clothianidin and thiacloprid

A comprehensive review of published and previously unpublished studies was performed to evaluate the neonicotinoid insecticides for evidence of developmental neurotoxicity (DNT).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

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

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

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

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

Co-occurrence of pesticide residues in food commodities raises a potential safety issue as their mixture effects on human health are largely unknown. In a previous study, we reported the toxicological effects (pathology and histopathology) of imazalil (IMZ), thiacloprid (THI), and clothianidin (CTD) alone and in binary mixtures in a 28-day oral gavage study in female Wistar rats. Five dose levels (up to 350 mg/kg body weight/day) ranging from a typical toxicological reference value to a clear effect dose were applied. In the present study, we undertook a transcriptomics analysis of rat livers by means of total RNA sequencing (RNA-Seq). Bioinformatic data analysis involving Ingenuity Pathway Analysis (IPA) was used to gain mechanistic information on hepatotoxicity-related pathways affected after treatment with the pesticides, alone and in mixtures. Our data show that 2986 genes were differentially regulated by CTD while IMZ and THI had effects on 194 and 225 genes, respectively. All three individual compounds shared a common subset of genes whose network is associated with xenobiotic metabolism and nuclear receptor activation. Similar networks were retrieved for the mixtures. Alterations in the expression of individual genes were in line with the assumption of dose addition. Our results bring new insight into the hepatotoxicity mechanisms of IMZ, THI, and CTD and their mixtures.

Topics: Animals; Chemical and Drug Induced Liver Injury; Dose-Response Relationship, Drug; Female; Gene Expression Profiling; Guanidines; Imidazoles; Neonicotinoids; Pesticides; Rats; Rats, Wistar; Sequence Analysis, RNA; 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

Humans are exposed to pesticide residues through various food products. As these residues can occur in mixtures, there is a need to investigate possible mixture effects on human health. Recent exposure studies revealed the preponderance of imazalil, thiacloprid, and clothianidin in food diets. In this study, we assessed their toxicity alone and in binary mixtures in a 28-day gavage study in female Wistar rats. Five dose levels (up to 350 mg/kg bw/day) ranging from a typical toxicological reference value to a clear effect dose were applied. Data show that the liver was a target organ of all pesticides and their mixtures. Increases in liver weight were observed and histopathological examination revealed centrilobular hepatocellular hypertrophy and cytoplasm degeneration for all treatment conditions. No accumulation of hepatic triglycerides was reported. Tissue residue analysis showed altered pesticide residues in the liver and the kidney when being in mixture as compared to the levels of pesticide residues for the single compound treatment, indicating possible toxicokinetic interactions. Overall, all mixtures appeared to follow the additivity concept, even though quantitative analysis was limited for some endpoints due to the semi-quantitative nature of the data, raising no specific concern for the risk assessment of the examined pesticides.

Topics: Animals; Body Weight; Chemical and Drug Induced Liver Injury; Female; Guanidines; Imidazoles; Kidney; Liver; Neonicotinoids; No-Observed-Adverse-Effect Level; Organ Size; Pesticides; Rats; Rats, Wistar; Risk Assessment; Thiazines; Thiazoles

Risk evaluations for agricultural chemicals are necessary to preserve healthy populations of honey bee colonies. Field studies on whole colonies are limited in behavioural research, while results from lab studies allow only restricted conclusions on whole colony impacts. Methods for automated long-term investigations of behaviours within comb cells, such as brood care, were hitherto missing. In the present study, we demonstrate an innovative video method that enables within-cell analysis in honey bee (Apis mellifera) observation hives to detect chronic sublethal neonicotinoid effects of clothianidin (1 and 10 ppb) and thiacloprid (200 ppb) on worker behaviour and development. In May and June, colonies which were fed 10 ppb clothianidin and 200 ppb thiacloprid in syrup over three weeks showed reduced feeding visits and duration throughout various larval development days (LDDs). On LDD 6 (capping day) total feeding duration did not differ between treatments. Behavioural adaptation was exhibited by nurses in the treatment groups in response to retarded larval development by increasing the overall feeding timespan. Using our machine learning algorithm, we demonstrate a novel method for detecting behaviours in an intact hive that can be applied in a versatile manner to conduct impact analyses of chemicals, pests and other stressors.

Topics: Animals; Bees; Behavior, Animal; Guanidines; Insecticides; Larva; Machine Learning; Neonicotinoids; Population Surveillance; Thiazines; Thiazoles; Video Recording

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 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

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

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

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

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

Queen health is crucial to colony survival of honeybees, since reproduction and colony growth rely solely on the queen. Queen failure is considered a relevant cause of colony losses, yet few data exist concerning effects of environmental stressors on queens. Here we demonstrate for the first time that exposure to field-realistic concentrations of neonicotinoid pesticides can severely affect the immunocompetence of queens of western honeybees (Apis mellifera L.). In young queens exposed to thiacloprid (200 µg/l or 2000 µg/l) or clothianidin (10 µg/l or 50 µg/l), the total hemocyte number and the proportion of active, differentiated hemocytes was significantly reduced. Moreover, functional aspects of the immune defence namely the wound healing/melanisation response, as well as the antimicrobial activity of the hemolymph were impaired. Our results demonstrate that neonicotinoid insecticides can negatively affect the immunocompetence of queens, possibly leading to an impaired disease resistance capacity.

Topics: Animals; Bees; Guanidines; Hemocytes; Hemolymph; Immune Tolerance; Immunocompetence; Insecticides; Lethal Dose 50; Neonicotinoids; Reproduction; Thiazines; 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

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

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

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

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

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

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

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

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

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

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

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

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

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

Three neonicotinyl insecticides, acetamiprid, thiacloprid and clothianidin, were evaluated for their impact on four species of lepidopteran pests of apple in Washington, the codling moth, Cydia pomonella (L.), the Pandemis leafroller, Pandemis pyrusana Kearfott, and the obliquebanded leafroller, Choristoneura rosaceana (Harris), and Lacanobia subjuncta (Grote & Robinson). None of the neonicotinyl insecticides demonstrated sufficient activity against P. pyrusana, C. rosaceana, or L. subjuncta to warrant field trials. Conversely, all had some activity against one or more stages of C. pomonella. Acetamiprid was highly toxic to larvae in laboratory bioassays, and had relatively long activity of field-aged residues (21 days). It also showed some toxicity to C. pomonella eggs (via topical exposure) and adults. Acetamiprid provided the highest level of fruit protection from C. pomonella attack in field trials conducted over five years in experimental orchards with extremely high codling moth pressure. Thiacloprid performed similarly in bioassays, but fruit protection in field trials was slightly lower than acetamiprid. Clothianidin showed moderate to high toxicity in bioassays, depending on the C. pomonella stage tested, but poor fruit protection from attack in field trials. None of the neonicotinyl insecticides were as toxic to larvae or effective in protecting fruit as the current standard organophosphate insecticide used for C. pomonella control, azinphosmethyl. However, both acetamiprid and thiacloprid should provide acceptable levels of C. pomonella control in commercial orchards where densities are much lower than in the experimental orchards used for our trials. The advantages and disadvantages of the neonicotinyl insecticides as replacements for the organophosphate insecticides and their role in a pest management system for Washington apple orchards are discussed.

Topics: Agriculture; Animals; Azinphosmethyl; Biological Assay; Dose-Response Relationship, Drug; Guanidines; Insect Control; Insecticides; Larva; Lepidoptera; Lethal Dose 50; Malus; Neonicotinoids; Ovum; Pyridines; Thiazines; Thiazoles; Time Factors; Washington

The effect of neonicotinyl insecticides on integrated mite control in Washington apple was examined from 0 In a series of 20 field trials (54 treatments) designed primarily to look at efficacy against the codling moth, Cydia pomonella, nearly half of the treatments using four or more applications of acetamiprid had peak mite densities exceeding the economic threshold of 5 mites per leaf. Overall, acetamiprid treatments had 4.6-fold higher mite densities than the standard organophosphate insecticide treatment. Of the treatments with high mite populations, Panonychus ulmi, the European red mite, and Tetranychus urticae, the twospotted spider mite, were the dominant species in roughly equal numbers of cases. Only 11.1% of the thiacloprid treatments exceeded 5 mites per leaf; these experimental treatments included eight applications, whereas the current label restricts the number of applications at the rate for C. pomonella to two applications. One out of six clothianidin treatments caused a significantly higher mite density than the standard treatment; however, this material appeared to suppress predatory mites. Neonicotinyl insecticides did not eliminate predatory mites, but they inhibited their ability to respond normally to increasing prey populations. In field trials designed specifically to examine mite population densities where neonicotinyl insecticides were used, significantly higher levels of tetranychid mites occurred in one or more acetamiprid treatments (one, two or four applications) in five out of six trials. In the sixth trial (in a commercial orchard), only two acetamiprid applications were made, and mite populations were low in all treatments. While elevated mite densities were more likely to occur with four applications, in one case it occurred following a single application. The predominant tetranychid mite species (either P. ulmi or T. urticae) varied from trial to trial; however, there was no apparent bias regarding stimulation of the two species. Horticultural mineral oil was used with acetamiprid in some trials in an attempt to mitigate mite outbreaks. However, the addition of oil did not counteract the tendency of acetamiprid to increase tetranychid mite populations, and in one trial, had a negative effect on predatory mite densities. Seasonal tetranychid mite density was positively related to the total grams AI (or number of applications) of acetamiprid, thus reducing the number of applications per season should lower the probability

Topics: Anabasine; Animals; Biological Assay; Dose-Response Relationship, Drug; Guanidines; Insect Control; Insecticides; Larva; Lepidoptera; Lethal Dose 50; Malus; Mineral Oil; Neonicotinoids; Ovum; Population Density; Pyridines; Seasons; Thiazines; Thiazoles; Time Factors; Washington