Guidance for the assessment of risks to bees from the use of plant protection products under the framework of Council Directive 91/414 and Regulation 1107/2009

About 15 years have passed since the implementation of Directive 91/414/EEC and the regulatory framework with regard to pesticide use and environmental protection aims, as well as the plant protection techniques involving pesticides, have evolved over this period. In the case of honey bees, further developments and updates with regards to larval testing have been undertaken. Experimental protocols for semi‐field and field testing have been reviewed in detail. Finally, a dedicated risk assessment scheme has been developed for seed coating and soil applications that involve systemic compounds. A revision of the original EPPO Standards: the Risk Assessment Scheme PP 3/10(2) Honeybees and the test guidelines PP1/170 (3) Side‐effects on honeybees has thus been undertaken in order to benefit from these improvements. This paper aims to provide the details of the reasoning on which the revised risk assessment scheme relies as well as providing the necessary references to the data which have been reviewed in the context of this revision.     

Risk assessment to honey bees: a scheme developed in France for non-sprayed systemic compounds

BACKGROUND: Directive 91/414/EEC envisages that the systemic properties of active substances, if any, are taken into account in evaluating the risk posed to the environment by plant protection products. Among others, honey bees may be exposed to substances via this route, which may pose problems when substances with high toxicity are ingested through pollen or nectar. The guidance documents in support of the risk assessment to bees within the framework of Directive 91/414/EEC do not provide detailed technical guidance on how to proceed in a risk assessment for substances with systemic properties. RESULTS: A stepwise approach aiming specifically to assess the risk posed by non-sprayed systemic substances to bees is therefore proposed. This approach first identifies substances with systemic properties, which should be quantified in plant material as pollen and nectar. Exposure estimates calculated for different categories of bees (e.g. foraging bees), based on expected concentrations of the product in pollen or nectar, may be compared with several toxicity endpoints for acute or chronic effects on adults and/or larvae with a toxicity/exposure ratio, which is a measurement of potential risks. CONCLUSION: Such a ratio is proposed to be used as a trigger for any further refined assessment that would focus on the measurement of effects at the colony level.     

中国主要蜜源作物上登记的农药品种及其中杀虫剂对蜜蜂的初级风险评估

对目前中国主要蜜源作物上登记的农药品种进行了梳理,并采用现有的风险评估标准方法,对其中毒死蜱、吡虫啉等共61种杀虫剂对蜜蜂的风险进行了初级评估。结果表明:在58种喷雾施用的杀虫剂中,35种对蜜蜂的风险商值均大于1,风险为不可接受;其余23种的风险商值小于1,风险为可接受;所评估的6种土壤或种子处理内吸性杀虫剂中,5种对蜜蜂的风险商值大于1,风险为不可接受,仅氯虫苯甲酰胺的风险商值小于1,风险为可接受。但由于文中是以药剂在所登记作物上的单次最高施药剂量为暴露量进行的初级评估,并未考虑农药在花粉、花蜜中的降解及降雨引起的淋洗损耗,以及施药时间与作物花期之间的关系等影响因素,因而使得评估结果具有较大的保守性。研究结果一方面可为这些农药的合理使用和管理提供参考,另一方面提示了目前中国关于农药对蜜蜂的初级风险评估程序需进一步优化。     

The relevance of sublethal effects in honey bee testing for pesticide risk assessment

The option of an evaluation and assessment of possible sublethal effects of pesticides on bees has been a subject of discussion by scientists and regulatory authorities. Effects considered included learning behaviour and orientation capacity. This discussion was enhanced by the French bee issue and allegations against systemic insecticides that were linked to the hypothesis that sublethal intoxication might even have led to reported colony losses. This paper considers whether and, if so, how sublethal effects should be incorporated into risk assessment, by addressing a number of questions: What is meant by a sublethal effect? Which sublethal effects should be measured, when and how? How are sublethal effects to be included in risk assessments? The authors conclude that sublethal studies may be helpful as an optional test to address particular, compound‐specific concerns, as a lower‐tier alternative to semi‐field or field testing, if the effects are shown to be ecologically relevant. However, available higher‐tier data (semi‐field, field tests) should make any additional sublethal testing unnecessary, and higher‐tier data should always override data of lower‐tier trials on sublethal effects. © Crown copyright 2007. Reproduced with the permission of Her Majesty's Stationery Office. Published by John Wiley & Sons, Ltd.     

农药对蜜蜂的风险评价技术进展

在对美国国家环保局关于农药对蜜蜂的风险评价程序及工作进行调查研究的基础上,详细介绍了该评价体系所涵盖的不同层次,包括蜜蜂的室内急性毒性研究、叶面残留农药对蜜蜂的毒性影响的半田间研究及田间风险评价技术研究方面的进展,旨在为我国建立农药对蜜蜂的风险评价技术体系提供一定的借鉴作用。     

Toxicity of diafenthiuron to honey bees in laboratory,semi‐field and field conditions

BACKGROUND: Cardamom, an important spice crop often attacked by many insect pests, is controlled mainly using synthetic insecticides. As honey bees play a vital role in pollination in cardamom, the impact of insecticides on honey bees needs to be explored to assess its safety. RESULTS: Risk assessment based on contact toxicity revealed diafenthiuron to be a non‐selective insecticide to bees with a low selectivity ratio (the ratio between the LD₅₀ for beneficial and pest species). A dose of diafenthiuron that killed 90% of cardamom borer, Conogethes punctiferalis Guenee, was found to kill 100% of Indian bees. Based on the hazard ratio (the ratio between the field‐recommended dose and the LD₅₀ for the beneficial), diafenthiuron was found to be slightly to moderately toxic to bees. Diafenthiuron, even at low concentrations of LC₁ (the concentration that killed 1% of bees), was found to affect the foraging and homing behaviour of Indian bees. Of bees fed with 30 µg mL^?1 of diafenthiuron, 40% were found missing on the third day after exposure. However, diafenthiuron did not affect bee visits to the cardamom fields. CONCLUSION: Diafenthiuron is more highly toxic to Apis cerana indica F. than to C. punctiferalis by contact, using selectivity ratio and probit substitution methods of risk assessment, but the hazard ratio revealed diafenthiuron to be a slightly to moderately toxic chemical. Diafenthiuron was found to affect the foraging and homing behaviour of bees at sublethal concentrations. Thus, sublethal effects are more relevant in risk assessment than lethal and acute effects. Copyright © 2010 Society of Chemical Industry     

An exposure study to assess the potential impact of fipronil in treated sunflower seeds on honey bee colony losses in Spain

BACKGROUND: There is great concern about the high losses and strong depopulation of honey bee colonies in some areas of Spain. Some beekeepers have suggested that sunflower seeds treated with the insecticide fipronil could be an important factor in causing those losses. Therefore, an in‐depth field study has been carried out in two regions of Spain where sunflower production is intense (Cuenca and Andalucía) and where, for some crops and varieties, fipronil has been used as seed insecticide. RESULTS: Samples of adult bees and pollen were analysed for bee pathogens and pesticide residues respectively. Neither fipronil residues nor its metabolites were detected in any of the samples analysed, indicating that short‐term or chronic exposure of bees to fipronil and/or its metabolites can be ruled out in the apiaries surveyed. Varroa destructor and Nosema ceranae were found to be very prevalent. CONCLUSION: The combination of the two pathogens could augment the risk of colony death in infected colonies, without fipronil residues exerting a significant effect in the given field conditions. Indeed, in this study the losses observed in apiaries located close to sunflower crops were similar to those in apiaries situated in forested areas with wild vegetation. Copyright © 2011 Society of Chemical Industry     

Is Apis mellifera more sensitive to insecticides than other insects?

BACKGROUND: Honey bees (Apis mellifera L.) are among the most important pollinators in natural and agricultural settings. They commonly encounter insecticides, and the effects of insecticides on honey bees have been frequently noted. It has been suggested that honey bees may be (as a species) uniquely sensitive to insecticides, although no comparative toxicology study has been undertaken to examine this claim. An extensive literature review was conducted, using data in which adult insects were topically treated with insecticides. The goal of this review was to summarize insecticide toxicity data between A. mellifera and other insects to determine the relative sensitivity of honey bees to insecticides. RESULTS: It was found that, in general, honey bees were no more sensitive than other insect species across the 62 insecticides examined. In addition, honey bees were not more sensitive to any of the six classes of insecticides (carbamates, nicotinoids, organochlorines, organophosphates, pyrethroids and miscellaneous) examined. CONCLUSIONS: While honey bees can be sensitive to individual insecticides, they are not a highly sensitive species to insecticides overall, or even to specific classes of insecticides. However, all pesticides should be used in a way that minimizes honey bee exposure, so as to minimize possible declines in the number of bees and/or honey contamination. Copyright © 2010 Society of Chemical Industry     

Presence of Brome mosaic virus in Barley Guttation Fluid and Its Association with Localized Cell Death Response

ABSTRACT Water exits from inside the leaf through transpiration or guttation. Under conditions to promote guttation, surface fluid (guttation fluid) from Brome mosaic virus (BMV)-infected barley, wheat, and maize plants was analyzed for the presence of the virus by biological and serological assays. We also investigated the route by which BMV exited infected cells to the intercellular space of the barley leaf. BMV was detected in guttation fluid from systemically infected barley leaves when the initial viral symptoms were observed on these leaves. The virus was also detected in guttation fluid from systemically infected wheat leaves, but not in maize leaves showing either systemic necrosis or chlorotic streaks. Interestingly, in BMV-infected barley leaves, but not in maize leaves showing chlorotic streaks, cell death occurred within and adjacent to veins. Staining of xylem and phloem networks in infected barley leaves with fluorescent dyes showed that xylem, and to a lesser extent phloem, were severely damaged and thus became leaky for dye transport. No such damage was observed in BMV-infected maize leaves showing chlorotic streaks. We propose that in infected barley leaves, BMV exits from damaged vein cells (especially the xylem elements), accumulates in intercellular spaces, and then reaches the surface of the leaves through stomata during guttation or transpiration. In nature, BMV may be carried to adjacent plants and cause infection by movement of vertebrate and invertebrate vectors among infected plants exuding guttation fluid.     

The significance of guttation in the secondary spread of Clavibacter michiganensis subsp. michiganensis in tomato greenhouses

Bacterial canker, caused by Clavibacter michiganensis subsp. michiganensis (Cmm), can spread in commercial tomato greenhouses causing epidemics. Results of greenhouse experiments with Cmm‐contaminated tools demonstrated disease spread for only a limited distance (<4 plants) from infected plants. However, touching symptomless infected plants bearing guttation droplets prior to touching nearby plants spread the pathogen over longer distances within rows (>22 plants). The pathogen was exuded in large numbers in the guttation fluid of infected plants; its presence in the guttation fluid was not influenced by the inoculation procedures, leaf age or the volume of the guttation droplets. Population size of Cmm and the incidence of leaflets with epiphytic bacteria were significantly higher in plants placed in a guttation‐induction chamber than in those kept in a growth chamber with high humidity, suggesting exudation through guttation contributed to the formation of epiphytic populations on leaflets. This new knowledge may provide a simple and environmentally friendly means for decreasing the spread of the disease by avoiding contact with plants during periods when they bear guttation droplets.     

Wind tunnel studies on spray deposition on leaves of tree species used for windbreaks and exposure of honey bees

A wind tunnel study was conducted to determine pesticide deposition on commonly used windbreak tree species used as spray drift barriers and associated exposure of honey bees. Although it has been known that windbreaks are effective in reducing chemical drift from agricultural applications, there is still an enormous information and data gap on details of the dependence of the mechanism on the biological materials of the barriers and on standardization of relevant assessment methods. Beneficial arthropods like honey bees are adversely affected by airborne drift of pesticides. A study was initiated by first establishing a wind tunnel to create a controlled environment for capture efficiency work. Suitable spray parameters were determined after a preliminary study to construct and develop a wind tunnel protocol. A tracer dye solution was sprayed onto the windbreak samples and honey bees located in the wind tunnel at various simulated wind speeds. Analysis of data from this work has shown that needle-like foliage of windbreak trees captures two to four times more spray than broad-leaves. In addition, it was determined that, at lower wind speeds, flying bees tend to capture slightly more spray than bees at rest.     

亚致死浓度噻虫嗪对意大利蜜蜂工蜂的生存风险分析

噻虫嗪是一类较第一代新烟碱类杀虫剂更广谱、高效、低毒的农药,但它对意大利蜜蜂工蜂的生存风险尚不明确。本研究通过糖水混毒法测定了噻虫嗪对意大利蜜蜂新出房蜂(1 d)、哺育蜂(7 d)和采集蜂(22 d)的48 h急性经口毒性,确定其各自的亚致死浓度LC5、LC15和LC25,并分别研究了其在3个亚致死浓度下的生存风险。结果表明,新出房蜂、哺育蜂和采集蜂48 h后LC50分别为248.13、64.18和44.20ng/g。新出房蜂经LC5和LC15处理后其平均存活寿命与对照均无显著差异,经LC25处理后其平均存活寿命与对照缩短19.0 d。哺育蜂经LC5处理后其平均存活寿命与对照均无显著差异,经LC15和LC25处理后其平均存活寿命与对照分别缩短9.2和15.0 d。采集蜂经LC5和LC15处理后其平均存活寿命与对照均无显著差异,经LC25处理后其平均存活寿命与对照缩短10.9 d。上述结果表明噻虫嗪的使用可能会对蜜蜂的生存存在较大风险,因此,建议相关管理部门加强噻虫嗪在中国的田间残留监测,开展半田间和田间条件下噻虫嗪对意大利蜜蜂的风险评价工作。     

Pesticidal natural products – status and future potential

There is a long history of using natural products as the basis for creating new pesticides but there is still a relatively low percentage of naturally derived pesticides relative to the number of pharmaceuticals derived from natural sources. Biopesticides as defined and regulated by the US Environmental Protection Agency (EPA) have been around for 70 years, starting with Bacillus thuringiensis, but they are experiencing rapid growth as the products have got better and more science‐based, and there are more restrictions on synthetic chemical pesticides. As such, biopesticides are still a small percentage (approximately US$3–4 billion) of the US$61.3 billion pesticide market. The growth of biopesticides is projected to outpace that of chemical pesticides, with compounded annual growth rates of between 10% and 20%. When integrated into crop production and pest management programs, biopesticides offer the potential for higher crop yields and quality than chemical‐only programs. Added benefits include reduction or elimination of chemical residues, therefore easing export, enabling delay in the development of resistance by pests and pathogens to chemicals and shorter field re‐entry, biodegradability and production using agricultural raw materials versus fossil fuels, and low risk to non‐target organisms, including pollinators. Challenges to the adoption of biopesticides include lack of awareness and education in how to deploy their unique modes of action in integrated programs, testing products alone versus in integrated programs, and lingering perceptions of cost and efficacy. © 2019 Society of Chemical Industry     

Lethal and sub-lethal effects of spinosad on bumble bees (Bombus impatiens Cresson)

Recent developments of new families of pesticides and growing awareness of the importance of wild pollinators for crop pollination have stimulated interest in potential effects of novel pesticides on wild bees. Yet pesticide toxicity studies on wild bees remain rare, and few studies have included long-term monitoring of bumble bee colonies or testing of foraging ability after pesticide exposure. Larval bees feeding on exogenous pollen and exposed to pesticides during development may result in lethal or sub-lethal effects during the adult stage. We tested the effects of a naturally derived biopesticide, spinosad, on bumble bee (Bombus impatiens Cresson) colony health, including adult mortality, brood development, weights of emerging bees and foraging efficiency of adults that underwent larval development during exposure to spinosad. We monitored colonies from an early stage, over a 10-week period, and fed spinosad to colonies in pollen at four levels: control, 0.2, 0.8 and 8.0 mg kg(-1), during weeks 2 through 5 of the experiment. At concentrations that bees would likely encounter in pollen in the wild (0.2-0.8 mg kg(-1)) we detected minimal negative effects to bumble bee colonies. Brood and adult mortality was high at 8.0 mg kg(-1) spinosad, about twice the level that bees would be exposed to in a 'worst case' field scenario, resulting in colony death two to four weeks after initial pesticide exposure. At more realistic concentrations there were potentially important sub-lethal effects. Adult worker bees exposed to spinosad during larval development at 0.8 mg kg(-1) were slower foragers on artificial complex flower arrays than bees from low or no spinosad treated colonies. Inclusion of similar sub-lethal assays to detect effects of pesticides on pollinators would aid in development of environmentally responsible pest management strategies.     

The effect of a piperonyl butoxide/tau-fluvalinate mixture on pollen beetle (Meligethes aeneus) and honey bees (Apis mellifera)

BACKGROUND: Previous work has characterised pyrethroid resistance in pollen beetle (Meligethes aeneus F.) as principally an oxidative mechanism. Piperonyl butoxide (PBO) can synergise this resistance in the field, but its effects on the honey bee are thought to be unacceptable. RESULTS: A field trial in Poland was conducted to show that a mixture of PBO and tau‐fluvalinate at the registered rate gave increased and longer‐lasting control of resistant pollen beetle. Four days after spraying with tau‐fluvalinate, only 20% of pollen beetles were controlled, compared with 70% if the tau‐fluvalinate/PBO mixture was used. No detriment to honey bee health was observed using the same mixture. CONCLUSIONS: PBO, if used in conjunction with a pyrethroid of relatively low bee toxicity, can successfully overcome pyrethroid resistance in pollen beetle without incurring an increased loss of honey bees, even if they are present at the time of spraying. Copyright © 2012 Society of Chemical Industry     

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. Copyright © 2012 Society of Chemical Industry     

农药对蜜蜂的影响及毒理学机制研究进展

农药在现代农业生产中发挥着积极作用,但是农药使用不当则会对重要授粉昆虫如蜜蜂等造成负面影响。本文主要围绕农药对蜜蜂的急性毒性、亚致死剂量农药对蜜蜂生长发育和行为的影响,农药对蜜蜂的联合作用,以及新烟碱类杀虫剂与蜜蜂烟碱型乙酰胆碱受体的相互作用、农药对蜜蜂毒性差异的机理、农药对蜜蜂解毒代谢相关酶活性及其他生理生化指标的影响、植物化学物质在调控蜜蜂对农药耐受性中的作用等方面的研究进展进行了综述,以期为农药的合理使用及提高其对蜜蜂的安全性提供参考。     

Pesticides and reduced‐risk insecticides,native bees and pantropical stingless bees: pitfalls and perspectives

Although invertebrates generally have a low public profile, the honey bee, Apis mellifera L., is a flagship species whose popularity likely derives from the products it provides and its perceived ecological services. Therefore, the raging debate regarding honey bee decline has surpassed the realm of beekeepers, academia, industry and regulatory agencies and now also encompasses non‐governmental agencies, media, fiction writers and the general public. The early interest and concern about honey bee colony collapse disorder (CCD) soon shifted to the bigger issue of pollinator decline, with a focus on the potential involvement of pesticides in such a phenomenon. Pesticides were previously recognised as the potential culprits of the reported declines, particularly the neonicotinoid insecticides owing to their widespread and peculiar use in agriculture. However, the evidence for the potential pivotal role of these neonicotinoids in honey bee decline remains a matter of debate, with an increased recognition of the multifactorial nature of the problem and the lack of a direct association between the noted decline and neonicotinoid use. The focus on the decline of honey bee populations subsequently spread to other species, and bumblebees became another matter of concern, particularly in Europe and the United States. Other bee species, ones that are particularly important in other regions of the world, remain the object of little concern (unjustifiably so). Furthermore, the continuous focus on neonicotinoids is also in need of revision, as the current evidence suggests that a broad spectrum of compounds deserve attention. Here we address both shortcomings. © 2015 Society of Chemical Industry     

福建省主产区普通白菜中农药残留水平及膳食暴露风险评估

为评估福建省主产区普通白菜中农药残留水平及对人体的膳食暴露风险,采用现有的标准检测方法,对在福建省主产区采集的88个普通白菜样品中的68种农药残留进行了检测分析,并对检出农药进行了短期和长期膳食暴露风险评估。结果显示：88个普通白菜样品中,共有68个样品检出了农药残留,总检出率为77.27%;共检出27种农药,其中杀虫剂18种,杀菌剂9种;检出2种及2种以上农药残留的样品占检出农药残留样品总量的83.82%,存在农药多残留现象。利用相关农药的毒理学数据——急性参考剂量(ARfD)和每日允许摄入量(ADI)、农药残留数据和普通白菜的膳食消费量数据,评估了所检出农药的短期和长期膳食暴露风险。其中,短期膳食暴露风险值(%ARfD)范围为0～90%,长期膳食暴露风险商贡献率(RQc%)范围为0～46%。研究表明,福建省主产区普通白菜中的农药残留水平较低,通过食用该地区生产的普通白菜对消费者的农药残留膳食暴露风险整体在可接受范围内。     

从经食、饮水和吸入途径评估8种经济林常用农药对鸟类的暴露风险

采用《农药登记环境风险评估指南第3部分:鸟类》中的暴露分析模型及美国环保署的SIP模型和STIR模型,分别从经食、饮水和吸入3种途径对残杀威、虫螨腈、敌敌畏、敌百虫、马拉硫磷、杀螟硫磷、茚虫威和甲氨基阿维菌素苯甲酸盐8种农药对鸟类的暴露风险进行了评估.农药的登记作物信息及其用药数据来源于"中国农药信息网",鸟类的毒理学...