in its closely related taxon, honeybee. The insertion sites of NUMTs in bumblebee are not random, with AT-rich regions harboring more NUMTs. In addition, NUMTs derived from the mitochondrial COX1 gene are most abundant in the nuclear genome. While the majority of NUMTs seem unfunctional in the bumblebee, some NUMTs show functional clues, which could fuse with their flanking sequences to form novel proteins. Our results shed light on the molecular features of NUMTs and uncover their contribution to genome innovation in the bumblebee. In eukaryotes, DNA of mitochondria is transferred into the nucleus and forms nuclear mitochondrial DNAs (NUMTs). Taking advantage of the abundant genomic resources for bumblebees, in this study, we de novo generated mitochondrial genomes (mitogenomes) for 11 bumblebee species. Then, we identified and characterized NUMTs in genus-wide bumblebee species. The number of identified NUMTs varies across those species, with numbers ranging from 32 to 72, and nuclear genome size is not positively related to NUMT number. The insertion sites of NUMTs in the nuclear genome are not random, with AT-rich regions harboring more NUMTs. In addition, our results suggest that NUMTs derived from the mitochondrial COX1 gene are most abundant in the bumblebee nuclear genome. Although the majority of NUMTs are found within intergenic regions, some NUMTs do reside within genic regions. Transcripts that contain both the NUMT sequence and its flanking non-NUMT sequences could be found in the bumblebee transcriptome, suggesting a potential domestication of NUMTs in the bumblebee. Taken together, our results shed light on the molecular features of NUMTs in the bumblebee and uncover their contribution to genome innovation. [ABSTRACT FROM AUTHOR]

Simple Summary: Here we discuss widespread changes in the community structure of bumble bees (Bombus spp.) found in the coastal-zone community of New England. One species in particular, Bombus impatiens Cresson, 1863, has increased in relative abundance nearly 45% since the 1990s to become the dominant species in the region, representing nearly 75% of all Bombus individuals collected in our studies. These changes in abundance may be, in part, due to differences in infection rates by microparasites, with B. impatiens having significantly fewer microparasites than several other less common and declining Bombus species. We discuss the possible role of microparasites in influencing the community composition of Bombus species in our region, and how these infections might be compounding declines in conjunction with habitat loss and climate change. Numerous studies have reported a diversity of stressors that may explain continental-scale declines in populations of native pollinators, particularly those in the genus Bombus. However, there has been little focus on the identification of the local-scale dynamics that may structure currently impoverished Bombus communities. For example, the historically diverse coastal-zone communities of New England (USA) now comprise only a few species and are primarily dominated by a single species, B. impatiens. To better understand the local-scale factors that might be influencing this change in community structure, we examined differences in the presence of parasites in different species of Bombus collected in coastal-zone communities. Our results indicate that Bombus species that are in decline in this region were more likely to harbor parasites than are B. impatiens populations, which were more likely to be parasite-free and to harbor fewer intense infections or co-infections. The contrasting parasite burden between co-occurring winners and losers in this community may impact the endgame of asymmetric contests among species competing for dwindling resources. We suggest that under changing climate and landscape conditions, increasing domination of communities by healthy, synanthropic Bombus species (such as B. impatiens) may be another factor hastening the further erosion of bumble bee diversity. [ABSTRACT FROM AUTHOR]

Simple Summary: Pollinators make a strong contribution to ecosystem stability. However, nowadays, they also need protection and sustainable habitat to live and develop. Not all regions can provide suitable habitats due to agricultural intensification, urbanization, climate changes and corresponding impacts. Our study was conducted in the late spring in south Estonia where arable lands were surrounded by forest patches and rural areas. For better performance, we used both light microscopy and DNA metabarcoding methods for pollen identification. We found that bumble bees foraged on the diverse food sources showing preferences for several main plant families. Additionally, in our case, land-use types did not show important effects on bumble bee food choices and foraging decisions. Various landscape features can provide diverse food sources at the early development stages and support nest longevity. Here, we can say that a better understanding of pollinators' food preferences can help in the application of more suitable measures for their conservation. Agricultural landscapes usually provide higher quantities of single-source food, which are noticeably lacking in diversity and might thus have low nutrient value for bumble bee colony development. Here, in this study, we analysed the pollen foraging preferences over a large territory of a heterogeneous agricultural landscape: southern Estonia. We aimed to assess the botanical diversity of bumble bee food plants in the spring time there. We looked for preferences for some food plants or signs of food shortage that could be associated with any particular landscape features. For this purpose, we took Bombus terrestris commercial hives to the landscape, performed microscopy analyses and improved the results with the innovative DNA metabarcoding technique to determine the botanical origin of bumble bee-collected pollen. We found high variability of forage plants with no strong relationship with any particular landscape features. Based on the low number of plant species in single flights, we deduce that the availability of main forage plants is sufficient indicating rich forage availabilities. Despite specific limitations, we saw strong correlations between microscopy and DNA metabarcoding data usable for quantification analyses. As a conclusion, we saw that the spring-time vegetation in southern Estonia can support bumble bee colony development regardless of the detailed landscape structure. The absence of clearly dominating food preference by the tested generalist bumble bee species B. terrestris makes us suggest that other bumble bee species, at least food generalists, should also find plenty of forage in their early development phase. [ABSTRACT FROM AUTHOR]

Simple Summary: Rearing insects in captivity is critical for both research and commercial purposes. One group of insects that stands out for their ecological and scientific importance are bumble bees. However, most research studies with bumble bees rely on commercial colonies due to the challenges associated with initiating colonies in the laboratory. This limits the ability to study the early stages of the colony life cycle and the ability to control for variables such as colony age and relatedness. To overcome these challenges, we tested several aspects related to the solitary phase of the North American bumble bee species, Bombus impatiens. In this species, queens emerge by the end of the season, mate, and enter a winter diapause. They then initiate a colony the next spring. We examined the optimal age for mating and how the timing of CO2 narcosis (a technique used to bypass diapause) and social cues post-mating affect queen egg laying. We demonstrate an optimum age for mating in males and females and the importance of worker and pupa presence to egg laying in queens. Overall, our findings indicate that the laboratory rearing of bumble bees can be improved by reducing challenges associated with rearing queens during the solitary phase. Bumble bees are key pollinators for wild and managed plants and serve as a model system in various research fields, largely due to their commercial availability. Despite their extensive use, laboratory rearing of bumble bees is often challenging, particularly during the solitary phase queens undergo before founding a colony. Using a literature survey, we demonstrate that most studies rely on commercially available species that are provided during the colony's social phase, limiting study on early phases of the life cycle and the ability to control for colony age and relatedness. Laboratory rearing is challenging since the queen solitary phase is less understood compared to the social phase. To overcome this barrier, we examined several aspects related to the queen solitary phase: the effect of age on likelihood of mating, how the timing of CO2 narcosis post-mating (a technique to bypass diapause) affects egg-laying, and whether different social cues affect the success of colony initiation. Our data show an optimum age for mating in both sexuals and decreased egg-laying latency in the presence of workers and pupae. The timing of CO2 narcosis did not significantly affect egg laying in queens. These findings can be incorporated to improve bumble bee rearing for research purposes. [ABSTRACT FROM AUTHOR]

Simple Summary: Blueberry is a crop that is increasing globally, both in terms of yield and extent, mostly thanks to the recent introduction of North American blueberry in the temperate areas of Europe, Asia, and South America. As blueberry depends largely on insects for pollination, farmers in these expansion areas face the challenge of adapting this crop to unexpected pollinator species, whose traits and features may not entirely fit the pollination needs of the introduced crop. Here we study the abundance, the behaviour, and the response to environmental conditions of different managed and wild native pollinators of blueberry in northern Spain. Our findings suggest the dominant role of native wild bumblebees and managed honeybee as suppliers of pollination service. Honeybee and bumblebees differed in both when and where they occurred, in how they responded to environmental conditions, and in how they behaved as an effective pollinator. The role of bumblebees and honeybee as blueberry pollinators thus seems complementary and additive. These results encourage the preservation of populations of native wild bees in order to ensure the effective management of introduced blueberry crops. The entomophilous pollination niche (abundance, phenotypic traits, foraging behaviours and environmental tolerances of insect pollinators) helps to understand and better manage crop pollination. We apply this niche approach to assess how an entomophilous crop (blueberry, Vaccinium ashei) can be expanded into new territories (i.e., northern Spain) far from their original area of domestication (North America). Insect visits to blueberry flowers were monitored in a plantation on 12 different days, at 8 different times during day and covering various weather conditions. Abundance, visitation rate, pollen gathering behaviour, and frequency of inter-plant and inter-row movements were recorded. The pollinator assemblage was basically composed of one managed honeybee species (50.8% of visits) and three native bumblebee species (48.3%). There was a marked pattern of seasonal segregation throughout bloom, with bumblebees dominating the early bloom and honeybee the late bloom. Pollinators also segregated along gradients of daily temperature and relative humidity. Finally, the two pollinator types differed in foraging behaviour, with bumblebees having a visitation rate double that of honeybee, collecting pollen more frequently and changing plant and row more frequently. The spatio-temporal and functional complementarity between honeybee and bumblebees suggested here encourages the consideration of an integrated crop pollination strategy for blueberries, based on the concurrence of both wild and managed bees. [ABSTRACT FROM AUTHOR]

Simple Summary: Bumblebees and honeybees are well known as the dominant and most important pollinators in natural and agricultural ecosystems. The quality characteristics of their colonies depend greatly on the reproductive ability/quality of the parents (queens and drones). Male bees, despite their exclusive reproductive role and ability to determine colony quality, have been less considered than female bees, especially bumblebees. We reviewed the current studies on environmental factors and inherent characteristics that affect the mating success and fecundity of male honeybees and bumblebees. Temperature, nutrients, pesticides, body size, weight and age affect reproduction in male bees and consequently the progeny colony quality. However, more studies, especially in male bumblebees, are still needed to address the impacts of these factors in detail to confront the requirements of agricultural pollination and declining wild bee pollinators worldwide. Bumblebees and honeybees are very important pollinators and play a vital role in agricultural and natural ecosystems. The quality of their colonies is determined by the queens and the reproductive drones of mother colonies, and mated drones transmit semen, including half of the genetic materials, to queens and enhance their fertility. Therefore, factors affecting drone fecundity will also directly affect progeny at the colony level. Here, we review environmental and bee-related factors that are closely related to drone reproductive ability. The environmental factors that mainly affect the sperm count and the viability of males include temperature, nutrients and pesticides. In addition, the inherent characteristics of male bees, such as body size, weight, age, seminal fluid proteins and proteins of the spermathecal fluid, contribute to mating success, sperm quality during long-term storage in the spermathecae and the reproductive behaviors of queens. Based on the results of previous studies, we also suggest that the effects of somatotype dimorphism in bumblebee males on sperm quality and queen fecundity and the indispensable and exploitable function of gland proteins in the fecundity of males and queens should be given more attention in further studies. [ABSTRACT FROM AUTHOR]

Simple Summary: Climate change is having a profound impact on pollination systems, yet we still do not know to what extent increasing concentrations of carbon dioxide (CO2) will directly affect the interactions between plants and their pollinators. We review all the existing published literature on the effect of elevated CO2 (eCO2) on flowering time, nectar and pollen production and plant–pollinator interactions. We also conduct a field experiment to test the effect of eCO2 on bluebells and their pollinators. We found that few studies have assessed the impact of eCO2 on pollination, and our field data found that bluebells flowered on average 6 days earlier under eCO2 conditions. Hoverflies and bumble bees were the main visitors to bluebell flowers, but insect activity was low early in the flowing period. Although we did not find a difference in the number of visits made by insects to bluebell flowers under eCO2, or the amount of seeds those flowers produced, the change in the timing of flowering could mean that a mismatch could develop between bluebells and their pollinators in the future, which would affect pollination success. The impact of elevated CO2 (eCO2) on plant–pollinator interactions is poorly understood. This study provides the first systematic review of this topic and identifies important knowledge gaps. In addition, we present field data assessing the impact of eCO2 (150 ppm above ambient) on bluebell (Hyacinthoides non-scripta)–pollinator interactions within a mature, deciduous woodland system. Since 1956, only 71 primary papers have investigated eCO2 effects on flowering time, floral traits and pollination, with a mere 3 studies measuring the impact on pollination interactions. Our field experiment documented flowering phenology, flower visitation and seed production, as well as the abundance and phenology of dominant insect pollinators. We show that first and mid-point flowering occurred 6 days earlier under eCO2, but with no change in flowering duration. Syrphid flies and bumble bees were the dominant flower visitors, with peak activity recorded during mid- and late-flowering periods. Whilst no significant difference was recorded in total visitation or seed set between eCO2 and ambient treatments, there were clear patterns of earlier flowering under eCO2 accompanied by lower pollinator activity during this period. This has implications for potential loss of synchrony in pollination systems under future climate scenarios, with associated long-term impacts on abundance and diversity. [ABSTRACT FROM AUTHOR]

Simple Summary: In the context of climate warming, modifications in plant pollination and reproductive success constitute a crucial issue. Modifications of both floral signals (display, size of flowers) and rewards (nectar and pollen) due to increased air temperatures may affect plant–pollinator interactions. However, relationships between modifications in floral traits and rewards caused by increased air temperatures and the associated effects on pollinator visitation rate and foraging behavior have not been thoroughly investigated. To explore the effects of temperature increase on plant–pollinator interactions, we chose the highly attractive bee-pollinated Borago officinalis and one of its pollinators, Bombus terrestris. We measured visual floral signals and rewards for plants cultivated at 21 °C or 26 °C and we investigated bumblebee behavior by tracking insect visits on plants in an indoor flight arena. Our results show that exposure to higher temperature during the flowering stages of B. officinalis negatively affects visual floral traits (e.g., by reducing the number of flowers) as well as floral rewards, affecting bumblebee visitation and foraging behavior. Bumblebees visited flowers from plants grown at 26 °C four times less frequently than they visited those from plants grown at 21 °C. Thus, the global increases in temperature caused by climate change could reduce plant pollination rates and reproductive success by reducing flower visitation. (1) Background: Plants attract pollinators using several visual signals, mainly involving the display, size, shape, and color of flowers. Each signal is relevant for pollinators foraging for floral rewards, pollen, and nectar. Changes in floral signals and rewards can be induced by an increase in temperature, drought, or other abiotic stresses and are expected to increase as global temperatures rise. In this study, we explored how pollinators respond to modified floral signals and rewards following an increase in temperature; (2) Methods: We tested the effects of warmer temperatures on bee-pollinated starflower (Borago officinalis, Boraginaceae) and determined the behavior of one of its main pollinators, the buff-tailed bumblebee (Bombus terrestris). We measured visual floral traits (display and size) and rewards (nectar and pollen) for plants cultivated at 21 °C or 26 °C. We investigated bumblebee behavior by tracking insect visits in a binary choice experiment in an indoor flight arena; (3) Results: Plants cultivated at 26 °C exhibited a smaller floral area (i.e., corolla sizes summed for all flowers per plant, 34.4 ± 2.3 cm2 versus 71.2 ± 2.7 cm2) and a greater flower height (i.e., height of the last inflorescence on the stem, 87 ± 1 cm versus 75 ± 1 cm) compared to plants grown at 21 °C. Nectar production per flower was lower in plants grown at 26 °C than in plants grown at 21 °C (2.67 ± 0.37 µL versus 4.15 ± 0.22 µL), and bumblebees visited flowers from plants grown at 26 °C four times less frequently than they visited those from plants grown at 21 °C; (4) Conclusions: These results show that warmer temperatures affect floral signals and reduce overall floral resources accessible to pollinators. Thus, the global increases in temperature caused by climate change could reduce plant pollination rates and reproductive success by reducing flower visitation. [ABSTRACT FROM AUTHOR]

Simple Summary: Pollinators are essential to produce fruits in apple production. Bumble bees are among the most effective pollinators in orchards during the blooming season, yet they are often threatened by the high levels of pesticide use in apple production. Hedgerows and flower strips are infrequently sprayed by pesticides and are thus potentially good shelter for bumble bees. This study evaluated the influence of landscaping in the form of hedgerows and flower strips on the abundance and number of bumble bee species found in apple orchards. The number of bumble bee species found in orchards with hedgerows or flower strips was higher than in orchards without such landscape enhancements. Similarly, three species were more abundant in orchards with landscaping than orchards without those enhancements. Our work provides additional evidence that landscaping in the form of hedgerows and/or flower strips improves bumble bee presence in apple orchards and should therefore be considered as a means to enhance and ensure pollination within farms. Bumble bees are among the most effective pollinators in orchards during the blooming period, yet they are often threatened by the high levels of pesticide use in apple production. This study aimed to evaluate the influence of landscape enhancements (e.g., hedgerows, flower strips) on bumble bee queens in apple orchards. Bumble bee queens from 12 orchards in southern Québec (Canada) were marked, released, and recaptured in the springs and falls of 2017 to 2019. Half of the 12 orchards had landscape enhancements. Apples were harvested in 2018 and 2019 to compare their quality (weight, diameter, sugar level, and seed number) in sites with and without landscape enhancements. Species richness, as well as the occurrence of three species out of eight, was higher in orchards with landscape enhancements than in orchards without such structures. The occurrence of Bombus ternarius was lower in orchards with high levels of pesticide use. Apples had fewer seeds when collected in orchards with landscape enhancements and were heavier in orchards that used more pesticides. Our work provides additional evidence that landscape enhancements improve bumble bee presence in apple orchards and should therefore be considered as a means to enhance pollination within farms. [ABSTRACT FROM AUTHOR]

Simple Summary: Behavioural specialisation of individuals may improve the performance of groups, but could also limit the ability to switch tasks (behavioural 'plasticity') in response to changing group needs. In bumble bee colonies, body size, which is fixed once the bees reach adulthood, influences the tasks that bees perform, meaning that large and small bees often act as specialists. We found that when we experimentally reduced the body-size variation of colonies, some performed less well than normal. Nonetheless, in other colonies, individuals increased task specialisation or effort, which apparently compensated for the absence of large and small workers. These results suggest that both behavioural specialisation and plasticity can be important in collective group performance. Specialisation and plasticity are important for many forms of collective behaviour, but the interplay between these factors is little understood. In insect societies, workers are often developmentally primed to specialise in different tasks, sometimes with morphological or physiological adaptations, facilitating a division of labour. Workers may also plastically switch between tasks or vary their effort. The degree to which developmentally primed specialisation limits plasticity is not clear and has not been systematically tested in ecologically relevant contexts. We addressed this question in 20 free-foraging bumble bee (Bombus terrestris) colonies by continually manipulating colonies to contain either a typically diverse, or a reduced ("homogeneous"), worker body size distribution while keeping the same mean body size, over two trials. Pooling both trials, diverse colonies produced a larger comb mass, an index of colony performance. The link between body size and task was further corroborated by the finding that foragers were larger than nurses even in homogeneous colonies with a very narrow body size range. However, the overall effect of size diversity stemmed mostly from one trial. In the other trial, homogeneous and diverse colonies showed comparable performance. By comparing behavioural profiles based on several thousand observations of individuals, we found evidence that workers in homogeneous colonies in this trial rescued colony performance by plastically increasing behavioural specialisation and/or individual effort, compared to same-sized individuals in diverse colonies. Our results are consistent with a benefit to colonies of large and small specialists under certain conditions, but also suggest that plasticity or effort can compensate for reduced (size-related) specialisation. Thus, we suggest that an intricate interplay between specialisation and plasticity is functionally adaptive in bumble bee colonies. [ABSTRACT FROM AUTHOR]

Simple Summary: Floral resource availability is one of the keys to preserving the health of bee communities. However, flowers also present a risk of pathogen transmission, as infected pollinators could deposit pathogens while foraging, exposing other pollinators to infection via the consumption of contaminated nectar or pollen. Here, we studied, over time, the prevalence of seven viruses in bee communities that share the same small surface of floral resource in order to assess the risk of virus spillover. In total, 2057 bee specimens from 30 species were caught, identified and checked for the presence of viruses. Specimens from the Halictidae family were the dominant wild bees. The prevalence of viruses was quite high: at least one virus was detected in 78% of the samples, and co-infections were frequent. The genetic diversity of the viruses was also investigated to look for the possible association of geographic origin or host with shared ancestry. Viruses are known to contribute to bee population decline. Possible spillover is suspected from the co-occurrence of viruses in wild bees and honey bees. In order to study the risk of virus transmission between wild and managed bee species sharing the same floral resource, we tried to maximize the possible cross-infections using Phacelia tanacetifolia, which is highly attractive to honey bees and a broad range of wild bee species. Virus prevalence was compared over two years in Southern France. A total of 1137 wild bees from 29 wild bee species (based on COI barcoding) and 920 honey bees (Apis mellifera) were checked for the seven most common honey bee RNA viruses. Halictid bees were the most abundant. Co-infections were frequent, and Sacbrood virus (SBV), Black queen cell virus (BQCV), Acute bee paralysis virus (ABPV) and Israeli acute paralysis virus (IAPV) were widespread in the hymenopteran pollinator community. Conversely, Deformed wing virus (DWV) was detected at low levels in wild bees, whereas it was highly prevalent in honey bees (78.3% of the samples). Both wild bee and honey bee virus isolates were sequenced to look for possible host-specificity or geographical structuring. ABPV phylogeny suggested a specific cluster for Eucera bees, while isolates of DWV from bumble bees (Bombus spp.) clustered together with honey bee isolates, suggesting a possible spillover. [ABSTRACT FROM AUTHOR]

Simple Summary: Pollinators are a major part of global biodiversity and they provide ecosystem services important for the production of many crops. Their abundance and diversity have declined steadily in recent years. Loss of foraging resources through degradation and fragmentation of natural habitats has been a major factor. Enhancing floral resources in the environment can mitigate this decline. Cowpea nectar has been reported to make the crop attractive to pollinators. We evaluated twenty-four cowpea varieties for pollinator abundance and diversity using pan traps, sticky traps, and direct visual counts. Sticky traps captured the highest number of pollinators and pan traps the least. The highest number of pollinators was recorded on Penny Rile cowpea and the lowest on Iron and Clay which had no flowers. Whippoorwill had the most flowers and ranked third in number of pollinators. Our findings indicate that cowpeas can be used to improve pollinator efficiency. Intercropping pollinator-dependent crops with cowpea varieties such as Penny Rile, Dixielee, and Whippoorwill will not only provide resources for the pollinators but can also be effective in increasing pollinator number and activity to increase crop yields. Pollinators are on the decline and loss of flower resources play a major role. This raises concerns regarding production of insect-pollinated crops and therefore food security. There is urgency to mitigate the decline through creation of farming systems that encourage flower-rich habitats. Cowpea is a crop that produces pollen and nectar attractive to pollinators. Twenty-four cowpea varieties were planted, and the number of pollinators were counted using three sampling methods: pan traps, sticky traps, and direct visual counts. Five pollinator types (honey bees, bumble bees, carpenter bees, wasps, and butterflies and moths), 11 and 16 pollinator families were recorded from direct visual counts, pan and sticky traps, respectively. Pollinator distribution varied significantly among varieties and sampling methods, with highest number on Penny Rile (546.0 ± 38.6) and lowest (214.8 ± 29.2) in Iron and Clay. Sticky traps accounted for 45%, direct visual counts (31%), and pan traps (23%) of pollinators. Pollinators captured by pan traps were more diverse than the other methods. The relationship between number of pollinators and number of flowers was significant (r2 = 0.3; p = 0.009). Cowpea can increase resources for pollinators and could be used to improve pollinator abundance and diversity in different farming systems. [ABSTRACT FROM AUTHOR]

Simple Summary: Bees are important pollinators for many agricultural crops. Compared with bumblebees, honeybees are less attracted to tomato flowers. Floral scent usually plays an important role in mediating the foraging behavior of bees, and tomato flowers release special scents. However, little is known about how tomato floral scent regulates the foraging behaviors of these two bee taxa. In the current study, we investigated the foraging behaviors of the widely used pollinator honeybee Apis mellifera and a native bumblebee, Bombus lantschouensis, on tomato flowers to evaluate the potential application of these two bee species for tomato pollination in solar greenhouses. Moreover, we determined whether honeybees and bumblebees show different responses to tomato floral scent and how innate biases and prior experience influence bee choice behavior. We found that naïve bees showed no preference for tomato floral scent but could develop such a preference after learning to associate tomato floral scent with a food reward on the basis of foraging experience or scent-learning procedures. We conclude that scent-learning experiences with food reward can change the innate bias of bees and could be utilized to improve the pollination service efficiency of bees for commercial crops. Bee responses to floral scent are usually influenced by both innate biases and prior experience. Honeybees are less attracted than bumblebees to tomato flowers. However, little is known about how tomato floral scent regulates the foraging behaviors of honeybees and bumblebees. In this study, the foraging behaviors of the honeybee Apis mellifera and the bumblebee Bombus lantschouensis on tomato flowers in greenhouses were investigated. Whether the two bee species exhibit different responses to tomato floral scent and how innate biases and prior experience influence bee choice behavior were examined. In the greenhouses, honeybees failed to collect pollen from tomato flowers, and their foraging activities decreased significantly over days. Additionally, neither naïve honeybees nor naïve bumblebees showed a preference for tomato floral scent in a Y-tube olfactometer. However, foraging experience in the tomato greenhouses helped bumblebees develop a strong preference for the scent, whereas honeybees with foraging experience continued to show aversion to tomato floral scent. After learning to associate tomato floral scent with a sugar reward in proboscis extension response (PER) assays, both bee species exhibited a preference for tomato floral scent in Y-tube olfactometers. The findings indicated that prior experience with a food reward strongly influenced bee preference for tomato floral scent. [ABSTRACT FROM AUTHOR]

Simple Summary: Olive cultivation is extensive throughout the Mediterranean region and essential for the rural economy, local heritage and the environment. To control the olive fruit fly, a major threat for this cultivation, pesticides, in the form of bait or cover sprays, are applied. These pesticide applications can potentially impact pollinators that forage in the nearby areas or live inside the olive orchards. Based on current practice, olive trees supply shadow, water, and still support some flowering plants at the period of the year with the highest temperature and minimum nectar and pollen flow, which can be beneficial for bees. In this study pesticide residues were monitored in honeybees, bumblebees, honey and olive oil, after placement of bee colonies in Greek olive orchards where applications to control the olive fruit fly took place. Variations of concentrations were evidenced, for the three active ingredients that were applied. In limited cases, concentrations in bees higher than the median lethal dose can possibly be attributed to bait dose rates or broad foraging of bees in nearby orchards with similar applications. Determined olive oil residues corroborated that those pesticides were applied in the olive orchards. In 2017 and 2018, a field survey was initiated on Greek olive orchards to investigate the attractiveness of bait spray applications and the impact of cover and bait sprays applied against the olive fruit fly Bactrocera oleae (Diptera: Tephritidae), on the honeybee, Apis mellifera L. and bumblebees Bombus terrestris, by investigating the pesticides' residual prevalence. Bee colonies were evenly distributed in three sites located on coastal areas of Western Crete and visited almost weekly between July and October. Samples collected, were analyzed using existing or developed-optimized liquid and gas chromatographic methods. In bee samples, concentrations varied from 0.0013 to 2.3 mg/kg for dimethoate, from 0.0013–0.059 mg/kg for its metabolite omethoate, and from 0.0035 to 0.63 mg/kg regarding the pyrethroids, β-cyfluthrin and λ-cyhalothrin. In one bee sample dimethoate concentration exceeded both acute oral and contact median lethal dose (LD50). Residue findings in bees, along with verified olive oil residues corroborated that those insecticides had been applied in the olive orchards and transferred to bees. The possibility of non-target effects of the bait sprays to the bees, as well as the impact of the contaminated olive to the bees are discussed. [ABSTRACT FROM AUTHOR]

Simple Summary: Social information use is a widespread phenomenon in animals affecting various important aspects of behavior. Animals observe and copy the behavioral patterns of conspecifics and other species on the same trophic level in their own decision-making. Copying others is adaptive only when it is selective, otherwise running the risk of copying bad behavior. Thus, it would be important to know when and which individuals should be copied. We showed that bumblebees Bombus terrestris learn to attend to specific types of bees based on their learned value as information-providers about reward. Learned information was transferred across foraging contexts, representing a useful pathway for bees to acquire relevant information about unfamiliar floral resources that goes beyond conspecific social information use, extending to heterospecifics. Our findings indicate that the transmission of social information across species can be highly selective in response to learned value of the information provider, plausibly making the phenomenon adaptive. Using social information can be an efficient strategy for learning in a new environment while reducing the risks associated with trial-and-error learning. Whereas social information from conspecifics has long been assumed to be preferentially attended by animals, heterospecifics can also provide relevant information. Because different species may vary in their informative value, using heterospecific social information indiscriminately can be ineffective and even detrimental. Here, we evaluated how selective use of social information might arise at a proximate level in bumblebees (Bombus terrestris) as a result of experience with demonstrators differing in their visual appearance and in their informative value as reward predictors. Bumblebees were first trained to discriminate rewarding from unrewarding flowers based on which type of "heterospecific" (one of two differently painted model bees) was next to each flower. Subsequently, these bumblebees were exposed to a novel foraging context with two live painted bees. In this novel context, observer bumblebees showed significantly more social information-seeking behavior towards the type of bees that had predicted reward during training. Bumblebees were not attracted by paint-marked small wooden balls (moved via magnets) or paint-marked non-pollinating heterospecifics (woodlice; Porcellio laevis) in the novel context, indicating that bees did not simply respond to conditioned color cues nor to irrelevant social cues, but rather had a "search image" of what previously constituted a valuable, versus invaluable, information provider. The behavior of our bumblebees suggests that their use of social information is governed by learning, is selective, and extends beyond conspecifics. [ABSTRACT FROM AUTHOR]

Simple Summary: Evidence of bumble bee population declines has led to an increase in conservation efforts to protect these important pollinators. However, effective conservation requires accurate species identification. We provide quantitative methods to accurately identify three cryptic species of bumble bees using morphometric measurements of the cheek length and width, and antennal segments. We validated the accuracy of our methods with DNA analysis. We predicted that these methods would reliably identify both the queens and worker bees of Bombus vagans and B. sandersoni. We expanded these methods to include an uncommon form of Bombus perplexus with all light hair on its thorax, rather than the more common light on top and dark below, that can mistakenly be identified as B. vagans or B. sandersoni. Although the species we consider here, Bombus vagans, B. sandersoni and B. perplexus, are not currently listed as species of concern in North America, there is uncertainty of their population status, some of which is due to difficulty in species identification, which we have resolved. Recent history informs us that some bumble bee species experience rapid declines within a few decades. Our methods to correctly identify these cryptic species is key to monitoring their status and population trends. Despite their large size and striking markings, the identification of bumble bees (Bombus spp.) is surprisingly difficult. This is particularly true for three North American sympatric species in the subgenus Pyrobombus that are often misidentified: B. sandersoni Franklin, B. vagans Smith B. perplexus Cresson. Traditionally, the identification of these cryptic species was based on observations of differences in hair coloration and pattern and qualitative comparisons of morphological characters including malar length. Unfortunately, these characteristics do not reliably separate these species. We present quantitative morphometric methods to separate these species based on the malar length to width ratio (MRL) and the ratios of the malar length to flagellar segments 1 (MR1) and 3 (MR3) for queens and workers, and validated our determinations based on DNA barcoding. All three measurements discriminated queens of B. sandersoni and B. vagans with 100% accuracy. For workers, we achieved 99% accuracy by combining both MR1 and MR3 measurements, and 100% accuracy differentiating workers using MRL. Moreover, measurements were highly repeatable within and among both experienced and inexperienced observers. Our results, validated by genetic evidence, demonstrate that malar measurements provide accurate identifications of B. vagans and B. sandersoni. There was considerable overlap in the measurements between B. perplexus and B. sandersoni. However, these species can usually be reliably separated by combining malar ratio measurements with other morphological features like hair color. The ability to identify bumble bees is key to monitoring the status and trends of their populations, and the methods we present here advance these efforts. [ABSTRACT FROM AUTHOR]

Simple Summary: Bumblebees are very important pollinators for many wild and agricultural flowers and play significant roles in natural and agricultural ecosystems. The distribution of bumblebees varies between the species; some species occupy narrow areas, while others have a varied distribution from low to high elevation. Bombus pyrosoma is one of the bumblebee species with a highly varied geomorphological distribution range in China. To answer why some bumblebee species have a varied distribution, we compared the transcriptomic and metabolomic data of B. pyrosoma from the low-altitude North China Plain and the high-altitude Tibet Plateau. The results showed that energy metabolism and innate immunity of the high-altitude B. pyrosoma had been enhanced in order to adapt to the extreme environment of hypoxia and low temperature, compared to the low-altitude bumblebees. This study highlights the ecological adaptation of bumblebees distributed from low- to high-altitude conditions. Bombus pyrosoma is one of the most abundant bumblebee species in China, with a distribution range of very varied geomorphology and vegetation, which makes it an ideal pollinator species for research into high-altitude adaptation. Here, we sequenced and assembled transcriptomes of B. pyrosoma from the low-altitude North China Plain and the high-altitude Tibet Plateau. Subsequent comparative analysis of de novo transcriptomes from the high- and low-altitude groups identified 675 common upregulated genes (DEGs) in the high-altitude B. pyrosoma. These genes were enriched in metabolic pathways and corresponded to enzyme activities involved in energy metabolism. Furthermore, according to joint analysis with comparative metabolomics, we suggest that the metabolism of coenzyme A (CoA) and the metabolism and transport of energy resources contribute to the adaptation of high-altitude B. pyrosoma. Meanwhile, we found many common upregulated genes enriched in the Toll and immune deficiency (Imd)signaling pathways that act as important immune defenses in insects, and hypoxia and cold temperatures could induce the upregulation of immune genes in insects. Therefore, we suppose that the Toll and Imd signaling pathways also participated in the high-altitude adaptation of B. pyrosoma. Like other organisms, we suggest that the high-altitude adaptation of B. pyrosoma is controlled by diverse mechanisms. [ABSTRACT FROM AUTHOR]

The brain-gut–microbiome axis is an emerging area of study, particularly in vertebrate systems. Existing evidence suggests that gut microbes can influence basic physiological functions and that perturbations to the gut microbiome can have deleterious effects on cognition and lead to neurodevelopmental disorders. While this relationship has been extensively studied in vertebrate systems, little is known about this relationship in insects. We hypothesized that because of its importance in bee health, the gut microbiota influences learning and memory in adult bumble bees. As an initial test of whether there is a brain-gut–microbiome axis in bumble bees, we reared microbe-inoculated and microbe-depleted bees from commercial Bombus impatiens colonies. We then conditioned experimental bees to associate a sucrose reward with a color and tested their ability to learn and remember the rewarding color. We found no difference between microbe-inoculated and microbe-depleted bumble bees in performance during the behavioral assay. While these results suggest that the brain-gut–microbiome axis is not evident in Bombus impatiens, future studies with different invertebrate systems are needed to further investigate this phenomenon. [ABSTRACT FROM AUTHOR]

Simple Summary: Populations of some bumble bee species have declined over the last decade. Recognizing the importance of bumble bees to agriculture and natural ecosystems, there has been an upwelling of research to better understand the underlying reasons for observed population declines. While most research has addressed the health of bumble bee females (i.e., workers and queens), males have been largely ignored. Here, we explore the available published literature on the role males play in improving queen health and reproductive fitness, as well as in overall nest success. We conclude that males serve a unique and important role in bumble bee colony success. Bumble bee population declines over the last decade have stimulated strong interest in determining causative factors and necessary conservation measures. Research attention has largely been directed toward bumble bee worker and queen health and their contributions to population stability, while male bees (i.e., drones) have typically been overlooked regarding their role in influencing colony fitness and longevity. In this review we assess existing literature on the diverse role of males within bumble bee nests and their importance to queen health and fitness, as well as to overall nest success. The implications of reproductive measures, including sperm transfer, mating behavior, mating plugs, and male immunity, among other topics, are examined. Overall, bumble bee males are found to drive colony function in a unique manner. Current knowledge gaps pertaining to the role of males are discussed. We highlight the importance of drones to queen success and fitness in many ways, and suggest future research exploring impacts of this often-neglected caste. [ABSTRACT FROM AUTHOR]

Bumble bees are important pollinators broadly used by farmers in greenhouses and under conditions in which honeybee pollination is limited. As such, bumble bees are increasingly being reared for commercial purposes, which brings into question whether individuals reared under laboratory conditions are fully capable of physiological adaptation to field conditions. To understand the changes in bumble bee organism caused by foraging, we compared the fundamental physiological and immunological parameters of Bombus terrestris workers reared under constant optimal laboratory conditions with workers from sister colonies that were allowed to forage for two weeks in the field. Nutritional status and immune response were further determined in wild foragers of B.terrestris that lived under the constant influence of natural stressors. Both wild and laboratory-reared workers subjected to the field conditions had a lower protein concentration in the hemolymph and increased antimicrobial activity, the detection of which was limited in the non-foragers. However, in most of the tested parameters, specifically the level of carbohydrates, antioxidants, total hemocyte concentration in the hemolymph and melanization response, we did not observe any significant differences between bumble bee workers produced in the laboratory and wild animals, nor between foragers and non-foragers. Our results show that bumble bees reared under laboratory conditions can mount a sufficient immune response to potential pathogens and cope with differential food availability in the field, similarly to the wild bumble bee workers. [ABSTRACT FROM AUTHOR]

Dietary macro-nutrients (i.e., carbohydrates, protein, and fat) are important for bee larval development and, thus, colony health and fitness. To which extent different diets (varying in macro-nutrient composition) affect adult bees and whether they can thrive on nectar as the sole amino acid source has, however, been little investigated. We investigated how diets varying in protein concentration and overall nutrient composition affected consumption, longevity, and breeding behavior of the buff-tailed bumble bee, Bombus terrestris (Hymenoptera: Apidae). Queenless micro-colonies were fed either natural nutrient sources (pollen), nearly pure protein (i.e., the milk protein casein), or sucrose solutions with low and with high essential amino acid content in concentrations as can be found in nectar. We observed micro-colonies for 110 days. We found that longevity was highest for pure pollen and lowest for pure sucrose solution and sucrose solution supplemented with amino acids in concentrations as found in the nectar of several plant species. Adding higher concentrations of amino acids to sucrose solution did only slightly increase longevity compared to sucrose alone. Consequently, sucrose solution with the applied concentrations and proportions of amino acids or other protein sources (e.g., casein) alone did not meet the nutritional needs of healthy adult bumble bees. In fact, longevity was highest and reproduction only successful in micro-colonies fed pollen. These results indicate that, in addition to carbohydrates and protein, adult bumble bees, like larvae, need further nutrients (e.g., lipids and micro-nutrients) for their well-being. An appropriate nutritional composition seemed to be best provided by floral pollen, suggesting that pollen is an essential dietary component not only for larvae but also for adult bees. [ABSTRACT FROM AUTHOR]

The pollen stores of bumble bees host diverse microbiota that influence overall colony fitness. Yet, the taxonomic identity of these symbiotic microbes is relatively unknown. In this descriptive study, we characterized the microbial community of pollen provisions within captive-bred bumble bee hives obtained from two commercial suppliers located in North America. Findings from 16S rRNA and ITS gene-based analyses revealed that pollen provisions from the captive-bred hives shared several microbial taxa that have been previously detected among wild populations. While diverse microbes across phyla Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteria, and Ascomycota were detected in all commercial hives, significant differences were detected at finer-scale taxonomic resolution based on the supplier source. The causative agent of chalkbrood disease in honey bees, Ascosphaera apis, was detected in all hives obtained from one supplier source, although none of the hives showed symptoms of infection. The shared core microbiota across both commercial supplier sources consisted of two ubiquitous bee-associated groups, Lactobacillus and Wickerhamiella/Starmerella clade yeasts that potentially contribute to the beneficial function of the microbiome of bumble bee pollen provisions. [ABSTRACT FROM AUTHOR]

Climate change poses a threat to global food security with extreme heat events causing drought and direct damage to crop plants. However, by altering behavioural or physiological responses of insects, extreme heat events may also affect pollination services on which many crops are dependent. Such effects may potentially be exacerbated by other environmental stresses, such as exposure to widely used agro-chemicals. To determine whether environmental stressors interact to affect pollination services, we carried out field cage experiments on the buff-tailed bumble bee (Bombus terrestris). Using a Bayesian approach, we assessed whether heat stress (colonies maintained at an ambient temperature of 25 °C or 31 °C) and insecticide exposure (5 ng g-1 of the neonicotinoid insecticide clothianidin) could induce behavioural changes that affected pollination of faba bean (Vicia faba). Only the bumble bee colonies and not the plants were exposed to the environmental stress treatments. Bean plants exposed to heat-stressed bumble bee colonies (31 °C) had a lower proportional pod set compared to colonies maintained at 25 °C. There was also weak evidence that heat stressed colonies caused lower total bean weight. Bee exposure to clothianidin was found to have no clear effect on plant yields, either individually or as part of an interaction. We identified no effect of either colony stressor on bumble bee foraging behaviours. Our results suggest that extreme heat stress at the colony level may impact on pollination services. However, as the effect for other key yield parameters was weaker (e.g. bean yields), our results are not conclusive. Overall, our study highlights the need for further research on how environmental stress affects behavioural interactions in plant-pollinator systems that could impact on crop yields. [ABSTRACT FROM AUTHOR]

This study compared the food plants, life cycle, colony development, and mating behaviour of the two Asian bumblebee species Bombus friseanus and B. breviceps, which are very important pollinators for many wild flowers and crops in local ecosystems. Both species were shown to be highly polylectic. Differences were observed in their life cycles and colony development patterns. The colony foundation rate of the field-collected queens was high in both species, 95.5% in B. friseanus and 86.5% in B. breviceps. The intervals from colony initiation to colony sizes of 30, 60, and 80 workers and to the first male and gyne emergence were significantly shorter in B. friseanus than in B. breviceps (p < 0.01). The development period of the first batch of workers showed no significant difference between the two species (p > 0.05). Compared with B. friseanus, B. breviceps produced remarkably higher numbers of workers (135 ± 30 workers/colony in B. friseanus and 318 ± 123 workers/colony in B. breviceps) and males (199 ± 46 males/colony in B. friseanus and 355 ± 166 males/colony in B. breviceps) (p < 0.01), with notable variation was found among the colonies in both species. With no significant difference in the mating rate between these two species, the copulation duration of B. breviceps (1.54 ± 0.63 min) was strikingly shorter than that of B. friseanus (27.44 ± 11.16 min) (p < 0.001). This study highlights the characteristics of the two Asian bumblebee species and will aid further studies on their conservation and agricultural pollination use. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The vitellogenin receptor (VgR) belongs to the low-density lipoprotein receptor (LDLR) family, responsible for mediating the endocytosis of vitellogenin (Vg) into the ovaries to promote ovarian growth and oviposition. Here, we cloned and measured VgR gene expression characteristics in the bumblebee Bombus lantschouensis. RNA interference was used to validate VgR function. The results showed that the full length of the BLVgR cDNA was 5519 bp, which included a 5280 bp open reading frame encoding 1759 amino acids (AAs). Sequence alignment revealed that the protein contained 12 LDLa, 5 EGF, 2 EGF-CA and 10 LY domains. Phylogenetic analysis showed that BLVgR and the VgR of Bombus terrestris clustered together and the tree of bumblebees (Bombus) appeared as one clade next to honeybees (Apis). Transcript expression analysis showed that BLVgR was expressed in all tested tissues and showed the highest abundance in the ovaries. BLVgR expression was present in all developmental stages. However, the expression level in larvae was extremely low. In addition, the expression of BLVgR was significantly upregulated after egg laying in both workers and queens. In new emerging workers injected with 5 µg of VgR dsRNA, the expression level of BLVgR was 4-fold lower than that in the GFP dsRNA-injected group after 72 h. Furthermore, BLVgR silencing significantly reduced the number of eggs laid (3.67 ± 1.96 eggs) and delayed the first egg-laying time (16.31 ± 2.07 days) in worker microcolonies when compared to dsGFP (37.31 ± 4.09 eggs, 8.15 ± 0.22 days) and DEPC-treated water injected controls (16.42 ± 2.24 eggs, 10.00 ± 0.37 days). In conclusion, the BLVgR gene and its reproductive function were explored in the bumblebee B. lantschouensis. This gene plays an important role in egg laying time and egg number. [ABSTRACT FROM AUTHOR]

Bumblebees are important insect pollinators for many wildflowers and crops. MicroRNAs (miRNAs) are endogenous non-coding small RNAs that regulate different biological functions in insects. In this study, the miRNAs in the heads of the three castes of the bumblebee Bombus lantschouensis were identified and characterized by small RNA deep sequencing. The significant differences in the expression of miRNAs and their target genes were analyzed. The results showed that the length of the small RNA reads from males, queens, and workers was distributed between 18 and 30 nt, with a peak at 22 nt. A total of 364 known and 89 novel miRNAs were identified from the heads of the three castes. The eight miRNAs with the highest expressed levels in males, queens, and workers were identical, although the order of these miRNAs based on expression differed. The male vs. queen, male vs. worker, and worker vs. queen comparisons identified nine, fourteen, and four miRNAs with significant differences in expression, respectively. The different castes were clustered based on the differentially expressed miRNAs (DE miRNAs), and the expression levels of the DE miRNAs obtained by RT-qPCR were consistent with the read counts obtained through Solexa sequencing. The putative target genes of these DE miRNAs were enriched in 29 Gene Ontology (GO) terms, and catalytic activity was the most enriched GO term, as demonstrated by its association with 2837 target genes in the male vs. queen comparison, 3535 target genes in the male vs. worker comparison, and 2185 target genes in the worker vs. queen comparison. This study highlights the characteristics of the miRNAs in the three B. lantschouensis castes and will aid further studies on the functions of miRNAs in bumblebees. [ABSTRACT FROM AUTHOR]