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Apis mellifera Linnaeus (Hymenoptera: Apis), honey bees, are the most widely used managed crop pollinators. However, their high rental cost and uncertain availability for North American orchard crops have motivated growers to explore alternative pollination options. We examined whether adding solitary, spring-flying Osmia lignaria Say (Hymenoptera: Megachilidae), blue orchard bees, as co-pollinators with A. mellifera in Washington sweet cherry and pear orchards enhances fruit set and yield compared to the use of A. mellifera alone. We added managed O. lignaria to orchard sites where A. mellifera hives were already present. Fruit set, fruit yield, and O. lignaria reproduction at O. lignaria-supplemented sites were compared to nearby, paired sites pollinated only by A. mellifera (3 paired cherry and 3 paired pear sites). For both crops, the addition of O. lignaria significantly increased fruit set but did not yield at harvest. Microscopic inspection of pollen grains from O. lignaria nest cell provisions confirmed that O. lignaria primarily visited orchard flowers. Mean retention of O. lignaria in cherry orchards was slightly higher (65%) than O. lignaria retention reported in other orchard crops (30%-60%). However, retention in pear orchards was much lower (≤20%). These results show that supplementing hives with O. lignaria in Washington spring orchard crops can increase overall pollination, but that trees fail to bear developing fruit to maturity. The strategy of using co-pollinators, O. lignaria and A. mellifera, in US orchards may act as "pollination insurance" when A. mellifera hives are in low supply or when the weather is not amenable for A. mellifera flight during the bloom period., (Published by Oxford University Press on behalf of Entomological Society of America 2024.)

The blue orchard bee, Osmia lignaria Say (Hymenoptera: Megachilidae), is a solitary, cavity-nesting species used for pollinating spring blooming crops. Commercial stocks are sourced from a few locations in the western United States but are sold across the country. However, the existence of local adaptations of these bees is unknown, such as the propensity to nest in nearby provided materials or to disperse broadly beyond release sites. In spring 2019, California- and Utah-sourced blue orchard bees were introduced into cherry orchards in both source and reciprocal states. Nest boxes were placed near (within 78 m) and far (500 m-1 km) from central bee release points. Paint-marked bees were released when floral resources were available. Observations of marked bees at nest boxes were used to evaluate female retention and dispersal pattern. Nesting bee counts in March-blooming California orchards revealed a significant difference in female retention by population source; over twice as many UT bees established nests than did CA bees. Few females were found at far nest sites. In May-blooming Utah orchards, counts of CA and UT bees were similar at near and far nest sites; neither female retention nor dispersal was significantly affected by bee origin. It is concerning that CA females were less likely to be retained in California orchards because the demand for commercial pollination is high for early-blooming California almond and cherry. Our results highlight the need to understand potential consequences of bee origin and their management on pollinator performance and reproduction in target crops., (Published by Oxford University Press on behalf of Entomological Society of America 2023.)

Melittobia acasta Walker is one among other hymenopterous parasitoids of Megachile rotundata F. Commercial M. rotundata populations are employed to pollinate North American alfalfa for seed production. This wasp can be prolific when using M. rotundata as a host and can reduce or destroy bee stocks. Hundreds of M. acasta female offspring can develop in a single M. rotundata cell and disperse to infest other cells, producing thousands of more parasitoids. In this study, we determined (i) upon what bee life stages M. acasta females choose to lay eggs and if those eggs ultimately become adults and (ii) M. acasta female longevity when exposed to various resources within M. rotundata cells. We found that M. acasta females lay eggs on M. rotundata prepupae and pupae and that those eggs can hatch and survive to adulthood. Eggs are not laid on early instar bee larvae; eggs laid on adults do not survive. Average female life span is 5 days without feeding, 8-9 days if a pollen-nectar provision is available while the bee develops through larval stages, and 34 days if the wasp can feed on prepupal hemolymph. Wasp females can emerge from bee cells several days after trays of cells are taken to fields. Therefore, adult females could survive long enough for new bee offspring to become prepupae. Our findings support a better understanding of host life stage preference and the longevity of M. acasta females that can inform the timing of the implementation of possible control measures., (Published by Oxford University Press on behalf of Entomological Society of America 2023.)

Osmia lignaria Say is used in combination with Apis mellifera L. to pollinate U.S. orchard crops. The deployment of O. lignaria requires artificial warming to synchronize adult bee emergence with crop bloom. However, current methods for emerging bees are time consuming and inefficient; the Hivetop Incubator (HTI) invention creates a space atop an A. mellifera hive whose heat flows through the screened bottom of the HTI to incubate cocooned O. lignaria adults therein. In response to HTI heat, O. lignaria adults chew out of cocoons, find an exit hole, and fly away to nest in provided nesting sites and, thereby, pollinate a crop. Objectives for studies performed in Utah and Washington were to: 1) determine whether HTIs inhibit A. mellifera thermoregulation or colony growth, 2) compare O. lignaria emergence duration from an HTI with and without hive heat, and 3) assess whether O. lignaria females leave HTIs located at the orchard edge to nest throughout the orchard. We found no significant differences between the internal temperatures of A. mellifera colonies with and without HTIs and no impact on A. mellifera food storage or brood production. Osmia lignaria in hive-heated HTIs emerged 3× faster than bees in unheated HTIs. Heated HTIs were significantly cooler than hive temperatures but significantly warmer than HTIs atop empty hive boxes. Osmia lignaria nest distribution was not correlated to the location of HTIs at the orchard edge. Overall, HTIs were effective for timely, on-site emergence of O. lignaria for orchard pollination without negatively impacting A. mellifera colonies., (Published by Oxford University Press on behalf of Entomological Society of America 2023.)

Maternal effects are an important source of phenotypic variance, whereby females influence offspring developmental trajectory beyond direct genetic contributions, often in response to changing environmental conditions. However, relatively little is known about the mechanisms by which maternal experience is translated into molecular signals that shape offspring development. One such signal may be maternal RNA transcripts (mRNAs and miRNAs) deposited into maturing oocytes. These regulate the earliest stages of development of all animals, but are understudied in most insects. Here we investigated the effects of female internal (body condition) and external (time of season) environmental conditions on maternal RNA in the maturing oocytes and 24-h-old eggs (24-h eggs) of alfalfa leafcutting bees. Using gene expression and WGCNA analysis, we found that females adjust the quantity of mRNAs related to protein phosphorylation, transcriptional regulation, and nuclease activity deposited into maturing oocytes in response to both poor body condition and shorter day lengths that accompany the late season. However, the magnitude of these changes was higher for time of season. Females also adjusted miRNA deposition in response to seasonal changes, but not body condition. We did not observe significant changes in maternal RNAs in response to either body condition or time of season in 24-h eggs, which were past the maternal-to-zygotic transition. Our results suggest that females adjust the RNA transcripts they provide for offspring to regulate development in response to both internal and external environmental cues. Variation in maternal RNAs may, therefore, be important for regulating offspring phenotype in response to environmental change., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Hagadorn, Hunter, DeLory, Johnson, Pitts-Singer and Kapheim.)

Comprehensive decisions on the management of commercially produced bees, depend largely on associated knowledge of genetic diversity. In this study, we present novel microsatellite markers to support the breeding, management, and conservation of the blue orchard bee, Osmia lignaria Say (Hymenoptera: Megachilidae). Native to North America, O. lignaria has been trapped from wildlands and propagated on-crop and used to pollinate certain fruit, nut, and berry crops. Harnessing the O. lignaria genome assembly, we identified 59,632 candidate microsatellite loci in silico, of which 22 were tested using molecular techniques. Of the 22 loci, 12 loci were in Hardy-Weinberg equilibrium (HWE), demonstrated no linkage disequilibrium (LD), and achieved low genotyping error in two Intermountain North American wild populations in Idaho and Utah, USA. We found no difference in population genetic diversity between the two populations, but there was evidence for low but significant population differentiation. Also, to determine if these markers amplify in other Osmia, we assessed 23 species across the clades apicata, bicornis, emarginata, and ribifloris. Nine loci amplified in three species/subspecies of apicata, 22 loci amplified in 11 species/subspecies of bicornis, 11 loci amplified in seven species/subspecies of emarginata, and 22 loci amplified in two species/subspecies of ribifloris. Further testing is necessary to determine the capacity of these microsatellite loci to characterize genetic diversity and structure under the assumption of HWE and LD for species beyond O. lignaria. These markers will inform the conservation and commercial use of trapped and managed O. lignaria and other Osmia species for both agricultural and nonagricultural systems., (Published by Oxford University Press on behalf of Entomological Society of America 2023.)

Seventy five percent of the world's food crops benefit from insect pollination. Hence, there has been increased interest in how global change drivers impact this critical ecosystem service. Because standardized data on crop pollination are rarely available, we are limited in our capacity to understand the variation in pollination benefits to crop yield, as well as to anticipate changes in this service, develop predictions, and inform management actions. Here, we present CropPol, a dynamic, open, and global database on crop pollination. It contains measurements recorded from 202 crop studies, covering 3,394 field observations, 2,552 yield measurements (i.e., berry mass, number of fruits, and fruit density [kg/ha], among others), and 47,752 insect records from 48 commercial crops distributed around the globe. CropPol comprises 32 of the 87 leading global crops and commodities that are pollinator dependent. Malus domestica is the most represented crop (32 studies), followed by Brassica napus (22 studies), Vaccinium corymbosum (13 studies), and Citrullus lanatus (12 studies). The most abundant pollinator guilds recorded are honey bees (34.22% counts), bumblebees (19.19%), flies other than Syrphidae and Bombyliidae (13.18%), other wild bees (13.13%), beetles (10.97%), Syrphidae (4.87%), and Bombyliidae (0.05%). Locations comprise 34 countries distributed among Europe (76 studies), North America (60), Latin America and the Caribbean (29), Asia (20), Oceania (10), and Africa (7). Sampling spans three decades and is concentrated on 2001-2005 (21 studies), 2006-2010 (40), 2011-2015 (88), and 2016-2020 (50). This is the most comprehensive open global data set on measurements of crop flower visitors, crop pollinators and pollination to date, and we encourage researchers to add more datasets to this database in the future. This data set is released for non-commercial use only. Credits should be given to this paper (i.e., proper citation), and the products generated with this database should be shared under the same license terms (CC BY-NC-SA)., (© 2021 The Ecological Society of America.)

Wild and managed bee populations are in decline, and one of many environmental causes is the impact of pesticides on developing bees. For solitary bees, delayed larval development could lead to asynchronous adult emergence, unhealthy and inefficient adult pollinators, and decreased brood production and survival. We examined a methodology for testing Osmia lignaria Say (Hymenoptera: Megachilidae) larval responses to pesticide exposure using a laboratory bioassay. We created two provision types: a homogenized blend of O. lignaria provisions from an apple orchard and homogenized almond pollen pellets collected by honey bees plus sugar water. Pesticides were administered to the provisions to compare toxic effects. We recorded larval developmental durations for second-fifth instar and for fifth instar to cocoon initiation for larvae fed provisions treated with water (control) or doses of three pesticides and a representative spray-tank mixture (acetamiprid, boscalid/pyraclostrobin, dimethoate, and acetamiprid plus boscalid/pyraclostrobin). All larvae survived to cocoon initiation when only water was added to provisions. Impacts of pesticide treatments significantly differed between the apple and almond homogenates. The greatest treatment effects occurred when the homogenized almond provision was mixed with acetamiprid alone and when combined with boscalid/pyraclostrobin. Optimizing bioassays through the use of appropriate larval food for exposing solitary bee larvae to agrochemicals is crucial for assessing risks for pollinators., (Published by Oxford University Press on behalf of Entomological Society of America 2021.)

Metrics to assess relative adult bee body size have included both mass and morphometrics, but these metrics may not equally or reliably estimate body size for all bee species and in all situations, due to bee age, diet, and/or environment. Understanding the relationships between different metrics and possible redundancies in the information they afford is important but not always known. Body size measurements provide valuable data for interpreting research outcomes for managed solitary bees, including Osmia lignaria Say and Megachile rotundata F. (Hymenoptera: Megachilidae). Applied studies of these important and readily available U.S. crop pollinators focus on refining commercial management practices, and basic empirical studies in various scientific disciplines (from genomics to ecology) employ them as model systems to study solitary bees. To examine common metrics of body size, we measured head capsule width (HCW), intertegular distance (ITD), and fresh and dry weights of newly emerged adults of both species. Using linear and exponential models, we determined relationships between these body size metrics. For M. rotundata , linear models best described relationships between ITD and all other metrics, and between HCW and fresh and dry weights. For O. lignaria , linear models best fit relationships between all metrics except for fresh weight with both ITD and HCW, which were fitted better with exponential models. For both species, model fits were strongest when males and females were pooled. Depending on the study question, knowing that only one metric may reliably measure body size can simplify evaluations of O. lignaria and M. rotundata responses to artificial or environmental variables., Competing Interests: The authors declare that they have no competing interests.

Most of the world's crops depend on pollinators, so declines in both managed and wild bees raise concerns about food security. However, the degree to which insect pollination is actually limiting current crop production is poorly understood, as is the role of wild species (as opposed to managed honeybees) in pollinating crops, particularly in intensive production areas. We established a nationwide study to assess the extent of pollinator limitation in seven crops at 131 locations situated across major crop-producing areas of the USA. We found that five out of seven crops showed evidence of pollinator limitation. Wild bees and honeybees provided comparable amounts of pollination for most crops, even in agriculturally intensive regions. We estimated the nationwide annual production value of wild pollinators to the seven crops we studied at over $1.5 billion; the value of wild bee pollination of all pollinator-dependent crops would be much greater. Our findings show that pollinator declines could translate directly into decreased yields or production for most of the crops studied, and that wild species contribute substantially to pollination of most study crops in major crop-producing regions.

Megachile rotundata F. populations are managed to pollinate alfalfa, Medicago sativa L. (Fabaceae), for seed production in western North America. Some progeny produced in summer by overwintered M. rotundata females enter diapause as prepupae to overwinter and become adults the following year. Other offspring avert diapause to undergo adult emergence in summer. The regulatory mechanism(s) of diapause induction or aversion is unknown; the process apparently involves multiple and integrated factors. This 4-yr study sought to determine effects of the exposure of mother bees to short, long, very long, and natural photoperiods on their production of non-diapausing and diapausing progeny. Just-emerged adult bees from both Utah and Canada sources were exposed to different photoperiods for 3 d in incubators and then released into field enclosures placed over blooming alfalfa. Control bees were those exposed to only outdoor conditions. Reproduction was monitored for each female bee, and offspring diapause outcome was recorded. Progeny outcome data were informative for only two of the four study years. Generally, progeny of Utah mothers exposed to long and very long photoperiods were more likely to be non-diapausers compared to progeny of mothers exposed to short and control photoperiods. Short and long (but not very long) photoperiod maternal exposures increased the likelihood of diapause aversion in Canada progeny. Performing multiyear field studies on geographically distinct populations is imperative for revealing environmental challenges and inconsistent bee performance that can impair analyses and interpretation. Future similar studies are needed to more fully evaluate photoperiod effects on diapause., (Published by Oxford University Press on behalf of Entomological Society of America 2020.)

Concerns over the availability of honeybees ( Apis mellifera L.) to meet pollination demands have elicited interest in alternative pollinators to mitigate pressures on the commercial beekeeping industry. The blue orchard bee, Osmia lignaria (Say), is a commercially available native bee that can be employed as a copollinator with, or alternative pollinator to, honeybees in orchards. To date, their successful implementation in agriculture has been limited by poor recovery of bee progeny for use during the next spring. This lack of reproductive success may be tied to an inadequate diversity and abundance of alternative floral resources during the foraging period. Managed, supplementary wildflower plantings may promote O. lignaria reproduction in California almond orchards. Three wildflower plantings were installed and maintained along orchard edges to supplement bee forage. Plantings were seeded with native wildflower species that overlapped with and extended beyond almond bloom. We measured bee visitation to planted wildflowers, bee reproduction, and progeny outcomes across orchard blocks at variable distances from wildflower plantings during 2015 and 2016. Pollen provision composition was also determined to confirm O. lignaria wildflower pollen use. Osmia lignaria were frequently observed visiting wildflower plantings during, and after, almond bloom. Most O. lignaria nesting occurred at orchard edges. The greatest recovery of progeny occurred along the orchard edges having the closest proximity (80 m) to managed wildflower plantings versus edges farther away. After almond bloom, O. lignaria nesting closest to the wildflower plantings collected 72% of their pollen from Phacelia spp., which supplied 96% of the managed floral area. Phacelia spp. pollen collection declined with distance from the plantings, but still reached 17% 800 m into the orchard. This study highlights the importance of landscape context and proximity to supplementary floral resources in promoting the propagation of solitary bees as alternative managed pollinators in commercial agriculture., Competing Interests: None declared., (© 2020 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Osmia lignaria is a commercially available, native solitary bee species recognized for its propensity to forage upon and pollinate tree fruit crops such as apple, almond and cherry. This study evaluated the implementation of O. lignaria co-pollination with honey bees in central Utah commercial tart cherry orchards during 2017 and 2018 bloom. Three paired 1.2 ha sites were selected for evaluation of cherry fruit set and yield with and without managed O. lignaria releases alongside the standard honey bee hive stocking rate of 2.5 hives/ha. Osmia lignaria supplementation did not measurably increase cherry fruit set, fruit per limb cross-sectional area or fruit weight. The lack of differences in yield is likely a consequence of local saturation of pollinator services supplied by managed honey bees throughout experimental orchards, such that no additive benefit of managed O. lignaria releases were measurable. An increase in managed O. lignaria populations was achieved in 2017 but not 2018, possibly due to unknown changes to orchard management or environmental factors. While flying O. lignaria in Utah tart cherries may support sustainable in-field bee propagation, their subsequent impacts on tart cherry yield were not detected when paired with standard stocking densities of honey bees., Competing Interests: The authors declare there are no competing interests.

Current pesticide risk assessment for bees relies on a single (social) species, the western honey bee, Apis mellifera L. (Hymenoptera: Apidae). However, most of the >20,000 bee species worldwide are solitary. Differences in life history traits between solitary bees (SB) and honey bees (HB) are likely to determine differences in routes and levels of pesticide exposure. The objectives of this review are to: 1) compare SB and HB life history traits relevant for risk assessment; 2) summarize current knowledge about levels of pesticide exposure for SB and HB; 3) identify knowledge gaps and research needs; 4) evaluate whether current HB risk assessment schemes cover routes and levels of exposure of SB; and 5) identify potential SB model species for risk assessment. Most SB exposure routes seem well covered by current HB risk assessment schemes. Exceptions to this are exposure routes related to nesting substrates and nesting materials used by SB. Exposure via soil is of particular concern because most SB species nest underground. Six SB species (Hymenoptera: Megachilidae - Osmia bicornis L., O. cornifrons Radoszkowski, O. cornuta Latreille, O. lignaria Say, Megachile rotundata F., and Halictidae - Nomia melanderi Cockerell) are commercially available and could be used in risk assessment. Of these, only N. melanderi nests underground, and the rest are cavity-nesters. However, the three Osmia species collect soil to build their nests. Life history traits of cavity-nesting species make them particularly suitable for semifield and, to a lesser extent, field tests. Future studies should address basic biology, rearing methods and levels of exposure of ground-nesting SB species.

Current pesticide risk assessment practices use the honey bee, Apis mellifera L., as a surrogate to characterize the likelihood of chemical exposure of a candidate pesticide for all bee species. Bees make up a diverse insect group that provides critical pollination services to both managed and wild ecosystems. Accordingly, they display a diversity of behaviors and vary greatly in their lifestyles and phenologies, such as their timing of emergence, degree of sociality, and foraging and nesting behaviors. Some of these factors may lead to disparate or variable routes of exposure when compared to honey bees. For those that possess life histories that are distinct from A. mellifera, further risk assessments may be warranted. In January 2017, 40 bee researchers, representative of regulatory agencies, academia, and agrochemical industries, gathered to discuss the current state of science on pesticide exposure to non-Apis bees and to determine how well honey bee exposure estimates, implemented by different regulatory agencies, may be protective for non-Apis bees. Workshop participants determined that although current risk assessment procedures for honey bees are largely conservative, several routes of exposure are unique to non-Apis bees and warranted further investigation. In this forum article, we discuss these key routes of exposure relevant to non-Apis bees and identify important research gaps that can help inform future bee risk assessment decisions.

Our understanding of the mechanisms controlling insect diapause has increased dramatically with the introduction of global gene expression techniques, such as RNA sequencing (RNA-seq). However, little attention has been given to how ecologically relevant field conditions may affect gene expression during diapause development because previous studies have focused on laboratory-reared and -maintained insects. To determine whether gene expression differs between laboratory and field conditions, prepupae of the alfalfa leafcutting bee, Megachile rotundata , entering diapause early or late in the growing season were collected. These two groups were further subdivided in early autumn into laboratory- and field-maintained groups, resulting in four experimental treatments of diapausing prepupae: early and late field, and early and late laboratory. RNA-seq and differential expression analyses were performed on bees from the four treatment groups in November, January, March and May. The number of treatment-specific differentially expressed genes (97 to 1249) outnumbered the number of differentially regulated genes common to all four treatments (14 to 229), indicating that exposure to laboratory or field conditions had a major impact on gene expression during diapause development. Principle component analysis and hierarchical cluster analysis yielded similar grouping of treatments, confirming that the treatments form distinct clusters. Our results support the conclusion that gene expression during the course of diapause development is not a simple ordered sequence, but rather a highly plastic response determined primarily by the environmental history of the individual insect., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2018. Published by The Company of Biologists Ltd.)

Studies of bee movement and activities across a landscape are important for developing an understanding of their behavior and their ability to withstand environmental stress. Recent research has shown that proteins, such as egg albumin, are effective for mass-marking bees. However, current protein mass-marking techniques require sacrificing individual bees during the data collection process. A nonlethal sampling method for protein mark-capture research is sorely needed, particularly for vulnerable, sensitive, or economically valuable species. This study describes a nonlethal sampling method, in which three non-Apis bee species (Bombus bifarius Cresson [Hymenoptera: Apidae], Osmia lignaria Say [Hymenoptera: Megachilidae], and Megachile rotundata Fabricius [Hymenoptera: Megachilidae]) were tested for a unique protein marker by immersing them momentarily in saline buffer and releasing them. Results showed that an egg albumin-specific enzyme-linked immunosorbent assay was 100% effective at detecting the protein on bees that were sampled nonlethally. Furthermore, this sampling method did not have an impact on bee survivorship, suggesting that immersing bees in buffer is a reliable and valid surrogate to traditional, destructive sampling methods for mark-capture bee studies.

Pollination services provided by managed bees are essential for California almond (Prunus dulcis Mill.; Rosales: Rosaceae) production. Currently, pollination needs are met by rented or owned Apis mellifera L. (Hymenoptera: Apidae; honey bee) colonies. Excessive demand on a challenged A. mellifera industry to provide strong colonies in early spring has caused sharp increases in rental prices over the past decade, inviting the consideration of alternative pollinators in addition to, or in place of, A. mellifera. Osmia lignaria Say (Hymenoptera: Megachilidae; the blue orchard bee) is an excellent pollinator of fruit and nut trees, but its pollination impacts when used in tandem with A. mellifera have yet to be evaluated in commercial almond orchards. A 2-yr study was conducted in California orchards to compare almond pollination and production using A. mellifera as sole pollinator to an alternative practice of adding O. lignaria as a co-pollinator with A. mellifera. Almond orchard managerial decisions, such as for pesticide use and irrigation intensity, vary between almond growing regions because of local climates. Therefore, both north-central and southern sites of California's San Joaquin Valley are represented. We compared bee visitation, nut set, and nut yield between orchards and between tree rows within orchards. Also, O. lignaria reproductive success was recorded to assure that these bees remained in the orchards as pollinators and to assess the ability to sustain these bees under regional orchard conditions. We demonstrated that augmenting large commercial almond orchards with O. lignaria can significantly increase nut set and sometimes nut yield in both regions evaluated.

Native managed bees can improve crop pollination, but a general framework for evaluating the associated economic costs and benefits has not been developed. We conducted a cost-benefit analysis to assess how managing blue orchard bees (Osmia lignaria Say [Hymenoptera: Megachildae]) alongside honey bees (Apis mellifera Linnaeus [Hymenoptera: Apidae]) can affect profits for almond growers in California. Specifically, we studied how adjusting three strategies can influence profits: (1) number of released O. lignaria bees, (2) density of artificial nest boxes, and (3) number of nest cavities (tubes) per box. We developed an ecological model for the effects of pollinator activity on almond yields, validated the model with published data, and then estimated changes in profits for different management strategies. Our model shows that almond yields increase with O. lignaria foraging density, even where honey bees are already in use. Our cost-benefit analysis shows that profit ranged from -US$1,800 to US$2,800/acre given different combinations of the three strategies. Adding nest boxes had the greatest effect; we predict an increase in profit between low and high nest box density strategies (2.5 and 10 boxes/acre). In fact, the number of released bees and the availability of nest tubes had relatively small effects in the high nest box density strategies. This suggests that growers could improve profits by simply adding more nest boxes with moderate number of tubes in each. Our approach can support grower decisions regarding integrated crop pollination and highlight the importance of a comprehensive ecological economic framework for assessing these decisions., (© The Author(s) 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Phenotypic plasticity involves adaptive responses to predictable environmental fluctuations and may promote evolutionary change. We studied the regulation of phenotypic plasticity in an important agricultural pollinator, the solitary alfalfa leafcutting bee (Megachile rotundata F.). Specifically, we investigated how larval nutrition affects M. rotundata diapause plasticity and how diapause plasticity affects adult female reproductive behavior. Field surveys and laboratory manipulations of aspects of larval diet demonstrated nutritional regulation of M. rotundata diapause plasticity. Manipulation of larval diet quality through the addition of royal jelly, the caste-determining substance of the honey bee Apis mellifera L., increased the probability of diapause in M. rotundata. We also found that larval nutrition and diapause status affected M. rotundata adult female reproductive behavior. Nutritional effects on larval diapause that also impact adult fitness have intriguing implications for the evolution of developmental plasticity in bees. In particular, as the solitary lifestyle of M. rotundata is considered to be the ancestral condition in bees, nutritionally regulated plasticity may have been an ancestral condition in all bees that facilitated the evolution of other forms of phenotypic plasticity, such as the castes of social bees., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The engineering of flowering agricultural field borders has emerged as a research and policy priority to mitigate threats to pollinators. Studies have, however, rarely addressed the potential that flowering field borders might compete with neighboring crops for pollinator visits if they both are in bloom at the same time, despite this being a concern expressed by growers. We evaluated how wildflower plantings added to orchard borders in a large (512 ha) commercial almond orchard affected honey bee and wild bee visitation to orchard borders and the crop. The study was conducted over two consecutive seasons using three large (0.48 ha) wildflower plantings paired with control orchard borders in a highly simplified agricultural landscape in California. Honey bee (Apis mellifera L.) and wild bee visitation to wildflower plots were at least an order of magnitude higher than to control plots, but increased honey bee visitation to wildflower plots did not lead to any detectable shifts in honey bee visitation to almond flowers in the neighboring orchard. Wild bees were rarely observed visiting almond flowers irrespective of border treatment, indicating a limited short-term potential for augmenting crop pollination using wild bees in highly simplified agricultural landscapes. Although further studies are warranted on bee visitation and crop yield from spatially independent orchards, this study indicates that growers can support bees with alternative forage in almond orchards without risking competition between the wildflower plantings and the crop., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Species-specific biochemistry, morphology, and function of the Dufour's gland have been investigated for social bees and some non-social bee families. Most of the solitary bees previously examined are ground-nesting bees that use Dufour's gland secretions to line brood chambers. This study examines the chemistry of the cuticle and Dufour's gland of cavity-nesting Megachile rotundata and Osmia lignaria, which are species managed for crop pollination. Glandular and cuticular lipid compositions were characterized and compared to each other and according to the nesting experience of adult females. Major lipid classes found were hydrocarbons, free fatty acids, and wax esters. Many components were common to the cuticle and Dufour's glands of each species, yet not identical in number or relative composition. Wax esters and fatty acids were more prevalent in Dufour's glands of M. rotundata than on cuticles. Wax esters were more abundant on cuticles of O. lignaria than in Dufour's glands. In both species, fatty acids were more prevalent in glands of field-collected females compared to any other sample type. Chemical profiles of cuticles and glands were distinct from each other, and, for O. lignaria, profiles of laboratory-maintained bees could be distinguished from those of field-collected bees. Comparison of percentiles of individual components of cuticular and glandular profiles of the same bee showed that the proportions of some cuticular components were predictive of the proportion of the same glandular components, especially for nesting females. Lastly, evidence suggested that Dufour's gland is the major source of nest-marking substances in M. rotundata, but evidence for this role in O. lignaria was less conclusive.

The blue orchard bee, Osmia lignaria (Say), is a solitary bee that is an excellent pollinator of tree fruit orchards. Due to the annual rising costs of honey bee hive rentals, many orchardists are eager to develop management tools and practices to support O. lignaria as an alternative pollinator. Establishing O. lignaria pollination as a sustainable industry requires careful consideration of both bee and orchard management. Here, we test the effect of artificial nest box distribution on in-orchard propagation of O. lignaria in Utah commercial tart cherry orchards. Two nest box distributions were compared across three paired, 1.2-ha plots. One distribution, traditionally employed by O. lignaria consultants, included a centrally located tote for mass-nesting with smaller, surrounding 'satellite' nest boxes at orchard margins. The other distribution was composed of smaller, more equally distributed nest boxes throughout the 1.2-ha plots. Significantly higher propagation of O. lignaria was observed in the latter nest box distribution, although all treatments resulted in bee return exceeding the number of bees initially released. These findings provide support for the use of O. lignaria in tart cherry orchards, and demonstrate how simple changes to bee set-up and management can influence propagation efforts., (Published by Oxford University Press on behalf of the Entomological Society of America 2017. This work is written by US Government employees and is in the public domain in the US.)

Background: The chitin synthesis inhibitor novaluron can suppress pests that affect alfalfa seed production, but can negatively affect reproductive success in the alfalfa pollinator Megachile rotundata. Novaluron is considered to be a reduced-risk insecticide because it disrupts ecdysis and is non-lethal to adult insects, but some exposed adults have fewer eggs and suppressed egg hatch. For this experiment, bees nested in field cages where they were exposed to alfalfa that had never been treated with novaluron, alfalfa that had recently been sprayed or alfalfa that had been sprayed 1 and 2 weeks earlier., Results: Compared with the control, greater proportions of dead eggs and larvae and lower proportions of live prepupae occurred when bees were exposed to recent novaluron sprays as well as one- or two-week old spray residues. Two possible routes of residual pesticide exposure were revealed. Mother bees become contaminated through ingestion or direct contact, or pollen-nectar provisions become contaminated with novaluron (1) on or within leaf pieces that surround provisions or (2) transferred from mother bees' bodies to provisions., Conclusion: We found strong immature mortality effects of novaluron and its residues on M. rotundata. Understanding all possible pesticide exposure routes for pollinating bees enhances decision-making for maintaining bee populations while protecting crops. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

There is a growing body of empirical evidence showing that wild and managed bees are negatively impacted by various pesticides that are applied in agroecosystems around the world. The lethal and sublethal effects of two widely used fungicides and one adjuvant were assessed in cage studies in California on blue orchard bees, Osmia lignaria, and in cage studies in Utah on alfalfa leafcutting bees, Megachile rotundata. The fungicides tested were Rovral 4F (iprodione) and Pristine (mixture of pyraclostrobin + boscalid), and the adjuvant tested was N-90, a non-ionic wetting agent (90% polyethoxylated nonylphenol) added to certain tank mixtures of fungicides to improve the distribution and contact of sprays to plants. In separate trials, we erected screened cages and released 20 paint-marked females plus 30-50 males per cage to document the behavior of nesting bees under treated and control conditions. For all females in each cage, we recorded pollen-collecting trip times, nest substrate-collecting trip times (i.e., mud for O. lignaria and cut leaf pieces for M. rotundata), cell production rate, and the number of attempts each female made to enter her own or to enter other nest entrances upon returning from a foraging trip. No lethal effects of treatments were observed on adults, nor were there effects on time spent foraging for pollen and nest substrates and on cell production rate. However, Rovral 4F, Pristine, and N-90 disrupted the nest recognition abilities of O. lignaria females. Pristine, N-90, and Pristine + N-90 disrupted nest recognition ability of M. rotundata females. Electroantennogram responses of antennae of O. lignaria females maintained in the laboratory did not differ significantly between the fungicide-exposed and control bees. Our results provide the first empirical evidence that two commonly used fungicides and a non-ionic adjuvant can disrupt nest recognition in two managed solitary bee species.

Population density may affect solitary bee maternal resource allocation. The number of Megachile rotundata (F.), alfalfa leafcutting bee, females released for seed production of Medicago sativa L., alfalfa, may limit flower availability for nest provisioning. In turn, pollinator abundance also may affect crop yield. The M. sativa pollination system presents an opportunity to test for effects of density dependence and maternal manipulation on M. rotundata reproduction. A multiyear study was performed on M. sativa fields upon which M. rotundata densities were altered to induce low, medium, and high density situations. Numbers of adult bees and open flowers were recorded weekly; bee reproduction variables were collected once. Fields varied in plant performance for each site and year, and the intended bee densities were not realized. Therefore, the variable density index (DI) was derived to describe the number of female bees per area of flowers over the study period. As DI increased, percentages of pollinated flowers, established females, and healthy brood significantly increased, and the number of pollinated flowers per female and of dead or diseased brood significantly decreased. Sex ratio was significantly more female biased as DI increased. Overwintered offspring weights were similar regardless of DI, but significantly differed by year for both sexes, and for males also by field and year × field interaction. Overall, resource limitation was not found in this field study. Other density-dependent factors may have induced a bee dispersal response soon after bees were released in the fields that circumvented the need for, or impact of, maternal manipulation., (Published by Oxford University Press on behalf of Entomological Society of America 2015. This work is written by a US Government employee and is in the public domain in the US.)

During the nesting season, adult females of the solitary bee Megachile rotundata (F.) face considerable physical and energy demands that could include increasing wear and tear on their bodies and decreasing lipid reserves. Consequently, their reproductive performance may be affected not only by extrinsic factors (e.g., weather and floral resource availability), but intrinsic changes in their own bodies. Because of the potential fitness effects of seasonal changes in body condition, our objectives were to determine how wing wear, lipid reserves, and oocyte sizes vary during nesting seasons, beginning when females emerge as adults. As nesting progressed, females in two populations experienced a steady increase in wing wear, which is known to reduce foraging efficiency and increase risk of mortality in other bees. Soon after emergence, females exhibited sharp declines in lipid content which remained low for the remainder of the season. Newly-emerged females ingested pollen, an activity known to be correlated with the initiation of egg maturation in this species. Additionally, the early summer drop in lipid stores was correlated with an increase in the size of the oocytes carried. However, by ∼6 weeks after emergence, oocytes began to decrease in length and volume, perhaps due to nutrient deficiencies related to loss of stored lipids. Our results suggest management of M. rotundata should include rearing bees at temperatures that maximize stored lipid reserves in adults and timing bee release so that significant pollen resources are available for both adults and offspring.

Intensively managed, commercial orchards offer resources for managed solitary bees within agricultural landscapes and provide a means to study bee dispersal patterns, spatial movement, nest establishment, and reproduction. In 2012, we studied the impact of 1) the color of nest boxes covaried with four nest box density treatments and 2) the number of bee release sites covaried with two nest box density treatments on the reproductive success of Osmia lignaria Say in a California almond orchard pollinated by a mixture of O. lignaria and Apis mellifera L. Nest box color influenced the number of nests, total cells, and cells with male and female brood. More nests and cells were produced in light blue nest boxes than in orange or yellow nest boxes. The covariate nest box density also had a significant effect on brood production. The number of release sites did not affect O. lignaria nesting and reproduction, but the number of cavities in nest boxes influenced reproduction. Overall, the color of nest boxes and their distribution, but not the number of release sites, can greatly affect O. lignaria nest establishment and reproductive success in a commercial almond orchard. The ability to locate nesting sites in a homogenous, large orchard landscape may also be facilitated by the higher frequency of nest boxes with low numbers of cavities, and by the ability to detect certain nest box colors that best contrast with the blooming trees., (© 2014 Entomological Society of America.)

Many insects, including some bees, have extensive subcontinental distributions that can differ in climatic conditions. Within and beyond these distributions, humans intentionally transport beneficial insects, including bees, to non-natal geographic locations. Insects also are experiencing unprecedented climatic change in their resident localities. For solitary bees, we know very little about the adaptive plasticity and geographic variation in developmental physiology that accommodates the different climates experienced within distributional ranges. Osmia lignaria Say (Hymenoptera: Megachilidae) is a widely distributed North American spring-emerging bee being developed as a managed pollinator for tree fruit crops, including almonds. We examined the development and survival of O. lignaria progeny that were descended from populations sourced from southern California, western Washington, and northern Utah, and then were reared together under an hourly and weekly temperature regime simulating those of a California almond-growing region. We found that developmental physiologies of Washington and Utah progeny were generally similar. However, California progeny developed slower, were more metabolically active, and survived better under California conditions than did populations native to regions at higher latitudes. Regardless of geographic origin, cocooned adults managed under prescribed thermal regimes emerged faster and lived longer after wintering. Progeny of parents from different regions exhibited some acclimatory plasticity in developmental phenologies to a novel climatic regime, but overall their responses reflected their geographic origins. This outcome is consistent with their developmental phenologies being largely heritable adaptations to regional climates., (Published by Elsevier Ltd.)

We conducted a broad geographic survey in the northwestern United States to quantify production losses in the alfalfa leafcutting bee (Megachile rotundata (F.), Hymenoptera: Megachilidae), a solitary pollinator used extensively in alfalfa seed production. Viable larvae were found in only 47.1% of the nest cells collected at the end of the season. Most of the rest of the cells contained pollen balls (typified by a provision but no larva; 16.7%), unknown causes of mortality (15.5%), or larvae killed by chalkbrood (8.0%). Prevalence of pollen balls was correlated positively with bee release rates and negatively with alfalfa stand age. The unknown mortality was correlated with the U.S. Department of Agriculture-Plant Hardiness Zone, and thus, some of the mortality may be caused by high temperature extremes, although the nesting season degree-days were not correlated with this mortality. Chalkbrood prevalence was correlated with possible nesting-resource or crowding-related factors, such as the number of bees released per hectare and the number of shelters used, but not with nesting board disinfection practices. Vapona is used to control parasitoids when the parent bees are incubated before release, and use of this fumigant was associated with an increase in both chalkbrood and diapausing offspring, although any reason for these correlations are unknown. This survey quantifies the variation in the quality of alfalfa leafcutting bee cocoons produced across much of the U.S. alfalfa seed production area.

In-nest observations of the solitary bee, Megachile rotundata (F.), revealed that nesting females apply olfactory cues to nests for nest recognition. On their way in and out of the nest, females drag the abdomen along the entire length of the nest, and sometimes deposit fluid droplets from the tip of the abdomen. The removal of bee-marked sections of the nest resulted in hesitation and searching behavior by females, indicating the loss of olfactory cues used for nest recognition. Chemical analysis of female cuticles and the deposits inside marked nesting tubes revealed the presence of hydrocarbons, wax esters, fatty aldehydes, and fatty alcohol acetate esters. Chemical compositions were similar across tube samples, but proportionally different from cuticular extracts. These findings reveal the importance of lipids as chemical signals for nest recognition and suggest that the nest-marking cues are derived from a source in addition to, or other than, the female cuticle.

The alfalfa leafcutting bee, Megachile rotundata F., is used extensively to pollinate alfalfa for seed production in western North America. However, it usually is not possible to sustain bee populations in the United States. In the managed pollination system, variable microenvironments are experienced by developing M. rotundata during the nesting season that may influence bee reproductive success. In this study, bee cells were produced in a small alfalfa field and collected from domiciles according to nesting boards and different portions of boards. Examination of cells showed that the production of live overwintering prepupae, diseased larvae, other dead eggs and larvae, and cells attacked by natural enemies varied according to board orientations and positions, and portions of the boards. Northeast-facing board samples were significantly heavier and had more pollen balls and chalkbrood compared with southwest-facing boards. Lower boards in stacks had significantly more chalkbrood than upper boards. Outer portions of boards had significantly more live prepupae and less chalkbrood than middle and inner portions, and outer and middle portions had less summer-emerging bees. These results suggest that reproductive success could be altered by changing the orientation of boards within domiciles or changing the designs of domiciles. To accurately assess the quality of bee populations managed in the commercial loose cell system, a sample of bees should be taken from a pooled sample from all boards in each domicile. Each domicile sample should then be evaluated according to its proportional contribution to the whole field's progeny production (e.g., by weight or volume).

Low, medium, and high stocking densities (15,000; 30,000; and 45,000-50,000 bees per acre, respectively) of Megachile rotundata (F.) (Hymenoptera: Megachilidae), the alfalfa leafcutting bee, were released over 4 yr in three research plots of Utah alfalfa, Medicago sativa L. (Fabaceae), planted at seed-production rates. A low percentage of bees (46-79% of released) survived the incubation and field-emergence processes; of those bees, the number of females that established at the nesting sites was 25-100%. Of the three field sites, one site consistently retained more females and produced more completed nests than the other sites, all of which usually had poor female establishment and progeny production. In addition, floral resources were depleted over time, but many flowers remained unvisited over the season. Nest production decreased over time, as numbers of flowers and female bees declined. Significant positive relationships were found between the intended stocking densities and 1) the number of females that actually survived incubation and field emergence and 2) the number of females that established nests. The number of females that established nests was positively affected by the number of females that survived to emerge in the field. The percentage of females that established was not significantly affected by the intended number of released bees, countering a prediction that the release of fewer bees would allow more females to establish nests and achieve high reproductive success. For growers, it may be more frugal to use modest numbers of M. rotundata for pollination, because many of the bees at medium and high stocking densities do not nest in grower-provided bee boards.

The impact of climate change on insect populations depends on specific life cycle traits and physiological adaptations. The solitary bee Osmia lignaria winters as a pre-emergent adult, and requires a period of cold temperature for winter diapause completion. It is a univoltine species, and diapause induction does not depend on photoperiod. To understand the potential effects of longer summers on O. lignaria populations, we exposed individuals to three treatments simulating early, mid and late winter arrivals, and measured respiration rates, metabolic expenditure, weight loss, fat body depletion, lipid levels and winter mortality. The early-winter treatment disrupted diapause development, but had no apparent negative effects on fitness. In contrast, late-winter bees had a greater energetic expenditure (1.5-fold), weight (1.4-fold) and lipid (2-fold) loss, greater fat body depletion, and a 19% increase in mortality compared to mid-winter bees. We also monitored adult eclosion and arrival of winter temperatures under natural conditions in four years. We found a positive correlation between mean degree-day accumulation during pre-wintering (a measure of asynchrony between adult eclosion and winter arrival) and yearly winter mortality. Individually, bees experiencing greater degree-day accumulations exhibited reduced post-winter longevity. Timing of adult eclosion in O. lignaria is dependent on the duration of the prepupal period, which occurs in mid-summer, is also diapause-mediated, and is longer in populations from southerly latitudes. In a global warming scenario, we expect long summer diapause phenotypes to replace short summer diapause phenotypes, effectively maintaining short pre-wintering periods in spite of delayed winter arrivals., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Osmia lignaria Say (Hymenoptera: Megachilidae) can be used to pollinate fruit trees. Populations are sometimes difficult to sustain because some female bees fail to establish at provided nesting sites. We address the hypothesis that rough handling of overwintered O. lignaria results in decreased establishment. We tested this by shaking (200 rpm for 2 min) overwintering bees as a proxy for rough handling. Bees were then released in an orchard, and nest establishment of shaken and unshaken bees was recorded. There was no significant difference in the proportion of shaken and unshaken females that nested, indicating that rough handling of overwintering bees does not discourage nest establishment.

The alfalfa leafcutting bee (ALCB), Megachile rotundata F. (Megachildae), was accidentally introduced into the United States by the 1940s. Nest management of this Eurasian nonsocial pollinator transformed the alfalfa seed industry in North America, tripling seed production. The most common ALCB management practice is the loose cell system, in which cocooned bees are removed from nesting cavities for cleaning and storage. Traits of ALCBs that favored their commercialization include gregarious nesting; use of leaves for lining nests; ready acceptance of affordable, mass-produced nesting materials; alfalfa pollination efficacy; and emergence synchrony with alfalfa bloom. The ALCB became a commercial success because much of its natural history was understood, targeted research was pursued, and producer ingenuity was encouraged. The ALCB presents a model system for commercializing other solitary bees and for advancing new testable hypotheses in diverse biological disciplines.

Alfalfa leafcutting bees, Megachile rotundata F. (Hymenoptera: Megachilidae), are the most common pollinators of alfalfa in the Pacific Northwest. Reports from users of M. rotundata in Idaho, Utah and Colorado have indicated exceptionally poor bee return from fields treated with novaluron to control Lygus spp. Our goal was to evaluate novaluron toxicity to immature M. rotundata using two different possible mechanisms of exposure. One goal was to assess immature mortality via treating nectar-pollen provisions and adults with novaluron. Immature M. rotundata mortality in all novaluron provision dosing treatments was significantly higher than the water or blank controls, providing evidence that novaluron is toxic to progeny in nest cells. The mean cumulative frequency showed that more eggs and 1st-2nd instars died compared to older instars. Female M. rotundata nested similarly in field cages during the field cage experiment; however, there was greater immature mortality in cages where females were fed sugar-water + novaluron compared to sugar-water only. Although females provided adequate provisions, there was a low percentage of egg hatch and larval development when females ingested novaluron before mating and nesting. Novaluron was also present in egg provision of bees collecting resources from novaluron-sprayed plants. At least 84% of progeny died when the females were allowed to mate and nest 24 hours after a novaluron application. Novaluron could be contributing to poor bee return in alfalfa grown for seed. Timely insecticide applications to suppress Lygus spp. is an important consideration to improve ongoing bee health.

The alfalfa leafcutting bee, Megachile rotundata (Fabricius), is used to pollinate alfalfa, Medicago sativa L., for seed production in the United States and Canada. It is difficult to reliably sustain commercial M. rotundata populations in the United States because of problems with disease, parasites, predators, and unexplained mortality. One possible explanation for early immature mortality is that, relative to floral availability, superfluous numbers of bees are released in alfalfa fields where resources quickly become limited. Our objective was to determine how M. rotundata density affects bee nesting, pollination efficiency, and reproductive success. Various numbers of bees were released into enclosures on an alfalfa field, but only 10-90% of released female bees established nests. Therefore, a "bee density index" was derived for each enclosure from the number of established females and number of open flowers over time. As the density index increased, significant reductions occurred in the number of pollinated flowers, number of nests, and number of cells produced per bee, as well as the percentage of cells that produced viable prepupae by summer's end and the percentage that produced adult bees. The percentage of cells resulting in early brood mortality (i.e., pollen balls) significantly increased as the density index increased. We conclude that bee nest establishment, pollination efficiency, and reproductive success are compromised when bee densities are high relative to floral resource availability. Open field studies are needed to determine commercial bee densities that result in sustainable bee populations and adequate pollination for profitable alfalfa seed production.

The alfalfa leafcutting bee, Megachile rotundata F. (Hymenoptera: Megachilidae), is widely used as a pollinator for production of alfalfa, Medicago sativa L., seed, and populations of these bees can be maintained by alfalfa seed growers or can be purchased from mostly Canadian bee providers. M. rotundata raised in Canada have higher survival rates during the incubation that occurs after winter storage than do bees produced in the northwestern United States, but no reason has been found for this difference. We investigated whether storing immature M. rotundata for various time periods at a warm temperature (16 degrees C) before winter or allowing them to remain unmanaged at ambient temperatures affects physiological aspects of prepupae during the winter as well as the survival and longevity of adult bees after spring or summer incubation. Our results show that the timing of the onset of winter storage and incubation does affect prepupal weights, prepupal lipid and water contents, adult emergence, and adult female longevity. Winter storage of prepupae in November or December with a late June incubation resulted in heavier adults that emerged more readily than bees incubated in late May. However, adult females incubated in May thrived longer than June-incubated bees if fed a honey-water diet. Thus, some prewinter management regimes for M. rotundata commercial stocks may be more effective than others for achieving optimal adult emergence synchrony, as well as adult survival and longevity for pollination of a summer crop.

The cuticular lipids of the cavity-nesting adult female solitary bees, Osmia lignaria Say and Megachile rotundata (F.) (Hymenoptera: Megachilidae), were analyzed by gas chromatography (GC) and combined GC-mass spectrometry. The cuticular lipids of these female bees are mainly consisted of hydrocarbons. For O. lignaria, nearly 64% of the cuticular lipids were C(25)-C(31) mono-alkenes. For M. rotundata, 48% of the cuticular lipids were C(23)-C(33) alkanes with nearly the same quantities of the same chain-length mono-alkenes (45%). For the mono-alkenes of O. lignaria, 14 mono-alkene constituents were identified, with two of these, 9-heptacosene and 7-nonacosene, comprising 67% of the total alkene distribution. For M. rotundata females, the mixtures of mono-alkenes were more complex with 26 constituents identified and quantified. For the M. rotundata mono-alkenes, 57% of the total composition consisted of the three alkenes, 7-pentacosene, 9-pentacosene and 7-heptacosene. For both bee species, small quantities of C(40)-C(48) wax esters were also characterized with the major components possessing a C(18) mono-unsaturated fatty acid (9-octadecenoate) moiety esterified to even-carbon number (C(22-30)) fatty alcohols. The possible role of these cuticular lipids as nest recognition chemicals is discussed in light of nesting behavior of managed crop pollinators.

Cavity-nesting alfalfa leafcutting bees, Megachile rotundata (F.) (Hymenoptera: Megachilidae), are excellent pollinators of alfalfa, Medicago savita L., for seed production. In commercial settings, artificial cavities are placed in field domiciles for nesting and, thereby, bee populations are sustained for future use. For this study, cells from leafcutting bee nests were collected in late summer from commercial seed fields. Over 3 yr (2003-2005), 39 samples in total of approximately equal to 1,000 cells each were taken from several northwestern U.S. states and from Manitoba, Canada. X-radiography of 500 cells from each sample was used to identify "pollen balls" (i.e., cells in which the pollen-nectar provision remained, but the egg or larva, if present, was not detectable on an x-radiograph). Most U.S. samples seemed to have higher proportions of pollen ball cells than Manitoba samples. Pollen ball cells were dissected to determine the moisture condition of the mass provision and true contents of each cell. Most pollen ball cells from Manitoba samples contained fungus, the frequency of which was positively correlated with cool, wet weather. In the United States, most pollen ball cells had moist provisions, and many of them lacked young brood. Correlation analysis revealed that pollen ball cells occurred in greater proportions in fields with more hot days (above 38 degrees C). Broodless pollen ball cells occurred in greater proportions under cool conditions, but dead small larvae (second-third instars) seemed to occur in greater proportions under hot conditions. Pollen ball cells with unhatched eggs and first instars (in the chorion) occurred in lesser proportions under hot conditions.

The megachilid bees Osmia lignaria Say, Megachile rotundata (F.), and M. pugnata Say were tested for attraction to various components associated with their old nest cavities, or chemical extracts of these components, using a Y-tube olfactory response bioassay. Female bees of these species are known to nest in or near old nest cavities, implying that remnant nest components are important cues for bees looking for nest cavities. Significant results show that female bees were attracted to components that may provide species-specific cues or indicate conspecific nesting activity. Specifically, O. lignaria females showed attraction only to the female cocoon. M. rotundata females were attracted to intact nest cells, the fecal material on the outside of a cocoon, leaf pieces used as nest cell lining, and the extract of leaf pieces. M. pugnata females were attracted to the whole nest cell, the paper straw nesting material with attached cocoon, and feces.

Samples of overwintering alfalfa leafcutting bee, Megachile rotundata (F.) (Hymenoptera: Megachilidae), cells were sent to the laboratory as loose cells or in nesting boards from bee managers in the United States and in Canada. X-radiographs of cells were used for determining cell contents. Cells containing live prepupae were incubated, and the sex of emerging adults was recorded daily. Cells from which no adult emerged were dissected to determine the developmental stage of dead bees and sex of dead pupae or adults. Bee cells incubated in commercial settings and placed in alfalfa fields by the same bee managers described above also were evaluated to determine adult emergence success. The proportion of live bees in wood nesting boards from the United States was much lower than the live proportion in polystyrene nesting boards from Canada and loose cells overwintered in the United States. For laboratory-incubated loose cells, survival and sex ratios of bees from Canadian sources were statistically higher than those of U.S. bees, but the onset and duration of emergence times were similar. Fewer bees survived in the commercial setting than in the laboratory. Prepupal mortality was significantly higher than pupal or adult mortality, but there was no significant difference between the sexes in the likelihood of survival during incubation. This study supports the commonly held belief that alfalfa leafcutting bees raised in Canada and then sold to the United States represent a more viable source of bees than most bees produced in the United States.

The alfalfa leafcutting bee, a solitary bee used to pollinate alfalfa seed crops, is seriously affected by chalkbrood, a larval disease caused by the fungus Ascosphaera aggregata. One attempt to control the disease includes removing nests from the nesting boards (the "loose cell" system). We report here that adults emerging from the loose cells are heavily contaminated with A. aggregata spores. The contamination levels are not as high as previously reported for bees emerging directly from the boards, but they are still a likely focus for disease spread and may need to be targeted in chalkbrood control strategies.