Honeybee colonies compensate for pesticide-induced effects on royal jelly composition and brood survival with increased brood production.

Sublethal doses of pesticides affect individual honeybees, but colony-level effects are less well understood and it is unclear how the two levels integrate. We studied the effect of the neonicotinoid pesticide clothianidin at field realistic concentrations on small colonies. We found that exposure to clothianidin affected worker jelly production of individual workers and created a strong dose-dependent increase in mortality of individual larvae, but strikingly the population size of capped brood remained stable. Thus, hives exhibited short-term resilience. Using a demographic matrix model, we found that the basis of resilience in dosed colonies was a substantive increase in brood initiation rate to compensate for increased brood mortality. However, computer simulation of full size colonies revealed that the increase in brood initiation led to severe reductions in colony reproduction (swarming) and long-term survival. This experiment reveals social regulatory mechanisms on colony-level that enable honeybees to partly compensate for effects on individual level.

Time-dependent effects on bumble bees of dietary exposures to farmland insecticides (imidacloprid, thiamethoxam and fipronil).

BACKGROUND: Farmland bees encounter insecticides in their diet when visiting the flowers of pesticide-treated crops with residues in nectar and pollen. A sustained exposure to even trace residues could be severely harmful if the ingested substance has a long biological half-life and its toxicity therefore intensifies over time, which is termed time-reinforced toxicity (TRT). Bumble bees are important farmland pollinators, but their susceptibility to TRT from insecticides has not been established previously. This study therefore investigated the potential for three farmland insecticides (imidacloprid, thiamethoxam and fipronil) to generate TRT in bumble bees (Bombus terrestris L.) by dietary exposure in the laboratory using lethality as an endpoint under both fixed-dose and pulsed-dose regimes. RESULTS: The insecticides varied in toxic effect. Fipronil exposures produced a dose-dependent reduction in longevity (days of exposure survived) and strong TRT with no evidence of clearance-based recovery. Thiamethoxam exposures also produced a dose-dependent reduction in longevity, but with moderate TRT and evidence of some clearance-based recovery. Imidacloprid exposures produced a hormesis with reduced longevity only at the highest doses and an absence of TRT. CONCLUSION: Our study further confirms the potential for certain dietary insecticides at trace levels to harm farmland bees during sustained exposures because of their capacity to cause time-reinforced toxicity. Our findings suggest that regulatory oversight of pesticides will better safeguard bee health by testing the active ingredients of farmland agrochemicals for their capacity to produce TRT in these ecologically important nontarget organisms. Our study demonstrates the potential for certain dietary insecticides at trace levels to harm farmland bumble bees during sustained exposures because of their capacity to cause time-reinforced toxicity. © 2020 Society of Chemical Industry.

Fipronil pesticide as a suspect in historical mass mortalities of honey bees.

Mass mortalities of honey bees occurred in France in the 1990s coincident with the introduction of two agricultural insecticides, imidacloprid and fipronil. Imidacloprid, a neonicotinoid, was widely blamed, but the differential potency of imidacloprid and fipronil has been unclear because of uncertainty over their capacity to bioaccumulate during sustained exposure to trace dietary residues and, thereby, cause time-reinforced toxicity (TRT). We experimentally quantified the toxicity of fipronil and imidacloprid to honey bees and incorporated the observed mortality rates into a demographic simulation of a honey bee colony in an environmentally realistic scenario. Additionally, we evaluated two bioassays from new international guidance for agrochemical regulation, which aim to detect TRT. Finally, we used analytical chemistry (GC-MS) to test for bioaccumulation of fipronil. We found in demographic simulations that only fipronil produced mass mortality in honey bees. In the bioassays, only fipronil caused TRT. GC-MS analysis revealed that virtually all of the fipronil ingested by a honey bee in a single meal was present 6 d later, which suggests that bioaccumulation is the basis of TRT in sustained dietary exposures. We therefore postulate that fipronil, not imidacloprid, caused the mass mortalities of honey bees in France during the 1990s because it is lethal to honey bees in even trace doses due to its capacity to bioaccumulate and generate TRT. Our results provide evidence that recently proposed laboratory bioassays can discriminate harmful bioaccumulative substances and, thereby, address evident shortcomings in a regulatory system that had formerly approved fipronil for agricultural use.

Dominance of cropland reduces the pollen deposition from bumble bees.

Intensive agricultural landscapes can be hostile for bees due to a lack of floral and nesting resources, and due to management-related stress such as pesticide use and soil tillage. This threatens the pollination services that bees deliver to insect-pollinated crops. We studied the effects of farming intensity (organic vs. conventional, number of insecticide applications) and availability of semi-natural habitats at the field and landscape scale on pollinator visits and pollen delivery to pumpkin in Germany. We found that wild bumble bees were the key pollinators of pumpkin in terms of pollen delivery, despite fivefold higher visitation frequency of honey bees. Critically, we observed that the area of cropland had stronger effects on bees' pollen deposition than the area of seminatural habitats. Specifically, a 10% increase of the proportion of cropland reduced pollen delivery by 7%. Pumpkin provides a striking example for a key role of wild pollinators in crop pollination even at high numerical dominance of honey bees. In addition, our findings suggest that habitat conversion to agricultural land is a driver of deteriorating pollination. This underlines the importance to maintain sufficient areas of non-crop habitats in agricultural landscapes.

Synergistic interactions between a variety of insecticides and an ergosterol biosynthesis inhibitor fungicide in dietary exposures of bumble bees (Bombus terrestris L.).

BACKGROUND: In recent years, concern has been raised over honey bee colony losses, and also among wild bees there is evidence for extinctions and range contractions in Europe and North America. Pesticides have been proposed as a potential cause of this decline. Bees are exposed simultaneously to a variety of agrochemicals, which may cause synergistically detrimental impacts, which are incompletely understood. We investigated the toxicity of the fungicide imazalil in mixture with four common insecticides: fipronil (phenylpyrazoid), cypermethrin (pyrethroid), thiamethoxam, and imidacloprid (neonicotinoids). Ergosterol biosynthesis inhibitor (EBI) fungicides like imazalil can inhibit P450 detoxification systems in insects and therefore fungicide - insecticide co-occurrence might produce synergistic toxicity in bees. We assessed the impact of dietary fungicide - insecticide mixtures on the mortality and feeding rates of laboratory bumble bees (Bombus terrestris L.). RESULTS: Regarding mortality, imazalil synergised the toxicity of fipronil, cypermethrin and thiamethoxam, but not imidacloprid. We found no synergistic effects on feeding rates. CONCLUSION: Our findings suggest that P450-based detoxification processes are differentially important in mitigating the toxicity of certain insecticides, even those of the same chemical class. Our evidence that cocktail effects can arise in bumble bees should extend concern about the potential impacts of agrochemical mixtures to include wild bee species in farmland. © 2017 Society of Chemical Industry.

The effect of dietary neonicotinoid pesticides on non-flight thermogenesis in worker bumble bees (Bombus terrestris).

For bumble bees (genus Bombus), the capacity for non-flight thermogenesis is essential for two fundamental processes undertaken by adult workers, namely recovery from torpor after chilling and brood incubation. Farmland bees can be widely exposed to dietary residues of neurotoxic neonicotinoid insecticides that appear in the nectar and pollen of treated bee-attractive crops, which may harm them. An earlier study shows that dietary neonicotinoids cause complex alterations to thermoregulation in honey bees, but their effect on the thermogenic capabilities of individual bumble bees has been untested previously. We therefore conducted laboratory trials involving separate dietary exposures of bumble bees to two neonicotinoids, imidacloprid and thiamethoxam, and we measured their effects on the thoracic temperatures of bees during recovery from chilling. Specifically, we used thermal imaging to measure the rates of rewarming by individual bees after chill-induced torpor and to quantify their equilibrated thoracic temperatures post-recovery. We found that both toxicants caused dose-dependent decreases in the rates of rewarming and in the equilibrated thoracic temperatures. As previously found in honey bees, the dose-response relationship for imidacloprid exhibited a biphasic hormesis with low-dose stimulation and high-dose inhibition, for which we propose a mechanism. Our present study is among the first to detect ecologically relevant effects on bees in neonicotinoid exposures involving dietary concentrations below 5 ppb. If the effects on thoracic temperatures that we observed over a short period were sustained, they could have ecologically significant impacts on farmland bumble bees.

Apomixis and Hybridization Drives Reticulate Evolution and Phyletic Differentiation in Sorbus L.: Implications for Conservation.

Hybridization and polyploidy are major forces in the evolution of plant diversity and the study of these processes is of particular interest to understand how novel taxa are formed and how they maintain genetic integrity. Sorbus is an example of a genus where active diversification and speciation are ongoing and, as such, represents an ideal model to investigate the roles of hybridization, polyploidy and apomixis in a reticulate evolutionary process. To elucidate breeding systems and evolutionary origins of a complex of closely related Sorbus taxa, we assessed genotypic diversity and population structure within and among taxa, combining data from nuclear DNA microsatellite markers and flow cytometry. Clonal analysis and low genotypic diversity within the polyploid taxa suggest apomixis is obligate. However, genetic variation has led to groups of 'clone-mates' within apomictic taxa that strongly suggest mutation is responsible for the genotypic diversity of these apomictic lineages. In addition, microsatellite profiles and site demographics suggest hybridization events among apomictic polyploid Sorbus may have contributed to the extant diversity of recognized taxa in this region. This research demonstrates that both macro- and micro-evolutionary processes are active within this reticulate Sorbus complex. Conservation measures should be aimed at maintaining this process and should therefore be prioritized for those areas of Sorbus species richness where the potential for interspecific gene flow is greatest.

Sensitivity of commercial pumpkin yield to potential decline among different groups of pollinating bees.

The yield of animal-pollinated crops is threatened by bee declines, but its precise sensitivity is poorly known. We therefore determined the yield dependence of Hokkaido pumpkin in Germany on insect pollination by quantifying: (i) the relationship between pollen receipt and fruit set and (ii) the cumulative pollen deposition of each pollinator group. We found that approximately 2500 pollen grains per flower were needed to maximize fruit set. At the measured rates of flower visitation, we estimated that bumblebees (21 visits/flower lifetime, 864 grains/visit) or honeybees (123 visits, 260 grains) could individually achieve maximum crop yield, whereas halictid bees are ineffective (11 visits, 16 grains). The pollinator fauna was capable of delivering 20 times the necessary amount of pollen. We therefore estimate that pumpkin yield was not pollination-limited in our study region and that it is currently fairly resilient to single declines of honeybees or wild bumblebees.

Breeding system and spatial isolation from congeners strongly constrain seed set in an insect-pollinated apomictic tree: Sorbus subcuneata (Rosaceae).

In plants, apomixis results in the production of clonal offspring via seed and can provide reproductive assurance for isolated individuals. However, many apomicts require pollination to develop functional endosperm for successful seed set (pseudogamy) and therefore risk pollination-limitation, particularly in self-incompatible species that require heterospecific pollen. We used microsatellite paternity analysis and hand pollinations to investigate pollen-limitation in Sorbus subcuneata, a threatened endemic tree that co-occurs with its congener, S. admonitor. We confirmed that S. subcuneata is an obligate pseudogamous apomict, but open-pollinated flowers rarely produced seed (flower-to-seed conversion < 1%) even though they rapidly accumulated pollen on their stigmas. Manual heterospecific pollination by S. admonitor resulted in a high flower-to-seed conversion rate (65%), however, we estimate that the ratio of self: heterospecific pollination in open-pollinated flowers was at least 22:1. Despite the efficacy of heterospecific pollination, the contribution of S. admonitor trees to paternity in seed from open-pollinated flowers of S. subcuneata decreased rapidly with the spatial separation between paternal and maternal trees. Conservation efforts aimed at maintaining species with this breeding system must therefore manage the congeners in tandem which will also maintain the potential for rare heterospecific fertilisation that typically cause rapid diversification in these lineages.

Effects of the neonicotinoid pesticide thiamethoxam at field-realistic levels on microcolonies of Bombus terrestris worker bumble bees.

Neonicotinoid pesticides are currently implicated in the decline of wild bee populations. Bumble bees, Bombus spp., are important wild pollinators that are detrimentally affected by ingestion of neonicotinoid residues. To date, imidacloprid has been the major focus of study into the effects of neonicotinoids on bumble bee health, but wild populations are increasingly exposed to alternative neonicotinoids such as thiamethoxam. To investigate whether environmentally realistic levels of thiamethoxam affect bumble bee performance over a realistic exposure period, we exposed queenless microcolonies of Bombus terrestris L. workers to a wide range of dosages up to 98 µgkg(-1) in dietary syrup for 17 days. Results showed that bumble bee workers survived fewer days when presented with syrup dosed at 98 µg thiamethoxamkg(-1), while production of brood (eggs and larvae) and consumption of syrup and pollen in microcolonies were significantly reduced by thiamethoxam only at the two highest concentrations (39, 98 µgkg(-1)). In contrast, we found no detectable effect of thiamethoxam at levels typically found in the nectars of treated crops (between 1 and 11 µgkg(-1)). By comparison with published data, we demonstrate that during an exposure to field-realistic concentrations lasting approximately two weeks, brood production in worker bumble bees is more sensitive to imidacloprid than thiamethoxam. We speculate that differential sensitivity arises because imidacloprid produces a stronger repression of feeding in bumble bees than thiamethoxam, which imposes a greater nutrient limitation on production of brood.

Clearance of ingested neonicotinoid pesticide (imidacloprid) in honey bees (Apis mellifera) and bumblebees (Bombus terrestris).

BACKGROUND: Bees in agricultural landscapes are exposed to dietary pesticides such as imidacloprid when they feed from treated mass-flowering crops. Concern about the consequent impact on bees makes it important to understand their resilience. In the laboratory, the authors therefore fed adult worker bees on dosed syrup (125 µg L(-1) of imidacloprid, or 98 µg kg(-1)) either continuously or as a pulsed exposure and measured their behaviour (feeding and locomotory activity) and whole-body residues. RESULTS: On dosed syrup, honey bees maintained much lower bodily levels of imidacloprid than bumblebees (<0.2 ng versus 2.4 ng of imidacloprid per bee). Dietary imidacloprid did not affect the behaviour of honey bees, but it reduced feeding and locomotory activity in bumblebees. After the pulsed exposure, bumblebees cleared bodily imidacloprid after 48 h and recovered behaviourally. CONCLUSION: The differential behavioural resilience of the two species can be attributed to the observed differential in bodily residues. The ability of bumblebees to recover may be environmentally relevant in wild populations that face transitory exposures from the pulsed blooming of mass-flowering crops.

Repression and recuperation of brood production in Bombus terrestris bumble bees exposed to a pulse of the neonicotinoid pesticide imidacloprid.

Currently, there is concern about declining bee populations and some blame the residues of neonicotinoid pesticides in the nectar and pollen of treated crops. Bumble bees are important wild pollinators that are widely exposed to dietary neonicotinoids by foraging in agricultural environments. In the laboratory, we tested the effect of a pulsed exposure (14 days 'on dose' followed by 14 days 'off dose') to a common neonicotinoid, imidacloprid, on the amount of brood (number of eggs and larvae) produced by Bombus terrestris L. bumble bees in small, standardised experimental colonies (a queen and four adult workers). During the initial 'on dose' period we observed a dose-dependent repression of brood production in colonies, with productivity decreasing as dosage increased up to 98 µg kg(-1) dietary imidacloprid. During the following 'off dose' period, colonies showed a dose-dependent recuperation such that total brood production during the 28-day pulsed exposure was not correlated with imidacloprid up to 98 µg kg(-1). Our findings raise further concern about the threat to wild bumble bees from neonicotinoids, but they also indicate some resilience to a pulsed exposure, such as that arising from the transient bloom of a treated mass-flowering crop.

Differential sensitivity of honey bees and bumble bees to a dietary insecticide (imidacloprid).

Currently, there is concern about declining bee populations and the sustainability of pollination services. One potential threat to bees is the unintended impact of systemic insecticides, which are ingested by bees in the nectar and pollen from flowers of treated crops. To establish whether imidacloprid, a systemic neonicotinoid and insect neurotoxin, harms individual bees when ingested at environmentally realistic levels, we exposed adult worker bumble bees, Bombus terrestris L. (Hymenoptera: Apidae), and honey bees, Apis mellifera L. (Hymenoptera: Apidae), to dietary imidacloprid in feeder syrup at dosages between 0.08 and 125µg l(-1). Honey bees showed no response to dietary imidacloprid on any variable that we measured (feeding, locomotion and longevity). In contrast, bumble bees progressively developed over time a dose-dependent reduction in feeding rate with declines of 10-30% in the environmentally relevant range of up to 10µg l(-1), but neither their locomotory activity nor longevity varied with diet. To explain their differential sensitivity, we speculate that honey bees are better pre-adapted than bumble bees to feed on nectars containing synthetic alkaloids, such as imidacloprid, by virtue of their ancestral adaptation to tropical nectars in which natural alkaloids are prevalent. We emphasise that our study does not suggest that honey bee colonies are invulnerable to dietary imidacloprid under field conditions, but our findings do raise new concern about the impact of agricultural neonicotinoids on wild bumble bee populations.

Comment on "A common pesticide decreases foraging success and survival in honey bees".

Henry et al. (Reports, 20 April, p. 348) used a model to predict that colony collapse in honey bees could be precipitated by pesticide-induced intoxication that disrupts navigation. Here, we show that collapse disappears when the model is recalculated with parameter values appropriate to the season when most pesticide-treated flowering crops bloom.

Effects of imidacloprid, a neonicotinoid pesticide, on reproduction in worker bumble bees (Bombus terrestris).

Bumble bees are important pollinators whose populations have declined over recent years, raising widespread concern. One conspicuous threat to bumble bees is their unintended exposure to trace residues of systemic neonicotinoid pesticides, such as imidacloprid, which are ingested when bees forage on the nectar and pollen of treated crops. However, the demographic consequences for bumble bees of exposure to dietary neonicotinoids have yet to be fully established. To determine whether environmentally realistic levels of imidacloprid are capable of making a demographic impact on bumble bees, we exposed queenless microcolonies of worker bumble bees, Bombus terrestris, to a range of dosages of dietary imidacloprid between zero and 125 µg L(-1) and examined the effects on ovary development and fecundity. Microcolonies showed a dose-dependent decline in fecundity, with environmentally realistic dosages in the range of 1 µg L(-1) capable of reducing brood production by one third. In contrast, ovary development was unimpaired by dietary imidacloprid except at the highest dosage. Imidacloprid reduced feeding on both syrup and pollen but, after controlling statistically for dosage, microcolonies that consumed more syrup and pollen produced more brood. We therefore speculate that the detrimental effects of imidacloprid on fecundity emerge principally from nutrient limitation imposed by the failure of individuals to feed. Our findings raise concern about the impact of neonicotinoids on wild bumble bee populations. However, we recognize that to fully evaluate impacts on wild colonies it will be necessary to establish the effect of dietary neonicotinoids on the fecundity of bumble bee queens.

Dietary traces of neonicotinoid pesticides as a cause of population declines in honey bees: an evaluation by Hill's epidemiological criteria.

BACKGROUND: Honey bees are important pollinators of both crops and wild plants. Pesticide regimes that threaten their sustainability should therefore be assessed. As an example, evidence that the agricultural use of neonicotinoid pesticides is a cause of the recently observed declines in honey bees is examined. The aim is to define exacting demographic conditions for a detrimental factor to precipitate a population decline, and Hill's epidemiological 'causality criteria' are employed as a structured process for making an expert judgement about the proposition that trace dietary neonicotinoids in nectar and pollen cause population declines in honey bees. RESULTS: In spite of the absence of decisive experimental results, the analysis shows that, while the proposition is a substantially justified conjecture in the context of current knowledge, it is also substantially contraindicated by a wide variety of circumstantial epidemiological evidence. CONCLUSION: It is concluded that dietary neonicotinoids cannot be implicated in honey bee declines, but this position is provisional because important gaps remain in current knowledge. Avenues for further investigations to resolve this longstanding uncertainty are therefore identified.

A meta-analysis of experiments testing the effects of a neonicotinoid insecticide (imidacloprid) on honey bees.

Honey bees provide important pollination services to crops and wild plants. The agricultural use of systemic insecticides, such as neonicotinoids, may harm bees through their presence in pollen and nectar, which bees consume. Many studies have tested the effects on honey bees of imidacloprid, a neonicotinoid, but a clear picture of the risk it poses to bees has not previously emerged, because investigations are methodologically varied and inconsistent in outcome. In a meta-analysis of fourteen published studies of the effects of imidacloprid on honey bees under laboratory and semi-field conditions that comprised measurements on 7073 adult individuals and 36 colonies, fitted dose-response relationships estimate that trace dietary imidacloprid at field-realistic levels in nectar will have no lethal effects, but will reduce expected performance in honey bees by between 6 and 20%. Statistical power analysis showed that published field trials that have reported no effects on honey bees from neonicotinoids were incapable of detecting these predicted sublethal effects with conventionally accepted levels of certainty. These findings raise renewed concern about the impact on honey bees of dietary imidacloprid, but because questions remain over the environmental relevance of predominantly laboratory-based results, I identify targets for research and provide procedural recommendations for future studies.

Conifer ovulate cones accumulate pollen principally by simple impaction.

In many pine species (Family Pinaceae), ovulate cones structurally resemble a turbine, which has been widely interpreted as an adaptation for improving pollination by producing complex aerodynamic effects. We tested the turbine interpretation by quantifying patterns of pollen accumulation on ovulate cones in a wind tunnel and by using simulation models based on computational fluid dynamics. We used computer-aided design and computed tomography to create computational fluid dynamics model cones. We studied three species: Pinus radiata, Pinus sylvestris, and Cedrus libani. Irrespective of the approach or species studied, we found no evidence that turbine-like aerodynamics made a significant contribution to pollen accumulation, which instead occurred primarily by simple impaction. Consequently, we suggest alternative adaptive interpretations for the structure of ovulate cones.

Effect of pollinator abundance on self-fertilization and gene flow: application to GM Canola.

Cross-pollination from fields of transgenic crops is of great public concern. Although cross-pollination in commercial canola (Brassica napus) fields has been empirically measured, field trials are expensive and do not identify the causes of cross-pollination. Therefore, theoretical models can be valuable because they can provide estimates of cross-pollination at any given site and time. We present a general analytical model of field-to-field gene flow due to the following competing mechanisms: the wind, bees, and autonomous pollination. We parameterize the model for the particular case of field-to-field cross-pollination of genetically modified (GM) canola via the wind and via bumble bees (Bombus spp.) and honey bees (Apis mellifera). We make extensive use of the large data set of bee densities collected during the recent U.K. Farm Scale Evaluations. We predict that canola approaches almost full seed set without pollinators and that autonomous pollination is responsible for > or = 25% of seed set, irrespective of pollinator abundance. We do not predict the relative contribution of bees vs. the wind in landscape-scale gene flow in canola. However, under model assumptions, we predict that the maximum field-to-field gene flow due to bumble bees is 0.04% and 0.13% below the current EU limit for adventitious GM presence for winter- and spring-sown canola, respectively. We predict that gene flow due to bees is approximately 3.1 times higher at 20% compared to 100% male-fertility, and due to the wind, 1.3 times higher at 20% compared to 100% male-fertility, for both winter- and spring-sown canola. Bumble bee-mediated gene flow is approximately 2.7 times higher and wind-mediated gene flow approximately 1.7 times lower in spring-sown than in winter-sown canola, regardless of the degree of male-sterility. The model of cross-pollination due to the wind most closely predicted three previously published observations: field-to-field gene flow is low; gene flow increases with the proportion of plants that are male-sterile; and gene flow is higher in winter- than in spring-sown canola. Our results therefore suggest that the wind, not bees, is the main vector of long-distance gene flow in canola.

The effect of wind direction on cross-pollination in wind-pollinated GM crops.

In Europe, regulatory thresholds restrict adventitious GM (genetically modified) presence in conventional crops. Minimum distances for the spatial separation of fields are often recommended to reduce field-to-field cross-pollination to an acceptable level. Field trials are typically the basis for setting separation distances. However, using records of wind direction and speed from weather stations across Europe, we predict theoretically that field-to-field windborne cross-pollination in maize, oilseed rape, sugar beet, and rice varies greatly according to the relative orientation of the GM and non-GM fields. Furthermore, at a given site and orientation from a GM field, we predict that the cross-pollination rate varies substantially from year to year. Consequently, even replicated field trials may inaccurately estimate typical levels of cross-pollination and therefore distort our perception of the separation distances required to achieve sub-threshold adventitious GM presence. We propose methods to predict the likely range in levels of cross-pollination based on the limited data typically available from field trials. Additionally, we suggest suitable time lags between peak flowering in adjacent fields that could be introduced to reduce cross-pollination to a specified level.