Pollen chemical and mechanical defences restrict host-plant use by bees.

Plants produce an array of chemical and mechanical defences that provide protection against many herbivores and pathogens. Putatively defensive compounds and structures can even occur in floral rewards: for example, the pollen of some plant taxa contains toxic compounds or possesses conspicuous spines. Yet little is known about whether pollen defences restrict host-plant use by bees. In other words, do bees, like other insect herbivores, tolerate the defences of their specific host plants while being harmed by non-host defences? To answer this question, we compared the effects of a chemical defence from Lupinus (Fabaceae) pollen and a putative mechanical defence (pollen spines) from Asteraceae pollen on larval survival of nine bee species in the tribe Osmiini (Megachilidae) varying in their pollen-host use. We found that both types of pollen defences reduce larval survival rate in some bee species. These detrimental effects were, however, mediated by host-plant associations, with bees being more tolerant of the pollen defences of their hosts, relative to the defences of plant taxa exploited by other species. This pattern strongly suggests that bees are adapted to the pollen defences of their hosts, and that host-plant use by bees is constrained by their ability to tolerate such defences.

Landscape and land use affect composition and nutritional values of bees' food.

Bees are primary pollinators across various terrestrial biomes and rely heavily on floral resources for sustenance. The composition of landscapes can influence bee foraging behavior, while human activities can directly affect both the composition and nutritional value of bee food. We aimed to assess how landscape structure and land use practices can impact the composition and nutritional value of food sources for two generalist social bee species, Apis mellifera and Scaptotrigona postica. Food samples were collected from twenty-five colonies of A. mellifera and thirteen of S. postica to examine how food composition and nutritional value may vary based on the extent of human land use and the composition of landscapes surrounding beekeeping sites. The pollen composition and nutritional value of A. mellifera were influenced by both land use practices and landscape heterogeneity. The number of patches determined total sugar and lipid content. Landscape heterogeneity affected pollen composition in S. postica, primarily due to the number of patches, while total sugar was affected by landscape diversity. Pollen nutritional value in S. postica was linked to land use, mainly meadow and vegetation, which influenced total sugar and dry matter. S. postica showed a higher sensitivity to land use changes compared to A. mellifera, which was more affected by landscape heterogeneity. Assuring landscape heterogeneity by preserving remaining forest patches around apiaries and meliponaries is crucial. Thoughtful land use planning is essential to support beekeeping activities and ensure an adequate quantity and quality of bee food resources.

Impact of landscape composition on honey bee pollen contamination by pesticides: A multi-residue analysis.

The honey bee is the most common and important managed pollinator of crops. In recent years, honey bee colonies faced high mortality for multiple causes, including land-use change and the use of plant protection products (hereafter pesticides). This work aimed to explore how contamination by pesticides of pollen collected by honey bees was modulated by landscape composition and seasonality. We placed two honey bee colonies in 13 locations in Northern Italy in contrasting landscapes, from which we collected pollen samples monthly during the whole flowering season in 2019 and 2020. We searched for almost 400 compounds, including fungicides, herbicides, insecticides, and acaricides. We then calculated for each pollen sample the Pollen Hazard Quotient (PHQ), an index that provides a measure of multi-residue toxicity of contaminated pollen. Almost all pollen samples were contaminated by at least one compound. We detected 97 compounds, mainly fungicides, but insecticides and acaricides showed the highest toxicity. Fifteen % of the pollen samples had medium-high or high levels of PHQ, which could pose serious threats to honey bees. Fungicides showed a nearly constant PHQ throughout the season, while herbicides and insecticides and acaricides showed higher PHQ values in spring and early summer. Also, PHQ increased with increasing cover of agricultural and urban areas from April to July, while it was low and independent of landscape composition at the end of the season. The cover of perennial crops, i.e., fruit trees and vineyards, but not of annual crops, increased PHQ of pollen samples. Our work highlighted that the potential toxicity of pollen collected by honey bees was modulated by complex interactions among pesticide category, seasonality, and landscape composition. Due to the large number of compounds detected, our study should be complemented with additional experimental research on the potential interactive effects of multiple compounds on honey bee health.

Landfill fire impact on bee health: beneficial effect of dietary supplementation with medicinal plants and probiotics in reducing oxidative stress and metal accumulation.

The honey bee is an important pollinator insect susceptible to environmental contaminants. We investigated the effects of a waste fire event on elemental content, oxidative stress, and metabolic response in bees fed different nutrients (probiotics, Quassia amara, and placebo). The level of the elements was also investigated in honey and beeswax. Our data show a general increase in elemental concentrations in all bee groups after the event; however, the administration of probiotics and Quassia amara help fight oxidative stress in bees. Significantly lower concentrations of Ni, S, and U for honey in the probiotic group and a general and significant decrease in elemental concentrations for beeswax in the probiotic group and Li in the Quassia amara group were observed after the fire waste event. The comparison of the metabolic profiles through pre- and post-event PCA analyses showed that bees treated with different feeds react differently to the environmental event. The greatest differences in metabolic profiles are observed between the placebo-fed bees compared to the others. This study can help to understand how some stress factors can affect the health of bees and to take measures to protect these precious insects.

Sterol composition in plants is specific to pollen, leaf, pollination and pollinator.

Sterols have several roles in planta, including as membrane components. Sterols are also essential nutrients for insects. Based on this, and the different functions of leaves and pollen, we tested the hypotheses that (a) the sterolome is different in leaves and pollen from the same plant, (b) pollens from wind- and insect pollinated plants comprise different sterols, and (c) sterol provision in pollen-rewarding angiosperms differs from nectar-rewarding species. A novel approach to sterolomics was developed, using LCMS to determine the sterol profile of leaf and pollen from a taxonomically diverse range of 36 plant species. Twenty-one sterols were identified unambiguously, with several more identified in trace amounts. C29 sterols dominated the sterolome in most plants. The sterol composition was significantly different in leaf and pollen and their main sterols evolved in different ways. The sterolome of pollen from animal- and wind-pollinated was also significantly different, but not between nectar- and pollen-rewarding species. Our results suggest that the sterol composition in different plant tissues is linked to their biological functions. Sterol composition in pollen might be driven by physical role rather than the nutrient needs of pollinating insects.

Decline in wild bee species richness associated with honey bee (Apis mellifera L.) abundance in an urban ecosystem.

The spatial heterogeneity of urban landscapes, relatively low agrochemical use, and species-rich floral communities often support a surprising diversity of wild pollinators in cities. However, the management of Western honey bees (Apis mellifera L.) in urban areas may represent a new threat to wild bee communities. Urban beekeeping is commonly perceived as an environmentally friendly practice or a way to combat pollinator declines, when high-density beekeeping operations may actually have a negative influence on native and wild bee populations through floral resource competition and pathogen transmission. On the Island of Montréal, Canada there has been a particularly large increase in beekeeping across the city. Over the years following a large bee diversity survey ending in 2013, there was an influx of almost three thousand honey bee colonies to the city. In this study, we examined the wild bee communities and floral resources across a gradient of honey bee abundances in urban greenspaces in 2020, and compared the bee communities at the same sites before and after the large influx of honey bees. Overall, we found a negative relationship between urban beekeeping, pollen availability, and wild bee species richness. We also found that honey bee abundance had the strongest negative effect on small (inter-tegular span <2.25 mm) wild bee species richness. Small bee species may be at higher risk in areas with abundant honey bee populations as their limited foraging range may reduce their access to floral resources in times of increased competition. Further research on the influence of urban beekeeping on native and wild pollinators, coupled with evidence-based beekeeping regulations, is essential to ensure cities contain sufficient resources to support wild bee diversity alongside managed honey bees.

Flower angle favors pollen export efficiency in the snowdrop Galanthus nivalis (Linnaeus, 1753) but not in the lesser celandine Ficaria verna (Huds, 1762).

Flower angle is crucially important for accurate pollination and flower protection against abiotic factors. Evolutionary factors shaping floral traits are particularly strong for bilaterally symmetric flowers because these flowers require more pollination accuracy than radially symmetrical flowers. We experimentally investigated the flower angle in the snowdrop's (Galanthus nivalis) radially symmetrical, early-blooming downward flowers. Bumblebees were able to gather significantly more pollen grains from downward flowers than from upward flowers, but female traits (fertility in the field) seem unaffected by flower angle. Similar experiments with radially symmetrical, later flowering Lesser celandine (Ficaria verna) upward flowers showed no differences in bees' abilities to gather pollen in upward vs downward-facing flowers. The downward angle of snowdrop flowers is an adaptation that increases the ability of insects to collect more pollen grains under unfavorable early spring weather conditions when pollinators are scarce.

The need for weeds: Man-made, non-cropped habitats complement crops and natural habitats in providing honey bees and bumble bees with pollen resources.

In Europe, honey bees and bumble bees are among the most important pollinators, and there is a growing interest in understanding the effects of floral resource availability on their survival. Yet, to date, data on nectar and pollen supplies available to bees in agricultural landscapes are still scarce. In this paper, we quantify species-, habitat- and landscape-scale pollen production in the Lublin Upland, SE Poland. The production per unit area was highest (mean = 2.2-2.6 g/m2) in non-forest woody vegetation, field margins and fallows, whilst significantly lower pollen amounts were shown to be available in road verges and railway embankments (mean = 1.3-1.6 g/m2). At landscape scale, natural and semi-natural areas (forests and meadows/pastures) offered ca. 44% of the total pollen resources during the year. Relatively high amounts of pollen (ca. 35% of the year-round total pollen resources) were from winter rape, but this resource was short-term. Man-made, non-cropped habitats added only ca. 18% of the total pollen mass offered for pollinators during flowering season. However, they provided 66-99% of pollen resources available from July to October. There exists an imbalance in the availability of pollen resources throughout the year. Hence, a diversity of natural, semi-natural and man-made, non-cropped areas is required to support the seasonal continuity of pollen resources for pollinators in an agricultural landscape. Efforts should be made to secure habitat heterogeneity to enhance the flower diversity and continual pollen availability for pollinators.

Fungicides and bees: a review of exposure and risk.

Fungicides account for more than 35% of the global pesticide market and their use is predicted to increase in the future. While fungicides are commonly applied during bloom when bees are likely foraging on crops, whether real-world exposure to these chemicals - alone or in combination with other stressors - constitutes a threat to the health of bees is still the subject of great uncertainty. The first step in estimating the risks of exposure to fungicides for bees is to understand how and to what extent bees are exposed to these active ingredients. Here we review the current knowledge that exists about exposure to fungicides that bees experience in the field, and link quantitative data on exposure to acute and chronic risk of lethal endpoints for honey bees (Apis mellifera). From the 702 publications we screened, 76 studies contained quantitative data on residue detections in honey bee matrices, and a further 47 provided qualitative information about exposure for a range of bee taxa through various routes. We compiled data for 90 fungicides and metabolites that have been detected in honey, beebread, pollen, beeswax, and the bodies of honey bees. The risks posed to honey bees by fungicide residues was estimated through the EPA Risk Quotient (RQ) approach. Based on residue concentrations detected in honey and pollen/beebread, none of the reported fungicides exceeded the levels of concern (LOC) set by regulatory agencies for acute risk, while 3 and 12 fungicides exceeded the European Food Safety Authority (EFSA) chronic LOC for honey bees and wild bees, respectively. When considering exposure to all bees, fungicides of most concern include many broad-spectrum systemic fungicides, as well as the widely used broad-spectrum contact fungicide chlorothalonil. In addition to providing a detailed overview of the frequency and extent of fungicide residue detections in the bee environment, we identified important research gaps and suggest future directions to move towards a more comprehensive understanding and mitigation of the risks of exposure to fungicides for bees, including synergistic risks of co-exposure to fungicides and other pesticides or pathogens.

Toxicological assessment of agrochemicals on bees using machine learning tools.

Machine learning (ML) is a branch of artificial intelligence (AI) that enables the analysis of complex multivariate data. ML has significant potential in risk assessments of non-target insects for modeling the multiple factors affecting insect health, including the adverse effects of agrochemicals. Here, the potential of ML for risk assessments of glyphosate (herbicide; formulation) and imidacloprid (insecticide, neonicotinoid; formulation) on the stingless bee Melipona quadrifasciata was explored. The collective behavior of forager bees was analyzed after in vitro exposure to agrochemicals. ML algorithms were applied to identify the agrochemicals that the bees have been exposed to based on multivariate behavioral features. Changes in the in situ detection of different proteins in the midgut were also studied. Imidacloprid exposure leads to the greatest changes in behavior. The ML algorithms achieved a higher accuracy (up to 91%) in identifying agrochemical contamination. The two agrochemicals altered the detection of cells positive for different proteins, which can be detrimental to midgut physiology. This study provides a holistic assessment of the sublethal effects of glyphosate and imidacloprid on a key pollinator. The procedures used here can be applied in future studies to monitor and predict multiple environmental factors affecting insect health in the field.

Toxicokinetics of three insecticides in the female adult solitary bee Osmia bicornis.

The worldwide decline of pollinators is of growing concern and has been related to the use of insecticides. Solitary bees are potentially exposed to many insecticides through contaminated pollen and/or nectar. The kinetics of these compounds in solitary bees is, however, unknown, limiting the use of these important pollinators in pesticide regulations. Here, the toxicokinetics (TK) of chlorpyrifos (as Dursban 480 EC), cypermethrin (Sherpa 100 EC), and acetamiprid (Mospilan 20 SP) was studied for the first time in Osmia bicornis females at sublethal concentrations (near LC20s). The TK of the insecticides was analysed in bees continuously exposed to insecticide-contaminated food in the uptake phase followed by feeding with clean food in the decontamination phase. The TK models differed substantially between the insecticides. Acetamiprid followed the classic one-compartment model with gradual accumulation during the uptake phase followed by depuration during the decontamination phase. Cypermethrin accumulated rapidly in the first two days and then its concentration decreased slowly. Chlorpyrifos accumulated similarly rapidly but no substantial depuration was found until the end of the experiment. Our study demonstrates that some insecticides can harm solitary bees when exposed continuously even at trace concentrations in food because of their constant accumulation leading to time-reinforced toxicity.

Pesticide residues in honeybee-collected pollen: does the EU regulation protect honeybees from pesticides?

Researchers globally identify pesticides as one of the main reasons for pollinator decline. In the European Union (EU), extensive legislation is implemented to protect pollinators from harmful pesticide exposure. The aim of our study was to discover whether the pesticide residue levels in honeybee matrices, such as nectar and pollen, exceeded the chronic or acute toxicity levels when beehives were located next to fields treated with specific insecticides. The insecticides were used according to the EU legislation and its national implementation. The experiments were conducted in turnip rape, oilseed rape, and caraway fields in southern Finland during the years 2019 and 2020. The pesticides used in the experiments contained the active substances lambda-cyhalothrin (2019), esfenvalerate (2020), and tau-fluvalinate (2020). However, the honeybee-collected pollen and nectar were analyzed for residues of more than 100 active substances. The results showed that the pesticide residue levels clearly remained under the oral acute toxicity for honeybees, although we found high levels of thiacloprid residues in the pollen collected in 2019. The pesticide residues in nectar were below LOQ values, which was most likely due to the rainy weather conditions together with the chosen sampling method. No statistically significant differences were observed between the insecticide-treated and untreated fields. In light of our research, the EU legislation protected honeybees from oral acute toxicity during the years 2019 and 2020. However, potential sublethal effects of thiacloprid and other pesticide compounds found in the collected pollen cannot be ruled out. In the future, constant monitoring of pesticide exposure of honeybees and wild pollinators should be established to ensure that pesticide legislation, and its implementation across the EU successfully protects pollinators and their services in agricultural environments.

Establishing realistic exposure estimates of solitary bee larvae via pollen for use in risk assessment.

Bees foraging in agricultural habitats can be exposed to plant protection products. To limit the risk of adverse events, a robust risk assessment is needed, which requires reliable estimates for the expected exposure. The exposure pathways to developing solitary bees in particular are not well described and, in the currently proposed form, rely on limited information. To build a scaling model predicting the amount of protein developing solitary bees need based on adult body weight, we used published data on the volume of pollen solitary bees provide for their offspring. This model was tested against and ultimately updated with additional literature data on bee weight and protein content of emerged bees. We rescaled this model, based on the known pollen protein content of bee-visited flowers, to predict the expected amount of pollen a generalist solitary bee would likely provide based on its adult body weight, and tested these predictions in the field. We found overall agreement between the models' predictions and the measured values in the field, but additional data are needed to confirm these initial results. Our study suggests that scaling models in the bee risk assessment could complement existing risk assessment approaches and facilitate the further development of accurate risk characterization for solitary bees; ultimately the models will help to protect them during their foraging activity in agricultural settings. Integr Environ Assess Manag 2022;18:308-313. © 2021 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Fragility of nocturnal interactions: Pollination intensity increases with distance to light pollution sources but decreases with increasing environmental suitability.

Light pollution represents a widespread long-established human-made disturbance and an important threat to nocturnal pollination. Distance from the niche centroid where optimal environmental conditions join may be related to species sensitivity to habitat change. We estimated the environmental suitability of the plant species Erythrostemon gilliesii and of its guild of hawkmoth pollinators. We considered the overlap of suitability maps of both partners as the environmental suitability of the interaction. We used a three-year record of ten E. gilliesii populations to calculate pollination intensity as the number of individuals that received pollen per population. In addition, for each population, we measured the distance to the high light pollution source around a buffer of 15 km radius. Finally, we predicted pollination intensity values for environmental suitability ranging from 0 to 1, and distance to high light pollution sources ranging from 0 to 56 Km. Pollination intensity decreased along an axis of increasing environmental suitability and increased with distance to sources of light pollution. The highest values of pollination intensity were observed at greatest distances to sources of light pollution and where environmental suitability of the interaction was lowest. The prediction model evidenced that, when environmental suitability was lowest, pollination intensity increased with distance to sources of high light pollution. However, when environmental suitability was intermediate or high, pollination intensity decreased away and until 28 km from the sources of high light pollution. Beyond 28 km from the sources of high light pollution, pollination intensity remained low and constant. Populations under conditions of low environmental suitability might be more likely to respond to disturbances that affect pollinators than populations under conditions of high environmental suitability.

Comparative effects of pollen limitation, floral traits and pollinators on reproductive success of Hedysarum scoparium Fisch. et Mey. in different habitats.

BACKGROUND: Reproduction in most flowering plants may be limited because of the decreased visitation or activity of pollinators in fragmented habitats. Hedysarum scoparium Fisch. et Mey. is an arid region shrub with ecological importance. We explored the pollen limitation and seed set of Hedysarum scoparium in fragmented and restored environments, and examined whether pollen limitation is a significant limiting factor for seed set. We also compared floral traits and pollinator visitation between both habitats, and we determined the difference of floral traits and pollinators influenced reproductive success in Hedysarum scoparium. RESULTS: Our results indicated that supplementation with pollen significantly increased seed set per flower, which is pollen-limited in this species. Furthermore, there was greater seed set of the hand cross-pollination group in the restored habitat compared to the fragmented environment. More visits by Apis mellifera were recorded in the restored habitats, which may explain the difference in seed production between the fragmented and restored habitats. CONCLUSIONS: In this study, a positive association between pollinator visitation frequency and open flower number was observed. The findings of this study are important for experimentally quantifying the effects of floral traits and pollinators on plant reproductive success in different habitats.

Fungicide and insecticide exposure adversely impacts bumblebees and pollination services under semi-field conditions.

Sulfoximines, the next generation systemic insecticides developed to replace neonicotinoids, have been shown to negatively impact pollinator development and reproduction. However, field-realistic studies on sulfoximines are few and consequences on pollination services unexplored. Moreover, the impacts of other agrochemicals such as fungicides, and their combined effects with insecticides remain poorly investigated. Here, we show in a full factorial semi-field experiment that spray applications of both the product Closer containing the insecticide sulfoxaflor and the product Amistar containing the fungicide azoxystrobin, negatively affected the individual foraging performance of bumblebees (Bombus terrestris). Insecticide exposure further reduced colony growth and size whereas fungicide exposure decreased pollen deposition. We found indications for resource limitation that might have exacerbated pesticide effects on bumblebee colonies. Our work demonstrates that field-realistic exposure to sulfoxaflor can adversely impact bumblebees and that applications before bloom may be insufficient as a mitigation measure to prevent its negative impacts on pollinators. Moreover, fungicide use during bloom could reduce bumblebee foraging performance and pollination services.

Herbivory and Time Since Flowering Shape Floral Rewards and Pollinator-Pathogen Interactions.

Herbivory can induce chemical changes throughout plant tissues including flowers, which could affect pollinator-pathogen interactions. Pollen is highly defended compared to nectar, but no study has examined whether herbivory affects pollen chemistry. We assessed the effects of leaf herbivory on nectar and pollen alkaloids in Nicotiana tabacum, and how herbivory-induced changes in nectar and pollen affect pollinator-pathogen interactions. We damaged leaves of Nicotiana tabacum using the specialist herbivore Manduca sexta and compared nicotine and anabasine concentrations in nectar and pollen. We then pooled nectar and pollen by collection periods (within and after one month of flowering), fed them in separate experiments to bumble bees (Bombus impatiens) infected with the gut pathogen Crithidia bombi, and assessed infections after seven days. We did not detect alkaloids in nectar, and leaf damage did not alter the effect of nectar on Crithidia counts. In pollen, herbivory induced higher concentrations of anabasine but not nicotine, and alkaloid concentrations rose and then fell as a function of days since flowering. Bees fed pollen from damaged plants had Crithidia counts 15 times higher than bees fed pollen from undamaged plants, but only when pollen was collected after one month of flowering, indicating that both damage and time since flowering affected interaction outcomes. Within undamaged treatments, bees fed late-collected pollen had Crithidia counts 10 times lower than bees fed early-collected pollen, also indicating the importance of time since flowering. Our results emphasize the role of herbivores in shaping pollen chemistry, with consequences for interactions between pollinators and their pathogens.

The Influence of Arbuscular Mycorrhizal Fungi on Plant Reproduction.

Arbuscular mycorrhizal (AM) fungi can influence all components of plant reproduction including pollen delivery, pollen germination, pollen tube growth, fertilization, and seed germination. AM fungi associate with plant roots, uptake nutrients, and prime plants for faster defense responses. Our literature review first identified four testable hypotheses describing how AM fungi could alter pollen delivery: (1) We hypothesize AM fungi promote floral display size. The influence of AM fungi on flower size and number is supported by literature, however there are no studies on floral color. (2) We hypothesize AM fungi promote pollen and nectar quality and quantity, and, as reported before, AM fungi promote male fitness over female fitness. (3) We hypothesize AM fungi promote both earlier and longer flowering times, but we found no consistent trend in the data for earlier or later or longer flowering times. (4) We hypothesize AM fungi alter floral secondary chemistry and VOCs, and find there is clear evidence for the alteration of floral chemistry but little data on VOCs. Second, we focus on how AM fungi could alter pollen germination, pollen tube growth, and fertilization, and present three testable hypotheses. We found evidence that AM fungi influence pollen germination and pollen tube growth, production of seeds, and seed germination. However, while most of these influences are positive they are not conclusive, because studies have been conducted in small numbers of systems and groups. Therefore, we conclude that the majority of research to date may not be measuring the influence of AM fungi on the most important components of plant reproduction: pollen germination, pollen tube growth, fertilization, and seed germination.

Nocturnal pollination: an overlooked ecosystem service vulnerable to environmental change.

Existing assessments of the ecosystem service of pollination have been largely restricted to diurnal insects, with a particular focus on generalist foragers such as wild and honey bees. As knowledge of how these plant-pollinator systems function, their relevance to food security and biodiversity, and the fragility of these mutually beneficial interactions increases, attention is diverting to other, less well-studied pollinator groups. One such group are those that forage at night. In this review, we document evidence that nocturnal species are providers of pollination services (including pollination of economically valuable and culturally important crops, as well as wild plants of conservation concern), but highlight how little is known about the scale of such services. We discuss the primary mechanisms involved in night-time communication between plants and insect pollen-vectors, including floral scent, visual cues (and associated specialized visual systems), and thermogenic sensitivity (associated with thermogenic flowers). We highlight that these mechanisms are vulnerable to direct and indirect disruption by a range of anthropogenic drivers of environmental change, including air and soil pollution, artificial light at night, and climate change. Lastly, we highlight a number of directions for future research that will be important if nocturnal pollination services are to be fully understood and ultimately conserved.