Floral Trait Preferences of Three Common wild Bee Species.

The interaction between bees and flowering plants is mediated by floral cues that enable bees to find foraging plants. We tested floral cue preferences among three common wild bee species: Lasioglossum villosulum, Osmia bicornis, and Bombus terrestris. Preferences are well studied in eusocial bees but almost unknown in solitary or non-eusocial generalist bee species. Using standardized artificial flowers altered in single cues, we tested preferences for color hue, achromatic contrast, scent complexity, corolla size, and flower depth. We found common attractive cues among all tested bees. Intensively colored flowers and large floral displays were highly attractive. No preferences were observed in scent complexity experiments, and the number of volatiles did not influence the behavior of bees. Differing preferences were found for color hue. The specific behaviors were probably influenced by foraging experience and depended on the flower choice preferences of the tested bee species. In experiments testing different flower depths of reward presentation, the bees chose flat flowers that afforded low energy costs. The results reveal that generalist wild bee species other than well-studied honeybees and bumblebees show strong preferences for distinct floral cues to find potential host plants. The diverse preferences of wild bees ensure the pollination of various flowering plants.

Diverse pollen nutrition can improve the development of solitary bees but does not mitigate negative pesticide impacts.

Floral resource loss and pesticide exposure are major threats to bees in intensively managed agroecosystems, but interactions among these drivers remain poorly understood. Altered composition and lowered diversity of pollen nutrition may reinforce negative pesticide impacts on bees. Here we investigated the development and survival of the solitary bee Osmia bicornis provisioned with three different pollen types, as well as a mixture of these types representing a higher pollen diversity. We exposed bees of each nutritional treatment to five pesticides at different concentrations in the laboratory. Two field-realistic concentrations of three nicotinic acetylcholine receptor (nAChR) modulating insecticides (thiacloprid, sulfoxaflor and flupyradifurone), as well as of two fungicides (azoxystrobin and tebuconazole) were examined. We further measured the expression of two detoxification genes (CYP9BU1, CYP9BU2) under exposure to thiacloprid across different nutrition treatments as a potential mechanistic pathway driving pesticide-nutrition interactions. We found that more diverse pollen nutrition reduced development time, enhanced pollen efficacy (cocoon weight divided by consumed pollen weight) and pollen consumption, and increased weight of O. bicornis after larval development (cocoon weight). Contrary to fungicides, high field-realistic concentrations of all three insecticides negatively affected O. bicornis by extending development times. Moreover, sulfoxaflor and flupyradifurone also reduced pollen efficacy and cocoon weight, and sulfoxaflor reduced pollen consumption and increased mortality. The expression of detoxification genes differed across pollen nutrition types, but was not enhanced after exposure to thiacloprid. Our findings highlight that lowered diversity of pollen nutrition and high field-realistic exposure to nAChR modulating insecticides negatively affected the development of O. bicornis, but we found no mitigation of negative pesticide impacts through increased pollen diversity. These results have important implications for risk assessment for bee pollinators, indicating that negative effects of nAChR modulating insecticides to developing solitary bees are currently underestimated.

Solitary bee behaviour and pollination service delivery is differentially impacted by neonicotinoid and pyrethroid insecticides.

Solitary bees are important pollinators of crops and wild plants, and their decline poses a risk to the sustained provision of the services they provide. Although evidence suggests that exposure to insecticides can affect bees, most pesticide research and risk assessment has focussed on social bees and mortality while solitary species are understudied. The ability to forage is critical for solitary bee reproduction, also in how they deliver pollination services, and we know little about how insecticides can impact these behaviours. We exposed solitary red mason bees (Osmia bicornis) to field realistic levels of two widely used insecticides with differing modes of action (lambda-cyhalothrin (pyrethroid) and acetamiprid (neonicotinoid)), in a semi-field setting over multiple rounds of exposure. We then tested impacts on bee behaviour and pollination in apples, an important global insect-pollinated crop. Pollination by insecticide-treated bees reduced apple production by up to 86 % depending on the compound and the number of exposures, but the underlying mechanism behind this remains unclear and should be investigated further. Other measurements of pollination services including number of seeds per apple and stigma pollen deposition showed no relationship with pesticide treatment. Bee foraging behaviour was also affected by treatment where both insecticides appear to induce an excitatory effect which was constant for acetamiprid and eventually ceased for lambda-cyhalothrin after multiple exposures. This suggests that both neonicotinoid and non­neonicotinoid insecticides can potentially affect behaviour and pollination services of solitary bees depending on how often they are exposed, which is particularly important given the changing usage patterns of these compound classes based on regulations around their use. This highlights the importance of moving insecticide risk assessment towards more field realistic scenarios and including sublethal effects on solitary and social bees, in addition to considering repeated exposures of bees to pesticides as is realistic in the field.

Pesticide residues in nectar and pollen of melon crops: Risk to pollinators and effects of a specific pesticide mixture on Bombus terrestris (Hymenoptera: Apidae) micro-colonies.

Residues detected in pollen collected by honey bees are often used to estimate pesticide exposure in ecotoxicological studies. However, for a more accurate assessment of pesticides effect on foraging pollinators, residues found directly on flowers are a more realistic exposure approximation. We conducted a multi-residue analysis of pesticides on pollen and nectar of melon flowers collected from five fields. The cumulative chronic oral exposure Risk Index (RI) was calculated for Apis mellifera, Bombus terrestris and Osmia bicornis to multiple pesticides. However, this index could underestimate the risk since sublethal or synergistic effects are not considered. Therefore, a mixture containing three of the most frequently detected pesticides in our study was tested for synergistic impact on B. terrestris micro-colonies through a chronic oral toxicity test. According to the result, pollen and nectar samples contained numerous pesticide residues, including nine insecticides, nine fungicides, and one herbicide. Eleven of those were not applied by farmers during the crop season, revealing that melon agroecosystems may be pesticide contaminated environments. The primary contributor to the chronic RI was imidacloprid and O. bircornis is at greatest risk for lethality resulting from chronic oral exposure at these sites. In the bumblebee micro-colony bioassay, dietary exposure to acetamiprid, chlorpyrifos and oxamyl at residue level concentration, showed no effects on worker mortality, drone production or drone size and no synergies were detected when pesticide mixtures were evaluated. In conclusion, our findings have significant implications for improving pesticide risk assessment schemes to guarantee pollinator conservation. In particular, bee pesticide risk assessment should not be limited to acute exposure effects to isolated active ingredients in honey bees. Instead, risk assessments should consider the long-term pesticide exposure effects in both pollen and nectar on a range of bees that reflect the diversity of natural ecosystems and the synergistic potential among pesticide formulations.

Effects of chronic exposure to the new insecticide sulfoxaflor in combination with a SDHI fungicide in a solitary bee.

The recent EU ban of the three most widely used neonicotinoids (imidacloprid, thiamethoxam and clothianidin) to all outdoors applications has stimulated the introduction of new insecticides into the market. Sulfoxaflor is a new systemic insecticide that, like neonicotinoids, acts as a modulator of nicotinic acetylcholine receptors. In agro-environments, bees can be exposed to this compound via contaminated pollen and nectar for long periods of time. Therefore, it is important to assess the potential effects of chronic exposure to sulfoxaflor, alone and in combination with fungicides, on pollinators. In this study, we tested the effects of chronic exposure to two field concentrations of sulfoxaflor (20 and 100 ppb) alone and in combination with four concentrations of the fungicide fluxapyroxad (7500, 15,000, 30,000 and 60,000 ppb) on syrup consumption and longevity in females of the solitary bee Osmia bicornis L. Exposure to 20 ppb of sulfoxaflor, alone and in combination with the fungicide, stimulated syrup consumption, but did not affect longevity. In contrast, syrup consumption decreased in bees exposed to 100 ppb, all of which died after 2-6 days of exposure. We found no evidence of synergism between the two compounds at any of the two sulfoxaflor concentrations tested. Comparison of our findings with the literature, confirms that O. bicornis is more sensitive to sulfoxaflor than honey bees. Our results highlight the need to include different bee species in risk assessment schemes.

No evidence for impaired solitary bee fitness following pre-flowering sulfoxaflor application alone or in combination with a common fungicide in a semi-field experiment.

Pesticide exposure is considered a major driver of pollinator decline and the use of neonicotinoid insecticides has been restricted by regulatory authorities due to their risks for pollinators. Impacts of new alternative sulfoximine-based compounds on solitary bees and their potential interactive effects with other commonly applied pesticides in agriculture remain unclear. Here, we conducted a highly replicated full-factorial semi-field experiment with the solitary bee Osmia bicornis, an important pollinator of crops and wild plants in Europe, and Phacelia tanacetifolia as a model crop. We show that spray applications of the insecticide sulfoxaflor (product Closer) and the fungicide azoxystrobin (product Amistar), both alone and combined, had no significant negative impacts on adult female survival or the production, mortality, sex ratio and body size of offspring when sulfoxaflor was applied five days before crop flowering. Our results indicate that for O. bicornis (1) the risk of adverse impacts of sulfoxaflor (Closer) on fitness is small when applied at least five days before crop flowering and (2) that azoxystrobin (Amistar) has a low potential of exacerbating sulfoxaflor effects under field-realistic conditions.

Wild bee larval food composition in five European cities.

Urbanization poses threats and opportunities for the biodiversity of wild bees. At the same time, cities can harbor diverse wild bee assemblages, partly due to the unique plant assemblages that provide resources. While bee dietary preferences have been investigated in various studies, bee dietary studies have been conducted mostly in nonurban ecosystems and data based on plant visitation observations or palynological techniques. This data set describes the larval food preferences of four wild bee species (i.e., Chelostoma florisomne, Hylaeus communis, Osmia bicornis, and O. cornuta) common in urban areas in five different European cities (i.e., Antwerp, Belgium; Paris, France; Poznan, Poland; Tartu, Estonia; and Zurich, Switzerland). In addition, the data set describes the larval food preferences of individuals from three wild bee genera (i.e., Chelostoma sp., Hylaeus sp., and Osmia sp.) that could not be identified to the species level. These data were obtained from a Europe-level study aimed at understanding the effects of urbanization on biodiversity across different cities and cityscapes and a Swiss project aimed at understanding the effects of urban ecosystems in wild bee feeding behavior. Wild bees were sampled using standardized trap nests at 80 sites (32 in Zurich and 12 in each of the remaining cities), selected following a double gradient of available habitat at local and landscape scales. Larval pollen was obtained from the bee nests and identified using DNA metabarcoding. The data provide the plant composition at the species or genus level preferred by each bee. These unique data can be used for a wide array of research questions, including urban ecology (e.g., diversity of food sources along urban gradients), bee ecology (characterization of bee feeding preferences), or comparative studies on the urban evolution of behavioral traits between urban and nonurban sites. In addition, the data can be used to inform urban planning and conservation strategies, particularly concerning flower resources (e.g., importance of exotic species and, thus, management activities). This data set can be freely used for noncommercial purposes, and this data paper should be cited if the data is used; we request that collaboration with the data set contact person to be considered if this data set represents an important part of the data analyzed in a study.

Bee Tracker-an open-source machine learning-based video analysis software for the assessment of nesting and foraging performance of cavity-nesting solitary bees.

The foraging and nesting performance of bees can provide important information on bee health and is of interest for risk and impact assessment of environmental stressors. While radiofrequency identification (RFID) technology is an efficient tool increasingly used for the collection of behavioral data in social bee species such as honeybees, behavioral studies on solitary bees still largely depend on direct observations, which is very time-consuming. Here, we present a novel automated methodological approach of individually and simultaneously tracking and analyzing foraging and nesting behavior of numerous cavity-nesting solitary bees. The approach consists of monitoring nesting units by video recording and automated analysis of videos by machine learning-based software. This Bee Tracker software consists of four trained deep learning networks to detect bees that enter or leave their nest and to recognize individual IDs on the bees' thorax and the IDs of their nests according to their positions in the nesting unit. The software is able to identify each nest of each individual nesting bee, which permits to measure individual-based measures of reproductive success. Moreover, the software quantifies the number of cavities a female enters until it finds its nest as a proxy of nest recognition, and it provides information on the number and duration of foraging trips. By training the software on 8 videos recording 24 nesting females per video, the software achieved a precision of 96% correct measurements of these parameters. The software could be adapted to various experimental setups by training it according to a set of videos. The presented method allows to efficiently collect large amounts of data on cavity-nesting solitary bee species and represents a promising new tool for the monitoring and assessment of behavior and reproductive success under laboratory, semi-field, and field conditions.

Development of a multiplex microsatellite marker set for the study of the solitary red mason bee, Osmia bicornis (Megachilidae).

BACKGROUND: Solitary bees, such as the red mason bee (Osmia bicornis), provide important ecosystem services including pollination. In the face of global declines of pollinator abundance, such haplodiploid Hymenopterans have a compounded extinction risk due to the potential for limited genetic diversity. In order to assess the genetic diversity of Osmia bicornis populations, we developed microsatellite markers and characterised them in two populations. METHODS AND RESULTS: Microsatellite sequences were mined from the recently published Osmia bicornis genome, which was assembled from DNA extracted from a single male bee originating from the United Kingdom. Sequences were identified that contained dinucleotide, trinucleotide, and tetranucleotide repeat regions. Seventeen polymorphic microsatellite markers were designed and tested, sixteen of which were developed into four multiplex PCR sets to facilitate cheap, fast and efficient genotyping and were characterised in unrelated females from Germany (n = 19) and England (n = 14). CONCLUSIONS: The microsatellite markers are highly informative, with a combined exclusion probability of 0.997 (first parent), which will enable studies of genetic structure and diversity to inform conservation efforts in this bee.

Interaction between warming and landscape foraging resource availability on solitary bee reproduction.

Solitary bees comprise around 90% of bee species, playing an essential role in both wild and crop plant pollination. Bee populations are jeopardized by different global change pressures such as climate change and landscape transformation. However, the interactive effects of global change components have been little explored, especially for solitary bees. We conducted a factorial experiment using artificial nest-traps to analyse the combined effect of climate warming and landscape transformation on Osmia bicornis reproduction and offspring body size. The number of bee cocoons increased with temperature and flower abundance in the landscape. However, the sex ratio was biased towards males with warming, especially at low flower abundances. Male body size increased with temperature. Conversely, female body sizes showed strong interactive responses, increasing in size with high flower abundance in the landscape, but only at low temperatures. The abortion rate of larvae and parasitization were not significantly affected by neither flower abundance nor temperature. Because the body size of females in O. bicornis is key for the next generation's progeny success, our results indicate that the simultaneous exposure to a shortage of floral resources and high temperatures may have adverse direct fitness effects.

Floral resource diversification promotes solitary bee reproduction and may offset insecticide effects - evidence from a semi-field experiment.

Pollinator declines in agricultural landscapes are driven by multiple stressors, but potential interactions of these remain poorly studied. Using a highly replicated semi-field study with 56 mesocosms of varying wild plant diversity (2-16 species) and oilseed rape treated with a neonicotinoid, we tested the interacting effects of resource diversity and insecticides on reproduction of a solitary wild bee. Compared to mesocosms with oilseed rape monocultures, availability of resources from wild plants complementing oilseed rape doubled brood cell production. In addition, bee reproduction increased due to plant diversity and identity effects. Exposure to neonicotinoid-treated oilseed rape reduced bee larval to adult development by 69%, but only in mesocosms with oilseed rape monocultures. Availability of complementary flower resources can thus offset negative effects of neonicotinoid-treated oilseed rape on wild bee reproduction. Policy should encourage the implementation of diverse floral resources mitigating negative effects of crop monocultures and insecticides, thereby sustaining solitary bee populations in agricultural landscapes.

Positive Correlation between Pesticide Consumption and Longevity in Solitary Bees: Are We Overlooking Fitness Trade-Offs?

The ubiquitous use of pesticides is one major driver for the current loss of biodiversity, and the common practice of simultaneously applying multiple agrochemicals may further contribute. Insect toxicology currently has a strong focus on survival to determine the potential hazards of a chemical routinely used in risk evaluations. However, studies revealing no effect on survival or even indicating enhanced survival are likely to be misleading, if potential trade-offs between survival and other physiological factors are overlooked. Here, we used standard laboratory experiments to investigate the sublethal (i.e., food consumption) and lethal (i.e., survival) effects of two common agricultural pesticides (Roundup® and clothianidin) on adult female solitary bees, Osmia bicornis. The data showed no significant effect of the treatment on cumulative survival; however, a significant positive correlation between herbicide and insecticide exposure and age was revealed, i.e., bees exposed to higher dosages lived longer. As no significant differences in daily food consumption were observed across treatment groups, increased food intake can be excluded as a factor leading to the prolonged survival. While this study does not provide data on fitness effects, two previous studies using solitary bees observed significant negative effects of neonicotinoid insecticides on fitness, yet not on survival. Thus, we conjecture that the observed non-significant effects on longevity may result from a trade-off between survival and reproduction. The data suggest that a focus on survival can lead to false-negative results and it appears inevitable to include fitness or at least tokens of fitness at the earliest stage in future risk assessments.

Susceptibility of Red Mason Bee Larvae to Bacterial Threats Due to Microbiome Exchange with Imported Pollen Provisions.

Solitary bees are subject to a variety of pressures that cause severe population declines. Currently, habitat loss, temperature shifts, agrochemical exposure, and new parasites are identified as major threats. However, knowledge about detrimental bacteria is scarce, although they may disturb natural microbiomes, disturb nest environments, or harm the larvae directly. To address this gap, we investigated 12 Osmia bicornis nests with deceased larvae and 31 nests with healthy larvae from the same localities in a 16S ribosomal RNA (rRNA) gene metabarcoding study. We sampled larvae, pollen provisions, and nest material and then contrasted bacterial community composition and diversity in healthy and deceased nests. Microbiomes of pollen provisions and larvae showed similarities for healthy larvae, whilst this was not the case for deceased individuals. We identified three bacterial taxa assigned to Paenibacillus sp. (closely related to P. pabuli/amylolyticus/xylanexedens), Sporosarcina sp., and Bacillus sp. as indicative for bacterial communities of deceased larvae, as well as Lactobacillus for corresponding pollen provisions. Furthermore, we performed a provisioning experiment, where we fed larvae with untreated and sterilized pollens, as well as sterilized pollens inoculated with a Bacillus sp. isolate from a deceased larva. Untreated larval microbiomes were consistent with that of the pollen provided. Sterilized pollen alone did not lead to acute mortality, while no microbiome was recoverable from the larvae. In the inoculation treatment, we observed that larval microbiomes were dominated by the seeded bacterium, which resulted in enhanced mortality. These results support that larval microbiomes are strongly determined by the pollen provisions. Further, they underline the need for further investigation of the impact of detrimental bacterial acquired via pollens and potential buffering by a diverse pollen provision microbiome in solitary bees.

Mating status and the evolution of eusociality: Oogenesis is independent of mating status in the solitary bee Osmia bicornis.

The fundamental trait underlying eusociality is the reproductive division of labour. In honeybees (Apis mellifera), queens lay eggs while workers forage, defend and care for brood. The division of labour is maintained by pheromones including queen mandibular pheromone (QMP) produced by the queen. QMP constrains reproduction in adult honeybee workers, but in the absence of their queen workers can activate their ovaries and, although they cannot mate, they lay haploid male eggs. The reproductive ground plan hypothesis suggests that reproductive constraint may have evolved by co-opting mechanisms of reproductive control in solitary ancestors. In many insects mating is required to activate or accelerate oogenesis. Here, we use the solitary bee Osmia bicornis (Megachilidae) to test whether reproductive constraint evolved from ancestral control of reproduction by mating status. We present a structural study of the O. bicornis ovary, and compare key stages of oogenesis with honeybee workers. Importantly, we show that mating does not affect any aspect of the reproductive physiology of O. bicornis. We therefore conclude that mechanisms governing reproductive constraint in honeybees were unlikely to have been co-opted from mechanisms pertaining to mating status.

Exposure of Larvae of the Solitary Bee Osmia bicornis to the Honey Bee Pathogen Nosema ceranae Affects Life History.

Wild bees are important pollinators of wild plants and agricultural crops and they are threatened by several environmental stressors including emerging pathogens. Honey bees have been suggested as a potential source of pathogen spillover. One prevalent pathogen that has recently emerged as a honey bee disease is the microsporidian Nosema ceranae. While the impacts of N. ceranae in honey bees are well documented, virtually nothing is known about its effects in solitary wild bees. The solitary mason bee Osmia bicornis is a common pollinator in orchards and amenable to commercial management. Here, we experimentally exposed larvae of O. bicornis to food contaminated with N. ceranae and document spore presence during larval development. We measured mortality, growth parameters, and timing of pupation in a semi-field experiment. Hatched individuals were assessed for physiological state including fat body mass, wing muscle mass, and body size. We recorded higher mortality in the viable-spore-exposed group but could only detect a low number of spores among the individuals of this treatment. Viable-spore-treated individuals with higher head capsule width had a delayed pupation start. No impact on the physiological status could be detected in hatched imagines. Although we did not find overt evidence of O. bicornis infection, our findings indicate that exposure of larvae to viable N. ceranae spores could affect bee development.

Combined exposure to sublethal concentrations of an insecticide and a fungicide affect feeding, ovary development and longevity in a solitary bee.

Pollinators in agroecosystems are often exposed to pesticide mixtures. Even at low concentrations, the effects of these mixtures on bee populations are difficult to predict due to potential synergistic interactions. In this paper, we orally exposed newly emerged females of the solitary bee Osmia bicornis to environmentally realistic levels of clothianidin (neonicotinoid insecticide) and propiconazole (fungicide), singly and in combination. The amount of feeding solution consumed was highest in bees exposed to the neonicotinoid, and lowest in bees exposed to the pesticide mixture. Ovary maturation and longevity of bees of the neonicotinoid and the fungicide treatments did not differ from those of control bees. By contrast, bees exposed to the pesticide mixture showed slow ovary maturation and decreased longevity. We found a synergistic interaction between the neonicotinoid and the fungicide on survival probability. We also found an interaction between treatment and emergence time (an indicator of physiological condition) on longevity. Longevity was negatively correlated to physiological condition only in the fungicide and the mixture treatments. Delayed ovary maturation and premature death imply a shortened nesting period (highly correlated to fecundity in Osmia). Our findings provide a mechanism to explain the observed dynamics of solitary bee populations exposed to multiple chemical residues in agricultural environments.

Effects of nonprotein amino acids on survival and locomotion of Osmia bicornis.

To investigate the effects of two nectar nonprotein amino acids, ß-alanine and γ-aminobutyric acid (GABA), on Osmia bicornis survival and locomotion, two groups of caged bees were fed with sugar syrup enriched with ß-alanine and GABA, respectively. A further control group was fed with sugar syrup. Five behavioural categories were chosen according to the principle of parsimony and intrinsic unitary consistency from start to end, and recorded by scan sampling: two states (remaining under paper or in tubes) and three events (walking on net, feeding from flower and flying). We also analysed the amino acid content of haemolymph sampled from an additional 45 bees fed the same diets (15 per diet type). Bees fed with ß-alanine had a significantly shorter survival time than those fed with the control and GABA diets. The GABA diet induced higher levels of locomotion than ß-alanine. The former nonprotein amino acid was only detected in the haemolymph of bees fed GABA. The results suggest that insects consuming nonprotein amino-acid-rich diets absorb and transfer these substances to the haemolymph and that nonprotein amino acids affect survival and locomotion. Ecological consequences are discussed in the framework of plant reproductive biology.

Effects of exposure to winter oilseed rape grown from thiamethoxam-treated seed on the red mason bee Osmia bicornis.

There has been increasing interest in the effects of neonicotinoid insecticides on wild bees. In solitary bee species the direct link between each individual female and reproductive success offers the opportunity to evaluate effects on individuals. The present study investigated effects of exposure to winter oilseed rape grown from thiamethoxam-treated seed on reproductive behavior and output of solitary red mason bees (Osmia bicornis) released in 6 pairs of fields over a 2-yr period and confined to tunnels in a single year. After adjustment to the number of females released, there was significantly lower production of cells and cocoons/female in tunnels than in open field conditions. This difference may be because of the lack of alternative forage within the tunnels. Under open field conditions, palynology of the pollen provisions within the nests demonstrated a maximum average of 31% oilseed rape pollen at any site, with Quercus (oak) contributing up to 86% of the pollen. There were no significant effects from exposure to oilseed rape grown from thiamethoxam-treated seed from nest establishment through cell production to emergence under tunnel or field conditions. Environ Toxicol Chem 2018;37:1071-1083. © 2017 SETAC.

Predation Cues in Solitary bee Nests.

Predation at the nesting site can significantly affect solitary bees' reproductive success. We tested female red mason bees' (Osmia bicornis L.) acceptance of potential nesting sites, some of which were marked with cues coming from predated conspecifics (crushed bees) or from a predator itself (rodent excreta). In our experiment, females did not avoid nests marked with either of the two predator cues. We suggest that bee females do not recognize these two cues as risky. Alternatively, costs of abandoning natal aggregation might be too high compared with any perceived predation risk of staying. Moreover, the presence of crushed bees can provide positive information about the presence of conspecifics and, possibly, information about a nesting aggregation that may be preferred by bees when choosing a nesting site.

The thermal environment of the nest affects body and cell size in the solitary red mason bee (Osmia bicornis L.).

Many ectotherms grow larger at lower temperatures than at higher temperatures. This pattern, known as the temperature-size rule, is often accompanied by plastic changes in cell size, which can mechanistically explain the thermal dependence of body size. However, the theory predicts that thermal plasticity in cell size has adaptive value for ectotherms because there are different optimal cell-membrane-to-cell-volume ratios at different temperatures. At high temperatures, the demand for oxygen is high; therefore, a large membrane surface of small cells is beneficial because it allows high rates of oxygen transport into the cell. The metabolic costs of maintaining membranes become more important at low temperatures than at high temperatures, which favours large cells. In a field experiment, we manipulated the thermal conditions inside nests of the red mason bee, a solitary bee that does not regulate the temperature in its nests and whose larvae develop under ambient conditions. We assessed the effect of temperature on body mass and ommatidia size (our proxy of cell size). The body and cell sizes decreased in response to a higher mean temperature and greater temperature fluctuations. This finding is in accordance with predictions of the temperature-size rule and optimal cell size theory and suggests that both the mean temperature and the magnitude of temperature fluctuations are important for determining body and cell sizes. Additionally, we observed that males of the red mason bee tend to have larger ommatidia in relation to their body mass than females, which might play an important role during mating flight.