Beyond the urgency: Pesticide Emergency Authorisations' exposure, toxicity, and risk for humans and the environment.

The global challenge to increase agricultural production goes along with the need of decreasing pesticide risks. The European Union (EU) therefore evaluates and controls the risks posed by pesticides by regulating their authorisation through the science-based Risk Assessment process. Member States can however act in derogation to this process and grant the Emergency Authorisation (EA) of pesticides that are currently non-authorised. To protect the health of humans and the environment, Emergency Authorisations are only permitted in exceptional circumstances of agricultural emergency: their use should be limited (i.e., cannot exceed 120 days and one growing season) and concurrent research on alternative strategies must be enforced. Here, we assessed the impact of the Emergency Authorisations process to human and environmental health. Bees, bioindicators of environmental health, were used as model species. Our research demonstrates that i) Emergency Authorisations are widely used throughout EU Member States (annually granted Emergency Authorisationsmin-max, 2017-2021 = 460-529); ii) 12 % of Emergency Authorisations granted the use of pesticides for longer than prescribed by EU regulations; iii) 37 % of Emergency Authorisations were repeatedly granted over time by the same Member State for the same agricultural purpose (i.e., to control the same pest on the same crop); iv) 21 % of Emergency Authorisations granted the use of Active Substances non-approved by the Risk assessment (EA-ASs Type3) which consequently contaminate the environment (44 % of environmental biomonitoring studies found EA-AS Type3) while being significantly more toxic to pollinators than regularly approved ASs. To facilitate the implementation of sustainable control strategies towards a safer environment for humans and other animals, we identified the most frequent agricultural emergencies and the key research needs. This first quantitative assessment of the Emergency Authorisation process unveils an enduring state of agricultural emergency that acts in derogation of the EU Regulation, leading to broad human, animal, and environmental implications.

Machine learning - based q-RASAR modeling to predict acute contact toxicity of binary organic pesticide mixtures in honey bees.

We have reported here a quantitative read-across structure-activity relationship (q-RASAR) model for the prediction of binary mixture toxicity (acute contact toxicity) in honey bees. Both the quantitative structure-activity relationship (QSAR) and the similarity-based read-across algorithms are used simultaneously for enhancing the predictability of the model. Several similarity and error-based parameters, obtained from the read-across prediction tool, have been put together with the structural and physicochemical descriptors to develop the final q-RASAR model. The calculated statistical and validation metrics indicate the goodness-of-fit, robustness, and good predictability of the partial least squares (PLS) regression model. Machine learning algorithms like ridge regression, linear support vector machine (SVM), and non-linear SVM have been used to further enhance the predictability of the q-RASAR model. The prediction quality of the q-RASAR models outperforms the previously reported quasi-SMILEs-based QSAR model in terms of external correlation coefficient (Q2F1 SVM q-RASAR: 0.935 vs. Q2VLD QSAR: 0.89). In this research, the toxicity values of several new untested binary mixtures have been predicted with the new models, and the reliability of the PLS predictions has been validated by the prediction reliability indicator tool. The q-RASAR approach can be used as reliable, complementary, and integrative to the conventional experimental approaches of pesticide mixture risk assessment.

Fungivorous mites enhance the survivorship and development of stingless bees even when exposed to pesticides.

Stingless bees are the largest group of eusocial bees in the world. They play an essential role as crop pollinators and have been considered for inclusion in pesticide risk assessments (RAs). Beyond the mutualism involving stingless bee larvae and fungi, the fungivorous mite Proctotydaeus (Neotydeolus) alvearii proved to be interesting for studies of associations with stingless bees. Their presence is related to colony strength and health, showing a permanent-host-association level. Here, we tested whether the coexistence with P. (N.) alvearii affects stingless bee larvae survivorship and development, including when fed pesticide-dosed food. We chose dimethoate, the reference standard for toxicity tests, and thiamethoxam, widely used in neotropical crops and listed to be reassessed in RAs. Bees associated with the mites showed higher larval survivorship rates, even in the dosed ones, and revealed changes in the developmental time and body size. Our study represents the first approach to stingless bee responses to the coexistence of fungivorous mites inside brood cells, leading us to believe that these mites play a beneficial role in stingless bees, including when they are exposed to pesticides.

Lethal, sublethal, and combined effects of pesticides on bees: A meta-analysis and new risk assessment tools.

Multiple stressors threaten bee health, a major one being pesticides. Bees are simultaneously exposed to multiple pesticides that can cause both lethal and sublethal effects. Risk assessment and most research on bee health, however, focus on lethal individual effects. Here, we performed a systematic literature review and meta-analysis that summarizes and re-interprets the available qualitative and quantitative information on the lethal, sublethal, and combined toxicity of a comprehensive range of pesticides on bees. We provide results (1970-2019) for multiple bee species (Bombus, Osmia, Megachile, Melipona, Partamona, Scaptotrigona), although most works focused on Apis mellifera L. (78 %). Our harmonised results document the lethal toxicity of pesticides in bees (n = 377 pesticides) and the types of sublethal testing methods and related effects that cause a sublethal effect (n = 375 sublethal experiments). We identified the most common combinations of pesticides and mode of actions tested, and summarize the experimental methods, magnitude of the interactions, and robustness of available data (n = 361 experiments). We provide open access searchable, comprehensive, and integrated list of pesticides and their levels causing lethal, sublethal, and combined effects. We report major data gaps related to pesticide's sublethal (71 %) and combined (e.g., ~99 %) toxicity. We identified pesticides and mode of actions of greatest concern in terms of sublethal (chlorothalonil, pymetrozine, glyphosate; neonicotinoids) and combined (tau-fluvalinate combinations; acetylcholinesterase inhibitors and neonicotinoids) effects. Although certain pesticides have faced regulatory restrictions in specific countries (chlorothalonil, pymetrozine, neonicotinoids), most are still widely used worldwide (e.g., glyphosate). This work aims at facilitating the implementation of more comprehensive and harmonised research and risk assessments, considering sublethal and combined effects. To ensure safeguarding pollinators and the environment, we advocate for a more refined and holistic assessment that do not only focus on lethality but uses harmonised methods to test sublethal and relevant combinations.

Evaluation of the risks for animal health related to the presence of hydroxymethylfurfural (HMF) in feed for honey bees.

The European Commission has asked the EFSA to evaluate the risk for animal health related to the presence of hydroxymethylfurfural (HMF) in honey bee feed. HMF is a degradation product of particular sugars and can be present in bee feed. HMF is of low acute toxicity in bees but causes increased mortality upon chronic exposure. A benchmark dose lower limit 10% (BMDL10) of 1.16 µg HMF per bee per day has been calculated from mortalities observed in a 20-day study and established as a Reference Point covering also mortality in larvae, drones and queens for which no or insufficient toxicity data were available. Winter bees have a much longer lifespan than summer bees and HMF shows clear time reinforced toxicity (TRT) characteristics. Therefore, additional Reference Point intervals of 0.21-3.1, 0.091-1.1 and 0.019-0.35 µg HMF/bee per day were calculated based on extrapolation to exposure durations of 50, 90 and 180 days, respectively. A total of 219 analytical data of HMF concentrations in bee feed from EU Member States and 88 from Industry were available. Exposure estimates of worker bees and larvae ranged between 0.1 and 0.48, and between 0.1 and 0.51 µg HMF/per day, respectively. They were well below the BMDL10 of 1.16 µg HMF/bee per day, and thus, no concern was identified. However, when accounting for TRT, the probability that exposures were below established reference point intervals was assessed to be extremely unlikely to almost certain depending on exposure duration. A concern for bee health was identified when bees are exposed to HMF contaminated bee feed for several months.

Thiamethoxam as an inadvertent anti-aphrodisiac in male bees.

Sexual reproduction is common to almost all multi-cellular organisms and can be compromised by environmental pollution, thereby affecting entire populations. Even though there is consensus that neonicotinoid insecticides can impact non-target animal fertility, their possible impact on male mating success is currently unknown in bees. Here, we show that sublethal exposure to a neonicotinoid significantly reduces both mating success and sperm traits of male bumblebees. Sexually mature male Bombus terrestris exposed to a field-realistic concentration of thiamethoxam (20 ng g-1) or not (controls) were mated with virgin gynes in the laboratory. The results confirm sublethal negative effects of thiamethoxam on sperm quantity and viability. While the latency to mate was reduced, mating success was significantly impaired in thiamethoxam-exposed males by 32% probably due to female choice. Gynes mated by exposed males revealed impaired sperm traits compared to their respective controls, which may lead to severe constraints for colony fitness. Our laboratory findings demonstrate for the first time that neonicotinoid insecticides can negatively affect male mating success in bees. Given that holds true for the field, this provides a plausible mechanism contributing to declines of wild bee populations globally. The widespread prophylactic use of neonicotinoids may therefore have previously overlooked inadvertent anti-aphrodisiac effects on non-target animals, thereby limiting conservation efforts.

CSI Pollen: Diversity of Honey Bee Collected Pollen Studied by Citizen Scientists.

A diverse supply of pollen is an important factor for honey bee health, but information about the pollen diversity available to colonies at the landscape scale is largely missing. In this COLOSS study, beekeeper citizen scientists sampled and analyzed the diversity of pollen collected by honey bee colonies. As a simple measure of diversity, beekeepers determined the number of colors found in pollen samples that were collected in a coordinated and standardized way. Altogether, 750 beekeepers from 28 different regions from 24 countries participated in the two-year study and collected and analyzed almost 18,000 pollen samples. Pollen samples contained approximately six different colors in total throughout the sampling period, of which four colors were abundant. We ran generalized linear mixed models to test for possible effects of diverse factors such as collection, i.e., whether a minimum amount of pollen was collected or not, and habitat type on the number of colors found in pollen samples. To identify habitat effects on pollen diversity, beekeepers' descriptions of the surrounding landscape and CORINE land cover classes were investigated in two different models, which both showed that both the total number and the rare number of colors in pollen samples were positively affected by 'urban' habitats or 'artificial surfaces', respectively. This citizen science study underlines the importance of the habitat for pollen diversity for bees and suggests higher diversity in urban areas.

Risk and protective indicators of beekeeping management practices.

Explaining the reasons for the high honey bee (Apis mellifera) colony loss rate in recent years has become a top global research priority in apicultural and agricultural sciences. Although there are indications of the role played by beekeeping management practices on honey bee health, very little information is currently available. Our study aimed to characterize the beekeeping management practices carried out in Belgium, and to determine the relationship between beekeeping management practices and colony losses. Variables obtained from face-to-face questioning of a representative randomized and stratified sample of Belgian beekeepers (n = 186) were integrated into a logistic regression model (univariate and multivariate) and correlated to the declared colony loss rates to identify risk and protective indicators. We used a classification tree analysis to validate the results. We present evidence of a relationship between poor beekeeping management practices and colony losses. The main factors protecting honey bee colonies are the aptitude of the beekeeper to change his management practices, the hive type, the equipment origin and hygiene, wintering in proper conditions (the use of divider boards, i.e. board blocks or space fillers off part of the hive body), the colony strength estimation before wintering, winter monitoring, and last but not least, appropriate integrated pest management. Proper estimation of the Varroa infestation level should be performed prior to treatment. The consequences of poor beekeeping practices on honey bee health can be addressed by proper training of beekeepers. An online tool was developed and published for beekeepers allowing them to evaluate the effect of their management practices on colony health.

Long-term field-realistic exposure to a next-generation pesticide, flupyradifurone, impairs honey bee behaviour and survival.

The assessment of pesticide risks to insect pollinators have typically focused on short-term, lethal impacts. The environmental ramifications of many of the world's most commonly employed pesticides, such as those exhibiting systemic properties that can result in long-lasting exposure to insects, may thus be severely underestimated. Here, seven laboratories from Europe and North America performed a standardised experiment (a ring-test) to study the long-term lethal and sublethal impacts of the relatively recently approved 'bee safe' butenolide pesticide flupyradifurone (FPF, active ingredient in Sivanto®) on honey bees. The emerging contaminant, FPF, impaired bee survival and behaviour at field-realistic doses (down to 11 ng/bee/day, corresponding to 400 µg/kg) that were up to 101-fold lower than those reported by risk assessments (1110 ng/bee/day), despite an absence of time-reinforced toxicity. Our findings raise concerns about the chronic impact of pesticides on pollinators at a global scale and support a novel methodology for a refined risk assessment.

A systems-based approach to the environmental risk assessment of multiple stressors in honey bees.

The European Parliament requested EFSA to develop a holistic risk assessment of multiple stressors in honey bees. To this end, a systems-based approach that is composed of two core components: a monitoring system and a modelling system are put forward with honey bees taken as a showcase. Key developments in the current scientific opinion (including systematic data collection from sentinel beehives and an agent-based simulation) have the potential to substantially contribute to future development of environmental risk assessments of multiple stressors at larger spatial and temporal scales. For the monitoring, sentinel hives would be placed across representative climatic zones and landscapes in the EU and connected to a platform for data storage and analysis. Data on bee health status, chemical residues and the immediate or broader landscape around the hives would be collected in a harmonised and standardised manner, and would be used to inform stakeholders, and the modelling system, ApisRAM, which simulates as accurately as possible a honey bee colony. ApisRAM would be calibrated and continuously updated with incoming monitoring data and emerging scientific knowledge from research. It will be a supportive tool for beekeeping, farming, research, risk assessment and risk management, and it will benefit the wider society. A societal outlook on the proposed approach is included and this was conducted with targeted social science research with 64 beekeepers from eight EU Member States and with members of the EU Bee Partnership. Gaps and opportunities are identified to further implement the approach. Conclusions and recommendations are made on a way forward, both for the application of the approach and its use in a broader context.

Pesticides in honey bee colonies: Establishing a baseline for real world exposure over seven years in the USA.

Honey bees Apis mellifera forage in a wide radius around their colony, bringing back contaminated food resources that can function as terrestrial bioindicators of environmental pesticide exposure. Evaluating pesticide exposure risk to pollinators is an ongoing problem. Here we apply five metrics for pesticide exposure risk (prevalence, diversity, concentration, significant pesticide prevalence, and hazard quotient (HQ)) to a nation-wide field study of honey bees, Apis mellifera in the United States. We examined samples from 1055 apiaries over seven years for 218 different pesticide residues and metabolites, determining that bees were exposed to 120 different pesticide products with a mean of 2.78 per sample. Pesticides in pollen were highly prevalent and variable across states. While pesticide diversity increased over time, most detections occurred at levels predicted to be of low risk to colonies. Varroacides contributed most to concentration, followed by fungicides, while insecticides contributed most to diversity above a toxicity threshold. High risk samples contained one of 12 different insecticides or varroacides. Exposures predicted to be low-risk were nevertheless associated with colony morbidity, and low-level fungicide exposures were tied to queen loss, Nosema infection, and brood diseases.

The neonicotinoid thiamethoxam impairs male fertility in solitary bees, Osmia cornuta.

The ongoing loss of global biodiversity is endangering ecosystem functioning and human food security. While environmental pollutants are well known to reduce fertility, the potential effects of common neonicotinoid insecticides on insect fertility remain poorly understood. Here, we show that field-realistic neonicotinoid exposure can drastically impact male insect fertility. In the laboratory, male and female solitary bees Osmia cornuta were exposed to four concentrations of the neonicotinoid thiamethoxam to measure survival, food consumption, and sperm traits. Despite males being exposed to higher dosages of thiamethoxam, females revealed an overall increased hazard rate for survival; suggesting sex-specific differences in toxicological sensitivity. All tested sublethal concentrations (i.e., 1.5, 4.5 and 10 ng g-1) reduced sperm quantity by 57% and viability by 42% on average, with the lowest tested concentration leading to a reduction in total living sperm by 90%. As the tested sublethal concentrations match estimates of global neonicotinoid pollution, this reveals a plausible mechanism for population declines, thereby reflecting a realistic concern. An immediate reduction in environmental pollutants is required to decelerate the ongoing loss of biodiversity.

Pesticide and veterinary drug residues in Belgian beeswax: Occurrence, toxicity, and risk to honey bees.

Pesticide and veterinary drug residues are one of the stress factors affecting bee health and mortality. To investigate the occurrence, the concentration and the toxicity risk to bees of pesticide residues in four different types of beeswax (brood comb wax, recycled comb wax, honey comb wax, and cappings wax), 182 samples were collected from apiaries located all over the Belgian territories, during spring 2016 and analysed by LC-MS/MS and GC-MS/MS for the presence of 294 chemical residues. The toxicity risk to bees expressed as the Hazard Quotient (HQ) was calculated for each wax sample, according to two scenarios with different tau-fluvalinate LD50 values. Residues showing the highest prevalence were correlated to bee mortality in a multivariate logistic regression model and a risk-based model was used to predict colony bee mortality. Altogether, 54 different pesticide and veterinary drug residues were found in the four types of beeswax. The residues with a higher likelihood to be retained in beeswax are applied in-hive or with a high lipophilic nature. The multivariate logistic regression model showed a statistically significant influence of chlorfenvinphos on bee mortality. All our results indicated that cappings wax was substantially less contaminated. This national survey on beeswax contamination provides guidelines on the re-use of beeswax by beekeepers and shows the necessity to introduce maximum residue levels for global trade in beeswax. An online tool was developed to enable beekeepers and wax traders to estimate the risk to honey bee health associated with contaminated wax.

Honeybee and consumer's exposure and risk characterisation to glyphosate-based herbicide (GBH) and its degradation product (AMPA): Residues in beebread, wax, and honey.

In order to assess bee and human exposure to residues of glyphosate-based herbicide (GBH) and its main degradation products aminomethylphosphonic acid (AMPA) and to characterise the risk posed by these substances, we analysed 3 different bee matrices; beebread (N = 81), wax (N = 100) and 10-paired samples of wax/honey collected in 2016/2017 from 379 Belgian apiaries. A high-performance liquid chromatography-electrospray ionisation tandem mass spectrometry (HPLC-ESI-MS-MS) was used as analytical method. Limit of quantification and detection (LOQ and LOD) for GBH residues and AMPA in the 3 matrices was respectively of 10 ng g-1 and 1 ng g-1. In beebread, 81.5% of the samples showed a residue concentration > LOQ and 9.9% of the samples a residue concentration < LOQ (detection without quantification); no significant difference in detection rate was found between the north and the south of the country. Glyphosate was detected in beeswax less frequently than in beebread (i.e. 26% >LOQ versus 81.5% >LOQ). The maximum GBH residues and AMPA concentration found in beebread (respectively 700 ng g-1 and 250 ng g-1) led to sub-lethal exposure to bees. The Hazard Quotient (HQ) for beebread and beeswax (7 and 3.2, respectively) were far below the "safety" oral and contact thresholds for bees. For human health, the highest exposure to GBH residues in pollen corresponded to 0.312% and 0.187% of the ADI and of the ARfD respectively and, to 0.002% and to 0.001% for beeswax. No transfer of glyphosate from wax to honey was detected. Considering our results and the available regulatory data on the glyphosate molecule considered solely, not including the adjuvants in GBH formulation, the consumption of these three contaminated matrices would not be a food safety issue. Nonetheless, caution should be taken in the interpretation of the results as new studies indicate possible glyphosate/GBH residues toxicity below regulatory limits and at chronic sub-lethal doses.

Investigating combined toxicity of binary mixtures in bees: Meta-analysis of laboratory tests, modelling, mechanistic basis and implications for risk assessment.

Bees are exposed to a wide range of multiple chemicals "chemical mixtures" from anthropogenic (e.g. plant protection products or veterinary products) or natural origin (e.g. mycotoxins, plant toxins). Quantifying the relative impact of multiple chemicals on bee health compared with other environmental stressors (e.g. varroa, viruses, and nutrition) has been identified as a priority to support the development of holistic risk assessment methods. Here, extensive literature searches and data collection of available laboratory studies on combined toxicity data for binary mixtures of pesticides and non-chemical stressors has been performed for honey bees (Apis mellifera), wild bees (Bombus spp.) and solitary bee species (Osmia spp.). From 957 screened publications, 14 publications provided 218 binary mixture toxicity data mostly for acute mortality (lethal dose: LD50) after contact exposure (61%), with fewer studies reporting chronic oral toxicity (20%) and acute oral LC50 values (19%). From the data collection, available dose response data for 92 binary mixtures were modelled using a Toxic Unit (TU) approach and the MIXTOX modelling tool to test assumptions of combined toxicity i.e. concentration addition (CA), and interactions (i.e. synergism, antagonism). The magnitude of interactions was quantified as the Model Deviation Ratio (MDR). The CA model applied to 17% of cases while synergism and antagonism were observed for 72% (MDR > 1.25) and 11% (MDR < 0.83) respectively. Most synergistic effects (55%) were observed as interactions between sterol-biosynthesis-inhibiting (SBI) fungicides and insecticide/acaricide. The mechanisms behind such synergistic effects of binary mixtures in bees are known to involve direct cytochrome P450 (CYP) inhibition, resulting in an increase in internal dose and toxicity of the binary mixture. Moreover, bees are known to have the lowest number of CYP copies and other detoxification enzymes in the insect kingdom. In the light of these findings, occurrence of these binary mixtures in relevant crops (frequency and concentrations) would need to be investigated. Addressing this exposure dimension remains critical to characterise the likelihood and plausibility of such interactions to occur under field realistic conditions. Finally, data gaps and further work for the development of risk assessment methods to assess multiple stressors in bees including chemicals and non-chemical stressors in bees are discussed.

Combined nutritional stress and a new systemic pesticide (flupyradifurone, Sivanto®) reduce bee survival, food consumption, flight success, and thermoregulation.

Flupyradifurone (FPF, Sivanto®) is a new butenolide insecticide that, like the neonicotinoids, is a systemic nicotinic acetylcholine receptor (nAChR) agonist. However, FPF is considered bee-safe (according to standard Risk Assessment tests), and is thus a potential solution to the adverse effects of other pesticides on beneficial insects. To date, no studies have examined the impact of nutritional stress (decreased food diversity and quality) and FPF exposure on bee health although both stressors can occur, especially around agricultural monocultures. We therefore tested the effects of a field-realistic FPF concentration (4 ppm, FPFdaily dose = 241 ±â€¯4 ng/bee/day, 1/12 of LD50) and nutritional stress (nectar with low-sugar concentrations) on honey bee (Apis mellifera L.) mortality, food consumption, thermoregulation, flight success (unsuccessful vs. successful), and flight ability (duration, distance, velocity). Flight and thermoregulation are critical to colony health: bees fly to collect food and reproduce, and they thermoregulate to increase flight efficiency and to rear brood. We studied the effects across seasons because seasonality can influence bee sensitivity to environmental stress. We demonstrate that, depending upon season and nutritional stress, FPF can reduce bee survival (-14%), food consumption (-14%), thermoregulation (-4%, i.e. hypothermia), flight success (-19%), and increase flight velocity (+13%). Because pesticide exposure and nutritional stress can co-occur, we suggest that future studies and pesticide risk assessments consider both seasonality and nutritional stress when evaluating pesticide safety for bees.

A 3-year survey of Italian honey bee-collected pollen reveals widespread contamination by agricultural pesticides.

Honey bee (Apis mellifera L.) health is compromised by complex interactions between multiple stressors, among which pesticides play a major role. To better understand the extent of honey bee colonies' exposure to pesticides in time and space, we conducted a survey by collecting corbicular pollen from returning honey bee foragers in 53 Italian apiaries during the active beekeeping season of 3 subsequent years (2012-2014). Of 554 pollen samples analysed for pesticide residues, 62% contained at least one pesticide. The overall rate of multiresidual samples (38%) was higher than the rate of single pesticide samples (24%), reaching a maximum of 7 pesticides per sample (1%). Over 3years, 18 different pesticides were detected (10 fungicides and 8 insecticides) out of 66 analysed. Pesticide concentrations reached the level of concern for bee health (Hazard Quotient (HQ) higher than 1000) at least once in 13% of the apiaries and exceeded the thresholds of safety for human dietary intake (Acute Reference Dose (ARfD), the Acceptable Daily Intake (ADI), and the Maximum Residue Limit (MRL)) in 39% of the analysis. The pesticide which was most frequently detected was the insecticide chlorpyrifos (30% of the samples overall, exceeding ARfD, ADI, or MRL in 99% of the positive ones), followed by the fungicides mandipropamid (19%), metalaxyl (16%), spiroxamine (15%), and the neonicotinoid insecticide imidacloprid (12%). Imidacloprid had also the highest HQ level (5054, with 12% of its positive samples with HQ higher than 1000). This 3year survey provides further insights on the contamination caused by agricultural pesticide use on honey bee colonies. Bee-collected pollen is shown to be a valuable tool for environmental monitoring, and for the detection of illegal uses of pesticides.

Lethal effects of Cr(III) alone and in combination with propiconazole and clothianidin in honey bees.

Several anthropogenic contaminants, including pesticides and heavy metals, can affect honey bee health. The effects of mixtures of heavy metals and pesticides are rarely studied in bees, even though bees are likely to be exposed to these contaminants in both agricultural and urban environments. In this study, the lethal toxicity of Cr alone and in combination with the neonicotinoid insecticide clothianidin and the ergosterol-biosynthesis-inhibiting fungicide propiconazole was assessed in Apis mellifera adults. The LD50 and lowest benchmark dose of Cr as Cr(NO3)3, revealed a low acute oral toxicity on honey bee foragers (2049 and 379 mg L-1, respectively) and the Cr retention (i.e. bee ability to retain the heavy metal in the body) was generally low compared to other metals. A modified method based on the binomial proportion test was developed to analyse synergistic and antagonistic interactions between the three tested contaminants. The combination of an ecologically-relevant field concentration of chromium with clothianidin and propiconazole did not increase bee mortality. On the contrary, the presence of Cr in mixture with propiconazole elicited a slight antagonistic effect.

Neonicotinoid pesticides and nutritional stress synergistically reduce survival in honey bees.

The honey bee is a major pollinator whose health is of global concern. Declines in bee health are related to multiple factors, including resource quality and pesticide contamination. Intensive agricultural areas with crop monocultures potentially reduce the quality and quantity of available nutrients and expose bee foragers to pesticides. However, there is, to date, no evidence for synergistic effects between pesticides and nutritional stress in animals. The neonicotinoids clothianidin (CLO) and thiamethoxam (TMX) are common systemic pesticides that are used worldwide and found in nectar and pollen. We therefore tested if nutritional stress (limited access to nectar and access to nectar with low-sugar concentrations) and sublethal, field-realistic acute exposures to two neonicotinoids (CLO and TMX at 1/5 and 1/25 of LD50) could alter bee survival, food consumption and haemolymph sugar levels. Bee survival was synergistically reduced by the combination of poor nutrition and pesticide exposure (-50%). Nutritional and pesticide stressors reduced also food consumption (-48%) and haemolymph levels of glucose (-60%) and trehalose (-27%). Our results provide the first demonstration that field-realistic nutritional stress and pesticide exposure can synergistically interact and cause significant harm to animal survival. These findings have implications for current pesticide risk assessment and pollinator protection.

A common neonicotinoid pesticide, thiamethoxam, impairs honey bee flight ability.

Pesticides can pose environmental risks, and a common neonicotinoid pesticide, thiamethoxam, decreases homing success in honey bees. Neonicotinoids can alter bee navigation, but we present the first evidence that neonicotinoid exposure alone can impair the physical ability of bees to fly. We tested the effects of acute or chronic exposure to thiamethoxam on the flight ability of foragers in flight mills. Within 1 h of consuming a single sublethal dose (1.34 ng/bee), foragers showed excitation and significantly increased flight duration (+78%) and distance (+72%). Chronic exposure significantly decreased flight duration (-54%), distance (-56%), and average velocity (-7%) after either one or two days of continuous exposure that resulted in bees ingesting field-relevant thiamethoxam doses of 1.96-2.90 ng/bee/day. These results provide the first demonstration that acute or chronic exposure to a neonicotinoid alone can significantly alter bee flight. Such exposure may impair foraging and homing, which are vital to normal colony function and ecosystem services.