The expansion of agriculture has shaped the recent evolutionary history of a specialized squash pollinator.

The expansion of agriculture is responsible for the mass conversion of biologically diverse natural environments into managed agroecosystems dominated by a handful of genetically homogeneous crop species. Agricultural ecosystems typically have very different abiotic and ecological conditions from those they replaced and create potential niches for those species that are able to exploit the abundant resources offered by crop plants. While there are well-studied examples of crop pests that have adapted into novel agricultural niches, the impact of agricultural intensification on the evolution of crop mutualists such as pollinators is poorly understood. We combined genealogical inference from genomic data with archaeological records to demonstrate that the Holocene demographic history of a wild specialist pollinator of Cucurbita (pumpkins, squashes, and gourds) has been profoundly impacted by the history of agricultural expansion in North America. Populations of the squash bee Eucera pruinosa experienced rapid growth in areas where agriculture intensified within the past 1,000 y, suggesting that the cultivation of Cucurbita in North America has increased the amount of floral resources available to these bees. In addition, we found that roughly 20% of this bee species' genome shows signatures of recent selective sweeps. These signatures are overwhelmingly concentrated in populations from eastern North America where squash bees were historically able to colonize novel environments due to human cultivation of Cucurbita pepo and now exclusively inhabit agricultural niches. These results suggest that the widespread cultivation of crops can prompt adaptation in wild pollinators through the distinct ecological conditions imposed by agricultural environments.

Plant virus impacts on yield and plant-pollinator interactions are phylogenetically modulated independently of domestication in Cucurbita spp.

Plant defenses are conserved among closely related species, but domestication can alter host genotypes through artificial selection with potential losses in host defenses. Therefore, both domestication and host phylogenetic structure may influence plant virus infection outcomes. Here, we examined the association of phylogeny and domestication with the fitness of infected plants. We inoculated three pairs of domesticated and wild/non-cultivated squash (Cucurbita spp.) with a combination of two viruses commonly found to co-infect cucurbits, zucchini yellow mosaic virus (ZYMV) and squash mosaic virus (SqMV), and recorded fitness traits related to flowers, pollination, fruit and seed viability in the field over two separate years. In an additional field experiment, we recorded the relative abundance of both viruses via RT-qPCR. We found a gradient of susceptibility across the 6 tested lineages and phylogenetic structure, but not domestication, contributed to differences in infection outcomes and impacts on several fitness traits including fruit number, fruit weight, and germination. Plant virus infection also impacted the quantity and quality of floral rewards and visitation rates of specialist bee pollinators. There were no detectable differences in viral load between the 6 host taxa for either virus individually or the ratio of ZYMV:SqMV. Our results highlight the importance of phylogenetic structure in predicting host susceptibility to disease across wild and domesticated plants and the ability of several hosts to maintain fitness in the field despite infection. Broader consequences of plant pathogens for beneficial insects, such as pollinators, should also be considered in future research.

Squash bees host high diversity and prevalence of parasites in the northeastern United States.

The squash bee Eucera (Peponapis) pruinosa is emerging as a model species to study how stressors impact solitary wild bees in North America. Here, we describe the prevalence of trypanosomes, microsporidians and mollicute bacteria in E. pruinosa and two other species, Bombus impatiens and Apis mellifera, that together comprise over 97% of the pollinator visitors of Cucurbita agroecosystems in Pennsylvania (United States). Our results indicate that all three parasite groups are commonly detected in these bee species, but E. pruinosa often exhibit higher prevalences. We further describe novel trypanosome parasites detected in E. pruinosa, however it is unknown how these parasites impact these bees. We suggest future work investigates parasite replication and infection outcomes.

Honey bee viruses are highly prevalent but at low intensities in wild pollinators of cucurbit agroecosystems.

Managed and wild bee populations are in decline around the globe due to several biotic and abiotic stressors. Pathogenic viruses associated with the Western honey bee (Apis mellifera) have been identified as key contributors to losses of managed honey bee colonies, and are known to be transmitted to wild bee populations through shared floral resources. However, little is known about the prevalence and intensity of these viruses in wild bee populations, or how bee visitation to flowers impacts viral transmission in agroecosystems. This study surveyed honey bee, bumble bee (Bombus impatiens) and wild squash bee (Eucera (Peponapis) pruinosa) populations in Cucurbita agroecosystems across Pennsylvania (USA) for the prevalence and intensity of five honey bee viruses: acute bee paralysis virus (ABPV), deformed wing virus (DWV), Israeli acute paralysis virus (IAPV), Kashmir bee virus (KBV), and slow bee paralysis virus (SBPV). We investigated the potential role of bee visitation rate to flowers on DWV intensity among species in the pollinator community, with the expectation that increased bee visitation to flowers would increase the opportunity for transmission events between host species. We found that honey bee viruses are highly prevalent but in lower titers in wild E. pruinosa and B. impatiens than in A. mellifera populations throughout Pennsylvania (USA). DWV was detected in 88% of B. impatiens, 48% of E. pruinosa, and 95% of A. mellifera. IAPV was detected in 5% of B. impatiens and 4% of E. pruinosa, compared to 9% in A. mellifera. KBV was detected in 1% of B. impatiens and 5% of E. pruinosa, compared to 32% in A. mellifera. Our results indicate that DWV titers are not correlated with bee visitation in Cucurbita fields. The potential fitness impacts of these low viral titers detected in E. pruinosa remain to be investigated.

A trait-based approach to predict population genetic structure in bees.

Understanding population genetic structure is key to developing predictions about species susceptibility to environmental change, such as habitat fragmentation and climate change. It has been theorized that life-history traits may constrain some species in their dispersal and lead to greater signatures of population genetic structure. In this study, we use a quantitative comparative approach to assess if patterns of population genetic structure in bees are driven by three key species-level life-history traits: body size, sociality, and diet breadth. Specifically, we reviewed the current literature on bee population genetic structure, as measured by the differentiation indices Nei's GST, Hedrick's G'ST , and Jost's D. We then used phylogenetic generalised linear models to estimate the correlation between the evolution of these traits and patterns of genetic differentiation. Our analyses revealed a negative and significant effect of body size on genetic structure, regardless of differentiation index utilized. For Hedrick's G'ST and Jost's D, we also found a significant impact of sociality, where social species exhibited lower levels of differentiation than solitary species. We did not find an effect of diet specialization on population genetic structure. Overall, our results suggest that physical dispersal or other functions related to body size are among the most critical for mediating population structure for bees. We further highlight the importance of standardizing population genetic measures to more easily compare studies and to identify the most susceptible species to landscape and climatic changes.

Physiological thermal limits predict differential responses of bees to urban heat-island effects.

Changes in community composition are an important, but hard to predict, effect of climate change. Here, we use a wild-bee study system to test the ability of critical thermal maxima (CTmax, a measure of heat tolerance) to predict community responses to urban heat-island effects in Raleigh, NC, USA. Among 15 focal species, CTmax ranged from 44.6 to 51.3°C, and was strongly predictive of population responses to urban warming across 18 study sites (r2 = 0.44). Species with low CTmax declined the most. After phylogenetic correction, solitary species and cavity-nesting species (bumblebees) had the lowest CTmax, suggesting that these groups may be most sensitive to climate change. Community responses to urban and global warming will likely retain strong physiological signal, even after decades of warming during which time lags and interspecific interactions could modulate direct effects of temperature.

Crop domestication facilitated rapid geographical expansion of a specialist pollinator, the squash bee Peponapis pruinosa.

Squash was first domesticated in Mexico and is now found throughout North America (NA) along with Peponapis pruinosa, a pollen specialist bee species of the squash genus Cucurbita The origin and spread of squash cultivation is well-studied archaeologically and phylogenetically; however, no study has documented how cultivation of this or any other crop has influenced species in mutualistic interactions. We used molecular markers to reconstruct the demographic range expansion and colonization routes of P. pruinosa from its native range into temperate NA. Populations east of the Rocky Mountains expanded from the wild host plant's range in Mexico and were established by a series of founder events. Eastern North America was most likely colonized from squash bee populations in the present-day continental Midwest USA and not from routes that followed the Gulf and Atlantic coasts from Mexico. Populations of P. pruinosa west of the Rockies spread north from the warm deserts much more recently, showing two genetically differentiated populations with no admixture: one in California and the other one in eastern Great Basin. These bees have repeatedly endured severe bottlenecks as they colonized NA, following human spread of their Cucurbita pollen hosts during the Holocene.

Reduced cellular immune response in social insect lineages.

Social living poses challenges for individual fitness because of the increased risk of disease transmission among conspecifics. Despite this challenge, sociality is an evolutionarily successful lifestyle, occurring in the most abundant and diverse group of organisms on earth--the social insects. Two contrasting hypotheses predict the evolutionary consequences of sociality on immune systems. The social group hypothesis posits that sociality leads to stronger individual immune systems because of the higher risk of disease transmission in social species. By contrast, the relaxed selection hypothesis proposes that social species have evolved behavioural immune defences that lower disease risk within the group, resulting in lower immunity at the individual level. We tested these hypotheses by measuring the encapsulation response in 11 eusocial and non-eusocial insect lineages. We built phylogenetic mixed linear models to investigate the effect of behaviour, colony size and body size on cellular immune response. We found a significantly negative effect of colony size on encapsulation response (Markov chain Monte Carlo generalized linear mixed model (mcmcGLMM) p < 0.05; phylogenetic generalized least squares (PGLS) p < 0.05). Our findings suggest that insects living in large societies may rely more on behavioural mechanisms, such as hygienic behaviours, than on immune function to reduce the risk of disease transmission among nest-mates.

Climate, physiological tolerance and sex-biased dispersal shape genetic structure of Neotropical orchid bees.

Understanding the impact of past climatic events on the demographic history of extant species is critical for predicting species' responses to future climate change. Palaeoclimatic instability is a major mechanism of lineage diversification in taxa with low dispersal and small geographical ranges in tropical ecosystems. However, the impact of these climatic events remains questionable for the diversification of species with high levels of gene flow and large geographical distributions. In this study, we investigate the impact of Pleistocene climate change on three Neotropical orchid bee species (Eulaema bombiformis, E. meriana and E. cingulata) with transcontinental distributions and different physiological tolerances. We first generated ecological niche models to identify species-specific climatically stable areas during Pleistocene climatic oscillations. Using a combination of mitochondrial and nuclear markers, we inferred calibrated phylogenies and estimated historical demographic parameters to reconstruct the phylogeographical history of each species. Our results indicate species with narrower physiological tolerance experienced less suitable habitat during glaciations and currently exhibit strong population structure in the mitochondrial genome. However, nuclear markers with low and high mutation rates show lack of association with geography. These results combined with lower migration rate estimates from the mitochondrial than the nuclear genome suggest male-biased dispersal. We conclude that despite large effective population sizes and capacity for long-distance dispersal, climatic instability is an important mechanism of maternal lineage diversification in orchid bees. Thus, these Neotropical pollinators are susceptible to disruption of genetic connectivity in the event of large-scale climatic changes.

Body mass, temperature, and pathogen intensity differentially affect critical thermal maxima and their population-level variation in a solitary bee.

Climate change presents a major threat to species distribution and persistence. Understanding what abiotic or biotic factors influence the thermal tolerances of natural populations is critical to assessing their vulnerability under rapidly changing thermal regimes. This study evaluates how body mass, local climate, and pathogen intensity influence heat tolerance and its population-level variation (SD) among individuals of the solitary bee Xenoglossa pruinosa. We assess the sex-specific relationships between these factors and heat tolerance given the differences in size between sexes and the ground-nesting behavior of the females. We collected X. pruinosa individuals from 14 sites across Pennsylvania, USA, that varied in mean temperature, precipitation, and soil texture. We measured the critical thermal maxima (CTmax) of X. pruinosa individuals as our proxy for heat tolerance and used quantitative PCR to determine relative intensities of three parasite groups-trypanosomes, Spiroplasma apis (mollicute bacteria), and Vairimorpha apis (microsporidian). While there was no difference in CTmax between the sexes, we found that CTmax increased significantly with body mass and that this relationship was stronger for males than for females. Air temperature, precipitation, and soil texture did not predict mean CTmax for either sex. However, population-level variation in CTmax was strongly and negatively correlated with air temperature, which suggests that temperature is acting as an environmental filter. Of the parasites screened, only trypanosome intensity correlated with heat tolerance. Specifically, trypanosome intensity negatively correlated with the CTmax of female X. pruinosa but not males. Our results highlight the importance of considering size, sex, and infection status when evaluating thermal tolerance traits. Importantly, this study reveals the need to evaluate trends in the variation of heat tolerance within and between populations and consider implications of reduced variation in heat tolerance for the persistence of ectotherms in future climate conditions.

Systemic pesticides in a solitary bee pollen food store affect larval development and increase pupal mortality.

Solitary bees are often exposed to various pesticides applied for pest control on farmland while providing pollination services to food crops. Increasing evidence suggests that sublethal toxicity of agricultural pesticides affects solitary bees differently than the social bees used to determine regulatory thresholds, such as honey bees and bumblebees. Studies on solitary bees are challenging because of the difficulties in obtaining large numbers of eggs or young larvae for bioassays. Here we show the toxic and sublethal developmental effects of four widely used plant systemic pesticides on the Japanese orchard bee (Osmia cornifrons). Pollen food stores of this solitary bee were treated with different concentrations of three insecticides (acetamiprid, flonicamid, and sulfoxaflor) and a fungicide (dodine). Eggs were transplanted to the treated pollen and larvae were allowed to feed on the pollen stores after egg hatch. The effects of chronic ingestion of contaminated pollen were measured until adult eclosion. This year-long study revealed that chronic exposure to all tested pesticides delayed larval development and lowered larval and adult body weights. Additionally, exposure to the systemic fungicide resulted in abnormal larval defecation and increased mortality at the pupal stage, indicating potential risk to bees from fungicide exposure. These findings demonstrate potential threats to solitary bees from systemic insecticides and fungicides and will help in making policy decisions to mitigate these effects.

Osmia taurus (Hymenoptera: Megachilidae): A Non-native Bee Species With Invasiveness Potential in North America.

Bees are important pollinators and are essential for the reproduction of many plants in natural and agricultural ecosystems. However, bees can have adverse ecological effects when introduced to areas outside of their native geographic ranges. Dozens of non-native bee species are currently found in North America and have raised concerns about their potential role in the decline of native bee populations. Osmia taurus Smith (Hymenoptera: Megachilidae) is a mason bee native to eastern Asia that was first reported in the United States in 2002. Since then, this species has rapidly expanded throughout the eastern part of North America. Here, we present a comprehensive review of the natural history of O. taurus, document its recent history of spread through the United States and Canada, and discuss the evidence suggesting its potential for invasiveness. In addition, we compare the biology and history of colonization of O. taurus to O. cornifrons (Radoszkowski), another non-native mason bee species now widespread in North America. We highlight gaps of knowledge and future research directions to better characterize the role of O. taurus in the decline of native Osmia spp. Panzer and the facilitation of invasive plant-pollinator mutualisms.

A longitudinal experiment demonstrates that honey bee colonies managed organically are as healthy and productive as those managed conventionally.

Honey bee colony management is critical to mitigating the negative effects of biotic and abiotic stressors. However, there is significant variation in the practices implemented by beekeepers, which results in varying management systems. This longitudinal study incorporated a systems approach to experimentally test the role of three representative beekeeping management systems (conventional, organic, and chemical-free) on the health and productivity of stationary honey-producing colonies over 3 years. We found that the survival rates for colonies in the conventional and organic management systems were equivalent, but around 2.8 times greater than the survival under chemical-free management. Honey production was also similar, with 102% and 119% more honey produced in conventional and organic management systems, respectively, than in the chemical-free management system. We also report significant differences in biomarkers of health including pathogen levels (DWV, IAPV, Vairimorpha apis, Vairimorpha ceranae) and gene expression (def-1, hym, nkd, vg). Our results experimentally demonstrate that beekeeping management practices are key drivers of survival and productivity of managed honey bee colonies. More importantly, we found that the organic management system-which uses organic-approved chemicals for mite control-supports healthy and productive colonies, and can be incorporated as a sustainable approach for stationary honey-producing beekeeping operations.

Six years of wild bee monitoring shows changes in biodiversity within and across years and declines in abundance.

Wild bees form diverse communities that pollinate plants in both native and agricultural ecosystems making them both ecologically and economically important. The growing evidence of bee declines has sparked increased interest in monitoring bee community and population dynamics using standardized methods. Here, we studied the dynamics of bee biodiversity within and across years by monitoring wild bees adjacent to four apple orchard locations in Southern Pennsylvania, USA. We collected bees using passive Blue Vane traps continuously from April to October for 6 years (2014-2019) amassing over 26,000 bees representing 144 species. We quantified total abundance, richness, diversity, composition, and phylogenetic structure. There were large seasonal changes in all measures of biodiversity with month explaining an average of 72% of the variation in our models. Changes over time were less dramatic with years explaining an average of 44% of the variation in biodiversity metrics. We found declines in all measures of biodiversity especially in the last 3 years, though additional years of sampling are needed to say if changes over time are part of a larger trend. Analyses of population dynamics over time for the 40 most abundant species indicate that about one third of species showed at least some evidence for declines in abundance. Bee family explained variation in species-level seasonal patterns but we found no consistent family-level patterns in declines, though bumble bees and sweat bees were groups that declined the most. Overall, our results show that season-wide standardized sampling across multiple years can reveal nuanced patterns in bee biodiversity, phenological patterns of bees, and population trends over time of many co-occurring species. These datasets could be used to quantify the relative effects that different aspects of environmental change have on bee communities and to help identify species of conservation concern.

The diversity, evolution, and development of setal morphologies in bumble bees (Hymenoptera: Apidae: Bombus spp.).

Bumble bees are characterized by their thick setal pile that imparts aposematic color patterns often used for species-level identification. Like all bees, the single-celled setae of bumble bees are branched, an innovation thought important for pollen collection. To date no studies have quantified the types of setal morphologies and their distribution on these bees, information that can facilitate understanding of their adaptive ecological function. This study defines several major setal morphotypes in the common eastern bumble bee Bombus impatiens Cresson, revealing these setal types differ by location across the body. The positions of these types of setae are similar across individuals, castes, and sexes within species. We analyzed the distribution of the two most common setal types (plumose and spinulate) across the body dorsum of half of the described bumble bee species. This revealed consistently high density of plumose (long-branched) setae across bumble bees on the head and mesosoma, but considerable variation in the amount of metasomal plumosity. Variation on the metasoma shows strong phylogenetic signal at subgeneric and smaller group levels, making it a useful trait for species delimitation research, and plumosity has increased from early Bombus ancestors. The distribution of these setal types suggests these setae may serve several functions, including pollen-collecting and thermoregulatory roles, and probable mechanosensory functions. This study further examines how and when setae of the pile develop, evidence for mechanosensory function, and the timing of pigmentation as a foundation for future genetic and developmental research in these bees.

Overlap of Ecological Niche Breadth of Euglossa cordata and Eulaema nigrita (Hymenoptera, Apidae, Euglossini) Accessed by Pollen Loads and Species Distribution Modeling.

Urban areas can serve as biodiversity refuges for pollinators because of the high diversity of available floral and nesting resources. However, it remains unclear what plant species commonly used for urban landscaping provide floral resources that pollinators actively use. Here, we integrate data from the pollen and species distribution models of two abundant euglossine bees-the large-bodied Eulaema nigrita (Lepeletier, 1841) and the small-bodied Euglossa cordata (Linnaeus, 1758)-in urban areas to investigate their overlap in diet breadth and distribution. We hypothesized that because bees with larger body sizes tend to have larger foraging areas, large-bodied bees would have a wider diet breath than small-bodied bees. Contrary to our hypothesis, we found that Eg. cordata has a wider diet breadth than El. nigrita with the former species showing higher diversity of pollen types collected (per pollen load and on average across pollen loads). Pollen grains from Solanum paniculatum and Tradescantia zebrina represented 63% of the diet of Eg. cordata, whereas pollen from S. paniculatum and Psidium guajava represented 87% of the diet of El. nigrita. After overlaying the distribution of both bee species and the three most important pollen resources, the distribution models revealed that these three plant species can co-occur with both euglossine bees throughout a large portion of eastern Brazil near the coast. Thus, we conclude S. paniculatum, T. zebrina, and P. guajava should be considered key plants for the maintenance of these two urban euglossine bee species. The results of this study provide important information for urban landscaping programs that aim to protect and preserve pollinators.

Defining Pollinator Health: A Holistic Approach Based on Ecological, Genetic, and Physiological Factors.

Evidence for global bee population declines has catalyzed a rapidly evolving area of research that aims to identify the causal factors and to effectively assess the status of pollinator populations. The term pollinator health emerged through efforts to understand causes of bee decline and colony losses, but it lacks a formal definition. In this review, we propose a definition for pollinator health and synthesize the available literature on the application of standardized biomarkers to assess health at the individual, colony, and population levels. We focus on biomarkers in honey bees, a model species, but extrapolate the potential application of these approaches to monitor the health status of wild bee populations. Biomarker-guided health measures can inform beekeeper management decisions, wild bee conservation efforts, and environmental policies. We conclude by addressing challenges to pollinator health from a One Health perspective that emphasizes the interplay between environmental quality and human, animal, and bee health.

Introduced bees (Osmia cornifrons) collect pollen from both coevolved and novel host-plant species within their family-level phylogenetic preferences.

Studying the pollen preferences of introduced bees allows us to investigate how species use host-plants when establishing in new environments. Osmia cornifrons is a solitary bee introduced into North America from East Asia for pollination of Rosaceae crops such as apples and cherries. We investigated whether O. cornifrons (i) more frequently collected pollen from host-plant species they coevolved with from their geographic origin, or (ii) prefer host-plant species of specific plant taxa independent of origin. To address this question, using pollen metabarcoding, we examined the identity and relative abundance of pollen in larval provisions from nests located in different landscapes with varying abundance of East-Asian and non-Asian plant species. Our results show that O. cornifrons collected more pollen from plant species from their native range. Plants in the family Rosaceae were their most preferred pollen hosts, but they differentially collected species native to East Asia, Europe, or North America depending on the landscape. Our results suggest that while O. cornifrons frequently collect pollen of East-Asian origin, the collection of pollen from novel species within their phylogenetic familial affinities is common and can facilitate pollinator establishment. This phylogenetic preference highlights the effectiveness of O. cornifrons as crop pollinators of a variety of Rosaceae crops from different geographic origins. Our results imply that globalization of non-native plant species may ease the naturalization of their coevolved pollinators outside of their native range.

Correction to 'Introduced bees (Osmia cornifrons) collect pollen from both coevolved and novel host-plant species within their family-level phylogenetic preferences'.

[This corrects the article DOI: 10.1098/rsos.200225.].

A new ingestion bioassay protocol for assessing pesticide toxicity to the adult Japanese orchard bee (Osmia cornifrons).

Adopting an Integrated Pest and Pollinator Management strategy requires an evaluation of pesticide risk for pollinator species. For non-Apid species, however, the standardized ingestion assays are difficult to implement. This hinders the consideration of non-Apid species in farm management strategies and government regulatory processes. We describe a new method for a mason bee, Osmia cornifrons, which is an important pollinator of apples and other fruit crops. Our approach overcomes high control mortality seen in other bioassay protocols and expands testing to include males as well as females. The new pesticide toxicity assessment protocol showed that (1) a group feeding method is optimum even though there is no trophallaxis, (2) males had better tolerance to pesticides although they are smaller, and (3) pesticides can cause additional mortality after the standard 48 h assessment time specified by European Food Safety Authority and U.S. Environmental Protection Agency.