Neonicotinoid Insecticide Imidacloprid Research Articles

Investigation on Heterostructured SeO2–TiO2 Nanofluoroprobe for Highly Selective and Sensitive Detection of a Neonicotinoid Insecticide, Imidacloprid in Soil and Water Matrixes

Heterostructured SeO2–TiO2 nanoparticles were used as a highly sensitive and selective fluoroprobe for the detection and determination of neonicotinoid insecticide imidacloprid. The sonication-assisted co-precipitation technique was used to create nanoparticles, which were then characterized using UV–Vis, SEM–EDS, HRTEM, XRD, and TGA. These analyses revealed that the nanoparticles had globular morphology, a crystallite size of 14 nm, a bandgap of 2.45 eV, and stability at high temperatures. Fluorescence emission at 548 nm wavelength exhibits high sensitivity and selectivity for imidacloprid over other pesticides with complete fluorescent quenching at 2 × 10–4 M concentration of imidacloprid. Stern–Volmer equation and thermodynamic parameters applied to the experimental data over a temperature range of 20–50 °C provide the Ksv value in the range of 1.56 × 105 to 2.69 × 105 M−1 and indicate strong interactions, dynamic, spontaneous, and endothermic quenching process. Furthermore, the excellent salient features and notably the unaffected performance even in the presence of various anions, fertilizer, and binary mixture of pesticides often sprayed on plants, will pave the way for the development of rapid, affordable, selective fluoroprobe for onsite monitoring of imidacloprid. The reliability for determination is also examined for spiked soil samples to develop a hybrid SeO2–TiO2 nanoparticles fluoroprobe as a simple alternative to existing techniques for the detection of imidacloprid.Graphical

Photocatalytic Degradation of Neonicotinoids—A Comparative Study of the Efficacy of Hybrid Photocatalysts

This study deals with the synthesis and characterization of a series of hybrid photocatalysts consisting of different loadings of TiO2, Cd, and Fe on mesoporous SBA-15 material. The prepared samples were characterized by Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), and scanning electron microscopy (SEM), and tested for the removal of the neonicotinoid insecticide imidacloprid. The results showed that uncalcined 10% Cd-SBA-15 catalyst exhibited the best photocatalytic activity. The photocatalytic degradation of the imidacloprid was carried out in a batch photoreactor at different pH values, and in the presence or absence of additional compounds such as peroxymonosulfate (PMS) and peroxydisulfate (PDS). The best degradation results were achieved at a pH value of 6.5 with 10% Cd/SBA-15. The degradation performance increased with the addition of PMS and PDS. Based on the results of the experimental measurements, Cd/SBA-15 is a good candidate that can show a reasonable degradation efficiency and reactivity, especially in the presence of PDS or PMS.

Development of a soybean leaf disc assay for determining oral insecticidal activity in the lepidopteran agricultural pest Helicoverpa armigera

Pest insects pose a heavy burden on global agricultural industries with small molecule insecticides being predominantly used for their control. Unwanted side effects and resistance development plagues most small molecule insecticides such as the neonicotinoids, which have been reported to be harmful to honeybees. Bioinsecticides like Bacillus thuringiensis (Bt) toxins can be used as environmentally-friendly alternatives. Arachnid venoms comprise another promising source of bioinsecticides, containing a multitude of selective and potent insecticidal toxins. Unfortunately, no standardised insect models are currently available to assess the suitability of insecticidal agents under laboratory conditions. Thus, we aimed to develop a laboratory model that closely mimics field conditions by employing a leaf disk assay (LDA) for oral application of insecticidal agents in a bioassay tray format. Neonate larvae of the cotton bollworm (Helicoverpa armigera) were fed with soybean (Glycine max) leaves that were treated with different insecticidal agents. We observed dose-dependent insecticidal effects for Bt toxin and the neonicotinoid insecticide imidacloprid, with imidacloprid exhibiting a faster response. Furthermore, we identified several insecticidal arachnid venoms that were active when co-applied with sub-lethal doses of Bt toxin. We propose the H. armigera LDA as a suitable tool for assessing the insecticidal effects of insecticidal agents against lepidopterans.

Non-native Rhizophora mangle as sinks for coastal contamination on Moloka'i, Hawai'i

Coastal mangrove forests provide a suite of environmental services, including sequestration of anthropogenic contamination. Yet, research lags on the environmental fate and potential human health risks of mangrove-sequestered contaminants in the context of mangrove removal for development and range shifts due to climate change. To address this, we conducted a study on Moloka'i, Hawai'i, comparing microplastic and pesticide contamination in coastal compartments both at areas modified by non-native red mangroves (Rhizophora mangle) and unmodified, open coastline. Sediment, porewater, and mangrove plant tissues were collected to quantify microplastic and pesticide concentrations across ecosystem type. Average microplastics were similar between mangrove (8.89 items/kg) and non-mangrove areas (9.01 items/kg) in sediment and porewater, but mangrove roots were a substantial reservoir of microplastics (2004 items/kg). Additionally, there was a strong relationship between proximity to urban development and microplastics detected. Six pesticides were detected, most commonly the insecticide bifenthrin, found in most sediment samples (11.3 ng/g), all root samples (243.3 ng/g), and one propagule sample (8.60 ng/g). Other pesticides detected with appreciable concentrations include the neonicotinoid insecticide imidacloprid and the legacy insecticide transformation product, p,p’-DDE. The other detections, all at concentrations < 1 ng/g, were p,p’-DDT, trifluralin, and permethrin. The high concentrations of bifenthrin in roots compared to lower concentrations detected in sediment suggest that mangrove roots strongly accumulate some pesticides, indicating mangrove roots as a sink for organic contaminants. Study methods could be applied to other Hawaiian Islands and other locations where mangroves have been introduced to further examine the observed trends. Additional information is needed to investigate the fate and cycling of pesticides and microplastics adhered to mangrove roots, to better inform non-native mangrove removal efforts on Moloka'i and elsewhere.

A cytochrome P450 insecticide detoxification mechanism is not conserved across the Megachilidae family of bees.

Recent work has demonstrated that many bee species have specific cytochrome P450 enzymes (P450s) that can efficiently detoxify certain insecticides. The presence of these P450s, belonging or closely related to the CYP9Q subfamily (CYP9Q-related), is generally well conserved across the diversity of bees. However, the alfalfa leafcutter bee, Megachile rotundata, lacks CYP9Q-related P450s and is 170-2500 times more sensitive to certain insecticides than bee pollinators with these P450s. The extent to which these findings apply to other Megachilidae bee species remains uncertain. To address this knowledge gap, we sequenced the transcriptomes of four Megachile species and leveraged the data obtained, in combination with publicly available genomic data, to investigate the evolution and function of P450s in the Megachilidae. Our analyses reveal that several Megachilidae species, belonging to the Lithurgini, Megachilini and Anthidini tribes, including all species of the Megachile genus investigated, lack CYP9Q-related genes. In place of these genes Megachile species have evolved phylogenetically distinct CYP9 genes, the CYP9DM lineage. Functional expression of these P450s from M. rotundata reveal they lack the capacity to metabolize the neonicotinoid insecticides thiacloprid and imidacloprid. In contrast, species from the Osmiini and Dioxyini tribes of Megachilidae have CYP9Q-related P450s belonging to the CYP9BU subfamily that are able to detoxify thiacloprid. These findings provide new insight into the evolution of P450s that act as key determinants of insecticide sensitivity in bees and have important applied implications for pesticide risk assessment.

Using dietary exposure to determine sub-lethal effects from imidacloprid in two springtail (Collembola) species

Standard toxicity tests expose springtails (Collembola) through soil, while dietary exposure tests with animals visible on a surface are less commonly applied. We refined a method for dietary chemical exposure for two widely distributed and abundant Collembola species: Folsomia quadrioculata and Hypogastrura viatica as existing methods were sub-optimal. Newly hatched Collembola were offered bark with a natural layer of Cyanobacteria that was either moistened with a solution of the neonicotinoid insecticide imidacloprid using a micropipette or soaked in the solution overnight. The first method was superior in producing a measured concentration close to the nominal (0.21 and 0.13 mg/kg dry bark, respectively), and resulting in sub-lethal effects as expected. The adult body size was reduced by 8% for both species, but egg production only in H. viatica. Contrastingly, soaked bark resulted in a measured concentration of 8 mg/kg dry bark, causing high mortality and no egg production in either species. Next, we identified the sub-lethal concentration-range by moistening the bark to expose H. viatica to 0, 0.01, 0.04, 0.13, 0.43 and 1.2 mg imidacloprid/kg dry bark. Only the highest concentration affected survival, causing a mortality of 77%. Imidacloprid reduced moulting rate and the body size at first reproduction. The age at first reproduction appeared delayed as some replicates did not reproduce within the experiment duration. The method of moistened bark for dietary exposure proved optimal to continuously study life history traits, such as growth and reproductive outcomes, which are important to understand effects on key events crucial for population viability and growth.

Direct Infusion Device for Molecule Delivery in Plants.

Testing the function of therapeutic compounds in plants is an important component of agricultural research. Foliar and soil-drench methods are routine but have drawbacks, including variable uptake and the environmental breakdown of tested molecules. Trunk injection of trees is well-established, but most methods for this require expensive, proprietary equipment. To screen various treatments for Huanglongbing, a simple, low-cost method to deliver these compounds to the vascular tissue of small greenhouse-grown citrus trees infected with the phloem-limited bacterium Candidatus Liberibacter asiaticus (CLas) or infested with the phloem-feeding CLas insect vector Diaphorina citri Kuwayama (D. citri) is needed. To meet these screening requirements, a direct plant infusion (DPI) device was designed that connects to the plant's trunk. The device is made using a nylon-based 3D-printing system and easily obtainable auxiliary components. The compound uptake efficacy of this device was tested in citrus plants using the fluorescent marker 5,6-carboxyfluorescein-diacetate. Uniform compound distribution of the marker throughout the plants was routinely observed. Furthermore, this device was used to deliver antimicrobial and insecticidal molecules to determine their effects on CLas and D. citri respectively. The aminoglycoside antibiotic streptomycin was delivered into CLas-infected citrus plants using the device, which resulted in a reduction in the CLas titer from 2 weeks to 4 weeks post treatment. Delivering the neonicotinoid insecticide imidacloprid into D. citri-infested citrus plants resulted in a significant increase in psyllid mortality after 7 days. These results suggest that this DPI device represents a useful system for delivering molecules into plants fortesting and facilitate research and screening purposes.

Effects of the neonicotinoid insecticides thiamethoxam and imidacloprid on metamorphosis of the toad Rhinella arenarum at environmentally-relevant concentrations

ABSTRACT The goal of the present study was to examine the effects of environmentally relevant concentrations of the neonicotinoid insecticides thiamethoxam and imidacloprid on the metamorphosis of the toad Rhinella arenarum. Tadpoles were exposed from stage 27 until completion of metamorphosis to concentrations of thiamethoxam ranging between 1.05 and 1050 µg/L and concentrations of imidacloprid varying between 3.4 and 3400 µg/L. The two neonicotinoids were found to act differently at the range of concentrations tested. Thiamethoxam did not markedly alter the final % tadpoles completing metamorphosis but extended by 6–20 days the time needed for tadpoles to complete metamorphosis. The extra number of days required to reach metamorphosis was concentration-dependent between 1.05 and 100.5 µg/L, and then stable at 20 days between 100.5 and 1005 µg/L. In contrast, imidacloprid did not significantly interfere with the overall time needed to complete metamorphosis but decreased success of metamorphosis at 3400 µg/L, the highest concentration tested. Both neonicotinoid concentrations did not markedly alter body size and weight of the newly metamorphosed toads. With a lowest observed effect concentration (LOEC) of 1.05 µg/L, thiamethoxam may be more likely to impact tadpole development in the wild compared to imidacloprid, which was without any apparent effect at concentrations up to 340 µg/L (no-observed effect concentration or NOEC). As the influence of thiamethoxam was triggered after tadpoles had reached Stage 39, when metamorphosis is strictly dependent upon thyroid hormones, this observed effect is attributed to result from actions of this neonicotinoid insecticide on the hypothalamic-pituitary-thyroid axis.

Developmental exposure of zebrafish to neonicotinoid pesticides: Long-term effects on neurobehavioral function

Neonicotinoid compounds are commonly used insecticides which have become increasingly used as replacements of older generations of insecticides, such as organophosphates. Given the established neurotoxicity of cholinergic toxicants, developmental neurotoxicity studies are needed to identify in vertebrate species the potential toxicity of these insecticides which act on nicotinic cholinergic receptors. Previously, developmental exposure to a neonicotinoid insecticide imidacloprid was shown to cause persisting neurobehavioral toxicity in zebrafish. The current study evaluated neurobehavioral effects of embryonic exposure to two other neonicotinoid insecticides, clothianidin (1–100 µM) and dinotefuran (1–100 µM) in zebrafish (5–120 h post-fertilization), concentrations below the threshold for increased lethality and overt dysmorphogenesis. Neurobehavioral tests were conducted at larval (6 days), adolescent (10 weeks) and adult (8 months) ages. Both compounds caused short-term behavioral effects on larval motility, although these effects were distinct from one another. At a lower concentration (1 µM) clothianidin increased dark-induced locomotor stimulation the second time the lights turned off, while a higher concentration (100 µM) reduced activity in the dark at its second presentation. By contrast, dinotefuran (10–100 µM) caused a general decrease in locomotion. Specific longer-term neurobehavioral toxicity after early developmental exposure was also seen. clothianidin (100 µM) reduced locomotor activity in the novel tank in adolescence and adulthood, as well as reduced baseline activity in the tap startle test (1–100 µM) and reduced activity early (1–10 µM) or throughout the predator avoidance test session (100 µM). In addition to locomotor effects, clothianidin altered the diving response in a dose-, age- and time-block-dependent manner (1 µM, 100 µM), causing fish to remain further away from a fast predator cue (100 µM) relative to controls. Dinotefuran produced comparatively fewer effects, increasing the diving response in adulthood (10 µM), but not adolescence, and suppressing initial locomotor activity in the predator avoidance test (1–10 µM). These data indicate that neonicotinoid insecticides may carry some of the same risks for vertebrates posed by other classes of insecticides, and that these adverse behavioral consequences of early developmental exposure are evident well into adulthood.

Do environmentally relevant concentrations of the neonicotinoid insecticide imidacloprid induce DNA damage and oxidative stress on Cnesterodon decemmaculatus (Jenyns, 1842)?

Lethal and sublethal effects of imidacloprid (IMI) were assessed on Cnesterodon decemmaculatus (Pisces: Poeciliidae) by acute exposure to environmentally relevant concentrations of the commercial formulation Punto 35® (Gleba S.A.) under laboratory conditions. Specimens were exposed for 96 h to 1, 10, 20, 25, 35, 75, 100, 125, 150 and 175 mg IMI L-1 from which an LC50 96 h value of 35.59 mg IMI L-1 was calculated. Moreover, sublethal concentrations 0.175, 0.35 and 1 mg IMI L-1 for 96 h were employed for the evaluation of the comet assay and the variation of activities of catalase (CAT) and glutathione content (GSH). Result demonstrated an increased genetic damage index and activity of CAT was observed. Conversely, no significant variation was observed in GSH activity. Total protein content decreased in treated organisms. These results represent the first report of acute effects induced by IMI on C. decemmaculatus exposed under laboratory conditions.

Potential Risk of Residues From Neonicotinoid-Treated Sugar Beet Flowering Weeds to Honey Bees (Apis mellifera L.).

In 2018 the European Union (EU) banned the three neonicotinoid insecticides imidacloprid, clothianidin (CLO), and thiamethoxam (TMX), but they can still be used if an EU Member State issues an emergency approval. Such an approval went into effect in 2021 for TMX-coated sugar beet seeds in Germany. Usually, this crop is harvested before flowering without exposing non-target organisms to the active ingredient or its metabolites. In addition to the approval, strict mitigation measures were imposed by the EU and the German federal states. One of the measures was to monitor the drilling of sugar beet and its impact on the environment. Hence we took residue samples from different bee and plant matrices and at different dates to fully map beet growth in the German states of Lower Saxony, Bavaria, and Baden-Württemberg. A total of four treated and three untreated plots were surveyed, resulting in 189 samples. Residue data were evaluated using the US Environmental Protection Agency BeeREX model to assess acute and chronic risk to honey bees from the samples, because oral toxicity data are widely available for both TMX and CLO. Within treated plots, we found no residues either in pools of nectar and honey crop samples (n = 24) or dead bee samples (n = 21). Although 13% of beebread and pollen samples and 88% of weed and sugar beet shoot samples were positive, the BeeREX model found no evidence of acute or chronic risk. We also detected neonicotinoid residues in the nesting material of the solitary bee Osmia bicornis, probably from contaminated soil of a treated plot. All control plots were free of residues. Currently, there are insufficient data on wild bee species to allow for an individual risk assessment. In terms of the future use of these highly potent insecticides, therefore, it must be ensured that all regulatory requirements are complied with to mitigate any unintentional exposure. Environ Toxicol Chem 2023;00:1-11. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

High sensitivity to dietary imidacloprid exposure in early life stages of Folsomia quadrioculata (Collembola) populations from contrasting climates

Survival of early life stages is critical for population growth. Understanding the vulnerability of these sensitive stages can therefore improve ecological risk predictions. The globally-used neonicotinoid insecticide imidacloprid does not degrade easily in soil and occurs at concentrations potentially toxic to non-target soil organisms, such as Collembola. Here, we studied the effects of dietary exposure to imidacloprid on the survival of juveniles in an arctic and a temperate population of Folsomia quadrioculata (Collembola), a commonly-occurring litter-dwelling species in the Northern Hemisphere. The exposure lasted from <24 h to 30 days since hatching at 15 °C. We assessed whether (1) the arctic population of the test species with faster development rates could be more vulnerable than the temperate population, (2) cumulative mortality could increase substantially with exposure time, and (3) the mortality risk experienced during the juvenile stage could be greater than that experienced by the adults. We found a strong concentration-dependent decline in survival in both populations, with small but significantly different LC50s, despite large differences in multiple life-history traits. Applying a General Unified Threshold model for Survival (GUTS) showed a low threshold for effects, slow damage dynamics, a 6–11-fold decrease in LC50 from 14 to 30 days of exposure, and saturation of damage at the three highest concentrations. In addition, exposure starting soon after hatching caused much more mortality than that starting in the adult stages. Our findings suggest that toxicity tests not emphasizing the neonate stages or not lasting long enough can underestimate the implications for populations of naturally abundant non-target organisms.

Assessment of Global DNA Methylation in SH-SY5Y Cells Exposed to the Neonicotinoid Insecticides Imidacloprid and Thiamethoxam.

Neonicotinoid insecticides, known for their selectivity and low mammalian toxicity, have been widely used in recent years as alternatives to organophosphate insecticides. Although neonicotinoids are generally considered to be safe, data show that they can cause harmful effects on human and environmental health. Due to the lack of information on their mechanism of toxicity, the effects of imidacloprid and thiamethoxam on DNA methylation as the most used marker for epigenetic effects were investigated in human neuroblastoma (SH-SY5Y) cells. The cells were exposed to imidacloprid and thiamethoxam in concentrations of 100, 200, and 500μM for 24 hours, then global DNA methylation and expression of genes involved in global DNA methylation (DNMT1, DNMT3a and DNMT3b) were investigated. Global DNA methylation significantly increased after imidacloprid exposure at 100μM, and thiamethoxam exposures at 200µM and 500μM (>1.5-fold). Imidacloprid significantly decreased the expression of DNMT1 and DNMT3a, whereas thiamethoxam did not cause any significant changes in the expression of DNMT genes. Our findings suggested that alteration in global DNA methylation may be involved in the toxic mechanisms of imidacloprid and thiametoxam.

The neonicotinoid insecticide imidacloprid has unexpected effects on the growth and development of soil amoebae

Neonicotinoid pesticides are the most widely used insecticides worldwide and have become a global environmental issue. Previous studies have shown that imidacloprid, the most used neonicotinoid, can negatively affect a wide range of organisms, including non-target insects, fish, invertebrates, and mammals. Imidacloprid can also accumulate and persist in soils, posing threats to the terrestrial ecosystem. However, we know little about one ecologically important group of organisms, the single-celled soil protists. In this study, we used a soil amoeba, Dictyostelium discoideum, to test whether and how imidacloprid affects the growth and development of soil amoebae. We provide the first empirical evidence that environmental concentrations of imidacloprid negatively impact the fitness and development of soil amoebae. In addition, the adverse effects did not show a dose-response relationship with increased imidacloprid concentrations, where no significant difference was observed among the treatment groups. Further transcriptome analyses showed that imidacloprid affected amoeba's key DEGs related to phagocytosis, cell division, morphogenesis, and cytochrome P450. Moreover, soil amoebae show both conserved and novel transcriptional responses to imidacloprid. In conclusion, this study has expanded the non-target list of imidacloprid from animals and plants to single-celled protists, and we believe the impact of neonicotinoid pesticides on the microbiome is significantly underestimated and deserves more studies.

Combining time-resolved transcriptomics and proteomics data for Adverse Outcome Pathway refinement in ecotoxicology

Conventional Environmental Risk Assessment (ERA) of pesticide pollution is based on soil concentrations and apical endpoints, such as the reproduction of test organisms, but has traditionally disregarded information along the organismal response cascade leading to an adverse outcome. The Adverse Outcome Pathway (AOP) framework includes response information at any level of biological organization, providing opportunities to use intermediate responses as a predictive read-out for adverse outcomes instead. Transcriptomic and proteomic data can provide thousands of data points on the response to toxic exposure. Combining multiple omics data types is necessary for a comprehensive overview of the response cascade and, therefore, AOP development. However, it is unclear if transcript and protein responses are synchronized in time or time lagged. To understand if analysis of multi-omics data obtained at the same timepoint reveal one synchronized response cascade, we studied time-resolved shifts in gene transcript and protein abundance in the springtail Folsomia candida, a soil ecotoxicological model, after exposure to the neonicotinoid insecticide imidacloprid. We analyzed transcriptome and proteome data every 12 h up to 72 h after onset of exposure. The most pronounced shift in both transcript and protein abundances was observed after 48 h exposure. Moreover, cross-correlation analyses indicate that most genes displayed the highest correlation between transcript and protein abundances without a time-lag. This demonstrates that a combined analysis of transcriptomic and proteomic data from the same time-point can be used for AOP improvement. This data will promote the development of biomarkers for the presence of neonicotinoid insecticides or chemicals with a similar mechanism of action in soils.

Determination of Neonicotinoid Insecticides in Environmental Water by the Enrichment of MIL-53 Mixed Matrix Membrane Coupled with High Performance Liquid Chromatography.

Metal organic framework based mixed matrix membranes (MOF-MMMs) were synthesized and applied for dispersive membrane extraction (DME) of four neonicotinoid insecticides (nitenpyram, thiacloprid, imidacloprid, and acetamiprid) in environmental water, combined with high performance liquid chromatography (HPLC) for determination. Several experimental conditions were optimized in detail, involving dosage percentage of MOF, extraction time, sample pH, salinity, type and volume of eluent, and elution time. High sensitivity with limits of detection and quantification were achieved as 0.013-0.064 μg L-1 and 0.038-0.190 μg L-1, respectively, and good precision with relative standard deviations were obtained as 3.07-12.78%. The proposed method has been successfully applied to determine four neonicotinoid insecticides in tap water, surface water, and seawater, satisfactory recoveries of spiked water samples were between 72.50 and 117.98%. Additionally, the MOF-MMMs showed good reusability with the extraction efficiencies almost remaining stable after 14 cycles. The MOF-MMMs based DME followed by the HPLC method can be a promising utility for the determination of neonicotinoid insecticides in environmental water samples, with high sensitivity and convenient operation.

Glassy-Winged Sharpshooters Cease Feeding and Avoid Plants Treated With Sub-Lethal Doses of the Neonicotinoid Insecticide Imidacloprid.

Insecticides are a primary means for suppressing populations of insects that transmit plant pathogens. Application of insecticides for limiting the spread of insect-transmitted plant pathogens is often most effective when applied on an area-wide scale. The glassy-winged sharpshooter is a vector of the bacterial pathogen Xylella fastidiosa, which causes numerous plant diseases including Pierce's disease of grapevine. The glassy-winged sharpshooter has been the subject of an area-wide suppression program in California for nearly two decades. Overreliance on a limited number of active ingredients including the neonicotinoid imidacloprid has resulted in increased levels of resistance to commonly applied products. In California, glassy-winged sharpshooters move between citrus, an important overwintering host, and vineyards. Accordingly, imidacloprid is routinely applied via the irrigation system in vineyards and citrus orchards. For soil applied applications, it may take days to weeks for concentrations in plants to increase to lethal doses. Further, as the dose of imidacloprid required to kill sharpshooters increases due to resistance, so too does the period that sharpshooters are exposed to sub-lethal doses. Response of glassy-winged sharpshooter to cowpea plants treated with sub-lethal doses of imidacloprid was evaluated by conducting no-choice and choice tests. In no-choice feeding assays, glassy-winged sharpshooters caged on plants treated with sub-lethal doses of imidacloprid ceased feeding and produced little excreta. Further, sub-lethal exposure to a range of doses over a 4-d period did not affect viability over a 9-wk post-exposure holding period on untreated plants. In choice-tests, glassy-winged sharpshooters avoided treated plants and were observed predominately on untreated plants. Results suggest that application of imidacloprid to vineyards and citrus orchards may push glassy-winged sharpshooters out of treated habitats rather than kill them.

Toxicity of the neonicotinoid insecticides thiamethoxam and imidacloprid to tadpoles of three species of South American amphibians and effects of thiamethoxam on the metamorphosis of Rhinella arenarum

ABSTRACT The present study examined the acute and chronic toxicity of the neonicotinoid insecticides imidacloprid (IMI) and thiamethoxam (TIA) on the neotropical amphibian species Rhinella arenarum, Rhinella fernandezae and Scinax granulatus. The median lethal concentration after 96 hr exposure (96 hr-LC50) ranged between 11.28 and >71.2 mg/L amongst all species and development stages tested, indicating that these pesticides are not likely to produce acute toxicity in the wild. The subchronic toxicity was also low, with 21 day-LC50 values ranging between 27.15 and >71.2 mg/L. However, tadpoles of Rhinella arenarum exposed to thiamethoxam from stage 27 until completion of metamorphosis presented a significantly lower metamorphic success rate together with a smaller size at metamorphosis, starting from the lowest concentration tested. Although a number of studies previously examined the effects of neonicotinoids on amphibian tadpoles, these investigations focused on the time to metamorphosis and reported a variety of results including retardation, acceleration or lack of effect. Here, data demonstrated that thiamethoxam predominantly impacts metamorphosis through reduction of the transformation success and body weight, rather than by affecting the timings of metamorphosis. By closely monitoring progression of tadpoles through the different stages, impairment of metamorphosis was demonstrated to occur during the transition from stage 39 to 42, suggesting an effect on the thyroid system. An asymmetry in the length of the arms was also observed in metamorphs treated with thiamethoxam. Overall, these results indicate that thiamethoxam, and conceivably other neonicotinoids, have the potential to significantly impair metamorphosis of amphibians and diminish their performance and survival in the wild.

Response of microbial community to the remediation of neonicotinoid insecticide imidacloprid contaminated wetland soil by Phanerochaete chrysosporium

Imidacloprid (IMI), a typic neonicotinoid insecticide, is widely used and persist in soils with long half-time causing serious threat to ecosystem and human health. It is urgent to develop suitable and effective methods to accelerate it degradation and alleviate its negative impacts in soil. In this study, the introduction of functional microbe white-rot fungus Phanerochaete chrysosporium to remediate IMI contaminated wetland soil was carried out. The remediation performance and the response of the soil microbial community were examined. The results showed that P. chrysosporium could improve the degradation of IMI in soil no matter the soil was sterilized or not. The bioaugmentation was especially observed in non-sterilized soil under the inoculation patterns of FE and SP with the maximum IMI degradation rate of 91% and 93% in 7 days, respectively. The invertase activity in soil was also enhanced with P. chrysosporium inoculation. Microbial community analysis revealed that P. chrysosporium inoculation could increase the diversity and richness of bacterial community, and stimulate some IMI degraders genera including Ochrobactrum, Leifsonia, Achromobacter, and Bacillus. Moreover, the xenobiotic degradation and metabolism pathway was generally enhanced with P. chrysosporium inoculation based on PICRUSt analysis. These obtained results demonstrated that the introduction of white-rot fungus is of great potentially enabling the remediation of neonicotinoids contaminated soil.

The Insecticide Imidacloprid Decreases Nannotrigona Stingless Bee Survival and Food Consumption and Modulates the Expression of Detoxification and Immune-Related Genes.

Stingless bees are ecologically and economically important species in the tropics and subtropics, but there has been little research on the characterization of detoxification systems and immune responses within them. This is critical for understanding their responses to, and defenses against, a variety of environmental stresses, including agrochemicals. Therefore, we studied the detoxification and immune responses of a stingless bee, Nanotrigona perilampoides, which is an important stingless bee that is widely distributed throughout Mexico, including urban areas, and has the potential to be used in commercial pollination. We first determined the LC50 of the neonicotinoid insecticide imidacloprid for foragers of N. perilampoides, then chronically exposed bees for 10 days to imidacloprid at two field-realistic concentrations, LC10 (0.45 ng/µL) or LC20 (0.74 ng/µL), which are respectively 2.7 and 1.3-fold lower than the residues of imidacloprid that have been found in honey (6 ng/g) in central Mexico. We found that exposing N. perilampoides stingless bees to imidacloprid at these concentrations markedly reduced bee survival and food consumption, revealing the great sensitivity of this stingless bee to the insecticide in comparison to honey bees. The expression of detoxification (GSTD1) and immune-related genes (abaecin, defensin1, and hymenopteacin) in N. perilampoides also changed over time in response to imidacloprid. Gene expression was always lower in bees after 8 days of exposure to imidacloprid (LC10 or LC20) than it was after 4 days. Our results demonstrate that N. perilampoides stingless bees are extremely sensitive to imidacloprid, even at low concentrations, and provide greater insight into how stingless bees respond to pesticide toxicity. This is the first study of its kind to look at detoxification systems and immune responses in Mexican stingless bees, an ecologically and economically important taxon.