clothianidin and imidacloprid

A comprehensive review of published and previously unpublished studies was performed to evaluate the neonicotinoid insecticides for evidence of developmental neurotoxicity (DNT). These insecticides have favorable safety profiles, due to their preferential affinity for nicotinic receptor (nAChR) subtypes in insects, poor penetration of the mammalian blood-brain barrier, and low application rates. Nevertheless, examination of this issue is warranted, due to their insecticidal mode of action and potential exposure with agricultural and residential uses. This review identified in vitro, in vivo, and epidemiology studies in the literature and studies performed in rats in accordance with GLP standards and EPA guidelines with imidacloprid, acetamiprid, thiacloprid, clothianidin, thiamethoxam, and dinotefuran, which are all the neonicotinoids currently registered in major markets. For the guideline-based studies, treatment was administered via the diet or gavage to primiparous female rats at three dose levels, plus a vehicle control (≥20/dose level), from gestation day 0 or 6 to lactation day 21. F1 males and females were evaluated using measures of motor activity, acoustic startle response, cognition, brain morphometry, and neuropathology. The principal effects in F1 animals were associated with decreased body weight (delayed sexual maturation, decreased brain weight, and morphometric measurements) and acute toxicity (decreased activity during exposure) at high doses, without neuropathology or impaired cognition. No common effects were identified among the neonicotinoids that were consistent with DNT or the neurodevelopmental effects associated with nicotine. Findings at high doses were associated with evidence of systemic toxicity, which indicates that these insecticides do not selectively affect the developing nervous system.

Topics: Animals; Central Nervous System; Disease Models, Animal; Dose-Response Relationship, Drug; Guanidine; Guanidines; Humans; Imidazoles; Insecta; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Pyridines; Randomized Controlled Trials as Topic; Receptors, Nicotinic; Reflex, Startle; Risk Assessment; Thiamethoxam; Thiazines; Thiazoles; United States; United States Environmental Protection Agency

The nitro-substituted neonicotinoid insecticides, which include imidacloprid, thiamethoxam and clothianidin, are widely used to control a range of important agricultural pests both by foliar applications and also as seed dressings and by soil application. Since they exhibit systemic properties, exposure of bees may occur as a result of residues present in the nectar and/or pollen of seed- or soil-treated crop plants and so they have been the subject of much debate about whether they cause adverse effects in pollinating insects under field conditions. Due to these perceived concerns, the use of the three neonicotinoids imidacloprid, clothianidin and thiamethoxam has been temporarily suspended in the European Union for seed treatment, soil application and foliar treatment in crops attractive to bees. Monitoring data from a number of countries are available to assess the presence of neonicotinoid residues in honey bee samples and possible impacts at the colony level and these are reviewed here together with a number of field studies which have looked at the impact of clothiandin on honey bees in relation to specific crop use and in particular with oilseed rape. Currently there is considerable uncertainty with regards to the regulatory testing requirements for field studies. Accordingly, a testing protocol was developed to address any acute and chronic risks from oilseed rape seeds containing a coating with 10 g clothianidin and 2 g beta-cyfluthrin per kg seeds (Elado

Topics: Animals; Bees; Crops, Agricultural; Environmental Monitoring; Environmental Pollutants; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Plant Nectar; Pollen; Pollination; Thiamethoxam; Thiazoles

The neonicotinoid insecticides imidacloprid, acetamiprid, dinotefuran, thiamethoxam and clothianidin are commonly used in greenhouses and/or interiorscapes (plant interiorscapes and conservatories) to manage a wide range of plant-feeding insects such as aphids, mealybugs and whiteflies. However, these systemic insecticides may also be harmful to natural enemies, including predators and parasitoids. Predatory insects and mites may be adversely affected by neonicotinoid systemic insecticides when they: (1) feed on pollen, nectar or plant tissue contaminated with the active ingredient; (2) consume the active ingredient of neonicotinoid insecticides while ingesting plant fluids; (3) feed on hosts (prey) that have consumed leaves contaminated with the active ingredient. Parasitoids may be affected negatively by neonicotinoid insecticides because foliar, drench or granular applications may decrease host population levels so that there are not enough hosts to attack and thus sustain parasitoid populations. Furthermore, host quality may be unacceptable for egg laying by parasitoid females. In addition, female parasitoids that host feed may inadvertently ingest a lethal concentration of the active ingredient or a sublethal dose that inhibits foraging or egg laying. There are, however, issues that require further consideration, such as: the types of plant and flower that accumulate active ingredients, and the concentrations in which they are accumulated; the influence of flower age on the level of exposure of natural enemies to the active ingredient; the effect of neonicotinoid metabolites produced within the plant. As such, the application of neonicotinoid insecticides in conjunction with natural enemies in protected culture and interiorscape environments needs further investigation.

Topics: Animals; Environmental Pollutants; Guanidines; Imidazoles; Insect Control; Insecta; Insecticides; Mites; Neonicotinoids; Nitro Compounds; Oxazines; Predatory Behavior; Pyridines; Thiamethoxam; Thiazoles

Neonicotinoids have been detected in many species of wild birds; however, few studies have quantified population-level exposure. We examined population-level exposure to 7 neonicotinoids in 2 species that use agricultural areas, sharp-tailed grouse (Tympanuchus phasianellus) and greater prairie-chickens (T. cupido). We sampled fecal pellets at leks in spring and collected livers from hunter-harvested birds in fall along an agricultural gradient throughout their respective ranges in Minnesota, USA. Most sharp-tailed grouse (93 %) and prairie-chicken (80 %) fecal pellets and livers (90 % and 76 %, respectively) had detectable concentrations of ≥1 neonicotinoid, with imidacloprid (IMI) and clothianidin (CLO) most commonly detected. Spring detections of IMI in both species increased with the proportion of a 2-km buffer in cultivation surrounding sampling locations and varied by year. A similar relationship with cultivation was not supported for CLO, which may reflect differences in the availability of seed types treated with IMI and CLO on the soil surface after planting. However, we also detected IMI and CLO from birds sampled in areas of low cultivation. Sharp-tailed grouse and prairie-chickens may select crop fields preferentially to forage, and thus have a higher risk of exposure than would be expected based only on the amount of cultivation. Year was important in models of IMI and CLO in both species and seasons, which likely reflects differences in planting and in the availability of natural foods among years. In contrast, the proportion of surrounding area in cultivation was not supported in models of fall neonicotinoid detections. Fewer crops are planted in the fall in Minnesota and grouse may be exposed through routes other than treated seeds. High detections, even in areas with little cultivation and during seasons with little planting, likely reflect prairie grouse selection of cultivated fields for food, but may also indicate that exposure risk extends beyond sites of application.

Topics: Animals; Chickens; Grassland; Neonicotinoids; Prevalence; Quail; Seasons

Neonicotinoid insecticides target insect nicotinic acetylcholine receptors (nAChRs) and their adverse effects on non-target insects are of serious concern. We recently found that cofactor TMX3 enables robust functional expression of insect nAChRs in Xenopus laevis oocytes and showed that neonicotinoids (imidacloprid, thiacloprid, and clothianidin) exhibited agonist actions on some nAChRs of the fruit fly (Drosophila melanogaster), honeybee (Apis mellifera) and bumblebee (Bombus terrestris) with more potent actions on the pollinator nAChRs. However, other subunits from the nAChR family remain to be explored. We show that the Dα3 subunit co-exists with Dα1, Dα2, Dβ1, and Dβ2 subunits in the same neurons of adult D. melanogaster, thereby expanding the possible nAChR subtypes in these cells alone from 4 to 12. The presence of Dα1 and Dα2 subunits reduced the affinity of imidacloprid, thiacloprid, and clothianidin for nAChRs expressed in Xenopus laevis oocytes, whereas the Dα3 subunit enhanced it. RNAi targeting Dα1, Dα2 or Dα3 in adults reduced expression of targeted subunits but commonly enhanced Dβ3 expression. Also, Dα1 RNAi enhanced Dα7 expression, Dα2 RNAi reduced Dα1, Dα6, and Dα7 expression and Dα3 RNAi reduced Dα1 expression while enhancing Dα2 expression, respectively. In most cases, RNAi treatment of either Dα1 or Dα2 reduced neonicotinoid toxicity in larvae, but Dα2 RNAi enhanced neonicotinoid sensitivity in adults reflecting the affinity-reducing effect of Dα2. Substituting each of Dα1, Dα2, and Dα3 subunits by Dα4 or Dβ3 subunit mostly increased neonicotinoid affinity and reduced efficacy. These results are important because they indicate that neonicotinoid actions involve the integrated activity of multiple nAChR subunit combinations and counsel caution in interpreting neonicotinoid actions simply in terms of toxicity.

Topics: Animals; Bees; Drosophila; Drosophila melanogaster; Insecta; Insecticides; Neonicotinoids; Receptors, Nicotinic

Topics: China; Female; Humans; Infant; Insecticides; Milk, Human; Neonicotinoids; Nitro Compounds; Thiamethoxam

Neonicotinoids (NEOs) and active pharmaceuticals ingredients (API) are contaminants widely diffused worldwide, causing increasing concern for potential adverse effects on wildlife. However, research on these contaminants have focused on target and non-target invertebrates, while information on potential effects in terrestrial mammals is lacking. We performed preliminary non-invasive monitoring of NEOs and API in a suburban and agricultural area using hair of the Red fox. The Red fox is a widely diffused mesopredator in Europe, and its plasticity in feeding habits makes it an excellent indicator for assessing exposure to environmental contamination. We observed the presence of NEOs in many Red fox hair samples (n = 11), including imidacloprid (IMI), acetamiprid (ACE), and clothianidin (CLO). The highest quantified concentrations were 6.4 ng g

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Antidepressive Agents; Foxes; Hair; Humans; Ibuprofen; Insecticides; Italy; Ketoprofen; Neonicotinoids; Pharmaceutical Preparations; Sertraline

Topics: 8-Hydroxy-2'-Deoxyguanosine; Animals; Antioxidants; Cyprinidae; Insecticides; Neonicotinoids; Nitro Compounds

As the most extensively used insecticides worldwide, neonicotinoid insecticides (NNIs) have received a growing global concern over their adverse health effects. This study aimed to assess the associations of urinary concentrations of NNIs in early pregnancy with gestational diabetes mellitus (GDM) and the mediation roles of oxidative DNA damage, RNA damage, and lipid peroxidation in the associations. With a prospective nested case-control study, 519 GDM cases and 519 controls were matched on the infant's sex and maternal age. Urinary biomarkers of NNIs exposure and oxidative stress were measured in early pregnancy. We estimated the associations of single and the mixture of NNIs and their metabolites with GDM by conditional logistic regression and quantile g-computation models, respectively. The mediating roles of oxidative stress were evaluated by the structural equation model. The odds of GDM significantly increased by 15 %, 18 %, 26 %, 42 %, 49 %, and 13 % in each unit increment of ln-transformed concentrations of urinary imidacloprid (IMI), imidacloprid-olefin (IMI-olefin), desnitro-imidacloprid (DN-IMI), thiamethoxam (THM), clothianidin, and desmethyl-clothianidin, respectively. Exposure to the mixture of NNIs was associated with increased odds of GDM (adjusted OR: 1.76; 95 %CI: 1.45, 2.13). Advanced maternal age enhanced the associations of 5-hydroxy-IMI, DN-IMI, and IMI-olefin with GDM (P < 0.05), and being overweight/obese before pregnancy strengthened the effects of IMI, IMI-olefin, and THM on GDM (P < 0.05). In the association of NNIs exposure and GDM, the proportions mediated by oxidative DNA damage, RNA damage, and overall oxidative stress were 9.8 %, 11.8 %, and 14.5 %, respectively (P < 0.05). Exposure to individual NNIs and a mixture of NNIs were associated with GDM, and maternal age and pre-pregnancy BMI may modify the association. The possible mechanism underlying the association between NNIs and GDM may involve oxidative damage to nucleic acids.

Topics: Alkenes; Case-Control Studies; Diabetes, Gestational; Drug-Related Side Effects and Adverse Reactions; Female; Humans; Infant; Insecticides; Mediation Analysis; Neonicotinoids; Oxidative Stress; Pregnancy; Prospective Studies; Thiamethoxam

The standard operating procedure for testing the susceptibility of adult mosquitoes to neonicotinoid or butenolide insecticides recommends using a vegetable oil ester (Mero) as a surfactant. However, there is growing evidence that this adjuvant contains surfactants that can enhance insecticide activity, mask resistance and bias the bioassay.. Using standard bioassays, we tested the effects of commercial formulations of vegetable oil-based surfactants similar to Mero on the activity of a spectrum of active ingredients including four neonicotinoids (acetamiprid, clothianidin, imidacloprid and thiamethoxam) and two pyrethroids (permethrin and deltamethrin). We found that three different brands of linseed oil soap used as cleaning products drastically enhanced neonicotinoid activity in Anopheles mosquitoes. At 1% (v/v), the surfactant reduced the median lethal concentration, LC50, of clothianidin more than 10-fold both in susceptible and in resistant populations of Anopheles gambiae. At 1% or 0.5% (v/v), linseed oil soap restored the susceptibility of adult mosquitoes fully to clothianidin, thiamethoxam and imidacloprid and partially to acetamiprid. By contrast, adding soap to the active ingredient did not significantly affect the level of resistance to permethrin or deltamethrin suggesting that vegetable oil-based surfactants specifically enhance the potency of some classes of insecticides.. Our findings indicate that surfactants are not inert ingredients, and their use in susceptibility testing may jeopardize the ability to detect resistance. Further research is needed to evaluate the potential, the limitations and the challenges of using some surfactants as adjuvants to enhance the potency of some chemicals applied in mosquito control.

Topics: Adjuvants, Immunologic; Animals; Culicidae; Insecticide Resistance; Insecticides; Linseed Oil; Mosquito Control; Mosquito Vectors; Neonicotinoids; Permethrin; Plant Oils; Pyrethrins; Soaps; Surface-Active Agents; Thiamethoxam

Recently, the high toxicity of neonicotinoids to the survival and reproduction of adult earthworms has become apparent in standard 56-day toxicity tests. The persistence of some neonicotinoids and/or their repeated application may lead to long-term exposure, possibly also affecting other parts of the life cycle of earthworms. The present study aimed at providing insight into the sublethal effects of imidacloprid, thiacloprid, and clothianidin on juvenile Eisenia andrei exposed for 16 weeks in Lufa 2.2 soil. Significant effects on growth and maturation were observed for all compounds. Exposure to 0.125 mg imidacloprid/kg dry soil and 0.03125 and 0.0625 mg thiacloprid/kg dry soil significantly affected the growth of the earthworms, while significant maturation effects were observed at 0.03125 mg/kg dry soil for imidacloprid and thiacloprid and 0.25 mg clothianidin/kg dry soil. The 16-week no-observed-effect concentrations (NOECs) found in the present study were lower than previously reported NOECs for effects on earthworm reproduction. Predicted environmental concentrations after a single application exceeded the observed NOECs for effects on earthworm maturation in the case of imidacloprid and thiacloprid and for effects on earthworm growth in the case of thiacloprid and clothianidin. Environ Toxicol Chem 2022;41:1686-1695. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Topics: Animals; Growth and Development; Guanidines; Insecticides; Neonicotinoids; Nitro Compounds; Oligochaeta; Soil; Soil Pollutants; Thiazines; Thiazoles

Neonicotinoids are widely used insecticides around the world and are preserved permanently in soils and appear in surface waters posing an increased threat to ecosystems. In the present study, we exposed adult specimens of amphipod Dikerogammarus villosus to environmentally relevant and higher concentrations of two widely used agricultural neonicotinoids, clothianidin (CLO) and imidacloprid (IMI), for 2 days. The acute effects were investigated at the behavioral (immobility time and swimming activity) and biochemical (glutathione S-transferase [GST] and acetylcholine esterase [AchE] activity) levels. All CLO concentrations used (64 nM, 128 nM, 192 nM) significantly decreased the immobility time and swimming activity. In the case of IMI, the immobility time decreased significantly only at the highest concentration applied (977 nM), but the distance travelled by the animals significantly decreased even at lower concentrations (78 nM and 313 nM). The GST enzyme activity did not change in the CLO-treated groups, however, the 626 nM and 977 nM IMI concentrations significantly increased the GST activity. Similarly, to the behavioral level, all CLO concentrations significantly decreased the AchE activity. In contrast, IMI has a significant stimulating effect on the AchE activity at the 313 nM, 626 nM, and 977 nM concentrations. Based on the authors' best knowledge, this is the first study to investigate the effects of CLO and IMI at environmentally-relevant concentrations on D. villosus. Our findings contribute to the understanding of the physiological effects of neonicotinoids.

Topics: Amphipoda; Animals; Ecosystem; Guanidines; Insecticides; Neonicotinoids; Nitro Compounds; Thiazoles

Neonicotinoid (NEO) pesticides have become a potential risk to ecological safety and human health after application. The combined use of biochar and organic fertilizer (OF) is a promising approach to reduce pesticide adverse effects and improve soil fertility in agricultural soils. However, the combined remediation effects of biochar and OF on immobilization and dissipation of NEOs in soils have not previously been systematically investigated. In this study, biochars derived from peanut shell prepared at low/high pyrolysis temperatures (PS400 and PS900) were combined with composted chicken manure (CCM) as an example for OF to remediate contaminated soils toward six typical NEOs, nitenpyram (NIT), thiamethoxam (THIA), clothianidin (CLO), imidacloprid (IMI), acetamiprid (ACE), thiacloprid (THI). Results shown that both biochars and CCM were effective in improving soil sorption capacity and immobilization efficiency. The Freundlich affinity parameters (K

Topics: Adsorption; Charcoal; Fertilizers; Guanidines; Humans; Manure; Neonicotinoids; Nitro Compounds; Nitrogen Dioxide; Pesticides; Soil; Soil Pollutants; Thiamethoxam; Thiazines; Thiazoles

Neonicotinoid insecticide use has increased over the last decade, including as agricultural seed treatments (application of chemical in a coating to the seed prior to planting). In California, multiple crops, including lettuce, can be grown using neonicotinoid treated seeds or receive a direct neonicotinoid soil application (drenching) at planting. Using research plots, this study compared pesticide runoff in four treatments: (1) imidacloprid seed treatment; (2) clothianidin seed treatment; (3) imidacloprid drench and an azoxystrobin seed treatment; and (4) a control with no pesticidal treatment. Neonicotinoid and azoxystrobin concentrations were measured in surface water runoff during six irrigations events in the 2020 growing seasons. Results showed runoff concentrations up to 1308 (±1200) ng L

Topics: Animals; Arthropods; Insecticides; Lactuca; Neonicotinoids; Nitro Compounds; Seeds; Soil

Neonicotinoids are systemic insecticides used since the 1990's , that possess renal tubular toxicity. We conducted a field-based descriptive study in the North Central Dry-zone of Sri Lanka, where chronic kidney disease (CKD) of unknown etiology has been increasing since the 1990's. To elucidate the relationship between renal tubular dysfunctions and urinary neonicotinoids concentrations, we collected spot urine samples from15 CKD patients, 15 family members, and 62 neighbors in 2015, analyzed two renal tubular biomarkers, Cystatin-C and L-FABP, quantified seven neonicotinoids and a metabolite N-desmethyl-acetamiprid by LC-MS/MS; and we investigated their symptoms using a questionnaire. Cystatin-C and L-FABP had a positive correlation (p < 0.001). N-Desmethyl-acetamiprid was detected in 92.4% of the urine samples, followed by dinotefuran (17.4%), thiamethoxam (17.4%), clothianidin (9.8%), thiacloprid and imidacloprid. Dinotefuran and thiacloprid have never been registered in Sri Lanka. In High Cystatin-C group (> 70 μg/gCre, n = 7), higher urinary concentration of dinotefuran (p = 0.009), and in Zero Cystatin-C group (< LOQ, n = 7), higher N-desmethyl-acetamiprid (p = 0.013), dinotefuran (p = 0.049), and thiacloprid (p = 0.035), and more complaints of chest pains, stomachache, skin eruption and diarrhea (p < 0.05) were found than in Normal Cystatin-C group (n = 78). Urinary neonicotinoids may be one of the potential risk factors for renal tubular dysfunction in this area.

Topics: Adult; Biomarkers; Chromatography, Liquid; Cystatin C; Farmers; Fatty Acid-Binding Proteins; Female; Geography; Guanidines; Humans; Insecticides; Kidney Tubules; Male; Middle Aged; Neonicotinoids; Nervous System Diseases; Nitro Compounds; Pyridines; Quality Control; Renal Insufficiency, Chronic; Sri Lanka; Surveys and Questionnaires; Tandem Mass Spectrometry; Thiamethoxam; Thiazines; Thiazoles

Globally, neonicotinoids are the most used insecticides, despite their well-documented sub-lethal effects on beneficial insects. Neonicotinoids are nicotinic acetylcholine receptor agonists. Memory, circadian rhythmicity and sleep are essential for efficient foraging and pollination and require nicotinic acetylcholine receptor signalling. The effect of field-relevant concentrations of the European Union-banned neonicotinoids: imidacloprid, clothianidin, thiamethoxam and thiacloprid were tested on Drosophila memory, circadian rhythms and sleep. Field-relevant concentrations of imidacloprid, clothianidin and thiamethoxam disrupted learning, behavioural rhythmicity and sleep whilst thiacloprid exposure only affected sleep. Exposure to imidacloprid and clothianidin prevented the day/night remodelling and accumulation of pigment dispersing factor (PDF) neuropeptide in the dorsal terminals of clock neurons. Knockdown of the neonicotinoid susceptible Dα1 and Dβ2 nicotinic acetylcholine receptor subunits in the mushroom bodies or clock neurons recapitulated the neonicotinoid like deficits in memory or sleep/circadian behaviour respectively. Disruption of learning, circadian rhythmicity and sleep are likely to have far-reaching detrimental effects on beneficial insects in the field.

Topics: Animals; Circadian Rhythm; Drosophila melanogaster; Guanidines; Insecticides; Memory; Neonicotinoids; Neuropeptides; Nicotinic Agonists; Nitro Compounds; Receptors, Nicotinic; Sleep; Thiamethoxam; Thiazines; Thiazoles

Topics: Acaricides; Animals; Guanidines; Mites; Neonicotinoids; Nitriles; Nitro Compounds; Pesticides; Pyrethrins; Spiders; Tetranychidae; Thiazoles

Neonicotinoids are pesticides used to protect crops but with known secondary influences at sublethal doses on bees. Honeybees use their sense of smell to identify the queen and nestmates, to signal danger and to distinguish flowers during foraging. Few behavioural studies to date have examined how neonicotinoid pesticides affect the ability of bees to distinguish odours. Here, we used a differential learning task to test how neonicotinoid exposure affects learning, memory and olfactory perception in foraging-age honeybees. Bees fed with thiamethoxam could not perform differential learning and could not distinguish odours during short- and long-term memory tests. Our data indicate that thiamethoxam directly impacts the cognitive processes involved in working memory required during differential olfactory learning. Using a combination of behavioural assays, we also identified that thiamethoxam has a direct impact on the olfactory perception of similar odours. Honeybees fed with other neonicotinoids (clothianidin, imidacloprid, dinotefuran) performed the differential learning task, but at a slower rate than the control. These bees could also distinguish the odours. Our data are the first to show that neonicotinoids have compound specific effects on the ability of bees to perform a complex olfactory learning task. Deficits in decision making caused by thiamethoxam exposure could mean that this is more harmful than other neonicotinoids, leading to inefficient foraging and a reduced ability to identify nestmates.

Topics: Animals; Bees; Flowers; Guanidines; Insecticides; Learning; Memory; Neonicotinoids; Nitro Compounds; Odorants; Olfactory Perception; Thiamethoxam; Thiazoles

This study measured both nutrient and pesticide concentrations at up to 13 different freshwater stream sites in New Brunswick, Nova Scotia and Prince Edward Island between 2013 and 2018. Up to 62 different pesticides were analysed in 248 discreet samples. A large majority of pesticides were below the detection limit of the laboratory while seven pesticides had at least 20% or more detections throughout the years of this study. The four pesticides that had the highest frequency of detection were the insecticides chlorantraniliprole, clothianidin, imidacloprid, and thiamethoxam of which the last three are categorised as neonicotinoid insecticides.

Topics: Canada; Environmental Monitoring; Guanidines; Neonicotinoids; Nitro Compounds; Pesticides; Rivers; Spatio-Temporal Analysis; Thiamethoxam; Thiazoles; Water Pollutants, Chemical

The projection of plant protection products' (PPPs) toxicity to non-target organisms at early stages of their development is challenging and demanding. Recent developments in bioanalytics, however, have facilitated the study of fluctuations in the metabolism of biological systems in response to treatments with bioactives and the discovery of corresponding toxicity biomarkers. Neonicotinoids are improved insecticides that target nicotinic acetylocholine receptors (nAChR) in insects which are similar to mammals. Nonetheless, they have sparked controversy due to effects on non-target organisms. Within this context, mammalian cell cultures represent ideal systems for the development of robust models for the dissection of PPPs' toxicity. Thus, we have investigated the toxicity of imidacloprid, clothianidin, and their mixture on primary mouse (Mus musculus) neural stem/progenitor (NSPCs) and mouse neuroblastoma-derived Neuro-2a (N2a) cells, and the undergoing metabolic changes applying metabolomics. Results revealed that NSPCs, which in vitro resemble those that reside in the postnatal and adult central nervous system, are five to seven-fold more sensitive than N2a to the applied insecticides. The energy equilibrium of NSPCs was substantially altered, as it is indicated by fluctuations of metabolites involved in energy production (e.g. glucose, lactate), Krebs cycle intermediates, and fatty acids, which are important components of cell membranes. Such evidence plausibly suggests a switch of cells' energy-producing mechanism to the direct metabolism of glucose to lactate in response to insecticides. The developed pipeline could be further exploited in the discovery of unintended effects of PPPs at early steps of development and for regulatory purposes.

Topics: Animals; Guanidines; Homeostasis; Insecticides; Metabolomics; Mice; Neonicotinoids; Nervous System; Nitro Compounds; Stem Cells; Thiazoles

Topics: Chromatography, Liquid; Guanidines; Honey; Insecticides; Neonicotinoids; Nitro Compounds; Thiamethoxam; Thiazines; Thiazoles

Systemic insecticides, such as neonicotinoids, are a major contributor towards beneficial insect declines. This has led to bans and restrictions on neonicotinoid use globally, most noticeably in the European Union, where four commonly used neonicotinoids (imidacloprid, thiamethoxam, clothianidin and thiacloprid) are banned from outside agricultural use. While this might seem like a victory for conservation, restrictions on neonicotinoid use will only benefit insect populations if newly emerging insecticides do not have similar negative impacts on beneficial insects. Flupyradifurone and sulfoxaflor are two novel insecticides that have been registered for use globally, including within the European Union. These novel insecticides differ in their chemical class, but share the same mode of action as neonicotinoids, raising the question as to whether they have similar sub-lethal impacts on beneficial insects. Here, we conducted a systematic literature search of the potential sub-lethal impacts of these novel insecticides on beneficial insects, quantifying these effects with a meta-analysis. We demonstrate that both flupyradifurone and sulfoxaflor have significant sub-lethal impacts on beneficial insects at field-realistic levels of exposure. These results confirm that bans on neonicotinoid use will only protect beneficial insects if paired with significant changes to the agrochemical regulatory process. A failure to modify the regulatory process will result in a continued decline of beneficial insects and the ecosystem services on which global food production relies.

Topics: 4-Butyrolactone; Animals; Ecosystem; Guanidines; Insecta; Insecticides; Neonicotinoids; Nitro Compounds; Pyridines; Sulfur Compounds; Thiamethoxam; Thiazines; Thiazoles

In this study, a method was developed for the determination of five neonicotinoid pesticides (acetamiprid, clothianidin, imidacloprid, thiacloprid, and thiamethoxam) in propolis. Two sample preparation methods were tested: solid-phase extraction and the quick, easy, cheap, effective, rugged, and safe (QuEChERS) method. The identities of analytes were confirmed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the selected reaction monitoring mode. Solid-phase extraction resulted in cleaner extracts; therefore, the SPE-LC-MS/MS method was validated according to the SANTE protocol in triplicate at two spiking levels (10 ng/g and 50 ng/g). The average recoveries of analytes ranged from 61% to 101%, except for clothianidin (10-20%). The LOD ranged from 0.2 ng/g to 4.4 ng/g, whereas the LOQ was in the range of 0.8 ng/g-14.7 ng/g. In order to compensate for the matrix effect, matrix-matched calibration was used. Good accuracy (relative error: 1.9-10.4%) and good linearity (R

Topics: Calibration; Chromatography, Liquid; Drug Contamination; Guanidines; Insecticides; Limit of Detection; Neonicotinoids; Nitro Compounds; Pesticide Residues; Propolis; Solid Phase Extraction; Tandem Mass Spectrometry; Thiamethoxam; Thiazines; Thiazoles

Among pesticides and foliar sprays involved in the treatment of seed, soil, and grass, also to crops, an important group is neonicotinoids. Neonicotinoid pesticides present similar properties with nicotine, but the mentioned compounds are less harmful for humans. Nevertheless, neonicotinoids are poisonous to insects and some invertebrates, which can act against insects' central nervous system, leading to their death. Moreover, neonicotinoids can affect the reproduction, foraging, and flying ability of honeybee and other insects including pollinators. In the present study, some neonicotinoids, such as imidacloprid, acetamiprid, clothianidin, thiacloprid, and thiamethoxam together with their toxic effects, have been presented. The Environmental Protection Agency (EPA) classifies these neonicotinoids as II and III class toxicity agents. Due to accumulation of these pesticides into the pollen of treated plants, especially due to their toxic effects against pollinators, the consequences of the occurrence of these insecticides have been discussed. Analytical aspects and methods involved in the isolation and determination of this class of pesticides have been presented in this contribution.

Topics: Animals; Bees; Guanidines; Humans; Insecta; Insecticides; Invertebrates; Neonicotinoids; Nitro Compounds; Soil; Thiamethoxam; Thiazines; Thiazoles

The effects of chronic exposure to two neonicotinoids (clothianidin and imidacloprid) and two organophosphates (chlorpyrifos and dimethoate) on survival, developmental rate and larval weight of honey bee larvae reared in vitro were determined. Diets containing chemicals were fed to larvae with the range of concentrations for each compound based on published acute toxicity experiments and residues found in pollen and nectar, both components of the larval diet.. Four concentrations of each compound and controls were tested: chlorpyrifos: 0.5, 0.8, 1.2, 8 mg/L; clothianidin: 0.1, 0.4, 2, 10 mg L. The treatment of larval diets with clothianidin, dimethoate and imidacloprid did not affect survival, developmental rate, or weight of immature honey bees; however, treatment with chlorpyrifos did. Overall, our results are valuable for evaluating the chronic toxicity of these pesticides to developing honey bees. © 2018 Society of Chemical Industry.

Topics: Animals; Bees; Chlorpyrifos; Dimethoate; Guanidines; In Vitro Techniques; Insecticides; Larva; Neonicotinoids; Nitro Compounds; Organophosphates; Thiazoles; Toxicity Tests, Chronic

Neonicotinoids have recently been demonstrated to cause direct negative impacts on birds from North America and Europe. To further understand the impact of these compounds on bird species and to improve risk assessment capacities, the current study determined the acute toxicities of imidacloprid, clothianidin, and thiamethoxam formulations on South American eared doves (Zenaida auriculata). Insecticides were administered by gavage to adult doves to determine median lethal doses (LD50) according to a standardized sequential procedure. The acute toxicity of formulated imidacloprid (LD50=59mg active ingredient, a.i./kg body weight, b.w.) was much higher than that of the tested formulations of clothianidin (LD50=4248mga.i./kg b.w.) and thiamethoxam (LD50=4366mga.i./kg b.w.). Imidacloprid also differed from the other two neonicotinoids in terms of the onset and intensity of intoxication signs and the times of death and recovery. All three insecticides induced a reduction in food consumption that led to body weight loss. An average weight dove of 127g would obtain a dose equivalent to the LD50 of imidacloprid by consuming 1.7g of treated sorghum seeds. As eared doves offered non-treated sorghum seeds 5h per day consumed on average 6.4±1.8g (mean±S.D.), it is concluded that these doves could feasibly be exposed to lethal doses in the field. This work is the first to describe intoxication signs and report oral neonicotinoid LD50s in a wild South-American bird species.

Topics: Animals; Columbidae; Dose-Response Relationship, Drug; Guanidines; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Risk Assessment; Thiamethoxam; Thiazoles; Toxicity Tests, Acute

The neurotoxicity of dinotefuran, thiamethoxam and imidacloprid against Chinese lizards (Eremias argus) were evaluated in acute oral exposure and 28d subchronic exposure. Dinotefuran was not easily metabolized and showed strong persistence in the lizard brain. Thiamethoxam and imidacloprid were rapidly absorbed and excreted in lizards, and were not easily enriched in the lizard brain. Dinotefuran and thiamethoxam could directly increase the concentrations of acetylcholine in the brain and blood by up-regulating the expression of the ach gene, which in turn enhanced the binding of acetylcholine and acetylcholinesterase receptors, eventually causing the release of dopamine. The effect of dinotefuran was more pronounced than thiamethoxam. Clothianidin was a major metabolite of thiamethoxam in the brain and aggravated the neurotoxic effects of thiamethoxam. Imidacloprid desnitro olefin was the only metabolite of imidacloprid that enriched in the brain. The protonation effect of imidacloprid desnitro olefin was stronger than that of the parent imidacloprid, which increased its binding ability to lizard acetylcholinesterase receptors. Competitive inhibition of imidacloprid desnitro olefin and acetylcholine led to the down-regulation of ach gene expression. Although neonicotinoids caused the opening of ligand-gated ion channel through the activation of acetylcholinesterase receptors, the body would alleviate these effects by the inhibition of voltage-dependent channel activity for compensatory mechanisms. This study provided a new perspective on the neotoxic effects of neonicotinoids.

Topics: Acetylcholine; Animals; Guanidines; Insecticides; Lizards; Neonicotinoids; Neurotoxicity Syndromes; Nitro Compounds; Thiamethoxam; Thiazoles

The purpose of the present study was to evaluate the presence of five neonicotinoid pesticides, acetamiprid, imidacloprid, clothianidin, thiacloprid, and thiamethoxam, in sheep and cow milk samples collected from animals bred in the Jordan Valley. In this area, numerous citrus plantations are present, and these insecticides are commonly used to protect plants from pests and diseases. Thirty-seven sheep milk samples and 31 cow milk samples were analysed. The analytical method, based on a single cleanup extraction step with SPE cartridges packed with diatomaceous earth material, together with analysis by LC-MS/MS, has guaranteed average recoveries between 75.1% and 88.3%, limits of detection (LOD) and quantification (LOQ) of 0.5 and 1 µg/kg, respectively, for all the five neonicotinoids. LOQ was much lower than the codex maximum residues limits for these pesticides in milks. No residues of the five neonicotinoids were found in any sample at a concentration level above LOD.

Topics: Animals; Cattle; Chromatography, Liquid; Female; Guanidines; Jordan; Limit of Detection; Milk; Neonicotinoids; Nitro Compounds; Pesticides; Reproducibility of Results; Sheep; Tandem Mass Spectrometry; Thiamethoxam; Thiazines; Thiazoles

External feather rinses and homogenized whole-carcass tissue matrix from two hummingbird species found in California (Calypte anna and Archilochus alexandri) were analyzed for the presence of nine insecticides commonly used in urban settings. Using a liquid chromatography-high-resolution mass spectrometry (LC-HRMS) analytical method, samples were quantitatively tested for the following neonicotinoids: dinotefuran, nitenpyram, thiamethoxam, acetamiprid, thiacloprid, clothianidin, imidacloprid, and sulfoxaflor. This analytical method was also used to qualitatively screen for the presence of approximately 150 other pesticides, drugs, and natural products. Feather rinsates from both hummingbird species had detectable concentrations of carbamate and neonicotinoid classes of insecticides. Combined results of the rinsate and homogenized samples (n = 64 individual hummingbirds) showed that 44 individuals (68.75%) were positive for one to four target compounds. This study documented that hummingbirds found in California are exposed to insecticides. Furthermore, feather rinsates and carcass homogenates are matrices that can be used for assessing pesticide exposure in small bird species. The small body size of hummingbirds limits traditional sampling methods for tissues and whole blood to evaluate for pesticide exposure. Thus, utilization of this analytical method may facilitate future research on small-sized avian species, provide insight into pesticide exposure, and ultimately lead to improved conservation of hummingbirds.

Topics: Animals; Birds; California; Chromatography, Liquid; Feathers; Guanidines; Insecticides; Neonicotinoids; Nitro Compounds; Pyridines; Sulfur Compounds; Tandem Mass Spectrometry; Thiamethoxam; Thiazines; Thiazoles

Nitenpyram is very effective in controlling Nilaparvata lugens (brown planthopper, BPH), and its resistance has been reported in field populations; however, the resistance mechanism remains unclear. In the present study, cross-resistance and resistance mechanisms in nitenpyram-resistant BPH were investigated. A resistant strain (NR) with a high resistance level (164.18-fold) to nitenpyram was evolved through successive selection for 42 generations from a laboratory susceptible strain (NS). The bioassay results showed that the NR exhibited cross-resistance to imidacloprid (37.46-fold), thiamethoxam (71.66-fold), clothianidin (149.17-fold), dinotefuran (98.13-fold), sulfoxaflor (47.24-fold), cycloxaprid (9.33-fold), etofenprox (10.51-fold) and isoprocarb (9.97-fold) but not to triflumezopyrim, chlorpyrifos and buprofezin. The NR showed a 3.21-fold increase in cytochrome P450 monooxygenase (P450) activity compared to that in the NS, while resistance was also synergized (4.03-fold) with the inhibitor piperonyl butoxide (PBO), suggesting a role of P450. Furthermore, the mRNA expression levels of cytochrome P450 (CYP) genes by quantitative real-time PCR results indicated that twelve P450 genes were significantly overexpressed in the NR strain, especially CYP6ER1 (203.22-fold). RNA interference (RNAi) suppression of CYP6ER1 through injection of dsCYP6ER1 led to significant susceptibility in the NR strain. The current study expands our understanding of the nitenpyram resistance mechanism in N. lugens, provides an important reference for integrated pest management (IPM), and enriches the theoretical system of insect toxicology.

Topics: Animals; Carbamates; Guanidines; Hemiptera; Insect Proteins; Insecticide Resistance; Neonicotinoids; Nitro Compounds; Pyrethrins; Pyridines; Pyrimidinones; RNA Interference; Thiazoles

The wetlands used for some agricultural activities constitute productive breeding sites for many mosquito species. Thus, the agricultural use of insecticide targeting other pests may select for insecticide resistance in malaria mosquitoes. The purpose of this study is to clarify some knowledge gaps on the role of agrochemicals in the development of insecticide resistance in malaria vectors is of utmost importance for vector control.. Using the CDC bottle test and the log-probit analysis, we investigated for the first time the resistance levels of Anopheles coluzzii mosquitoes to neonicotinoids, insecticides used exclusively for crop protection in Côte d'Ivoire. The study was conducted in two agricultural regions (Tiassale and Gagnoa) and one non-agricultural region (Vitre) between June and August 2017 using clothianidin, acetamiprid and imidacloprid.. Mosquito populations from Tiassale and Gagnoa (agricultural settings) were determined to be resistant to acetamiprid with mortality rates being < 85% at 24 h post-exposure. In Vitre (non-agricultural area) however, the mosquito population was susceptible to acetamiprid. In all three localities, mosquito populations were resistant to imidacloprid (mortality rates were 60% in Vitre, 37% in Tiassale, and 13% in Gagnoa) and completely susceptible to clothianidin (100% mortality). An. coluzzii represented 100% of mosquito collected in Gagnoa, 86% in Tiassale and 96% in Vitre.. This study provides strong evidence that agricultural use of insecticides can cause insecticide resistance in malaria vector populations. Insecticide resistance driven by agrochemical usage should be considered when vector control strategies are developed.

Topics: Animals; Anopheles; Cote d'Ivoire; Guanidines; Insecticide Resistance; Insecticides; Mosquito Vectors; Neonicotinoids; Nitro Compounds; Selection, Genetic; Thiazoles

Neonicotinoid insecticides have been shown to have high chronic toxicity relative to acute toxicity, and therefore short-term toxicity tests ≤96 h in duration may underestimate their environmental risks. Among nontarget aquatic invertebrates, insects of the orders Diptera and Ephemeroptera have been found to be the most sensitive to neonicotinoids. To undertake a more accurate assessment of the risks posed by neonicotinoids to freshwater ecosystems, more data are needed from long-term tests employing the most sensitive taxa. Using nymphs of the common New Zealand mayfly genus Deleatidium spp., we performed 28-d static-renewal exposures with the widely used neonicotinoids imidacloprid, clothianidin, and thiamethoxam. We monitored survival, immobility, impairment, and mayfly moulting propensity at varying time points throughout the experiment. Imidacloprid and clothianidin exerted strong chronic toxicity effects on Deleatidium nymphs, with 28-d median lethal concentrations (LC50s) of 0.28 and 1.36 µg/L, respectively, whereas thiamethoxam was the least toxic, with a 28-d LC50 > 4 µg/L (highest concentration tested). Mayfly moulting propensity was also negatively affected by clothianidin (during 3 of 4 wk), imidacloprid (2 of 4 wk), and thiamethoxam (1 of 4 wk). Comparisons with published neonicotinoid chronic toxicity data for other mayfly taxa and larvae of the midge genus Chironomus showed similar sensitivities for mayflies and midges, suggesting that experiments using these taxa provide reliable assessments of the threats of neonicotinoids to the most vulnerable freshwater species. Environ Toxicol Chem 2019;38:2459-2471. © 2019 SETAC.

Topics: Animals; Environmental Exposure; Ephemeroptera; Guanidines; Insecticides; Linear Models; Molting; Neonicotinoids; New Zealand; Nitro Compounds; Nymph; Survival Analysis; Thiamethoxam; Thiazoles; Toxicity Tests, Chronic; Water Pollutants, Chemical

Surface waters are sometimes contaminated with neonicotinoids: a widespread, persistent, systemic class of insecticide with leaching potential. Previous ecotoxicological investigations of this chemical class in aquatic ecosystems have largely focused on the impacts of the neonicotinoid imidacloprid; few empirical, manipulative studies have investigated the effect on invertebrate abundances of two other neonicotinoids which are now more widely used: clothianidin and thiamethoxam. In this study, we employ a simple microcosm semi-field design, incorporating a one-off contamination event, to investigate the effect of these pesticides at field-realistic levels (ranging from 0 to 15 ppb) on invertebrate colonisation and survival in small ephemeral ponds. In line with previous research on neonicotinoid impacts on aquatic invertebrates, significant negative effects of both neonicotinoids were found. There were clear differences between the two chemicals, with thiamethoxam generally producing stronger negative effects than clothianidin. Populations of Chironomids (Diptera) and Ostracoda were negatively affected by both chemicals, while Culicidae appeared to be unaffected by clothianidin at the doses used. Our data demonstrate that field-realistic concentrations of neonicotinoids are likely to reduce populations of invertebrates found in ephemeral ponds, which may have knock on effects up the food chain. We highlight the importance of developing pesticide monitoring schemes for European surface waters.

Topics: Animals; Guanidines; Insecticides; Invertebrates; Neonicotinoids; Nitro Compounds; Pesticides; Thiamethoxam; Thiazoles

Neonicotinoid insecticides are environmentally persistent and highly water-soluble, and thus are prone to leaching into surface waters where they may negatively affect non-target aquatic insects. Most of the research to date has focused on imidacloprid, and few data are available regarding the effects of other neonicotinoids or their proposed replacements (butenolide insecticides). The objective of this study was to assess the toxicity of six neonicotinoids (imidacloprid, thiamethoxam, acetamiprid, clothianidin, thiacloprid, and dinotefuran) and one butenolide (flupyradifurone) to Hexagenia spp. (mayfly larvae). Acute (96-h), water-only tests were conducted, and survival and behaviour (number of surviving mayflies inhabiting artificial burrows) were assessed. Acute sublethal tests were also conducted with imidacloprid, acetamiprid, and thiacloprid, and in addition to survival and behaviour, mobility (ability to burrow into sediment) and recovery (survival and growth following 21 d in clean sediment) were measured. Sublethal effects occurred at much lower concentrations than survival: 96-h LC50s ranged from 780 μg/L (acetamiprid) to >10,000 μg/L (dinotefuran), whereas 96-h EC50s ranged from 4.0 μg/L (acetamiprid) to 630 μg/L (thiamethoxam). Flupyradifurone was intermediate in toxicity, with a 96-h LC50 of 2000 μg/L and a 96-h EC50 of 81 μg/L. Behaviour and mobility were impaired significantly and to a similar degree in sublethal exposures to 10 μg/L imidacloprid, acetamiprid, and thiacloprid, and survival and growth following the recovery period were significantly lower in mayflies exposed to 10 μg/L acetamiprid and thiacloprid, respectively. A suite of effects on mayfly swimming behaviour/ability and respiration were also observed, but not quantified, following exposures to imidacloprid, acetamiprid, and thiacloprid at 1 μg/L and higher. Imidacloprid concentrations measured in North American surface waters have been found to meet or exceed those causing toxicity to Hexagenia, indicating that environmental concentrations may adversely affect Hexagenia and similarly sensitive non-target aquatic species.

Topics: 4-Butyrolactone; Animals; Ephemeroptera; Guanidines; Imidazoles; Insecticides; Larva; Neonicotinoids; Nitro Compounds; Oxazines; Pyridines; Thiamethoxam; Thiazines; Thiazoles

Widespread agricultural use of neonicotinoid insecticides has resulted in frequent detection of mixtures of these compounds in global surface waters. Recent evidence suggests that neonicotinoid mixtures can elicit synergistic toxicity in aquatic insects under acute exposure conditions, however this has not been validated for longer exposures more commonly encountered in the environment. Therefore, we aimed to characterize the chronic (28-day) toxicity of imidacloprid, clothianidin, and thiamethoxam mixtures under different doses and mixture ratios to determine if the assumption of synergistic toxicity would hold under more environmentally realistic exposure settings. The sensitive aquatic insect Chironomus dilutus was used as a representative test species, and successful emergence was used as a chronic endpoint. Applying the MIXTOX modeling approach, predictive parametric models were fitted using single-compound toxicity data and statistically compared to observed toxicity in subsequent mixture tests. Imidacloprid-clothianidin, clothianidin-thiamethoxam and imidacloprid-clothianidin-thiamethoxam mixtures did not significantly deviate from concentration-additive toxicity. However, the cumulative toxicity of the imidacloprid-thiamethoxam mixture deviated from the concentration-additive reference model, displaying dose-ratio dependent synergism and resulting in up to a 10% greater reduction in emergence from that predicted by concentration addition. Furthermore, exposure to select neonicotinoid mixtures above 1.0 toxic unit tended to shift sex-ratios toward more male-dominated populations. Results indicate that, similar to acute exposures, the general assumption of joint additivity cannot adequately describe chronic cumulative toxicity of all neonicotinoid mixtures. Indeed, our observations of weak synergism and sex-ratio shifts elicited by some mixture combinations should be considered in water quality guideline development and environmental risk assessment practices for neonicotinoid insecticides, and explored in further investigations of the effects of neonicotinoid mixtures on aquatic communities.

Topics: Animals; Chironomidae; Female; Guanidines; Insecticides; Male; Neonicotinoids; Nitro Compounds; Oxazines; Thiamethoxam; Thiazoles; Toxicity Tests, Chronic; Water Quality

Because of the persistence and solubility of neonicotinoid insecticides (NNIs), there is concern that these compounds may contaminate sources of drinking water. The objective of this project was to evaluate the distribution of NNIs in raw and treated drinking water from selected municipalities that draw their water from the lower Great Lakes in areas of southern Ontario, Canada where there is high intensity agriculture. Sites were monitored using Polar Organic Chemical Integrative Samplers (POCIS) and by collecting grab samples at six drinking water treatment plants. Thiamethoxam, clothianidin and imidacloprid were detected in both POCIS and grab samples of raw water. The frequency of detection of NNIs was much lower in treated drinking water, but some compounds were still detected at estimated concentrations in the low ng L

Topics: Agriculture; Drinking Water; Guanidines; Humans; Neonicotinoids; Nitro Compounds; Ontario; Oxazines; Pesticides; Thiamethoxam; Thiazoles; Water Pollutants, Chemical

Neonicotinoids (neonics), a class of systemic insecticides, have been frequently detected in pollen, vegetables, and fruits. Recently, an increasing concern has been aroused for human exposure to neonics. However, biological monitoring for quantifying body burden of neonics has rarely been reported. In this study, we developed an isotope-dilution ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) method to simultaneously quantify nine neonics, including acetamiprid (ACE), thiamethoxam (THIAM), imidacloprid (IMIP), clothianidin (CLO), flonicamid (FLO), thiacloprid (THIAC), dinotefuran (DIN), nitenpyram (NIT), and imidaclothiz (IMIT) in urine. The limits of quantification were 0.1 μg/L for ACE, FLO, DIN, NIT and IMIT, and 0.2 μg/L for THIAM, IMIP, CLO, and THIAC. The overall recoveries were 80.8-103%, 81.5-91.7% and 83.0-92.3% for QA/QC samples fortifying at 1, 25, and 100 μg/L levels, respectively. UPLC/MS/MS method was used to analyze urine samples obtained from 10 children in Hangzhou, China. The detection frequencies were 80% for ACE and IMIP, 70% for THIAM and CLO, 20% for DIN and IMIT and 10% for THIAC. FLO and NIT were not detected in those urine samples. The data provided here will be helpful for conducting biological monitoring of neonics exposure in the future.

Topics: Child; China; Chromatography, High Pressure Liquid; Environmental Monitoring; Guanidines; Humans; Indicator Dilution Techniques; Insecticides; Isotopes; Neonicotinoids; Niacinamide; Nitro Compounds; Oxazines; Pyridines; Tandem Mass Spectrometry; Thiamethoxam; Thiazines; Thiazoles

Neonicotinoids are implicated in the decline of honey bees, but the molecular basis underlying adverse effects is poorly known. Here we describe global transcriptomic profiles in the brain of honey bee workers exposed for 48 h at one environmentally realistic and one sublethal concentration of 0.3 and 3.0 ng/bee clothianidin and imidacloprid, respectively, and 0.1 and 1.0 ng/bee thiamethoxam (1-30 ng/mL sucrose solution) by high-throughput RNA-sequencing (RNA-seq). All neonicotinoids led to significant alteration (mainly down-regulation) of gene expression, generally with a concentration-dependent effect. Among many others, genes related to metabolism and detoxification were differently expressed. Gene ontology (GO) enrichment analysis of biological processes revealed catabolic carbohydrate metabolism (regulation of enzyme activities such as amylase), lipid metabolism, and transport mechanisms as shared terms between all neonicotinoids at high concentrations. KEGG pathway analysis indicated that at least two neonicotinoids induced changes in expression of various metabolic pathways: pentose phosphate pathways, starch and sucrose metabolism, and sulfur metabolism, in which glucose 1-dehydrogenase and alpha-amylase were down-regulated and 3'(2'), 5'-bisphosphate nucleotidase was up-regulated. RT-qPCR analysis confirmed the down-regulation of major royal jelly proteins, hbg3, and cyp9e2 found by RNA-seq. Our study highlights the comparative molecular effects of neonicotinoid exposure to bees. Further studies should link these effects with physiological outcomes for a better understanding of effects of neonicotinoids.

Topics: Animals; Bees; Brain; Guanidines; Insecticides; Neonicotinoids; Nitro Compounds; Thiamethoxam; Thiazoles; Transcriptome

Easy-to-use commercial kit-based enzyme-linked immunosorbent assays (ELISAs) have been used to detect neonicotinoid dinotefuran, clothianidin and imidacloprid in Chinese chives, which are considered a troublesome matrix for chromatographic techniques. Based on their high water solubility, water was used as an extractant. Matrix interference could be avoided substantially just diluting sample extracts. Average recoveries of insecticides from spiked samples were 85-113%, with relative standard deviation of <15%. The concentrations of insecticides detected from the spiked samples with the proposed ELISA methods correlated well with those by the reference high-performance liquid chromatography (HPLC) method. The residues analyzed by the ELISA methods were consistently 1.24 times that found by the HPLC method, attributable to loss of analyte during sample clean-up for HPLC analyses. It was revealed that the ELISA methods can be applied easily to pesticide residue analysis in troublesome matrix such as Chinese chives.

Topics: Chive; Chromatography, High Pressure Liquid; Enzyme-Linked Immunosorbent Assay; Food Analysis; Food Contamination; Guanidines; Insecticides; Neonicotinoids; Nitro Compounds; Pesticide Residues; Thiazoles

Freshwater mussels contribute important ecological functions to aquatic systems. The water filtered by mussel assemblages can improve water quality, and the mixing of sediments by burrowing mussels can improve oxygen content and release nutrients. However, nearly 70% of North American freshwater mussel species are listed as either endangered, threatened, or in decline. In Ontario, 28 species are in decline or in need of protection. Even though freshwater mussels have a heightened sensitivity to some contaminants, few studies have investigated the risks that various pesticide classes pose to one freshwater mussel species or among life stages. Lampsilis siliquoidea and Villosa iris were the focus of the present study, with the latter currently listed as of "special concern" in Canada. A potential risk to the recovery of freshwater mussel species is the presence and persistence of pesticides in Ontario surface waters. Acute (48 h) toxicity tests were performed with V. iris glochidia to determine the effect on viability (surrogate for survival) following exposure to 4 fungicides (azoxystrobin, boscalid, metalaxyl, and myclobutanil), 3 neonicotinoids (clothianidin, imidacloprid, and thiamethoxam), 2 carbamates (carbaryl and malathion), 1 organophosphate (chlorpyrifos), and 1 butenolide (flupyradifurone). Juvenile and adult L. siliquoidea were also exposed to azoxystrobin, clothianidin, imidacloprid (juvenile only), and carbaryl (adult only). Our study found in general that all life stages were insensitive to the pesticides tested, with median effect and lethal concentrations >161 µg/L. The pesticides tested likely represent a minimal risk (hazard quotients <5.4 × 10

Topics: Animals; Bivalvia; Chlorpyrifos; Environmental Exposure; Fresh Water; Guanidines; Life Cycle Stages; Neonicotinoids; Nitro Compounds; Ontario; Pesticides; Pyrimidines; Strobilurins; Thiazoles; Toxicity Tests; Water Pollutants, Chemical; Water Quality

Pesticide residues (112 compounds) were quantified by GC-MS/MS or LC-MS/MS in 85 bee bread samples and 154 pollen samples obtained from five apiaries each with three or four colonies (genotype Buckfast) in Luxembourg over the period 2011-2013. Thiacloprid, chlorfenvinphos, tebuconazole, and methiocarb were found most frequently in bee bread while thiacloprid, permethrin-cis, and permethrin-trans were detected most frequently in the pollen samples. Three neonicotinoid insecticides (clothianidin, imidacloprid, and thiamethoxam) that were restricted by an EU regulation in 2013 after our sampling campaign was finished were each found in less than 8% of the pollen or bee bread samples. The maximum concentrations of thiacloprid, metazachlor, and methiocarb measured in the pollen collected by a group of honeybee colonies (n = 5) without survivors within the 3-year period of observation were 86.20 ± 10.74 ng/g, 2.80 ± 1.26 ng/g, and below the limit of quantification, respectively. The maximum concentrations of the same compounds measured in the pollen collected by a group of honeybee colonies with significantly (P = 0.02) more survivors (7 out of 9) than expected, if the survivors had been distributed randomly among the groups of colonies, were 11.98 ± 2.28 ng/g, 0.44 ± 0.29 ng/g, and 8.49 ± 4.13 ng/g, respectively. No honeybee colony that gathered pollen containing more than 23 ng/g thiacloprid survived the 3-year project period. There was no statistically significant association between pesticide residues in the bee bread and the survival of the colonies. Actions already taken or planned and potential further actions to protect bees from exposure to pesticides are discussed.

Topics: Animals; Bees; Chromatography, Gas; Chromatography, Liquid; Environmental Monitoring; Guanidines; Insecticides; Luxembourg; Neonicotinoids; Nitro Compounds; Pesticide Residues; Pesticides; Pollen; Propolis; Tandem Mass Spectrometry; Thiazines; Thiazoles

Neonicotinoid insecticide mixtures are frequently detected in aquatic environments in agricultural regions. Recent laboratory studies have indicated that neonicotinoid mixtures can elicit greater-than-additive toxicity in sensitive aquatic insects (e.g. Chironomus dilutus). However, this has yet to be validated under field conditions. In this study, we compared the chronic (28- and 56-day) toxicity of three neonicotinoids (imidacloprid, clothianidin, and thiamethoxam) and their mixtures to natural aquatic insect communities. Using experimental in-situ enclosures (limnocorrals), we exposed wetland insects to single-compounds and binary mixtures at equitoxic concentrations (1 toxic unit under the principle of Concentration Addition). We assessed the composition of all emerged insect taxa and the cumulative Chironomidae emergence and biomass over time. In treated limnocorrals, there were subtle shifts in community composition, with greater mean proportions of emergent Trichoptera and Odonata. Cumulative emergence and biomass increased over time and there was a significant interaction between time and treatment. At 28 days, cumulative Chironomidae emergence and biomass were not significantly different between neonicotinoid treatments and controls. However, cumulative emergence in the imidacloprid, clothianidin, and clothianidin-thiamethoxam treatments were 42%, 20%, and 44% lower than predicted from applied doses. At 56 days, effects on cumulative emergence and biomass were significant for imidacloprid, clothianidin, and the clothianidin-thiamethoxam mixture. Contrary to laboratory predictions, mixtures were not more toxic than single compounds under semi-controlled field settings. Furthermore, only clothianidin significantly shifted sex-ratios towards female-dominated populations. Results showed that the responses of natural Chironomidae populations to neonicotinoids and their mixtures cannot be adequately predicted from laboratory-derived single-species models, and although occasional overdosing may have influenced the magnitude of effects, reductions in Chironomidae emergence and biomass can occur at average neonicotinoid concentrations below some current water quality guidelines. Therefore, neonicotinoid guidelines should be revised to ensure that Chironomidae and other sensitive aquatic insects inhabiting agricultural wetlands are adequately protected.

Topics: Animals; Chironomidae; Female; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Thiamethoxam; Thiazoles; Water Pollutants, Chemical; Water Quality; Wetlands

With the widespread adoption of no-tillage technology, outbreaks of thrips have caused serious damage to summer corn fields in China. Therefore, effective control of pest populations is often essential for cost-effective crop production. In this study, experiments were conducted in 2014 and 2015 to determine the control efficacy of seven neonicotinoid insecticide seed treatments against corn thrips and the effects of these treatments on natural enemy population densities and emergence rates, seedling characteristics, and yield of corn. The results showed that among the tested neonicotinoid seed treatments, thiamethoxam (1.0 and 2.0 g active ingredient (AI)/kg of seeds), clothianidin (1.0 and 2.0 g AI/kg of seeds), and imidacloprid (2.0 g AI/kg of seeds) showed the highest control efficacy against corn thrips throughout the corn growing season. Seed treatments with acetamiprid, nitenpyram, dinotefuran, and thiacloprid at rates of 1.0 and 2.0 g AI/kg of seeds were difficult to effectively control thrips on summer corn. Neonicotinoid seed treatments showed no adverse effects on the numbers of spiders and lady beetles. Furthermore, treatments did not negatively influence the seedling growth or development of corn but did prevent yield losses. Therefore, treating corn seeds with thiamethoxam, clothianidin, and imidacloprid can provide effective protection against early-season thrips and reduce yield losses under field conditions.

Topics: Animals; Guanidines; Insect Control; Insecticides; Neonicotinoids; Nitro Compounds; Seeds; Thiamethoxam; Thiazoles; Thysanoptera; Zea mays

Neonicotinoids are the most widely used class of insecticides in the world, and there are increasing concerns about their effects on non-target organisms. Analytical methods to diagnose exposure to neonicotinoids in wildlife are still very limited, particularly for small animals such as songbirds. Blood can be used as a non-lethal sampling matrix, but the sample volume is limited by body size. Neonicotinoids have a low bioaccumulation potential and are rapidly metabolized, therefore, sensitive assays are critically needed to reliably detect their residues in blood samples. We developed an efficient LC-MS/MS method at a part-per-trillion (pg/ml) level to measure eight neonicotinoid related insecticides (acetamiprid, clothianidin, dinotefuran, flonicamid, imidacloprid, nitenpyram, thiacloprid and thiamethoxam) plus one metabolite (6-chloronicotinic acid) in small volumes (50 μL) of avian plasma. The average recovery of target compounds ranged from 95.7 to 101.3%, and relative standard deviations were between 0.82 and 2.13%. We applied the method to screen blood samples from 36 seed-eating songbirds (white-crowned sparrows; Zonotrichia leucophrys) at capture, and detected imidacloprid in 78% (28 of 36), thiamethoxam in 22% (8 of 36), thiacloprid in 11% (4 of 36), and acetamiprid in 11% (4 of 36) of wild-caught sparrows. 6 h after capture, birds were orally dosed with 0 (control), 1.2 or 3.9 mg of imidacloprid/kg bw, test results using this method indicated that plasma imidacloprid was significantly elevated (low 26-times, high 316-times) in exposed groups. This is the first study to confirm neonicotinoid exposure in small free-living songbirds through non-lethal blood sampling, and to demonstrate that environmentally realistic doses significantly elevate circulating imidacloprid concentrations. This sensitive method could be applied to characterize exposure to neonicotinoids in free-living wildlife and in toxicological studies.

Topics: Animals; Chromatography, Liquid; Environmental Monitoring; Guanidines; Insecticides; Neonicotinoids; Nitro Compounds; Plasma; Songbirds; Tandem Mass Spectrometry; Thiazines; Thiazoles

Uptake and translocation of imidacloprid (IMI), thiamethoxam (THX) and difenoconazole (DFZ) in rice plants (Oryza sativa L.) were investigated with a soil-treated experiment at two application rates: field rate (FR) and 10*FR under laboratory conditions. The dissipation of the three compounds in soil followed the first-order kinetics and DFZ showed greater half-lives than IMI and THX. Detection of the three compounds in rice tissues indicated that rice plants could take up and accumulate these pesticides. The concentrations of IMI and THX detected in leaves (IMI, 10.0 and 410 mg/kg dw; THX, 23.0 and 265 mg/kg dw) were much greater than those in roots (IMI, 1.37 and 69.3 mg/kg dw; THX, 3.19 and 30.6 mg/kg dw), which differed from DFZ. The DFZ concentrations in roots (15.6 and 79.1 mg/kg dw) were much greater than those in leaves (0.23 and 3.4 mg/kg dw). The bioconcentration factor (BCF), representing the capability of rice to accumulate contaminants from soil into plant tissues, ranged from 1.9 to 224.3 for IMI, from 2.0 to 72.3 for THX, and from 0.4 to 3.2 for DFZ at different treated concentrations. Much higher BCFs were found for IMI and THX at 10*FR treatment than those at FR treatment, however, the BCFs of DFZ at both treatments were similar. The translocation factors (TFs), evaluating the capability of rice to translocate contaminants from the roots to the aboveground parts, ranged from 0.02 to 0.2 for stems and from 0.02 to 9.0 for leaves. The tested compounds were poorly translocated from roots to stems, with a TF below 1. However, IMI and THX were well translocated from roots to leaves. Clothianidin (CLO), the main metabolite of THX, was detected at the concentrations from 0.02 to 0.5 mg kg

Topics: Dioxolanes; Guanidines; Imidazoles; Neonicotinoids; Nitro Compounds; Oryza; Oxazines; Plant Leaves; Plant Roots; Soil; Soil Pollutants; Thiamethoxam; Thiazoles; Triazoles

Garden centres frequently market nectar- and pollen-rich ornamental plants as "pollinator-friendly", however these plants are often treated with pesticides during their production. There is little information on the nature of pesticide residues present at the point of purchase and whether these plants may actually pose a threat to, rather than benefit, the health of pollinating insects. Using mass spectrometry analyses, this study screened leaves from 29 different 'bee-friendly' plants for 8 insecticides and 16 fungicides commonly used in ornamental production. Only two plants (a Narcissus and a Salvia variety) did not contain any pesticide and 23 plants contained more than one pesticide, with some species containing mixtures of 7 (Ageratum houstonianum) and 10 (Erica carnea) different agrochemicals. Neonicotinoid insecticides were detected in more than 70% of the analysed plants, and chlorpyrifos and pyrethroid insecticides were found in 10% and 7% of plants respectively. Boscalid, spiroxamine and DMI-fungicides were detected in 40% of plants. Pollen samples collected from 18 different plants contained a total of 13 different pesticides. Systemic compounds were detected in pollen samples at similar concentrations to those in leaves. However, some contact (chlorpyrifos) and localised penetrant pesticides (iprodione, pyroclastrobin and prochloraz) were also detected in pollen, likely arising from direct contamination during spraying. The neonicotinoids thiamethoxam, clothianidin and imidacloprid and the organophosphate chlorpyrifos were present in pollen at concentrations between 6.9 and 81 ng/g and at levels that overlap with those known to cause harm to bees. The net effect on pollinators of buying plants that are a rich source of forage for them but simultaneously risk exposing them to a cocktail of pesticides is not clear. Gardeners who wish to gain the benefits without the risks should seek uncontaminated plants by growing their own from seed, plant-swapping or by buying plants from an organic nursery.

Topics: Agrochemicals; Animals; Bees; Environmental Monitoring; Fungicides, Industrial; Gardening; Guanidines; Imidazoles; Insecta; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Pesticide Residues; Pesticides; Plant Nectar; Pollen; Pollination; Seeds; Thiamethoxam; Thiazoles

Extensive agricultural use of neonicotinoid insecticide products has resulted in the presence of neonicotinoid mixtures in surface waters worldwide. Although many aquatic insect species are known to be sensitive to neonicotinoids, the impact of neonicotinoid mixtures is poorly understood. In the present study, the cumulative toxicities of binary and ternary mixtures of select neonicotinoids (imidacloprid, clothianidin, and thiamethoxam) were characterized under acute (96-h) exposure scenarios using the larval midge Chironomus dilutus as a representative aquatic insect species. Using the MIXTOX approach, predictive parametric models were fitted and statistically compared with observed toxicity in subsequent mixture tests. Single-compound toxicity tests yielded median lethal concentration (LC50) values of 4.63, 5.93, and 55.34 μg/L for imidacloprid, clothianidin, and thiamethoxam, respectively. Because of the similar modes of action of neonicotinoids, concentration-additive cumulative mixture toxicity was the predicted model. However, we found that imidacloprid-clothianidin mixtures demonstrated response-additive dose-level-dependent synergism, clothianidin-thiamethoxam mixtures demonstrated concentration-additive synergism, and imidacloprid-thiamethoxam mixtures demonstrated response-additive dose-ratio-dependent synergism, with toxicity shifting from antagonism to synergism as the relative concentration of thiamethoxam increased. Imidacloprid-clothianidin-thiamethoxam ternary mixtures demonstrated response-additive synergism. These results indicate that, under acute exposure scenarios, the toxicity of neonicotinoid mixtures to C. dilutus cannot be predicted using the common assumption of additive joint activity. Indeed, the overarching trend of synergistic deviation emphasizes the need for further research into the ecotoxicological effects of neonicotinoid insecticide mixtures in field settings, the development of better toxicity models for neonicotinoid mixture exposures, and the consideration of mixture effects when setting water quality guidelines for this class of pesticides. Environ Toxicol Chem 2017;36:3091-3101. © 2017 SETAC.

Topics: Animals; Chironomidae; Guanidines; Insecticides; Larva; Neonicotinoids; Nitro Compounds; Oxazines; Thiamethoxam; Thiazoles; Toxicity Tests, Acute; Water Pollutants, Chemical

Acute toxicities (LD50s) of imidacloprid and clothianidin to Apis mellifera and A. cerana were investigated. Changing patterns of immune-related gene expressions and the activities of four enzymes between the two bee species were compared and analyzed after exposure to sublethal doses of insecticides. Results indicated that A. cerana was more sensitive to imidacloprid and clothianidin than A. mellifera. The acute oral LD50 values of imidacloprid and clothianidin for A. mellifera were 8.6 and 2.0ng/bee, respectively, whereas the corresponding values for A. cerana were 2.7 and 0.5ng/bee. The two bee species possessed distinct abilities to mount innate immune response against neonicotinoids. After 48h of imidacloprid treatment, carboxylesterase (CCE), prophenol oxidase (PPO), and acetylcholinesterase (AChE) activities were significantly downregulated in A. mellifera but were upregulated in A. cerana. Glutathione-S-transferase (GST) activity was significantly elevated in A. mellifera at 48h after exposure to imidacloprid, but no significant change was observed in A. cerana. AChE was downregulated in both bee species at three different time points during clothianidin exposure, and GST activities were upregulated in both species exposed to clothianidin. Different patterns of immune-related gene expression and enzymatic activities implied distinct detoxification and immune responses of A. cerana and A. mellifera to imidacloprid and clothianidin.

Topics: Animals; Bees; Guanidines; Histocompatibility Antigens; Insecticides; Neonicotinoids; Nitro Compounds; Species Specificity; Thiazoles

Growing evidence for global pollinator decline is causing concern for biodiversity conservation and ecosystem services maintenance. Neonicotinoid pesticides have been identified or suspected as a key factor responsible for this decline. We assessed the global exposure of pollinators to neonicotinoids by analyzing 198 honey samples from across the world. We found at least one of five tested compounds (acetamiprid, clothianidin, imidacloprid, thiacloprid, and thiamethoxam) in 75% of all samples, 45% of samples contained two or more of these compounds, and 10% contained four or five. Our results confirm the exposure of bees to neonicotinoids in their food throughout the world. The coexistence of neonicotinoids and other pesticides may increase harm to pollinators. However, the concentrations detected are below the maximum residue level authorized for human consumption (average ± standard error for positive samples: 1.8 ± 0.56 nanograms per gram).

Topics: Animals; Bees; Biodiversity; Food Contamination; Guanidines; Honey; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Pollination; Thiamethoxam; Thiazines; Thiazoles

Topics: Agrochemicals; Animals; Bees; Dissent and Disputes; European Union; Guanidines; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Plants; Pollination; Thiamethoxam; Thiazines; Thiazoles; Toxicity Tests; Uncertainty; United States; United States Environmental Protection Agency

A quantitative weight of evidence (QWoE) methodology was used to assess higher-tier studies on the effects of imidacloprid (IMI) on honeybees. Assessment endpoints were population size and viability of commercially managed bees and quantity of hive products. A colony-level no-observed-adverse effect concentration (NOAEC) of 25 µg IMI/kg syrup, equivalent to an oral no-observed-adverse-effect-dose of 7.3 ng/bee/d for all responses, was measured. The overall weight of evidence indicates that there is minimal risk to honeybees from exposure to IMI from its use as a seed treatment. Exposures via dusts from currently used seed coatings present a de minimis risk to honeybees when the route of exposure is via uptake in plants that are a source of pollen or nectar for honeybees. There were few higher-tier observational (ecoepidemiological) studies conducted with IMI. Considering all lines of evidence, the quality of the studies included in this analysis was variable, but the results of the studies were consistent and point to the same conclusion - that IMI had no adverse effects on viability of the honeybee colony. Thus, the overall conclusion is that IMI, as currently used as a seed treatment and with good agricultural practices, does not present a significant risk to honeybees at the level of the colony.

Topics: Animals; Bees; Environmental Monitoring; Guanidines; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Risk Assessment; Thiamethoxam; Thiazoles; Toxicity Tests

A quantitative weight of evidence (QWoE) methodology was used to assess higher tier studies on the effects of clothianidin (CTD) on honeybees. Assessment endpoints were population size and viability of commercially managed bees and quantity of hive products. A colony-level no-observed-adverse effect concentration (NOAEC) of 25 µg CTD/kg syrup, equivalent to an oral no-observed-adverse effect-dose (NOAED) of 7.3 ng/bee/d for all responses measured. Based on a NOAEC of 19.7 µg/kg pollen, the NOAED for honeybee larvae was 2.4 ng/bee larva/d. For exposures via dust, a no-observed-adverse effect rate of 4 g CTD/ha was used to assess relevance of exposures via deposition of dust. The overall weight of evidence suggested that there is minimal risk to honeybees from exposure to CTD from its use as a seed treatment. For exposures via dust, dust/seed and dust/foliar applications, there were no exposures greater than the NOAED for CTD in nectar and pollen, indicating a de minimis risk to honeybees when the route of exposure was via uptake in plants. Analysis of effect studies in the field indicated a consistent lack of relevant effects, regardless of the way CTD was applied. For exposures via dust, there were no adverse effects because of these applications and there were no exposures greater than the NOAED for CTD in nectar and pollen. The overall weight of evidence based on many studies indicated no adverse effects on colony viability or survival of the colony. Thus, the overall conclusion is that clothianidin, as currently used in good agricultural practices, does not present a significant risk to honeybees at the level of the colony.

Topics: Animals; Bees; Environmental Monitoring; Guanidines; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Risk Assessment; Thiamethoxam; Thiazoles; Toxicity Tests

This paper is a postscript to the four companion papers in this issue of the Journal (Solomon and Stephenson 2017a , 2017b ; Stephenson and Solomon 2017a , 2017b ). The first paper in the series described the conceptual model and the methods of the QWoE process. The other three papers described the application of the QWoE process to studies on imidacloprid (IMI), clothianidin (CTD), and thiamethoxam (TMX). This postscript was written to summarize the utility of the methods used in the quantitative weight of evidence (QWoE), the overall relevance of the results, and the environmental implications of the findings. Hopefully, this will be helpful to others who wish to conduct QWoEs and use these methods in assessment of risks.

Topics: Animals; Bees; Environmental Monitoring; Guanidines; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Risk Assessment; Seeds; Thiamethoxam; Thiazoles; Toxicity Tests

A quantitative weight of evidence (QWoE) methodology was used to assess several higher-tier studies on the effects of thiamethoxam (TMX) on honeybees. Assessment endpoints were population size and viability of commercially managed honeybee colonies and quantity of hive products. A higher-tier field toxicology study indicated a no-observed-adverse effect concentration (NOAEC) of 29.5 µg TMX/kg syrup, equivalent to an oral no-observed-adverse-effect-dose (NOAED) of 8.6 ng/bee/day for all responses measured. For exposures via deposition of dust, a conservative no-observed-adverse-effect-rate at the level of the colony was 0.1 g TMX/ha. There was minimal risk to honeybees from exposure to TMX via nectar and pollen from its use as a seed-treatment. For exposures via dust and dust/seed applications, there were no concentrations above the risk values for TMX in nectar and pollen. Although some risks were identified for potential exposures via guttation fluid, this route of exposure is incomplete; no apparent adverse effects were observed in field studies. For exposures via dust/seed and dust/foliar applications, few adverse effects were observed. Considering all lines of evidence, the quality of the studies included in this analysis was variable. However, the results of the studies were consistent and point to the same conclusion. The overall weight of evidence based on many studies indicates that TMX has no adverse effects on viability or survival of the colony. Thus, the overall conclusion is that the treatment of seeds with thiamethoxam, as currently used in good agricultural practices, does not present a significant risk to honeybees at the level of the colony.

Topics: Animals; Bees; Environmental Monitoring; Guanidines; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Risk Assessment; Thiamethoxam; Thiazoles; Toxicity Tests

A quantitative weight of evidence (QWoE) methodology was developed and used to assess many higher-tier studies on the effects of three neonicotinoid insecticides: clothianidin (CTD), imidacloprid (IMI), and thiamethoxam (TMX) on honeybees. A general problem formulation, a conceptual model for exposures of honeybees, and an analysis plan were developed. A QWoE methodology was used to characterize the quality of the available studies from the literature and unpublished reports of studies conducted by or for the registrants. These higher-tier studies focused on the exposures of honeybees to neonicotinoids via several matrices as measured in the field as well as the effects in experimentally controlled field studies. Reports provided by Bayer Crop Protection and Syngenta Crop Protection and papers from the open literature were assessed in detail, using predefined criteria for quality and relevance to develop scores (on a relative scale of 0-4) to separate the higher-quality from lower-quality studies and those relevant from less-relevant results. The scores from the QWoEs were summarized graphically to illustrate the overall quality of the studies and their relevance. Through mean and standard errors, this method provided graphical and numerical indications of the quality and relevance of the responses observed in the studies and the uncertainty associated with these two metrics. All analyses were conducted transparently and the derivations of the scores were fully documented. The results of these analyses are presented in three companion papers and the QWoE analyses for each insecticide are presented in detailed supplemental information (SI) in these papers.

Topics: Animals; Bees; Environmental Monitoring; Guanidines; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Research Design; Risk Assessment; Thiamethoxam; Thiazoles; Toxicity Tests

Nontarget aquatic insects are susceptible to chronic neonicotinoid insecticide exposure during the early stages of development from repeated runoff events and prolonged persistence of these chemicals. Investigations on the chronic toxicity of neonicotinoids to aquatic invertebrates have been limited to a few species and under different laboratory conditions that often preclude direct comparisons of the relative toxicity of different compounds. In the present study, full life-cycle toxicity tests using Chironomus dilutus were performed to compare the toxicity of 3 commonly used neonicotinoids: imidacloprid, clothianidin, and thiamethoxam. Test conditions followed a static-renewal exposure protocol in which lethal and sublethal endpoints were assessed on days 14 and 40. Reduced emergence success, advanced emergence timing, and male-biased sex ratios were sensitive responses to low-level neonicotinoid exposure. The 14-d median lethal concentrations for imidacloprid, clothianidin, and thiamethoxam were 1.52 μg/L, 2.41 μg/L, and 23.60 μg/L, respectively. The 40-d median effect concentrations (emergence) for imidacloprid, clothianidin, and thiamethoxam were 0.39 μg/L, 0.28 μg/L, and 4.13 μg/L, respectively. Toxic equivalence relative to imidacloprid was estimated through a 3-point response average of equivalencies calculated at 20%, 50%, and 90% lethal and effect concentrations. Relative to imidacloprid (toxic equivalency factor [TEF] = 1.0), chronic (lethality) 14-d TEFs for clothianidin and thiamethoxam were 1.05 and 0.14, respectively, and chronic (emergence inhibition) 40-d TEFs were 1.62 and 0.11, respectively. These population-relevant endpoints and TEFs suggest that imidacloprid and clothianidin exert comparable chronic toxicity to C. dilutus, whereas thiamethoxam induced comparable effects only at concentrations an order of magnitude higher. However, the authors caution that under field conditions, thiamethoxam readily degrades to clothianidin, thereby likely enhancing toxicity. Environ Toxicol Chem 2017;36:372-382. © 2016 SETAC.

Topics: Animals; Chironomidae; Dose-Response Relationship, Drug; Guanidines; Imidazoles; Insecticides; Invertebrates; Neonicotinoids; Nitro Compounds; Oxazines; Thiamethoxam; Thiazoles; Time Factors; Toxicity Tests; Water Pollutants, Chemical

Among the many factors responsible for the decline of bee populations are plant protection products such as neonicotinoids. In general, bees are exposed to not only one but mixtures of such chemicals. At environmental realistic concentrations neonicotinoids may display negative effects on the immune system, foraging activity, learning and memory formation of bees. Neonicotinoids induce alterations of gene transcripts such as nicotinic acetylcholine receptor (nAChR) subunits, vitellogenin, genes of the immune system and genes linked to memory formation. While previous studies focused on individual compounds, the effect of neonicotinoid mixtures in bees is poorly known. Here we investigated the effects of neonicotinoids acetamiprid, clothianidin, imidacloprid and thiamethoxam as single compounds, and binary mixtures thereof in honeybees. We determined transcriptional changes of nAChR subunits and vitellogenin in the brain of experimentally exposed honeybees after exposure up to 72 h. Exposure concentrations were selected on the basis of lowest effect concentrations of the single compounds. Transcriptional induction of nAChRs and vitellogenin was strongest for thiamethoxam, and weakest for acetamiprid. To a large extent, binary mixtures did not show additive transcriptional inductions but they were less than additive. Our data suggest that the joint transcriptional activity of neonicotinoids cannot be explained by concentration addition. The in vivo effects are not only governed by agonistic interaction with nAChRs alone, but are more complex as a result of interactions with other pathways as well. Further studies are needed to investigate the physiological joint effects of mixtures of neonicotinoids and other plant protection products on bees to better understand their joint effects.

Topics: Animals; Bees; Drug Interactions; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Pyridines; Receptors, Nicotinic; Thiamethoxam; Thiazoles; Transcription, Genetic

The widespread use of neonicotinoid insecticides and recent increased regulatory scrutiny requires the generation of monitoring data with sufficient scope and resolution to provide decision makers with a better understanding of occurrence and distribution in the environment. This study presents a wide-scale investigation of neonicotinoid insecticides used across the range of agricultural activities from fifteen surface water sites in southern Ontario. Using statistical analysis, the correlation of individual compounds with land use was investigated, and the relationship between neonicotinoid occurrence and hydrologic parameters in calibrated water courses was also assessed. Of the five neonicotinoids studied, imidacloprid, clothianidin and thiamethoxam exhibited detection rates above 90% at over half the sites sampled over a three year period (2012-2014). At two sites in southwestern Ontario, the Canadian Federal freshwater guideline value for imidacloprid (230 ng/L) was exceeded in roughly 75% of the samples collected. For some watersheds, there were correlations between the occurrence of neonicotinoids and precipitation and/or stream discharge. Some watersheds exhibited seasonal maxima in concentrations of neonicotinoids in spring and fall, particularly for those areas where row crop agriculture is predominant; these seasonal patterns were absent in some areas characterized by a broad range of agricultural activities.

Topics: Agriculture; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Ontario; Oxazines; Rivers; Seasons; Solubility; Thiamethoxam; Thiazoles; Water Pollutants, Chemical; Weather

Wheat aphid (Hemiptera: Aphididae) is one of the major pests of winter wheat and has posed a significant threat to winter wheat production in China. Although neonicotinoid insecticidal seed treatments have been suggested to be a control method, the season-long efficacy on pests and the impact on their natural enemies are still uncertain. Experiments were conducted to determine the efficacy of imidacloprid and clothianidin on the control of aphids, the number of their natural enemies and the emergence rate and yield of wheat during 2011-2014.. Imidacloprid and clothianidin seed treatments had no effect on the emergence rate of winter wheat and could prevent yield losses and wheat aphid infestations throughout the winter wheat growing season. Furthermore, their active ingredients were detected in winter wheat leaves up to 200 days after sowing. Imidacloprid and clothianidin seed treatments had no adverse effects on ladybirds, hoverflies or parasitoids, and instead increased the spider-aphid ratios.. Wheat seeds treated with imidacloprid and clothianidin were effective against wheat aphids throughout the winter wheat growing season and reduced the yield loss under field conditions. Imidacloprid and clothianidin seed treatments may be an important component of the integrated management of wheat aphids on winter wheat. © 2015 Society of Chemical Industry.

Topics: Animals; Aphids; Crop Protection; Guanidines; Imidazoles; Insect Control; Insecticides; Neonicotinoids; Nitro Compounds; Seeds; Thiazoles; Triticum

Seed treatment insecticides have become a popular management option for early-season insect control. This study investigated the total uptake and translocation of seed-applied [(14) C]imidacloprid, [(14) C]clothianidin and [(14) C]flupyradifurone into different plant parts in three soybean vegetative stages (VC, V1 and V2). The effects of soil moisture stress on insecticide uptake and translocation were also assessed among treatments. We hypothesized that (1) uptake and translocation would be different among the insecticides owing to differences in water solubility, and (2) moisture stress would increase insecticide uptake and translocation.. Uptake and translocation did not follow a clear trend in the three vegetative stages. Initially, flupyradifurone uptake was greater than clothianidin uptake in VC soybeans. In V1 soybeans, differences in uptake among the three insecticides were not apparent and unaffected by soil moisture stress. Clothianidin was negatively affected by soil moisture stress in V2 soybeans, while imidacloprid and flupyradifurone were unaffected. Specifically, soil moisture stress had a positive effect on the distribution of flupyradifurone in leaves. This was not observed with the neonicotinoids.. This study enhances our understanding of the uptake and distribution of insecticides used as seed treatments in soybean. The uptake and translocation of these insecticides differed in response to soil moisture stress. © 2015 Society of Chemical Industry.

Topics: 4-Butyrolactone; Glycine max; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Pyridines; Seeds; Soil; Thiazoles; Water

The water-foraging activity of honey bees (Apis mellifera L.) on guttation fluid of seed-coated crops, such as winter oilseed rape (WOR; Brassica napus L.), has not yet been evaluated. We analyzed the uptake of active substances (a.s.) in guttation fluid by evaluating residues of honey-sac contents. In autumn, insecticide residues of up to 130 µg a.s. per liter were released in WOR guttation fluid; this concentration is noticeably lower than levels reported in guttation fluid of seed-coated maize. Until winter dormancy, the concentrations declined to <30 µg a.s. per liter. In spring, residues were linked to prewintered plants and declined steadily until flowering. The maximum release of residues in guttation fluid of seed-coated WOR occurs on the first leaves in autumn when the colonies' water demand decreases. For the first time, proof for the uptake of guttation fluid from seed-coated WOR by honey bees was provided by measuring residues in individual honey-sac contents. In total, 38 out of 204 samples (19%) showed residues of thiamethoxam at concentrations ranging from 0.3 to 0.95 µg per liter while the corresponding concentrations in guttation fluid of WOR varied between 3.6 to 12.9 µg thiamethoxam per liter. The amounts of thiamethoxam we found in the honey sacs of water-foraging honey bees were therefore below the thresholds in nectar and pollen that are considered to have negative effects on honey bees after chronic exposure.

Topics: Animals; Bees; Brassica napus; Chromatography, High Pressure Liquid; Feeding Behavior; Germany; Guanidines; Imidazoles; Insecticides; Mass Spectrometry; Neonicotinoids; Nitro Compounds; Oxazines; Pesticide Residues; Plant Leaves; Plant Nectar; Pollen; Seasons; Thiamethoxam; Thiazoles; Water

A study was carried out to evaluate the possible presence of thiamethoxam, clothianidin and imidacloprid, as well as the metabolic breakdown products of these three neonicotinoids in pollen and honey obtained from brood chamber combs of honeybee colonies located next to sunflower and maize crops from coated seeds. Samples were analyzed by liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry detector, in combination with accurate mass tools such as diagnostic ions by exact mass, chlorine mass filters, and MS/MS experiments. The presence of thiamethoxam and clothianidin was confirmed in some of the pollen samples analyzed. Moreover, different metabolites of neonicotinoids were tentatively detected in the pollen and honey samples collected. The results suggested that four metabolites were found in the honey samples, while for pollen samples eleven metabolites were identified; among these, five were considered for the first time as metabolic breakdown products in sunflower and maize plants.

Topics: Animals; Chemistry Techniques, Analytical; Chromatography, Liquid; Crops, Agricultural; Guanidines; Helianthus; Honey; Imidazoles; Neonicotinoids; Nitro Compounds; Oxazines; Pollen; Reproducibility of Results; Seeds; Tandem Mass Spectrometry; Thiamethoxam; Thiazoles; Zea mays

Northern bobwhite (quail) (Colinus virginianus) and scaled quail (Callipepla squamata) populations have declined dramatically in the Rolling Plains ecoregion of Texas and Oklahoma (USA). There is rising concern about potential toxicity of neonicotinoids to birds. To investigate this concern, the authors examined crops of 81 northern bobwhite and 17 scaled quail to determine the presence or absence of seeds treated with 3 neonicotinoids (clothianidin, imidacloprid, and thiamethoxam). No treated seeds were found in the 98 crops examined. Liver samples from all 98 quail were collected and analyzed for neonicotinoid residues. Analysis revealed very low concentrations of neonicotinoids within the quail liver samples. The results suggest there is little to no risk of direct toxicity to quail from neonicotinoids. Environ Toxicol Chem 2016;35:1511-1515. © 2015 SETAC.

Topics: Animals; Colinus; Drug Residues; Environmental Exposure; Guanidines; Imidazoles; Insecticides; Liver; Mass Spectrometry; Neonicotinoids; Nitro Compounds; Oklahoma; Oxazines; Quail; Texas; Thiamethoxam; Thiazoles

Neonicotinoid insecticides (NIs) and their transformation products were detected in honey, pollen and honey bees, (Apis mellifera) from hives located within 30 km of the City of Saskatoon, Saskatchewan, Canada. Clothianidin and thiamethoxam were the most frequently detected NIs, found in 68 and 75% of honey samples at mean concentrations of 8.2 and 17.2 ng g(-1) wet mass, (wm), respectively. Clothianidin was also found in >50% of samples of bees and pollen. Concentrations of clothianidin in bees exceed the LD50 in 2 of 28 samples, while for other NIs concentrations were typically 10-100-fold less than the oral LD50. Imidaclorpid was detected in ∼30% of samples of honey, but only 5% of pollen and concentrations were

Topics: Animals; Bees; Guanidines; Honey; Imidazoles; Insecticides; Limit of Detection; Neonicotinoids; Nitro Compounds; Oxazines; Pollen; Saskatchewan; Seasons; Thiamethoxam; Thiazoles

A validated analytical method to determine seven neonicotinoids (dinotefuran, nitenpyram, thiamethoxam, clothianidin, imidacloprid, acetamiprid and thiacloprid) in sunflower seeds (hull and kernel) using HPLC coupled to electrospray ionisation mass spectrometry (ESI-MS) is presented. Sample clean-up based on a solid-liquid extraction, and the removal of lipid fraction, in the case of kernels, is proposed and optimised. Low limits of detection and quantification were obtained, ranging from 0.3 × 10(-3) to 1.2 × 10(-3) µg g(-1) and from 1.0 × 10(-3) to 4.0 × 10(-3) µg g(-1), with good precision, and recovery values ranged from 90% to 104% for hulls and kernels. The method was applied for the analysis of five thiamethoxam-dressed sunflower seeds and four non-treated seeds, where, besides thiamethoxam, residues of the other neonicotinoid, clothianidin, were also detected and confirmed via tandem mass spectrometry (LC-ESI-MS/MS). Finally, the presence of residues of thiamethoxam and clothianidin in collected sunflower seeds (hulls) coming from coated seeds confirmed the translocation of these neonicotinoids through the plant up to these seeds.

Topics: Chromatography, High Pressure Liquid; Guanidines; Helianthus; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Pesticide Residues; Pyridines; Seeds; Tandem Mass Spectrometry; Thiamethoxam; Thiazines; Thiazoles

Neonicotinoids, which are novel pesticides, have entered into usage around the world because they are selectively toxic to arthropods and relatively non-toxic to vertebrates. It has been suggested that several neonicotinoids cause neurodevelopmental toxicity in mammals. The aim was to establish the relationship between oral intake and urinary excretion of neonicotinoids by humans to facilitate biological monitoring, and to estimate dietary neonicotinoid intakes by Japanese adults.. Deuterium-labeled neonicotinoid (acetamiprid, clothianidin, dinotefuran, and imidacloprid) microdoses were orally ingested by nine healthy adults, and 24 h pooled urine samples were collected for 4 consecutive days after dosing. The excretion kinetics were modeled using one- and two-compartment models, then validated in a non-deuterium-labeled neonicotinoid microdose study involving 12 healthy adults. Increased urinary concentrations of labeled neonicotinoids were observed after dosing. Clothianidin was recovered unchanged within 3 days, and most dinotefuran was recovered unchanged within 1 day. Around 10% of the imidacloprid dose was excreted unchanged. Most of the acetamiprid was metabolized to desmethyl-acetamiprid. Spot urine samples from 373 Japanese adults were analyzed for neonicotinoids, and daily intakes were estimated. The estimated average daily intake of these neonicotinoids was 0.53-3.66 μg/day. The highest intake of any of the neonicotinoids in the study population was 64.5 μg/day for dinotefuran, and this was <1% of the acceptable daily intake.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Environmental Monitoring; Female; Guanidines; Humans; Imidazoles; Male; Middle Aged; Neonicotinoids; Nitro Compounds; Pesticides; Pyridines; Tandem Mass Spectrometry; Thiazoles; Young Adult

Concentrations of the neonicotinoid insecticides clothianidin, thiamethoxam and imidacloprid were determined in honey collected in Spring 2013 from a variety of locations in England. The honey was produced before the moratorium in the EU on the use of neonicotinoids in pollinator-attractive crops became effective.. Neither imidacloprid nor its metabolites were detected in any honey samples. Concentrations of clothianidin ranged from <0.02 to 0.82 µg kg(-1) , and thiamethoxam concentrations were between <0.01 and 0.79 µg kg(-1) .. Neonicotinoid concentrations were below those likely to cause any chronic mortality. The concentrations detected should provide a useful baseline against which the effectiveness of the moratorium in reducing exposure of honeybees can be measured. © 2016 Society of Chemical Industry.

Topics: Animals; Bees; England; Environmental Monitoring; Guanidines; Honey; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Pesticide Residues; Thiamethoxam; Thiazoles

A strong immune defense is vital for honey bee health and colony survival. This defense can be weakened by environmental factors that may render honey bees more vulnerable to parasites and pathogens. Honey bees are frequently exposed to neonicotinoid pesticides, which are being discussed as one of the stress factors that may lead to colony failure. We investigated the sublethal effects of the neonicotinoids thiacloprid, imidacloprid, and clothianidin on individual immunity, by studying three major aspects of immunocompetence in worker bees: total hemocyte number, encapsulation response, and antimicrobial activity of the hemolymph. In laboratory experiments, we found a strong impact of all three neonicotinoids. Thiacloprid (24h oral exposure, 200 μg/l or 2000 μg/l) and imidacloprid (1 μg/l or 10 μg/l) reduced hemocyte density, encapsulation response, and antimicrobial activity even at field realistic concentrations. Clothianidin had an effect on these immune parameters only at higher than field realistic concentrations (50-200 μg/l). These results suggest that neonicotinoids affect the individual immunocompetence of honey bees, possibly leading to an impaired disease resistance capacity.

Topics: Animals; Bees; Guanidines; Hemocytes; Hemolymph; Imidazoles; Immunocompetence; Neonicotinoids; Nitro Compounds; Pyridines; Thiazines; Thiazoles

The increasing structural diversity of the neonicotinoid class of insecticides presently used in crop protection calls for a more detailed analysis of their mode of action at their cellular targets, the nicotinic acetylcholine receptors.. Comparative radioligand binding studies using membranes of Myzus persicae (Sulzer) and representatives of the chloropyridyl subclass (imidacloprid), the chlorothiazolyl subclass (thiamethoxam), the tetrahydrofuranyl subclass (dinotefuran), as well as the novel sulfoximine type (sulfoxaflor), which is not a neonicotinoid, reveal significant differences in the number of binding sites, the displacing potencies and the mode of binding interference. Furthermore, the mode of interaction of [. There is no single conserved site or mode of binding of neonicotinoids and related nicotinic ligands to their target receptor, but a variety of binding pockets depending on the combination of receptor subunits, the receptor subtype, its functional state, as well as the structural flexibility of both the binding pockets and the ligands. © 2016 Society of Chemical Industry.

Topics: Animals; Aphids; Guanidines; Imidazoles; Insect Proteins; Insecticides; Neonicotinoids; Nicotinic Agonists; Nicotinic Antagonists; Nitro Compounds; Oxazines; Receptors, Nicotinic; Thiamethoxam; Thiazoles

There is growing concern over the risk to bee populations from neonicotinoid insecticides and the long-term consequences of reduced numbers of insect pollinators to essential ecosystem services and food security. Our knowledge of the risk of neonicotinoids to bees is based on studies of imidacloprid and thiamethoxam and these findings are extrapolated to clothianidin based on its higher potency at nicotinic acetylcholine receptors. This study addresses the specificity and consequences of all three neonicotinoids to determine their relative risk to bumblebees at field-relevant levels (2.5 ppb). We find compound-specific effects at all levels (individual cells, bees and whole colonies in semi-field conditions). Imidacloprid and clothianidin display distinct, overlapping, abilities to stimulate Kenyon cells, indicating the potential to differentially influence bumblebee behavior. Bee immobility was induced only by imidacloprid, and an increased vulnerability to clothianidin toxicity only occurred following chronic exposure to clothianidin or thiamethoxam. At the whole colony level, only thiamethoxam altered the sex ratio (more males present) and only clothianidin increased queen production. Finally, both imidacloprid and thiamethoxam caused deficits in colony strength, while no detrimental effects of clothianidin were observed. Given these findings, neonicotinoid risk needs to be considered independently for each compound and target species.

Topics: Animals; Bees; Behavior, Animal; Brain; Cells, Cultured; Chromatography, High Pressure Liquid; Guanidines; Insecticides; Neonicotinoids; Neurons; Nitro Compounds; Oxazines; Receptors, Nicotinic; Risk; Sex Ratio; Tandem Mass Spectrometry; Thiamethoxam; Thiazoles

Liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) is a primary tool for analysis of low volatility compounds in complex matrices. However, complex matrices, such as different types of tea, complicate analysis through ionization suppression or enhancement. In this study, sample preparation by a refined QuEChERS method combined with a dilution strategy removed almost all matrix effects caused by six types of tea. Tea samples were soaked with water and extracted with acetonitrile, cleaned up with a combination of PVPP (160mg) and GCB (20mg), and dried. Dried extracts were diluted with 20mL acetonitrile/water (15:85, v/v) before analysis by UPLC-MS/MS. The average recoveries of eight neonicotinoid insecticides (dinotefuran, nitenpyram, thiamethoxam, imidacloprid, clothianidin, imidaclothiz, acetamiprid, and thiacloprid) ranged from 66.3 to 108.0% from tea samples spiked at 0.01-0.5mgkg(-1). Relative standard deviations were below 16% for all recovery tests. The limit of quantification ranged from 0.01 to 0.05mgkg(-1).

Topics: Chromatography, Liquid; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Pyridines; Tandem Mass Spectrometry; Tea; Thiamethoxam; Thiazines; Thiazoles

This study evaluated the applicability of commercially available kit-based enzyme-linked immunosorbent assay (ELISA) to simple, quick, and quantitative detection for three water-extractable (phytoavailable) neonicotinoid insecticides: dinotefuran, clothianidin, and imidacloprid in soils. ELISA showed excellent analytical sensitivity for determination, but with cross-reaction to structurally related neonicotinoid analogues, which might produce false positives. To analyze insecticides in soil samples of diverse physicochemical properties, they were extracted with water. The aqueous soil extracts were assayed directly with ELISA. No matrix interference was observed without additional dilution with water. Recovery experiments for the insecticides from aqueous soil extracts spiked at 2-10 ng/mL showed good accuracy (72-126%) and precision (<16%). Kit-based ELISAs were used to estimate soil-water distribution coefficients (Kd). Values estimated using this method showed positive correlation between organic carbon contents in soil and those for evaluated insecticides. Results indicate that the evaluated kit-based ELISA has applicability for simple, quick, and reliable detection of phytoavailable insecticides in soils and for estimating Kd values in soil.

Topics: Crops, Agricultural; Cross Reactions; Environmental Monitoring; Enzyme-Linked Immunosorbent Assay; Guanidines; Imidazoles; Insecticides; Japan; Neonicotinoids; Nitro Compounds; Soil; Soil Pollutants; Thiazoles

Neonicotinoid insecticides can be transported from agricultural fields, where they are used as foliar sprays or seed treatments, to surface waters by surface or sub-surface runoff. Few studies have investigated the toxicity of neonicotinoid or the related butenolide insecticides to freshwater mollusk species. The current study examined the effect of neonicotinoid and butenolide exposures to the early-life stages of the ramshorn snail, Planorbella pilsbryi, and the wavy-rayed lampmussel, Lampsilis fasciola. Juvenile P. pilsbryi were exposed to imidacloprid, clothianidin, or thiamethoxam for 7 or 28 d and mortality, growth, and biomass production were measured. The viability of larval (glochidia) L. fasciola was monitored during a 48 h exposure to six neonicotinoids (imidacloprid, thiamethoxam, clothianidin, acetamiprid, thiacloprid, or dinotefuran), or a butenolide (flupyradifurone). The 7-d LC50s of P. pilsbryi for imidacloprid, clothianidin, and thiamethoxam were ≥4000 μg/L and the 28-d LC50s were ≥182 μg/L. Growth and biomass production were considerably more sensitive endpoints than mortality with EC50s ranging from 33.2 to 122.0 μg/L. The 48-h LC50s for the viability of glochidia were ≥456 μg/L for all seven insecticides tested. Our data indicate that neonicotinoid and butenolide insecticides pose less of a hazard with respect to mortality of the two species of mollusk compared to the potential hazard to other non-target aquatic insects.

Topics: 4-Butyrolactone; Animals; Environmental Monitoring; Fresh Water; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Pyridines; Thiamethoxam; Thiazines; Thiazoles; Unionidae; Water Pollutants, Chemical

The brown planthopper is a serious rice pest in China. Chemical insecticides have been considered a satisfactory means of controlling the brown planthopper. In the present study, we determined the susceptibility of twenty-one populations of Nilaparvata lugens to eleven insecticides by a rice-stem dipping method from 2012 to 2014 in eight provinces of China. These field-collected populations of N. lugens had developed high levels of resistance to imidacloprid (resistant ratio, RR=233.3-2029-fold) and buprofezin (RR=147.0-1222). Furthermore, N. lugens showed moderate to high levels of resistance to thiamethoxam (RR=25.9-159.2) and low to moderate levels of resistance to dinotefuran (RR=6.4-29.1), clothianidin (RR=6.1-33.6), ethiprole (RR=11.5-71.8), isoprocarb (RR=17.1-70.2), and chlorpyrifos (RR=7.4-30.7). In contrast, the susceptibility of N. lugens to etofenprox (RR=1.1-4.9), thiacloprid (RR=2.9-8.2) and acetamiprid (RR=2.7-26.2) remained susceptible to moderate levels of resistance. Significant correlations were detected between the LC50 values of imidacloprid and thiamethoxam, dinotefuran, buprofezin, and etofenprox, as well as between clothianidin and thiamethoxam, dinotefuran, ethiprole, acetamiprid, and thiacloprid. Similarly, significant correlations were observed between chlorpyrifos and etofenprox, acetamiprid and thiacloprid. Additionally, the activity of the detoxification enzymes of N. lugens showed a significant correlation with the log LC50 values of imidacloprid, dinotefuran and ethiprole. These results will be beneficial for effective insecticide resistance management strategies to prevent or delay the development of insecticide resistance.

Topics: Animals; Carbamates; China; Chlorpyrifos; Guanidines; Hemiptera; Imidazoles; Insecticide Resistance; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Pyrazoles; Pyrethrins; Pyridines; Thiadiazines; Thiamethoxam; Thiazines; Thiazoles

A survey of insecticide resistance in over 150 Canadian populations of Colorado potato beetle was completed between 2008 and 2011. Three neonicotinoid and two anthranilic diamide insecticides were tested at a discriminating concentration (DC) with second-instar larvae in a leaf-disc bioassay.. The mean mortality for the imidacloprid (Admire) DC was 46-67% between 2008 and 2011 respectively. Over the 4 years, 10-46% and 26-40% of the populations were classified as resistant or showed reduced susceptibility to imidacloprid. The mean mortality for thiamethoxam (Actara) and clothianidin (Poncho/Titan) ranged from 56-76% in 2008 to 81-84% in 2010 for each insecticide respectively, indicating continuous susceptibility to clothianidin but reduced susceptibility to thiamethoxam. In 2008 and 2009, susceptibility to chlorantraniliprole (Coragen) was observed in 85% of populations. Similarly, cyantraniliprole (Cyazypyr) affected 93% of the 2009 and 74% of the 2010 populations. There was a significant (P < 0.05) and high positive correlation (R = 0.4-0.84) between the three neonicotinoids, indicating the potential for cross-resistance.. The trend observed in decreasing susceptibility for thiamethoxam and clothianidin will continue unless resistance management practices are followed.

Topics: Animals; Canada; Coleoptera; Guanidines; Imidazoles; Insecticide Resistance; Insecticides; Larva; Neonicotinoids; Nitro Compounds; ortho-Aminobenzoates; Oxazines; Pyrazoles; Thiamethoxam; Thiazoles

The objective of this study was to quantify whether the presence of three different neonicotinoid insecticides (imidacloprid, thiamethoxam or clothianidin) in sucrose solution results in antifeedant effects in individual worker bumblebees (Bombus terrestris), and, if so, whether this effect is reversible if bees are subsequently offered untreated feed.. Bees exposed to imidacloprid displayed a significant dose-dependent reduction in consumption at 10 and 100 µg L(-1), which was reversed when untreated feed was offered. No consistent avoidance/antifeedant response to nectar substitute with thiamethoxam was detected at the more field-realistic dose rates of 1 and 10 µg L(-1), and exposure to the very high 100 µg L(-1) dose rate was followed by 100% mortality of experimental insects. No reduction in food intake was recorded at 1 µg clothianidin L(-1), reduced consumption was noted at 10 µg clothianidin L(-1) and 100% mortality occurred when bees were exposed to rates of 100 µg clothianidin L(-1).. This study provides evidence of a direct antifeedant effect of imidacloprid and clothianidin in individual bumblebees but highlights that this may be a compound-specific effect.

Topics: Animals; Bees; Eating; Feeding Behavior; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Sucrose; Thiamethoxam; Thiazoles

Systemic neonicotinoid insecticides are used to control turfgrass insect pests. The authors tested their transference into nectar of flowering lawn weeds or grass guttation droplets, which, if high enough, could be hazardous to bees or other insects that feed on such exudates. The authors applied imidacloprid or clothianidin to turf with white clover, followed by irrigation, and used liquid chromatography-tandem mass spectrometry (LC-MS/MS) to analyze residues in clover blooms that were directly sprayed during application or that formed after the first mowing. Imidacloprid residues in guttation fluid from field-grown creeping bentgrass were assessed similarly. The authors used Orius insidiosus, a small anthrocorid bug that is sensitive to dietary neonicotinoids, as a bioindicator of the exudates' toxicity. Nectar from directly sprayed clover blooms contained 5493 ng/g to 6588 ng/g imidacloprid or 2882 ng/g to 2992 ng/g clothianidin and was acutely toxic to Orius. Residues were 99.4% to 99.8% lower in nectar of blooms formed after mowing, and nontoxic to Orius. Imidacloprid residues in turfgrass guttation averaged 88 ng/g at 1 wk after treatment, causing some intoxication of Orius, but declined to 23 ng/g within 3 wk. Systemic transference of neonicotinoids into white clover nectar and creeping bentgrass guttation appears relatively low and transitory. The hazard to nontarget insects via nectar of flowering weeds in treated lawns can be mitigated by adhering to label precautions and mowing to remove blooms if they are inadvertently sprayed.

Topics: Animals; Flowers; Guanidines; Hemiptera; Imidazoles; Insect Control; Insecticides; Medicago; Neonicotinoids; Nitro Compounds; Pesticide Residues; Plant Nectar; Poaceae; Reproduction; Tandem Mass Spectrometry; Thiazoles

High sensitivity to neonicotinoid insecticides have been reported in the miridbug Cyrtorhinus lividipennis, an important predatory enemy of rice planthoppers, such as Nilaparvata lugens (brown planthopper). In the present study, the sensitivity of neonicotinoid insecticides between C. lividipennis and N. lugens were detected and compared. The results showed that neonicotinoid insecticides were much more toxic to the miridbug than to the brown planthopper. A nicotinic acetylcholine receptor subunit was cloned from the miridbug and denoted as α8 subunit (Clα8) according to sequence similarities and important functional motifs. Key amino acid differences were found in specific loops from α8 subunits between C. lividipennis (Clα8) and N. lugens (Nlα8). In order to understand the roles of key amino acid differences in insecticide sensitivities, the different amino acid residues in specific loops of Nlα8 were introduced into the corresponding sites in Clα8 to construct several subunit mutants. Clα8 or subunit mutants were co-expressed with rat β2 to obtain the functional receptors in Xenopus oocytes. The single mutation N191F in loop B reduced imidacloprid sensitivity, with EC50 value in Clα8(N191F)/β2 of 15.21μM and 5.74μM in Clα8/β2. Interestingly, although the single mutation E240T in loop C did not cause the significant change in imidacloprid sensitivity, it could enhance the effects of N191F and cause more decrease in imidacloprid sensitivity. The results indicated that E240T might contribute to neonicotinoid sensitivity in an indirect way.

Topics: Amino Acid Sequence; Animals; Female; Guanidines; Hemiptera; Heteroptera; Imidazoles; Insect Proteins; Insecticides; Molecular Sequence Data; Mutation; Neonicotinoids; Nitro Compounds; Oocytes; Protein Subunits; Pyridines; Rats; Receptors, Cholinergic; Species Specificity; Thiazines; Thiazoles; Xenopus

The global decline in the abundance and diversity of insect pollinators could result from habitat loss, disease, and pesticide exposure. The contribution of the neonicotinoid insecticides (e.g., clothianidin and imidacloprid) to this decline is controversial, and key to understanding their risk is whether the astonishingly low levels found in the nectar and pollen of plants is sufficient to deliver neuroactive levels to their site of action: the bee brain. Here we show that bumblebees (Bombus terrestris audax) fed field levels [10 nM, 2.1 ppb (w/w)] of neonicotinoid accumulate between 4 and 10 nM in their brains within 3 days. Acute (minutes) exposure of cultured neurons to 10 nM clothianidin, but not imidacloprid, causes a nicotinic acetylcholine receptor-dependent rapid mitochondrial depolarization. However, a chronic (2 days) exposure to 1 nM imidacloprid leads to a receptor-dependent increased sensitivity to a normally innocuous level of acetylcholine, which now also causes rapid mitochondrial depolarization in neurons. Finally, colonies exposed to this level of imidacloprid show deficits in colony growth and nest condition compared with untreated colonies. These findings provide a mechanistic explanation for the poor navigation and foraging observed in neonicotinoid treated bumblebee colonies.

Topics: Acetylcholinesterase; Animals; Apoptosis; Bees; Brain; Cell Proliferation; Cells, Cultured; Chromatography, Liquid; Guanidines; Imidazoles; Insecticides; Isotope Labeling; Membrane Potential, Mitochondrial; Mitochondria; Neonicotinoids; Neurons; Nitro Compounds; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Thiazoles

The impact of neonicotinoid insecticides on insect pollinators is highly controversial. Sublethal concentrations alter the behaviour of social bees and reduce survival of entire colonies. However, critics argue that the reported negative effects only arise from neonicotinoid concentrations that are greater than those found in the nectar and pollen of pesticide-treated plants. Furthermore, it has been suggested that bees could choose to forage on other available flowers and hence avoid or dilute exposure. Here, using a two-choice feeding assay, we show that the honeybee, Apis mellifera, and the buff-tailed bumblebee, Bombus terrestris, do not avoid nectar-relevant concentrations of three of the most commonly used neonicotinoids, imidacloprid (IMD), thiamethoxam (TMX), and clothianidin (CLO), in food. Moreover, bees of both species prefer to eat more of sucrose solutions laced with IMD or TMX than sucrose alone. Stimulation with IMD, TMX and CLO neither elicited spiking responses from gustatory neurons in the bees' mouthparts, nor inhibited the responses of sucrose-sensitive neurons. Our data indicate that bees cannot taste neonicotinoids and are not repelled by them. Instead, bees preferred solutions containing IMD or TMX, even though the consumption of these pesticides caused them to eat less food overall. This work shows that bees cannot control their exposure to neonicotinoids in food and implies that treating flowering crops with IMD and TMX presents a sizeable hazard to foraging bees.

Topics: Animals; Bees; Chemoreceptor Cells; Diet; Female; Flowers; Food Preferences; Guanidines; Imidazoles; Insecticides; Male; Neonicotinoids; Nitro Compounds; Oxazines; Plant Nectar; Pollen; Pollination; Reproduction; Survival Analysis; Taste; Thiamethoxam; Thiazoles

Topics: Animals; Bees; Evaluation Studies as Topic; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Pollination; Population Dynamics; Thiamethoxam; Thiazoles; Uncertainty

Topics: Agriculture; Animals; Bees; Conflict of Interest; European Union; Evaluation Studies as Topic; Female; Guanidines; Homing Behavior; Humans; Imidazoles; Insecticides; Male; Mammals; Neonicotinoids; Nitro Compounds; Orientation; Oxazines; Pollination; Reproducibility of Results; Spatial Navigation; Thiamethoxam; Thiazoles; Uncertainty; United States; Varroidae

To accurately estimate exposure of bees to pesticides, analytical methods are needed to enable quantification of nanogram/gram (ng/g) levels of contaminants in small samples of pollen or the individual insects. A modified QuEChERS extraction method coupled with ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) analysis was tested to quantify residues of 19 commonly used neonicotinoids and fungicides and the synergist, piperonyl butoxide, in 100 mg samples of pollen and in samples of individual bumblebees (Bombus terrestris). Final recoveries ranged from 71 to 102 % for most compounds with a repeatability of below 20 % for both pollen and bumblebee extracts spiked at 5 and 40 ng/g. The method enables the detection of all compounds at sub-ng/g levels in both matrices and the method detection limits (MDL) ranged from 0.01 to 0.84 ng/g in pollen and 0.01 to 0.96 ng/g in individual bumblebees. Using this method, mixtures of neonicotinoids (thiamethoxam, clothianidin, imidacloprid and thiacloprid) and fungicides (carbendazim, spiroxamine, boscalid, tebuconazole, prochloraz, metconazole, fluoxastrobin, pyraclostrobin and trifloxystrobin) were detected in pollens of field bean, strawberry and raspberry at concentrations ranging from MDL, and in some bees, the fungicides carbendazim, boscalid, tebuconazole, flusilazole and metconazole were present at concentrations between 0.80 to 30 ng/g. This new method allows the analysis of mixtures of neonicotinoids and fungicides at trace levels in small quantities of pollen and individual bumblebees and thus will facilitate exposure assessment studies.

Topics: Animals; Bees; Chromatography, High Pressure Liquid; Crops, Agricultural; Fragaria; Fungicides, Industrial; Guanidines; Imidazoles; Insecticides; Limit of Detection; Neonicotinoids; Nitro Compounds; Oxazines; Pollen; Pyridines; Rubus; Tandem Mass Spectrometry; Thiamethoxam; Thiazines; Thiazoles

Protection of green ash trees (Fraxinus pennsylvanica Marshall) from the emerald ash borer (EAB), Agrilus planipennis Fairmaire, by soil applications of neonicotinoids (imidacloprid, clothianidin, and dinotefuran) was tested at five locations between 2005 and 2013. Application rate and spring versus fall application dates were evaluated in tests with neighborhood street trees and in one plantation of 65 ash trees. Insecticide treatments of ash trees at all five sites were initiated as the leading edge of the EAB invasion began to kill the first ash trees at each location. Trees were treated and evaluated at each site for 4 to 7 yr. Spring applications of imidacloprid were more efficacious than fall applications. Application rates of 0.8 g a.i./cm dbh or greater per year gave a higher level of protection and were more consistent than rates of 0.56 g a.i./cm dbh per year or less. The number of years between the first observation of canopy loss due to EAB and death of most of the control trees varied from three to seven years among test sites, depending on how many non-treated ash trees were nearby.

Topics: Animals; Coleoptera; Fraxinus; Guanidines; Imidazoles; Insect Control; Insecticides; Larva; Neonicotinoids; Nitro Compounds; Ohio; Soil; Thiazoles

Over the last two decades, usage of neonicotinoid (NEO) insecticides has increased due to their high selectivity for insects versus mammals and their effectiveness for extermination of insects resistant to conventional pesticides such as pyrethroids and organophosphates (OPs). However, historical change of the NEO exposure level in humans is poorly understood. The aim of this study is to reveal changes in the levels of NEO and OP exposure in the human body over the last two decades using biomonitoring technique. We quantified urinary concentrations of 7 NEOs (acetamiprid, clothianidin, dinotefuran, imidacloprid, nitenpyram, thiacloprid, and thiamethoxam) and 4 metabolites of OPs (dimethylphosphate, dimethylthiophosphate, diethylphosphate, and diethylthiophosphate) in 95 adult females aged 45-75 in 1994, 2000, 2003, 2009, and 2011 (n = 17-20 different individuals in each year). The results show that the detection rates of urinary NEOs in Japanese women increased significantly between 1994 and 2011, suggesting that intakes of NEOs into the human body rose during that period. In contrast, exposure to OPs having O,O-dimethyl moieties decreased steadily according to a finding that geometric means of urinary dimethylphosphate concentrations kept diminishing considerably. These changes may reflect the amounts of NEOs and OPs used as insecticides in Japan.

Topics: Adult; Aged; Animals; Environmental Monitoring; Environmental Pollutants; Female; Guanidines; Humans; Imidazoles; Insecticides; Japan; Middle Aged; Neonicotinoids; Nitro Compounds; Organophosphates; Organophosphorus Compounds; Oxazines; Pyridines; Thiamethoxam; Thiazines; Thiazoles

Increasing and widespread use of neonicotinoid insecticides all over the world, together with their environmental persistence mean that surface and ground waters need to be monitored regularly for their residues. However, current multi-residue analytical methods for waters are inadequate for trace residue analysis of these compounds, while passive sampling devices are unavailable. A new method using UltraPerformance Liquid Chromatography provided good separation of the five most common neonicotinoid compounds, with limits of quantitation in the range 0.6-1.0ng. The method was tested in a survey of rivers around Sydney (Australia), where 93% of samples contained two or more neonicotinoids in the range 0.06-4.5μgL(-1). Styrenedivinylbenzene-reverse phase sulfonated Empore™ disks were selected as the best matrix for use in passive samplers. Uptake of clothianidin, imidacloprid and thiacloprid in a flow-through laboratory system for 3weeks was linear and proportional to their water concentrations over the range 1-10μgL(-1). Sampling rates of 8-15mLd(-1) were correlated to the hydrophobicity of the individual compounds. The passive samplers and analytical methods presented here can detect trace concentrations of neonicotinoids in water.

Topics: Chromatography, Liquid; Environmental Monitoring; Guanidines; Imidazoles; Insecticides; Neonicotinoids; New South Wales; Nitro Compounds; Pyridines; Rivers; Thiazines; Thiazoles; Water Pollutants, Chemical

Topics: Animals; Bees; Conservation of Natural Resources; Drug Evaluation; Europe; Guanidines; Imidazoles; Legislation, Drug; Neonicotinoids; Nitro Compounds; Oxazines; Pesticides; Risk Assessment; Thiamethoxam; Thiazoles

In an 11-week greenhouse study, caged queenright colonies of Bombus impatiens Cresson, were fed treatments of 0 (0 ppb actual residue I, imidacloprid; C, clothianidin), 10 (14 I, 9 C), 20 (16 I, 17C), 50 (71 I, 39 C) and 100 (127 I, 76 C) ppb imidacloprid or clothianidin in sugar syrup (50%). These treatments overlapped the residue levels found in pollen and nectar of many crops and landscape plants, which have higher residue levels than seed-treated crops (less than 10 ppb, corn, canola and sunflower). At 6 weeks, queen mortality was significantly higher in 50 ppb and 100 ppb and by 11 weeks in 20 ppb-100 ppb neonicotinyl-treated colonies. The largest impact for both neonicotinyls starting at 20 (16 I, 17 C) ppb was the statistically significant reduction in queen survival (37% I, 56% C) ppb, worker movement, colony consumption, and colony weight compared to 0 ppb treatments. Bees at feeders flew back to the nest box so it appears that only a few workers were collecting syrup in the flight box and returning the syrup to the nest. The majority of the workers sat immobilized for weeks on the floor of the flight box without moving to fed at sugar syrup feeders. Neonicotinyl residues were lower in wax pots in the nest than in the sugar syrup that was provided. At 10 (14) ppb I and 50 (39) ppb C, fewer males were produced by the workers, but queens continued to invest in queen production which was similar among treatments. Feeding on imidacloprid and clothianidin can cause changes in behavior (reduced worker movement, consumption, wax pot production, and nectar storage) that result in detrimental effects on colonies (queen survival and colony weight). Wild bumblebees depending on foraging workers can be negatively impacted by chronic neonicotinyl exposure at 20 ppb.

Topics: Animals; Bees; Exploratory Behavior; Feeding Behavior; Female; Guanidines; Imidazoles; Insecticides; Longevity; Male; Neonicotinoids; Nesting Behavior; Nitro Compounds; Plant Nectar; Thiazoles

Three neonicotinoids, imidacloprid, clothianidin and thiacloprid, agonists of the nicotinic acetylcholine receptor in the central brain of insects, were applied at non-lethal doses in order to test their effects on honeybee navigation. A catch-and-release experimental design was applied in which feeder trained bees were caught when arriving at the feeder, treated with one of the neonicotinoids, and released 1.5 hours later at a remote site. The flight paths of individual bees were tracked with harmonic radar. The initial flight phase controlled by the recently acquired navigation memory (vector memory) was less compromised than the second phase that leads the animal back to the hive (homing flight). The rate of successful return was significantly lower in treated bees, the probability of a correct turn at a salient landscape structure was reduced, and less directed flights during homing flights were performed. Since the homing phase in catch-and-release experiments documents the ability of a foraging honeybee to activate a remote memory acquired during its exploratory orientation flights, we conclude that non-lethal doses of the three neonicotinoids tested either block the retrieval of exploratory navigation memory or alter this form of navigation memory. These findings are discussed in the context of the application of neonicotinoids in plant protection.

Topics: Animals; Bees; Exploratory Behavior; Feeding Behavior; Flight, Animal; Guanidines; Homing Behavior; Imidazoles; Memory; Neonicotinoids; Nicotinic Agonists; Nitro Compounds; Orientation; Pyridines; Radar; Receptors, Nicotinic; Spatial Navigation; Thiazines; Thiazoles

Neonicotinoid insecticides act on nicotinic acetylcholine receptor and are particularly effective against sucking pests. They are widely used in crops protection to fight against aphids, which cause severe damage. In the present study we evaluated the susceptibility of the pea aphid Acyrthosiphon pisum to the commonly used neonicotinoid insecticides imidacloprid (IMI), thiamethoxam (TMX) and clothianidin (CLT). Binding studies on aphid membrane preparations revealed the existence of high and low-affinity binding sites for [3H]-IMI (Kd of 0.16 ± 0.04 nM and 41.7 ± 5.9 nM) and for the nicotinic antagonist [125I]-α-bungarotoxin (Kd of 0.008 ± 0.002 nM and 1.135 ± 0.213 nM). Competitive binding experiments demonstrated that TMX displayed a higher affinity than IMI for [125I]-α-bungarotoxin binding sites while CLT affinity was similar for both [125I]-α-bungarotoxin and [3H]-IMI binding sites. Interestingly, toxicological studies revealed that at 48 h, IMI (LC50 = 0.038 µg/ml) and TMX (LC50 = 0.034 µg/ml) were more toxic than CLT (LC50 = 0.118 µg/ml). The effect of TMX could be associated to its metabolite CLT as demonstrated by HPLC/MS analysis. In addition, we found that aphid larvae treated either with IMI, TMX or CLT showed a strong variation of nAChR subunit expression. Using semi-quantitative PCR experiments, we detected for all insecticides an increase of Apisumα10 and Apisumβ1 expressions levels, whereas Apisumβ2 expression decreased. Moreover, some other receptor subunits seemed to be differently regulated according to the insecticide used. Finally, we also demonstrated that nAChR subunit expression differed during pea aphid development. Altogether these results highlight species specificity that should be taken into account in pest management strategies.

Topics: Animals; Aphids; Binding Sites; Bungarotoxins; Cell Membrane; Chromatography, High Pressure Liquid; Guanidines; Imidazoles; Insecticides; Larva; Neonicotinoids; Nicotinic Antagonists; Nitro Compounds; Oxazines; Protein Binding; Protein Subunits; Receptors, Nicotinic; Tandem Mass Spectrometry; Thiamethoxam; Thiazoles

Concentrations of the neonicotinoid insecticides clothianidin, thiamethoxam and imidacloprid were determined in arable soils from a variety of locations in England.. In soil samples taken from the central area of fields, concentrations of clothianidin ranged from 0.02 to 13.6 µg kg(-1) . Thiamethoxam concentrations were between <0.02 and 1.50 µg kg(-1) , and imidacloprid concentrations between <0.09 and 10.7 µg kg(-1) . Concentrations of clothianidin and thiamethoxam were lower in soil samples taken from the edges of fields than from the centres of fields, but this difference was less pronounced for imidacloprid.. This work gives a clear indication of the levels of neonicotinoids in arable soils after typical use of these compounds as seed dressings in the United Kingdom. There was evidence that imidacloprid was more persistent in the soils studied than clothianidin and thiamethoxam. As clothianidin and thiamethoxam have largely superseded imidacloprid in the United Kingdom, neonicotinoid levels were lower than suggested by predictions based on imidacloprid alone.

Topics: England; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Pesticide Residues; Soil Pollutants; Thiamethoxam; Thiazoles

Research was done during 2012 to evaluate the potential exposure of pollinators to neonicotinoid insecticides used as seed treatments on corn, cotton, and soybean. Samples were collected from small plot evaluations of seed treatments and from commercial fields in agricultural production areas in Arkansas, Mississippi, and Tennessee. In total, 560 samples were analyzed for concentrations of clothianidin, imidacloprid, thiamethoxam, and their metabolites. These included pollen from corn and cotton, nectar from cotton, flowers from soybean, honey bees, Apis mellifera L., and pollen carried by foragers returning to hives, preplanting and in-season soil samples, and wild flowers adjacent to recently planted fields. Neonicotinoid insecticides were detected at a level of 1 ng/g or above in 23% of wild flower samples around recently planted fields, with an average detection level of about 10 ng/g. We detected neonicotinoid insecticides in the soil of production fields prior to planting at an average concentration of about 10 ng/g, and over 80% of the samples having some insecticide present. Only 5% of foraging honey bees tested positive for the presence of neonicotinoid insecticides, and there was only one trace detection (< 1 ng/g) in pollen being carried by those bees. Soybean flowers, cotton pollen, and cotton nectar contained little or no neonicotinoids resulting from insecticide seed treatments. Average levels of neonicotinoid insecticides in corn pollen ranged from less than 1 to 6 ng/g. The highest neonicotinoid concentrations were found in soil collected during early flowering from insecticide seed treatment trials. However, these levels were generally not well correlated with neonicotinoid concentrations in flowers, pollen, or nectar. Concentrations in flowering structures were well below defined levels of concern thought to cause acute mortality in honey bees. The potential implications of our findings are discussed.

Topics: Animals; Arkansas; Bees; Environmental Monitoring; Flowers; Glycine max; Gossypium; Guanidines; Imidazoles; Insecticides; Mississippi; Neonicotinoids; Nitro Compounds; Oxazines; Plant Nectar; Pollen; Pollination; Seeds; Soil Pollutants; Tennessee; Thiamethoxam; Thiazoles; Zea mays

Systemic pesticides such as neonicotinoids are commonly used on flowering crops visited by pollinators, and their use has been implicated in the decline of insect pollinator populations in Europe and North America. Several studies show that neonicotinoids affect navigation and learning in bees but few studies have examined whether these substances influence their basic motor function. Here, we investigated how prolonged exposure to sublethal doses of four neonicotinoid pesticides (imidacloprid, thiamethoxam, clothianidin, dinotefuran) and the plant toxin, nicotine, affect basic motor function and postural control in foraging-age worker honeybees. We used doses of 10 nM for each neonicotinoid: field-relevant doses that we determined to be sublethal and willingly consumed by bees. The neonicotinoids were placed in food solutions given to bees for 24 h. After the exposure period, bees were more likely to lose postural control during the motor function assay and fail to right themselves if exposed to imidacloprid, thiamethoxam, clothianidin. Bees exposed to thiamethoxam and nicotine also spent more time grooming. Other behaviours (walking, sitting and flying) were not significantly affected. Expression of changes in motor function after exposure to imidacloprid was dose-dependent and affected all measured behaviours. Our data illustrate that 24 h exposure to sublethal doses of neonicotinoid pesticides has a subtle influence on bee behaviour that is likely to affect normal function in a field setting.

Topics: Animals; Bees; Behavior, Animal; Guanidines; Imidazoles; Insecticides; Motor Activity; Neonicotinoids; Nitro Compounds; Oxazines; Posture; Thiamethoxam; Thiazoles; Toxicity Tests, Acute

Polymorphisms are sometimes observed in native insect nicotinic acetylcholine receptor (nAChR) subunits, which are important insecticide targets, yet little is known of their impact on insecticide actions. Here we investigated the effects of a polymorphism involving the substitution of histidine108 by leucine in the Drosophila melanogaster Dα1 subunit on the agonist actions of the neurotransmitter acetylcholine (ACh) and two commercial neonicotinoid insecticides (imidacloprid and clothianidin). There was no significant impact of the H108L substitution on either the ACh EC50, the concentration leading to a half maximal ACh response, or the maximum current amplitude in response at 10 μM ACh, of the Dα1-chicken β2 nAChR expressed in Xenopus laevis oocytes. However, the response amplitudes to imidacloprid and clothianidin were significantly enhanced, indicating a role of His108 in the selective interactions of Dα1 with these neonicotinoids.

Topics: Acetylcholine; Animals; Chickens; Drosophila melanogaster; Drosophila Proteins; Female; Gene Expression; Guanidines; Imidazoles; Models, Molecular; Neonicotinoids; Nitro Compounds; Oocytes; Polymorphism, Genetic; Protein Conformation; Receptors, Nicotinic; Recombinant Fusion Proteins; Thiazoles; Xenopus laevis

Neonicotinoid insecticides are of environmental concern, but little is known about their occurrence in surface water. An area of intense corn and soybean production in the Midwestern United States was chosen to study this issue because of the high agricultural use of neonicotinoids via both seed treatments and other forms of application. Water samples were collected from nine stream sites during the 2013 growing season. The results for the 79 water samples documented similar patterns among sites for both frequency of detection and concentration (maximum:median) with clothianidin (75%, 257 ng/L:8.2 ng/L) > thiamethoxam (47%, 185 ng/L:<2 ng/L) > imidacloprid (23%, 42.7 ng/L: <2 ng/L). Neonicotinoids were detected at all nine sites sampled even though the basin areas spanned four orders of magnitude. Temporal patterns in concentrations reveal pulses of neonicotinoids associated with rainfall events during crop planting, suggesting seed treatments as their likely source.

Topics: Agriculture; Animals; Environmental Monitoring; Glycine max; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Rivers; Seeds; Thiamethoxam; Thiazoles; Water Pollutants, Chemical; Zea mays

Agricultural use of neonicotinoid (NEO) insecticides has been increasing in recent years, but their biological monitoring methods have been scarcely reported. In this study, we developed and validated a rapid and sensitive method for quantifying urinary NEO concentrations using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS).. After phosphate-induced acidification of a urine sample, urinary NEOs were trapped by a solid-phase extraction column and eluted with methanol for acetamiprid, imidacloprid, thiacloprid, thiamethoxam, clothianidin and dinotefuran and with an acetonitrile and methanol solution (1:1, v/v) containing 5% NH3 for nitenpyram. A separation analysis was performed by LC-MS/MS within 10 minutes for the sample. This method was applied to first morning urine obtained from 52 Japanese (40.9 ± 10.5 years old, mean ± standard deviation) without occupational NEO exposure.. The linear dynamic ranges and their limit of quantification (LOQ, signal to noise ratio=10) levels were 0.3-20 or 50 µg/l (r=0.998-0.999) and 0.05-0.36 µg/l, respectively. The absolute recovery was 64-95%, and the intra- and inter-day precisions were less than 16.4% (relative standard deviation, %RSD). This method was successfully applied for analysis of NEOs in human urine samples obtained from 52 adults. The frequencies of individuals who showed more than LOD levels was above 90% for imidacloprid, thiamethoxam, clothianidin and dinotefuran, more than 50% for acetamiprid and thiacloprid and 29% for nitenpyram.. These results indicated that our new method could be applied to biological monitoring of NEO exposure even at environmental exposure levels in Japanese adults without occupational spraying histories.

Topics: Acetonitriles; Adult; Chromatography, High Pressure Liquid; Environmental Monitoring; Female; Guanidines; Humans; Imidazoles; Insecticides; Japan; Male; Methanol; Middle Aged; Neonicotinoids; Nitro Compounds; Occupational Exposure; Oxazines; Pyridines; Solid Phase Extraction; Tandem Mass Spectrometry; Thiamethoxam; Thiazines; Thiazoles

Competitive enzyme-linked immunosorbent assay was used to quantify the amounts of the neonicotinoids clothianidin and imidacloprid in Poa annua L. clippings from treated golf course fairways. Average clothianidin residues 7 d after application ranged from 674 to 1,550 ng/g tissue in 2012 and 455-2,220 ng/g tissue in 2013. Average clothianidin residues the day of application ranged from 17,100-38,800 ng/g tissue in 2014. Average imidacloprid residues 7 d after treatment ranged from 1,950-3,030 ng/g tissue in 2012 and 7,780-9,230 ng/g tissue in 2013. Average imidacloprid residues the day of application ranged from 31,500-40,400 ng/g tissue in 2014. Neonicotinoid or bifenthrin-neonicotinoid combination products applied in field plots in 2012 did not significantly reduce the numbers of larvae relative to the untreated control. However, in 2013, statistically significant reductions in the numbers of larvae recovered from treated field plots were associated with the presence of bifenthrin alone or when used in combination with neonicotinoid active ingredients. Listronotus maculicollis (Kirby) adults caged on neonicotinoid-, bifenthrin-, and bifenthrin-neonicotinoid-treated P. annua turf plugs fed on P. annua leaves, but mortality was only highly significantly different between treated and untreated foliage when weevils were placed on treated foliage the day after treatment and allowed to feed for 7 d. The modest degree of population suppression with bifenthrin in these experiments may not be adequate to justify the continued use of these products due to the increased risk of insecticide resistance and disruption of biological control.

Topics: Animals; Guanidines; Imidazoles; Larva; Neonicotinoids; Nitro Compounds; Pesticide Residues; Poa; Thiazoles; Weevils

Neonicotinoid insecticides are generally efficacious against many turfgrass pests, including several important phloem-feeding insects. However, inconsistencies in control of western chinch bugs, Blissus occiduus, have been documented in field efficacy studies. This research investigated the efficacy of three neonicotinoid insecticides (clothianidin, imidacloprid and thiamethoxam) against B. occiduus in buffalograss under field conditions and detected statistically significant differences in B. occiduus numbers among treatments. A subsequent study documented the relative quantity and degradation rate of these insecticides in buffalograss systemic leaf tissues, using HPLC.. Neonicotinoid insecticides initially provided significant reductions in B. occiduus numbers, but mortality diminished over the course of the field studies. Furthermore, while all three neonicotinoids were present in the assayed buffalograss leaf tissues, imidacloprid concentrations were significantly higher than those of clothianidin and thiamethoxam. Over the course of the 28 day study, thiamethoxam concentrations declined 700-fold, whereas imidacloprid and clothianidin declined only 70-fold and 60-fold respectively.. Field studies continued to verify inconsistencies in B. occiduus control with neonicotinoid insecticides. This is the first study to document the relative concentrations of topically applied neonicotinoid insecticides in buffalograss systemic leaf tissues.

Topics: Animals; Heteroptera; Imidazoles; Insect Control; Insecticides; Neonicotinoids; Nitro Compounds; Plant Diseases; Plant Leaves; Poaceae

Topics: Animals; Bees; Conservation of Natural Resources; Environmental Policy; European Union; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Policy Making; Population Dynamics; Thiamethoxam; Thiazoles; Truth Disclosure; United Kingdom

In Québec, as observed globally, abnormally high honey bee mortality rates have been reported recently. Several potential contributing factors have been identified, and exposure to pesticides is of increasing concern. In maize fields, foraging bees are exposed to residual concentrations of insecticides such as neonicotinoids used for seed coating. Highly toxic to bees, neonicotinoids are also reported to increase AChE activity in other invertebrates exposed to sub-lethal doses. The purpose of this study was therefore to test if the honey bee's AChE activity could be altered by neonicotinoid compounds and to explore possible effects of other common products used in maize fields: atrazine and glyphosate. One week prior to pollen shedding, beehives were placed near three different field types: certified organically grown maize, conventionally grown maize or non-cultivated. At the same time, caged bees were exposed to increasing sub-lethal doses of neonicotinoid insecticides (imidacloprid and clothianidin) and herbicides (atrazine and glyphosate) under controlled conditions. While increased AChE activity was found in all fields after 2 weeks of exposure, bees close to conventional maize crops showed values higher than those in both organic maize fields and non-cultivated areas. In caged bees, AChE activity increased in response to neonicotinoids, and a slight decrease was observed by glyphosate. These results are discussed with regard to AChE activity as a potential biomarker of exposure for neonicotinoids.

Topics: Acetylcholinesterase; Animals; Atrazine; Bees; Glycine; Glyphosate; Guanidines; Herbicides; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Pollen; Quebec; Thiazoles; Zea mays

Topics: Animals; Bees; Environmental Exposure; Environmental Monitoring; Europe; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Risk Assessment; Thiamethoxam; Thiazoles; Uncertainty

The objective of this study was to develop analytical method based on optimized dispersive liquid-liquid microextraction (DLLME) as a pretreatment procedure combined with reversed phase liquid chromatographic separation on C18 column and isocratic elution for simultaneous MS/MS determination of selected neonicotinoid insecticides in honey. The LC-MS/MS parameters were optimized to unequivocally provide good chromatographic separation, low detection (LOD, 0.5-1.0 μg kg(-1)) and quantification (LOQ, 1.5-2.5 μg kg(-1)) limits for acetamiprid, clothianidin, thiamethoxam, imidacloprid, dinotefuran, thiacloprid and nitenpyram in honey samples. Using different types (chloroform, dichloromethane) and volumes of extraction (0.5-3.0 mL) and dispersive (acetonitrile; 0.0-1.0 mL) solvent and by mathematical modeling it was possible to establish the optimal sample preparation procedure. Matrix-matched calibration and blank honey sample spiked in the concentration range of LOQ-100.0 μg kg(-1) were used to compensate the matrix effect and to fulfill the requirements of SANCO/12495/2011 for the accuracy (R 74.3-113.9%) and precision (expressed in terms of repeatability (RSD 2.74-11.8%) and within-laboratory reproducibility (RSDs 6.64-16.2%)) of the proposed method. The rapid (retention times 1.5-9.9 min), sensitive and low solvent consumption procedure described in this work provides reliable, simultaneous, and quantitative method applicable for the routine laboratory analysis of seven neonicotinoid residues in real honey samples.

Topics: Anabasine; Chromatography, Liquid; Guanidines; Honey; Imidazoles; Insecticides; Liquid Phase Microextraction; Molecular Structure; Neonicotinoids; Nitro Compounds; Oxazines; Pyridines; Reproducibility of Results; Tandem Mass Spectrometry; Thiamethoxam; Thiazines; Thiazoles

Japanese beetle, Popillia japonica Newman (Coleoptera: Scarabaeidae), is an important quarantine pest of nurseries. Nursery plant movement from P. japonica-infested regions is regulated by the U.S. Domestic Japanese Beetle Harmonization Plan (DJHP), which classifies states by risk categories. Treatments for category 2 states include preharvest soil surface treatment of nursery plants grown in field soil using Discus SC, Marathon (1G or 60 WP), or Flagship (0.22G or 25 WG). In this study, Discus, Marathon 60 WP, or Flagship 0.22G DJHP standards were compared with labeled rates of non-DJHP-approved insecticides, including neonicotinoids clothianidin (Arena 50WDG), generic imidacloprid (Quali-Pro Imidacloprid 2 F T&O Insecticide, Mallet 2 F T&O Insecticide, and Lada 2 F Insecticide), and imidacloprid + bifenthrin (Allectus SC), as well as the anthranilic diamide, chlorantraniliprole (Acelepryn Insecticide). Arena provided 100% P. japonica control in May, June, and July over four test years, but had one larva recovered during August in two of those 4 yr. Acelepryn did not provide DJHP-acceptable P. japonica control. During July, Allectus provided 100% P. japonica control in three of four test years, but had four larvae in one test year. Other treatments tested only during July, which provided 100% P. japonica control, included Discus (five tests); Marathon, Quali-Pro, and Mallet (two tests); and Lada and Flagship (one test). Generic imidacloprid 2 F formulations were equivalent in P. japonica control to DJHP-approved insecticides. Insecticides generally performed poorly on other scarabs or curculionid larvae. The study supports Arena, Allectus, and generic imidacloprid 2 F products as suitable candidates for the DJHP.

Topics: Animals; Coleoptera; Guanidines; Imidazoles; Insect Control; Insecticides; Larva; Neonicotinoids; Nitro Compounds; ortho-Aminobenzoates; Plant Roots; Quarantine; Random Allocation; Tennessee; Thiazoles; Trees

Topics: Animals; Bees; Crops, Agricultural; Diet; Environmental Policy; European Union; Feeding Behavior; Government Regulation; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Species Specificity; Thiazoles

A microextraction procedure based on vortex-assisted surfactant-enhanced-emulsification liquid-liquid microextraction with solidification of floating organic droplet (VSLLME-SFO) for preconcentration of neonicotinoid pesticides, including acetamiprid, clotianidin, nitenpyram, imidacloprid, and thiamethoxam, has been developed. In VSLLME-SFO process, the addition of surfactant (as an emulsifier), could be enhance the mass-transfer from the aqueous solution into the extraction solvent. The extraction solvent could be dispersed into the aqueous by vortex process. Other experimental parameters affected the extraction efficiency, including the kind and concentration of salt, concentration and volume of HCl, kind and concentration of surfactant and its volume, kind and volume of extraction solvent, vortex time and the centrifugation extraction time, were also optimized. The optimum extraction conditions of VSLLME-SFO were 10.00 mL of sample, 0.3% (w/v) Na2SO4, 50 µL of 0.050 mol L(-1) SDS, 1.0 mol L(-1) HCl (400 µL), 150 µL of octanol, vortex time 1 min and centrifugation time 10 min. The sediment phase was analyzed by subjecting it to HPLC using a mobile phase of 25% acetonitrile in water, at a flow rate of 1.0 mL min(-1), and photodiode array detection at 254 nm. Under the optimum extraction conditions, high enrichment factors (20-100 fold) and low limit of detection (0.1-0.5 μg L(-1)) could be obtained. This method provided high sensitivity, low toxic organic solvents used, and simplicity of the extraction processes. The proposed method was successfully applied in the analysis of neonicotinoids in fruit juice and water samples.

Topics: Beverages; Chromatography, High Pressure Liquid; Emulsions; Fresh Water; Fruit; Guanidines; Humans; Imidazoles; Liquid Phase Microextraction; Neonicotinoids; Nitro Compounds; Oxazines; Pesticides; Pyridines; Sensitivity and Specificity; Surface-Active Agents; Thiamethoxam; Thiazoles; Water Pollutants, Chemical

A simple, cheap and rugged method was developed for simultaneous deter mination of 6 neonicotinoid residues in soil, including imidacloprid, acetamiprid, thiamethoxam, thiacloprid, clothianidin and nitenpyram. The soil sample was produced by dispersive liquid-liquid micro-extraction (DLLME) after extracted by the mixed solution of acetonitrile and CH2Cl2 (2:1, phi). The analytes were separated by HPLC with Alltima C18 column (4.6 mm x 250 mm, 5 microm) and detected by PDA at 260 nm. External standard method was used for quantification. The results showed that good linearity was obtained with correlation coefficients between 0.9982 and 0.9999 in the range of 0.5-200 microg x L(-1). The limits of detection (LODs) were in the range between 0.0005 and 0.003 microg x mL(-1) (S/N = 3). The method was validated with five soil samples spiked at three fortification levels (0.05, 0.1, 1.0 mg x kg(-1)) and recoveries were in the range of 55.3%-95.6% with RSD of 1.4%-7.0%. The effect of clean-up was evaluated by UV spectra and demonstrated that the method established is effective. In conclusion, this method is competent for the simultaneous analysis of 6 neonicotinoid residues in soil.

Topics: Chromatography, High Pressure Liquid; Guanidines; Imidazoles; Limit of Detection; Neonicotinoids; Nitro Compounds; Oxazines; Pesticide Residues; Pyridines; Soil; Thiamethoxam; Thiazines; Thiazoles

Bioassays were designed to evaluate Beauveria bassiana (Balsamo) Vuillemin strain GHA against Listronotus maculicollis (Kirby) adults. B. bassiana and its "inert" carrier oil in the product BotaniGard and the inert carrier oil alone provided 99 and 96% mortality, respectively, in petri dish assays 1 d after treatment when applied in 1 ml water. When the same treatments were applied in 0.5 ml of carrier water, mortality was only 1.4 and 0.7%, respectively, 1 d after treatment. After 10 d in petri dishes, B. bassiana and its inert carrier oil and the inert carrier oil alone applied in 0.5 ml water showed 77 and 9% mortality, respectively. When one-tenth the label dosage of B. bassiana and inert carrier oil was combined with neonicotinoids applied in 1 ml water, there were significant increases in weevil mortality over the neonicotinoids alone 1 d after treatment. When 88.7% of one-tenth the label dosage of inert carrier oil alone was combined with neonicotinoids clothianidin, imidacloprid, and dinotefuran applied in 1 ml water, there was also a significant increases (38%) with clothianidin in weevil mortality over clothianidin alone 1 d after treatment. B. bassiana and its inert carrier oil provided 28, 50, and 78% mortality at the highest label dosage and 47, 76, and 89% mortality at 4x the highest label dosage in turf plug assays at 7, 10, and 14 d after treatment. Addition of 5 or 20% MycoMax (a nutrient source for B. bassiana) did not significantly increase mortality.

Topics: Animals; Beauveria; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Pest Control, Biological; Poaceae; Thiazoles; Weevils

Recently, legislative decisions withdrew or temporarily suspended the use of neonicotinoids and fipronil as seeds tanning in many countries because of their endocrine-disrupting activity imputable to the bees' toxicity. In this study, the occurrence of acetamiprid, fipronil, clothianidin, flonicamid, imidacloprid, nitenpyram, thiacloprid and thiamethoxam was detected in 66 samples of commercial treated corn seeds, collected in the Italian market in the frame of ministerial institutional quality control activity. Because of the lack of a validated analytical protocol for neonicotinoid detection in seeds, a routinely suitable liquid chromatography-tandem mass spectroscopy (LC-MS/MS) analytical method was developed and statistically validated on fortified corn seeds. Survey results demonstrated that 88% of the investigated seed samples showed the presence of residues of clothianidin, fipronil, thiamethoxam and thiacloprid, either individually or simultaneously, with values that ranged from about 0.002 to 20 mg kg(-1), which evidenced the alarming illicit use of these pesticides in seed treatments.

Topics: Agriculture; Anabasine; Animals; Bees; Chromatography, Liquid; Endocrine Disruptors; Guanidines; Humans; Imidazoles; Insecticides; Italy; Legislation, Drug; Neonicotinoids; Nitro Compounds; Oxazines; Pesticide Residues; Pyrazoles; Pyridines; Reproducibility of Results; Seeds; Tandem Mass Spectrometry; Thiamethoxam; Thiazines; Thiazoles; Zea mays

In the understanding of colony loss phenomena, a worldwide crisis of honeybee colonies which has serious consequences for both apiculture and bee-pollination-dependent farm production, analytical chemistry can play an important role. For instance, rapid and accurate analytical procedures are currently required to better assess the effects of neonicotinoid insecticides on honeybee health. Since their introduction in agriculture, neonicotinoid insecticides have been blamed for being highly toxic to honeybees, possibly at the nanogram per bee level or lower. As a consequence, most of the analytical methods recently optimized have focused on the analysis of ultratraces of neonicotinoids using liquid chromatography-mass spectrometry techniques to study the effects of sublethal doses. However, recent evidences on two novel routes-seedling guttations and seed coating particulate, both associated with corn crops-that may expose honeybees to huge amounts of neonicotinoids in the field, with instantly lethal effects, suggest that selected procedures need optimizing. In the present work, a simplified ultra-high-performance liquid chromatography-diode-array detection method for the determination of neonicotinoids in single bees has been optimized and validated. The method ensures good selectivity, good accuracy, and adequate detection limits, which make it suitable for the purpose, while maintaining its ability to evaluate exposure variability of individual bees. It has been successfully applied to the analysis of bees in free flight over an experimental sowing field, with the bees therefore being exposed to seed coating particulate released by the pneumatic drilling machine.

Topics: Agriculture; Animals; Bees; Chromatography, High Pressure Liquid; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Pyridines; Thiamethoxam; Thiazines; Thiazoles

The development of insecticides requires valid risk assessment procedures to avoid causing harm to beneficial insects and especially to pollinators such as the honeybee Apis mellifera. In addition to testing according to current guidelines designed to detect bee mortality, tests are needed to determine possible sublethal effects interfering with the animal's vitality and behavioral performance. Several methods have been used to detect sublethal effects of different insecticides under laboratory conditions using olfactory conditioning. Furthermore, studies have been conducted on the influence insecticides have on foraging activity and homing ability which require time-consuming visual observation. We tested an experimental design using the radiofrequency identification (RFID) method to monitor the influence of sublethal doses of insecticides on individual honeybee foragers on an automated basis. With electronic readers positioned at the hive entrance and at an artificial food source, we obtained quantifiable data on honeybee foraging behavior. This enabled us to efficiently retrieve detailed information on flight parameters. We compared several groups of bees, fed simultaneously with different dosages of a tested substance. With this experimental approach we monitored the acute effects of sublethal doses of the neonicotinoids imidacloprid (0.15-6 ng/bee) and clothianidin (0.05-2 ng/bee) under field-like circumstances. At field-relevant doses for nectar and pollen no adverse effects were observed for either substance. Both substances led to a significant reduction of foraging activity and to longer foraging flights at doses of ≥0.5 ng/bee (clothianidin) and ≥1.5 ng/bee (imidacloprid) during the first three hours after treatment. This study demonstrates that the RFID-method is an effective way to record short-term alterations in foraging activity after insecticides have been administered once, orally, to individual bees. We contribute further information on the understanding of how honeybees are affected by sublethal doses of insecticides.

Topics: Animals; Bees; Behavior, Animal; Feeding Behavior; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Radio Waves; Thiazoles; Time Factors

Neonicotinoid insecticides act selectively on insect nicotinic acetylcholine receptors (nAChRs). Recent studies revealed that their efficiency was altered by the phosphorylation/dephosphorylation process and the intracellular signaling pathway involved in the regulation of nAChRs. Using whole-cell patch-clamp electrophysiology adapted for dissociated cockroach dorsal unpaired median (DUM) neurons, we demonstrated that intracellular factors involved in the regulation of nAChR function modulated neonicotinoid sensitivity. DUM neurons were known to express two α-bungarotoxin-insensitive nAChR subtypes: nAChR1 and nAChR2. Whereas nAChR1 was sensitive to imidacloprid, nAChR2 was insensitive to this insecticide. Here, we demonstrated that, like nicotine, acetamiprid and clothianidin, other types of neonicotinoid insecticides, acted as agonists on the nAChR2 subtype. Using acetamiprid, we revealed that both steady-state depolarization and hyperpolarization affected nAChR2 sensitivity. The measurement of the input membrane resistance indicated that change in the acetamiprid-induced agonist activity was related to the receptor conformational state. Using cadmium chloride, ω-conotoxin GVIA, and (R,S)-(3,4-dihydro-6,7-dimethoxy-isoquinoline-1-yl)-2-phenyl-N,N-di-acetamide (LOE 908), we found that inhibition of calcium influx through high voltage-activated calcium channels and transient receptor potential γ (TRPγ) activated by both depolarization and hyperpolarization increased nAChR2 sensitivity to acetamiprid. Finally, using N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride (W7), forskolin, and cAMP, we demonstrated that adenylyl cyclase sensitive to the calcium/calmodulin complex regulated internal cAMP concentration, which in turn modulated TRPγ function and nAChR2 sensitivity to acetamiprid. Similar TRPγ-induced modulatory effects were also obtained when clothianidin was tested. These findings bring insights into the signaling pathway modulating neonicotinoid efficiency and open novel strategies for optimizing insect pest control.

Topics: Animals; Bungarotoxins; Calcium; Calcium Channels; Calmodulin; Cell Membrane; Cholinergic Agonists; Cockroaches; Cyclic AMP; Guanidines; Imidazoles; Insecta; Insecticides; Male; Membrane Potentials; Neonicotinoids; Neurons; Nicotine; Nicotinic Agonists; Nitro Compounds; Phosphorylation; Pyridines; Receptors, Nicotinic; Signal Transduction; Thiazoles; Transient Receptor Potential Channels; Tubocurarine

This paper assessed the potential exposure of bees (Apis mellifera L.) to pesticides during maize (Zea mays L.) sowing with pneumatic drills. Data were derived from tests carried out in field tests, comparing two configurations of a pneumatic precision drill: conventional drill; drill with air deflectors. In addition, static tests simulating the sowing under controlled conditions, were performed on the drill equipped with an innovative system developed at CRA-ING. During the field tests, the concentrations in the air of the active ingredients of four insecticides used in maize seed dressing (imidacloprid, clothianidin, thiamethoxam and fipronil) were recorded. The concentrations of active ingredients in the air were used for assessing the quantities of active ingredients that a bee might intercept as it flies in a sort of virtual tunnel, the dimensions of which were dependent upon the bee body cross-section and the length of flight. The results of the field tests show that the air deflectors were not completely effective in reducing the amount of active ingredients dispersed in the air. The results of the static tests with drill equipped with the prototype indicated reductions of the active ingredient air concentrations ranging from 72 % up to 95 %, with reference to the conventional drill. Such ratios were applied to the amounts of active ingredients intercepted by the bees in the virtual tunnel contributing to a consistent reduction of the probability that sub-lethal effects can occur.

Topics: Animals; Bees; Environmental Exposure; Environmental Monitoring; Guanidines; Imidazoles; Neonicotinoids; Nitro Compounds; Oxazines; Particulate Matter; Pesticide Residues; Pesticides; Seeds; Thiamethoxam; Thiazoles; Zea mays

This research focused on photocatalytic degradation of imidacloprid, thiamethoxam and clothianidin employing a tailor-made photoreactor with six polychromatic fluorescent UVA (broad maximum at 355 nm) lamps and immobilised titanium dioxide (TiO(2)) on glass slides. The disappearance was followed by high pressure liquid chromatography (HPLC-DAD) analyses, wherein the efficiency of mineralization was monitored by measurements of total organic carbon (TOC). Within 2h of photocatalysis, all three neonicotinoids were degraded following first order kinetics with rate constants k=0.035 ± 0.001 min(-1) for imidacloprid, k=0.019 ± 0.001 min(-1) for thiamethoxam and k=0.021 ± 0.000 min(-1) for clothianidin. However, the rate of mineralization was low, i.e. 19.1 ± 0.2% for imidacloprid, 14.4 ± 2.9% for thiamethoxam and 14.1 ± 0.4% for clothianidin. This indicates that several transformation products were formed instead. Some of them were observed within HPLC-DAD analyses and structures were proposed according to the liquid chromatography-electro spray ionization tandem mass spectrometry analyses (LC-ESI-MS/MS). The formation of clothianidin, as thiamethoxam transformation product, was reported for the first time.

Topics: Catalysis; Chromatography, High Pressure Liquid; Chromatography, Liquid; Guanidines; Imidazoles; Insecticides; Kinetics; Neonicotinoids; Nitro Compounds; Oxazines; Photolysis; Tandem Mass Spectrometry; Thiamethoxam; Thiazoles; Titanium

Imidacloprid has been a major neonicotinoid insecticide for controlling Aphis gossypii (Glover) (Homoptera: Aphididae) and other piercing-sucking pests. However, the resistance to imidacloprid has been recorded in many target insects. At the same time, cross-resistance of imidacloprid and other insecticides, especially neonicotinoid insecticides, has been detected.. Results showed that the level of cross-resistance was different between imidacloprid and tested neonicotinoid insecticides (no cross-resistance: dinotefuran, thiamethoxam and clothianidin; a 3.68-5.79-fold cross-resistance: acetamiprid, nitenpyram and thiacloprid). In the study of sublethal effects, imidacloprid at LC(20) doses could suppress weight gain and honeydew excretion, but showed no significant effects on longevity and fecundity of the imidacloprid-resistant cotton aphid, A. gossypii. However, other neonicotinoid insecticides showed significant adverse effects on biological characteristics (body weight, honeydew excretion, longevity and fecundity) in the order of dinotefuran > thiamethoxam and clothianidin > nitenpyram > thiacloprid and acetamiprid.. The results indicated that dinotefuran is the most effective insecticide for use against imidacloprid-resistant A. gossypii. To avoid further resistance development, the use of nitenpyram, acetamiprid and thiacloprid should be avoided on imidacloprid-resistant populations of A. gossypii.

Topics: Animals; Aphids; Body Weight; Fertility; Guanidines; Imidazoles; Insecticide Resistance; Longevity; Neonicotinoids; Nitriles; Nitro Compounds; Oxazines; Pyrimidines; Reproduction; Survival Analysis; Thiamethoxam; Thiazoles; Toxicity Tests

The western chinch bug, Blissus occiduus Barber (Hemiptera: Blissidae), has emerged as a serious pest of buffalograss, Buchlod dactyloides (Nuttall) Engelmann. In general, neonicotinoid insecticides effectively control a variety of turfgrass insects, particularly phloem-feeding pests. However, because of well documented inconsistencies in control, these compounds are generally not recommended for chinch bugs. This study was designed to document the contact and systemic toxicity of three neonicotinoid insecticides (clothianidin, imidacloprid, and thiamethoxam) to B. occiduus. In contact bioassays, thiamethoxam was approximately 20-fold less toxic than clothianidin or imidacloprid to B. occiduus nymphs and three-fold more toxic to adults. In adult systemic bioassays, thiamethoxam was up to five-fold more toxic than clothianidin or imidacloprid. Interestingly, thiamethoxam was significantly more toxic to adults than to nymphs in both contact and systemic bioassays. This was not observed with clothianidin or imidacloprid. Bifenthrin, used for comparative purposes, exhibited 1844-fold and 122-fold increase in toxicity to nymphs and adults, respectively. These results provide the first documentation of the relative toxicity of these neonicotinoid insecticides to B. occiduus.

Topics: Animals; Dose-Response Relationship, Drug; Guanidines; Heteroptera; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Thiamethoxam; Thiazoles; Toxicity Tests

Regarding the hypothesis that neonicotinoid insecticides used for seed coating of agricultural crops - mainly corn, sunflower and seed rape - are related to the extensive death of honey bees, the phenomenon of corn seedling guttation has been recently considered as a possible route of exposure of bees to these systemic insecticides. In the present study, guttation drops of corn plants obtained from commercial seeds coated with thiamethoxam, clothianidin, imidacloprid and fipronil have been analyzed by an optimized fast UHPLC-DAD procedure showing excellent detection limits and accuracy, both adequate for the purpose. The young plants grown both in pots - in greenhouse - and in open field from coated seeds, produced guttation solutions containing high levels of the neonicotinoid insecticides (up to 346 mg L(-1) for imidacloprid, 102 mg L(-1) for clothianidin and 146 mg L(-1) for thiamethoxam). These concentration levels may represent lethal doses for bees that use guttation drops as a source of water. The neonicotinoid concentrations in guttation drops progressively decrease during the first 10-15 days after the emergence of the plant from the soil. Otherwise fipronil, which is a non-systemic phenylpyrazole insecticide, was never detected into guttation drops. Current results confirm that the physiological fluids of the corn plant can effectively transfer neonicotinoid insecticides from the seed onto the surface of the leaves, where guttation drops may expose bees and other insects to elevated doses of neurotoxic insecticides.

Topics: Environmental Monitoring; Guanidines; Imidazoles; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Seedlings; Seeds; Thiamethoxam; Thiazoles; Xylem; Zea mays

Neonicotinoids are synthetic, nicotine-derived insecticides used for agricultural and household pest control. Though highly effective at activating insect nicotinic receptors, many neonicotinoids are also capable of directly activating and/or modulating the activation of vertebrate nicotinic receptors. In this study, we have investigated the actions of the neonicotinoids clothianidin (CTD) and imidacloprid (IMI) on human neuronal α4β2 nicotinic acetylcholine receptors. The data demonstrate that the compounds are weak agonists of the human receptors with relative peak currents of 1-4% of the response to 1 mM acetylcholine (ACh). Coapplication of IMI strongly inhibited currents elicited by ACh. From Schild plot analysis, we estimate that the affinity of IMI for the human α4β2 receptor is 18 μM. The application of low concentrations of CTD potentiated responses to low concentrations of ACh, suggesting that receptors occupied by one ACh and one CTD molecule have a higher gating efficacy than receptors with one ACh bound. Interestingly, subunit stoichiometry affected inhibition by CTD, with (α4)(2) (β2)(3) receptors significantly more strongly inhibited than the (α4)(3) (β2)(2) receptors.

Topics: Cholinergic Agents; Electrophysiological Phenomena; Guanidines; HEK293 Cells; Humans; Imidazoles; Neonicotinoids; Neurons; Nitro Compounds; Receptors, Nicotinic; Thiazoles

The utilization of dressed seed for spring sowing is a widespread practice to control some pests with reduced doses of chemical products. However some insecticides employed in maize seed dressing, namely belonging to the neonicotinoid family and fipronil, have been claimed to play a role in the decline of honeybees (Apis mellifera L.). Pneumatic drills used in maize sowing are charged with contributing to the dispersion of the abrasion dust produced by dressed seeds, favoring the contamination of the honeybee habitat. Different devices similar to air deflectors have been introduced on pneumatic drills in order to reduce dust drift. During previous field tests carried out by the authors during recent years reduction of dust concentration both in the air and at soil surface has been shown as a consequence of their application. As field tests are affected by the variability of environmental parameters (namely wind speed and direction) the results are not always reliable, comparable and of a general validity. This paper refers to a sowing simulation test system in which pneumatic drills can be tested at a fixed point under controlled conditions of the main environmental parameters. In the test area, protected by external influences, artificial wind conditions are created by means of a fan. The drill, suitably placed in the test area, operates the seed distribution "sur place" by means of an electric engine connected to the drill's driving wheel. A 22.5 m long sampling area, leeward with respect to the drill position, has been identified. Along the sampling area a series of Petri dishes has been placed, with the aim of capturing the depositing dust and providing the concentration of the active ingredients (a.i.) at ground level. At the same time, three air samplers with PTFE diskette filters have been used for the detection of the a.i. The test system has been used for the test of a pneumatic drill, equipped with and without air deflectors, using maize seed dressed with four a. i. (imidacloprid, clothianidin, thiametoxam, fipronil). The results showed regularly decreasing of the concentrations as distance increased, both in the air and at ground level. Moreover, the difference determined by the adoption of the drift reducing device (air deflectors) resulted clear and it can be quantified at around 50 % of the a.i. amounts observed without deflectors. Finally, the paper proposes a data processing method that, from the values observed at fixed point, provides th

Topics: Agriculture; Animals; Bees; Dust; Guanidines; Imidazoles; Insecticides; Models, Theoretical; Neonicotinoids; Nitro Compounds; Seeds; Soil; Thiazoles; Wind; Zea mays

Systemic insecticides and application methods were examined for the control of the vector psyllid of citrus greening disease, Diaphorina citri, on grown king mandarin trees in an orchard in southern Vietnam from May 2007 to September 2008. Leaf spraying of imidacloprid, thiamethoxam, and clothianidin attained about 50 % to 70 % mortality of the psyllid for one month after the application and showed decreased efficacy thereafter. Imidacloprid was more effective than the other two insecticides, but the efficacy on grown trees was still much lower than that following application to young seedlings. Trunk injection of these insecticides accomplished similar mortality, about 50 %, and the efficacy of the insecticides continued for one month. An adjuvant was used with the goal of protecting the insecticide applied on leaves from precipitation, and mineral oil was used for the same reason, as well as its potential to control the psyllid. Neither the adjuvant nor the mineral oil played an evident role in the increase of insecticide efficacy or longevity. Application of systemic insecticides at even 50 times the dose described above did not show an apparent increase in psyllid mortality. The insecticides commonly used for the control of the psyllid were not as effective on this insect on grown trees as we had expected they would be.

Topics: Animals; Citrus; Guanidines; Hemiptera; Imidazoles; Insect Vectors; Insecticides; Neonicotinoids; Nitro Compounds; Oxazines; Plant Diseases; Reproducibility of Results; Thiamethoxam; Thiazoles; Time Factors; Vietnam

Neonicotinoid insecticides control crop pests based on their action as agonists at the insect nicotinic acetylcholine receptor, which accepts chloropyridinyl- and chlorothiazolyl-analogs almost equally well. In some cases, these compounds have also been reported to enhance plant vigor and (a)biotic stress tolerance, independent of their insecticidal function. However, this mode of action has not been defined. Using Arabidopsis thaliana, we show that the neonicotinoid compounds, imidacloprid (IMI) and clothianidin (CLO), via their 6-chloropyridinyl-3-carboxylic acid and 2-chlorothiazolyl-5-carboxylic acid metabolites, respectively, induce salicylic acid (SA)-associated plant responses. SA is a phytohormone best known for its role in plant defense against pathogens and as an inducer of systemic acquired resistance; however, it can also modulate abiotic stress responses. These neonicotinoids effect a similar global transcriptional response to that of SA, including genes involved in (a)biotic stress response. Furthermore, similar to SA, IMI and CLO induce systemic acquired resistance, resulting in reduced growth of a powdery mildew pathogen. The action of CLO induces the endogenous synthesis of SA via the SA biosynthetic enzyme ICS1, with ICS1 required for CLO-induced accumulation of SA, expression of the SA marker PR1, and fully enhanced resistance to powdery mildew. In contrast, the action of IMI does not induce endogenous synthesis of SA. Instead, IMI is further bioactivated to 6-chloro-2-hydroxypyridinyl-3-carboxylic acid, which is shown here to be a potent inducer of PR1 and inhibitor of SA-sensitive enzymes. Thus, via different mechanisms, these chloropyridinyl- and chlorothiazolyl-neonicotinoids induce SA responses associated with enhanced stress tolerance.

Topics: Arabidopsis; Ascomycota; Chromatography, High Pressure Liquid; Gene Expression Regulation, Plant; Guanidines; Imidazoles; Insecticides; Mass Spectrometry; Molecular Structure; Neonicotinoids; Nitro Compounds; Reverse Transcriptase Polymerase Chain Reaction; Salicylic Acid; Thiazoles

The cabbage seedpod weevil, Ceutorhynchus obstrictus (Marsham), is a major pest in the production of canola (Brassica napus L.) in North America and Europe, and effective population control is often essential for economical crop production. In North America, neonicotinoid insecticides have been used for several years in canola as seed treatments for reducing herbivory by flea beetles. The neonicotinoids clothianidin and imidacloprid were investigated to determine their effects on preimaginal development and on emergence of new-generation adults of C. obstrictus in comparison with effects of lindane, a chlorinated hydrocarbon seed treatment.. Mean numbers of second- and third-instar larvae were significantly higher in plants seed-treated with lindane than in plants treated with the neonicotinoid compounds, even though weevil oviposition was similar for all treatments. Emergence of new-generation adults was reduced by 52 and 39% for plants seed-treated with clothianidin and imidacloprid, respectively, compared with emergence from plants treated with lindane.. Seed treatment with both clothianidin and imidacloprid produced systemic insecticidal effects on larvae of C. obstrictus, with clothianidin slightly more effective than imidacloprid. Use of clothianidin or imidacloprid as seed treatments can comprise an important component in the integrated management of cabbage seedpod weevil in canola.

Topics: Animals; Brassica napus; Guanidines; Imidazoles; Insect Control; Insecticides; Neonicotinoids; Nitro Compounds; Oviposition; Plant Diseases; Seeds; Thiazoles; Weevils

Nitropolychlorobutadienes are valuable precursors for highly functionalized acyclic or (hetero)cyclic compounds. In this 8th part of our synthetically oriented series we focus on the application of these versatile starting materials in the synthesis of analogs of the heterocyclic insecticides imidacloprid and thiacloprid, and the acyclic counterpart clothianidin. In addition to the main synthetic part, leading to imidazolidines or oxazolidines, further promising types of compounds derived by subsequent chemical modifications, are introduced. Most of the new compounds show high insecticidal activity.

Topics: Butadienes; Guanidines; Halogenation; Imidazoles; Imidazolidines; Insecticides; Neonicotinoids; Nitro Compounds; Oxazoles; Pyridines; Thiazines; Thiazoles

Neonicotinoid insecticide metabolism involves considerable substrate specificity and regioselectivity of the relevant CYP450, aldehyde oxidase, and phase II enzymes. Human CYP450 recombinant enzymes carry out the following conversions: CYP3A4, 2C19, and 2B6 for thiamethoxam (TMX) to clothianidin (CLO); 3A4, 2C19, and 2A6 for CLO to desmethyl-CLO; 2C19 for TMX to desmethyl-TMX. Human liver aldehyde oxidase reduces the nitro substituent of CLO to nitroso much more rapidly than it does that of TMX. Imidacloprid (IMI), CLO, and several of their metabolites do not give detectable N-glucuronides but 5-hydroxy-IMI, 4,5-diol-IMI, and 4-hydroxythiacloprid are converted to O-glucuronides in vitro with mouse liver microsomes and UDP-glucuronic acid or in vivo in mice. Mouse liver cytosol with S-adenosylmethionine converts desmethyl-CLO to CLO but not desmethyl-TMX to TMX. Two organophosphorus CYP450 inhibitors partially block IMI, thiacloprid, and CLO metabolism in vivo in mice, elevating brain and liver levels of the parent compounds while reducing amounts of the hydroxylated metabolites.

Topics: Aldehyde Oxidase; Animals; Cytochrome P-450 Enzyme Inhibitors; Cytochrome P-450 Enzyme System; Enzyme Inhibitors; Guanidines; Humans; Imidazoles; Insecticides; Kinetics; Male; Microsomes, Liver; Neonicotinoids; Nitro Compounds; Oxazines; Substrate Specificity; Thiamethoxam; Thiazoles

We examined lethal and sublethal effects of imidacloprid on Osmia lignaria (Cresson) and clothianidin on Megachile rotundata (F.) (Hymenoptera: Megachilidae). We also made progress toward developing reliable methodology for testing pesticides on wild bees for use in pesticide registration by using field and laboratory experiments. Bee larvae were exposed to control, low (3 or 6 ppb), intermediate (30 ppb), or high (300 ppb) doses of either imidacloprid or clothianidin in pollen. Field experiments on both bee species involved injecting the pollen provisions with the corresponding pesticide. Only O. lignaria was used for the laboratory experiments, which entailed both injecting the bee's own pollen provisions and replacing the pollen provision with a preblended pollen mixture containing imidacloprid. Larval development, emergence, weight, and mortality were monitored and analyzed. There were no lethal effects found for either imidacloprid or clothianidin on O. lignaria and M. rotundata. Minor sublethal effects were detected on larval development for O. lignaria, with greater developmental time at the intermediate (30 ppb) and high doses (300 ppb) of imidacloprid. No similar sublethal effects were found with clothianidin on M. rotundata. We were successful in creating methodology for pesticide testing on O. lignaria and M. rotundata; however, these methods can be improved upon to create a more robust test. We also identified several parameters and developmental stages for observing sublethal effects. The detection of sublethal effects demonstrates the importance of testing new pesticides on wild pollinators before registration.

Topics: Animals; Bees; Dose-Response Relationship, Drug; Female; Guanidines; Hymenoptera; Imidazoles; Insecticides; Male; Neonicotinoids; Nitro Compounds; Pollen; Thiazoles

The grub Tomarus subtropicus Blatchley (Coleoptera: Scarabaeidae) is one of the most destructive pests of turfgrass and sugarcane (Saccharum spp.) in southern Florida, but its life cycle has only been described in sugarcane production. Preventive insecticides applied against T. subtropicus in turfgrass when adult activity and oviposition were expected in sugarcane have necessitated subsequent curative treatments. Thus, the seasonal activity of T. subtropicus was monitored using UV blacklight traps and by sampling soil in infested St. Augustinegrass lawns in Cape Coral and Punta Gorda, FL, during 2005-2006. Several preventive and curative insecticides and entomopathogenic nematode species were evaluated in laboratory, greenhouse and field tests. T. subtropicus is univoltine in both sugarcane and St. Augustinegrass, but adult flight peaked approximately 1 mo later in St. Augustinegrass than in sugarcane. Halofenozide, clothianidin, and imidacloprid were effective preventive control products, and carbaryl and trichlorfon were satisfactory curative insecticides. The nematodes Steinernema scarabei (Stock & Koppenhöfer), S. glaseri (Steiner), and Heterorhabditis bacteriophora (Poinar) were most effective against T. subtropicus grubs, and second instars were more susceptible compared with third instars. Improved knowledge of the biology and effectiveness of different management options for T. subtropicus will help minimize excessive pesticide use, especially in coastal areas of Florida.

Topics: Animals; Benzoates; Coleoptera; Female; Florida; Guanidines; Hydrazines; Imidazoles; Insect Control; Insecticides; Larva; Male; Nematoda; Neonicotinoids; Nitro Compounds; Pest Control, Biological; Poaceae; Saccharum; Seasons; Thiazoles

An investigation was carried out to determine whether new neonicotinoid-nematode combinations and earlier applications against younger larval stages could increase the efficacy of synergistic neonicotinoid-entomopathogenic nematode combinations against white grubs.. In the laboratory, combinations of the neonicotinoids imidacloprid and clothianidin and the nematodes Heterorhabditis bacteriophora Poinar and H. zealandica Poinar against third instars were compared. In Anomala orientalis (Waterhouse) and Popillia japonica Newman, H. bacteriophora-imidacloprid combinations showed the most consistent synergism but did not cause significantly higher mortality than H. zealandica-imidacloprid combinations. In Cyclocephala borealis Arrow, there was no clear trend as to which combinations caused the most consistent synergism, but H. zealandica-imidacloprid combinations tended to cause the highest mortalities. In the laboratory, imidacloprid-H. bacteriophora combinations provided more consistent synergism against third-instar than against second-instar A. orientalis, but mortality was higher in second instars. In field experiments, imidacloprid-H. bacteriophora combinations against A. orientalis and P. japonica provided more consistent synergism when applied in mid-September but more consistent control when applied in late August.. Imidacloprid is a better synergist for entomopathogenic nematodes than clothianidin. Imidacloprid-nematode combinations are more effective against second instars than against third instars, allowing rate reductions of both agents to make this approach more competitive with full-rate applications of neonicotinoid alone.

Topics: Animals; Coleoptera; Guanidines; Imidazoles; Insect Control; Insecticides; Larva; Neonicotinoids; Nitro Compounds; Pest Control, Biological; Rhabditida; Thiazoles; Time Factors

Neonicotinoid insecticides, which act on nicotinic acetylcholine receptors (nAChRs) in a variety of ways, have extremely low mammalian toxicity, yet the molecular basis of such actions is poorly understood. To elucidate the molecular basis for nAChR-neonicotinoid interactions, a surrogate protein, acetylcholine binding protein from Lymnaea stagnalis (Ls-AChBP) was crystallized in complex with neonicotinoid insecticides imidacloprid (IMI) or clothianidin (CTD). The crystal structures suggested that the guanidine moiety of IMI and CTD stacks with Tyr185, while the nitro group of IMI but not of CTD makes a hydrogen bond with Gln55. IMI showed higher binding affinity for Ls-AChBP than that of CTD, consistent with weaker CH-pi interactions in the Ls-AChBP-CTD complex than in the Ls-AChBP-IMI complex and the lack of the nitro group-Gln55 hydrogen bond in CTD. Yet, the NH at position 1 of CTD makes a hydrogen bond with the backbone carbonyl of Trp143, offering an explanation for the diverse actions of neonicotinoids on nAChRs.

Topics: Animals; Binding Sites; Guanidines; Imidazoles; Lymnaea; Models, Molecular; Molecular Conformation; Molecular Sequence Data; Neonicotinoids; Nitro Compounds; Receptors, Nicotinic; Structure-Activity Relationship; Thiazoles

Laboratory bioassays and field trials were conducted to evaluate an "attract-and-kill" system using methyl eugenol (ME) with neonicotinoid insecticides against male oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae). In laboratory bioassays, mortality of male flies resulting from the conventional toxicant, naled was 98.3-100% at 24 through 72 h after treatment, whereas the neonicotinoid insecticides imidacloprid and acetamiprid caused only approximately 60-80% at 24 through 72 h after treatment. In the assays of residual effect, naled was persistent up to 96 wk, whereas imidacloprid or acetamiprid was persistent up to 150 wk, resulting in 38.9 or 61.2% male mortality, respectively. Imidacloprid, in particular, caused a delayed lethal effect on flies. In another experiment, male mortality within 28 wk from clothianidin, another neonicotinoid insecticide, was approximately 80% after exposure for 24 h, suggesting a delayed lethal effect similar to those treated with imidacloprid, and mortality was up to 91.8%, if observed, 72 h after treatment. In field trials, attractiveness was similar between ME alone and ME incorporated with naled or neonicotinoids, indicating that addition of these insecticides to ME in traps is not repellent to B. dorsalis males. Using an improved wick-typed trap with longer attractiveness for simulating field application, addition of imidacloprid or acetamiprid maintained 40.1 or 64.3% male mortality, respectively, when assayed once every 2 wk from traps placed in orchards for 42 wk without changing the poison, whereas incorporation with naled resulted in as high as 98.1% after 34 wk and approximately 80% at 42 wk, indicating that persistence is increased compared with sugarcane fiberboard blocks for carrying poison attractants. This study also suggests that neonicotinoid insecticides could be used as an alternative for broad-spectrum insecticides as toxicants in fly traps.

Topics: Animals; Eugenol; Guanidines; Imidazoles; Insect Control; Insecticides; Male; Neonicotinoids; Nitro Compounds; Pyridines; Tephritidae; Thiazoles

One of the major challenges in managing the Colorado potato beetle, Leptinotarsa decemlineata (Say) is its remarkable ability to develop resistance to virtually every insecticide that has ever been used against it. Resistance is particularly common throughout northeastern USA as far north as Maine. The first instances of resistance to imidacloprid have already been reported from several locations in New York, Delaware and southern Maine. Rotating insecticides with different modes of action may delay insecticide resistance, but successful implementation of this technique depends on a good understanding of resistance and cross-resistance patterns in populations of target pests. LC(50) values were measured for imidacloprid and thiamethoxam in Colorado potato beetle populations from a variety of locations in the USA and Canada using diet incorporation bioassays. The field performance of imidacloprid, thiamethoxam and clothianidin against imidacloprid-resistant beetles on a commercial potato farm in southern Maine was also evaluated. Correlation between LC(50) values for imidacloprid and thiamethoxam was highly significant, even when populations previously exposed to thiamethoxam were excluded from the analysis. There was no statistically detectable difference in the LC(50) values between populations exposed to both insecticides and to imidacloprid alone. Applications of neonicotinoid insecticides at planting delayed build-up of imidacloprid-resistant beetle populations on field plots by 1-2 weeks but failed to provide adequate crop protection. Consistently with bioassay results, there was also substantial cross-resistance among the three tested neonicotinoid insecticides. Results of the present study support the recommendation to avoid rotating imidacloprid with thiamethoxam as a part of a resistance management plan.

Topics: Animals; Coleoptera; Guanidines; Imidazoles; Insecticide Resistance; Insecticides; Interviews as Topic; Lethal Dose 50; Neonicotinoids; Nitro Compounds; Oxazines; Solanum tuberosum; Thiamethoxam; Thiazoles

The efficacies of four systemic neonicotinoid insecticides applied to potted avocado trees at manufacturer-recommended rates were assessed against the avocado thrips, Scirtothrips perseae Nakahara. At the time of treatment, fully expanded first-flush young leaves were tagged for identification, and a proportion of these leaves was used in bioassays with second-instar thrips. At 7 weeks post-treatment, a second flush of leaves had fully expanded on the trees, and these leaves were included in additional bioassays comparing avocado thrips mortality on both first- and second-flush leaves. In bioassays with first-flush leaves, imidacloprid (273 mg AI pot(-1)) was the most effective insecticide, providing at least 70% mortality of thrips for 14 weeks. Thiamethoxam (137 mg AI pot(-1)), clothianidin (109 mg AI pot(-1)) and dinotefuran (241 mg AI pot(-1)) provided good control in bioassays that were conducted within 4 weeks of treatment, but thereafter their efficacies were inconsistent. In bioassays with second-flush leaves, imidacloprid provided at least 70% mortality up to 9 weeks after the insecticide application. Thereafter, mortality declined to 30% or lower. Bioassays with second-flush leaves collected from trees treated with thiamethoxam, clothianidin and dinotefuran resulted in unacceptably low thrips mortality. Monitoring of imidacloprid and thiamethoxam residues by ELISA showed that the greater persistence of imidacloprid in both first and second leaf flushes was due to a steadier uptake of this material. Although thiamethoxam residues rose quickly within the first leaf flush, levels had already begun to dissipate by the time the second leaf flush had started to develop.

Topics: Animals; Biological Assay; Guanidines; Imidazoles; Insecta; Insecticides; Mortality; Neonicotinoids; Nitro Compounds; Oxazines; Persea; Thiamethoxam; Thiazoles

The soil bacterium Bacillus thuringiensis Berliner subsp. israelensis (Bti), the neonicotinoid insecticides dinotefuran, imidacloprid, thiamethoxam and clothianidin and the insect growth regulator pyriproxyfen were evaluated to determine their efficacy against the larval stages of the fungus gnat Bradysia sp nr. coprophila (Lintner) in the laboratory. Treatments were applied as a drench to the growing medium in polypropylene deli containers. The Bti treatments had no effect on either instar tested, whereas all the other compounds negatively affected both the second and third instars. This study demonstrates that the soil bacterium B. thuringiensis var. israelensis may not be active on these larval stages, whereas the neonicotinoid insecticides and the insect growth regulator pyriproxyfen are effective on these stages. The fact that Bti is not effective on the second and third instars of the fungus gnat means that greenhouse producers using this insecticide must make applications before fungus gnat populations build up and before overlapping generations develop.

Topics: Animals; Bacillus thuringiensis; Diptera; Guanidines; Imidazoles; Insecticides; Juvenile Hormones; Larva; Neonicotinoids; Nitro Compounds; Oxazines; Pest Control, Biological; Pyridines; Thiamethoxam; Thiazoles

Nicotinic acetylcholine receptors (nAChRs) are present in high density in insect nervous tissue and are targeted by neonicotinoid insecticides. Improved understanding of the actions of these insecticides will assist in the development of new compounds. Here, we have used whole-cell patch-clamp recording of cholinergic neurons cultured from the central nervous system of 3rd instar Drosophila larvae to examine the actions of acetylcholine (ACh) and nicotine, as well as the neonicotinoids imidacloprid, clothianidin and P-CH-clothianidin on native nAChRs of these neurons. Dose-response data yield an EC(50) value for ACh of 19 microm. Both nicotine and imidacloprid act as low efficacy agonists at native nAChRs, evoking maximal current amplitudes 10-14% of those observed for ACh. Conversely, clothianidin and P-CH-clothianidin evoke maximal current amplitudes up to 56% greater than those evoked by 100 microm ACh in the same neurons. This is the first demonstration of 'super' agonist actions of an insecticide on native insect nAChRs. Cell-attached recordings indicate that super agonism results from more frequent openings at the largest (63.5 pS) conductance state observed.

Topics: Acetylcholine; Animals; Animals, Genetically Modified; Binding, Competitive; Cells, Cultured; Central Nervous System; Dose-Response Relationship, Drug; Drosophila melanogaster; Guanidines; Imidazoles; Insecticides; Larva; Membrane Potentials; Molecular Structure; Neonicotinoids; Neurons; Nicotine; Nicotinic Agonists; Nitro Compounds; Patch-Clamp Techniques; Receptors, Nicotinic; Thiazoles

As part of the ongoing evaluation of different systemic insecticides for prophylactic treatment of trees, responses of the beetle Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae) to different doses of four systemic neonicotinyl insecticides were studied. Adult beetles were provided with twigs or leaves of trees treated with different concentrations of imidacloprid to evaluate the toxicity of the insecticide through ingestion or contact or through both. Adult beetles also were provided with twigs of host plant treated with clothianidin, dinotefuran, and thiamethoxam to establish dose response of the beetle to these insecticides. Levels of individual insecticides in twigs and leaves were determined by using the "parent" method with high-performance liquid chromatography, and these levels were compared with the applied concentrations to determine their relationship. The LC50 values for detected level of each insecticide in twigs was 5.1 ppm at 24 h, 2.9 at 48 h, and 1.9 ppm at 72 h for imidacloprid; 1.1 ppm at 72 h for clothianidin; 2.2 ppm at 72 h for dinotefuran; and 1.0 ppm at 72 h for thiamethoxam. Our results indicate that mortality of adult beetles resulted not only from the ingestion and contact toxicity but also possibly from the antifeedant effect of imidacloprid.

Topics: Animals; Coleoptera; Dose-Response Relationship, Drug; Guanidines; Imidazoles; Insecticides; Lethal Dose 50; Neonicotinoids; Nitro Compounds; Oxazines; Thiamethoxam; Thiazoles

To compare the actions of clothianidin, a neonicotinoid acting on insect nicotinic acetylcholine receptors, and related compounds with that of imidacloprid, the compounds were tested on the Drosophila SAD-chicken beta2 nicotinic acetylcholine receptor expressed in Xenopus laevis oocytes using two-electrode voltage-clamp electrophysiology. The maximum response of the SAD beta 2 nicotinic receptor to clothianidin was larger than that observed for acetylcholine. Ring breakage of the imidazolidine ring of imidacloprid resulting in the generation of a guanidine group was critical for this super agonist action.

Topics: Acetylcholine; Anabasine; Animals; DNA, Complementary; DNA, Recombinant; Drosophila Proteins; Electrophysiology; Guanidines; Imidazoles; Molecular Structure; Neonicotinoids; Nicotinic Agonists; Nitro Compounds; Oocytes; Receptors, Nicotinic; Thiazoles; Transfection; Xenopus laevis

Two alpha-bungarotoxin-sensitive nicotinic receptor subtypes in cockroach neurons are identified as desensitizing (nAChD), selectively inhibitable with 100 nM imidacloprid, and non-desensitizing (nAChN), selectively inhibitable with 100 pM methyllycaconitine. Although the desensitization rate of nAChD receptors is highly variable, pharmacology is largely independent of desensitization rate. Because desensitized states tightly bind agonists, nAChD receptors are potently inhibited by neonicotinoids and specifically measured in radiolabeled imidacloprid binding assays. However, they are not usually detected in binding assays with radiolabeled alpha-bungarotoxin, which has a Kd for the resting state of 21 nM, but binds poorly to desensitized states often present in binding assays. In contrast, nAChN receptors are specifically measured in binding assays with radiolabeled alpha-bungarotoxin, which binds them with a Kd of 1.3 nM. nAChN receptors are activated by neonicotinoids at micromolar concentrations, and allosterically by spinosyn A, with an EC50 of 27 nM. Spinosyn A weakly antagonizes nAChD receptors -23% at 10 microM. The roles of the two nAChR subtypes in insecticide poisoning are discussed.

Topics: Acetylcholine; Aconitine; Animals; Atropine; Bungarotoxins; Cockroaches; Dihydro-beta-Erythroidine; Electrophysiology; Guanidines; Imidazoles; Inhibitory Concentration 50; Kinetics; Macrolides; Neonicotinoids; Neurons; Nicotinic Antagonists; Nitro Compounds; Receptors, Nicotinic; Thiazoles

The insect nicotinic acetylcholine receptor (nAChR) is a major target for insecticide action. The rapidly expanding use of neonicotinoid insecticides of varied structures makes it increasingly important to define similarities and differences in their action, particularly for the first-generation chloropyridinyl compounds versus the second-generation chlorothiazolyl derivatives. We have shown with Musca domestica that a convenient and relevant determination of the neonicotinoid insecticide target is a binding site assay with [(3)H]imidacloprid ([(3)H]IMI). This study uses membranes from the aphids MYZUS: persicae and Aphis craccivora and from heads of the flies Drosophila melanogaster and Musca domestica to characterize the [(3)H]IMI binding sites relative to their number and possible species variation in structure-activity relationships. With emphasis on commercial neonicotinoids, six potent chloropyridinyl compounds are compared with the corresponding six chlorothiazolyl analogues (syntheses are given for chemicals prepared differently than previously described). The preference for chloropyridinyl versus chlorothiazolyl is not dependent on the insect species examined but instead on other structural features of the molecule. The chlorothiazolyl substituent generally confers higher potency in the clothianidin and desmethylthiamethoxam series and the chloropyridinyl moiety in the imidacloprid, thiacloprid, acetamiprid, and nitenpyram series. Two chlorothiazolyl compounds compete directly with the chloropyridinyl [(3)H]IMI for the same binding sites in Myzus and Drosophila membranes. This study shows conserved neonicotinoid specificity of the [(3)H]IMI binding site in each of the four insect species examined.

Topics: Amino Acid Sequence; Animals; Aphids; Binding Sites; Cell Membrane; Conserved Sequence; Drosophila; Houseflies; Imidazoles; Insecta; Insecticides; Kinetics; Neonicotinoids; Nitro Compounds; Receptors, Nicotinic; Species Specificity; Structure-Activity Relationship; Tritium