Recent and future declines of a historically widespread pollinator linked to climate, land cover, and pesticides.

The acute decline in global biodiversity includes not only the loss of rare species, but also the rapid collapse of common species across many different taxa. The loss of pollinating insects is of particular concern because of the ecological and economic values these species provide. The western bumble bee (Bombus occidentalis) was once common in western North America, but this species has become increasingly rare through much of its range. To understand potential mechanisms driving these declines, we used Bayesian occupancy models to investigate the effects of climate and land cover from 1998 to 2020, pesticide use from 2008 to 2014, and projected expected occupancy under three future scenarios. Using 14,457 surveys across 2.8 million km2 in the western United States, we found strong negative relationships between increasing temperature and drought on occupancy and identified neonicotinoids as the pesticides of greatest negative influence across our study region. The mean predicted occupancy declined by 57% from 1998 to 2020, ranging from 15 to 83% declines across 16 ecoregions. Even under the most optimistic scenario, we found continued declines in nearly half of the ecoregions by the 2050s and mean declines of 93% under the most severe scenario across all ecoregions. This assessment underscores the tenuous future of B. occidentalis and demonstrates the scale of stressors likely contributing to rapid loss of related pollinator species throughout the globe. Scaled-up, international species-monitoring schemes and improved integration of data from formal surveys and community science will substantively improve the understanding of stressors and bumble bee population trends.

Comparability of comparative toxicity: insect sensitivity to imidacloprid reveals huge variations across species but also within species.

Pesticides have been identified as major drivers of insect biodiversity loss. Thus, the study of their effects on non-pest insect species has attracted a lot of attention in recent decades. In general toxicology, the 'gold standard' to assess the toxicity of a substance is to measure mass-specific LD50 (i.e. median lethal dose per unit body mass). In entomology, reviews attempting to compare these data across all available studies are lacking. To fill this gap in knowledge, we performed a systematic review of the lethality of imidacloprid for adult insects. Imidacloprid is possibly the most extensively studied insecticide in recent times, yet we found that little is comparable across studies, owing to both methodological divergence and missing estimates of body mass. By accounting for body mass whenever possible, we show how imidacloprid sensitivity spans across an apparent range of approximately six orders of magnitude across insect species. Very high variability within species can also be observed owing to differences in exposure methods and observation time. We suggest that a more comparable and comprehensive approach has both biological and economic relevance. Ultimately, this would help to identify differences that could direct research towards preventing non-target species from being negatively affected.

Neonicotinoids differentially modulate nicotinic acetylcholine receptors in immature and antral follicles in the mouse ovary1.

Neonicotinoids are the most widely used insecticides in the world. They are synthetic nicotine derivatives that act as nicotinic acetylcholine receptor (nAChR) agonists. Although parent neonicotinoids have low affinity for the mammalian nAChR, they can be activated in the environment and the body to positively charged metabolites with high affinity for the mammalian nAChR. Imidacloprid (IMI), the most popular neonicotinoid, and its bioactive metabolite desnitro-imidacloprid (DNI) differentially interfere with ovarian antral follicle physiology in vitro, but their effects on ovarian nAChR subunit expression are unknown. Furthermore, ovarian nAChR subtypes have yet to be characterized in the ovary. Thus, this work tested the hypothesis that ovarian follicles express nAChRs and their expression is differentially modulated by IMI and DNI in vitro. We used PCR, RNA in situ hybridization, and immunohistochemistry to identify and localize nAChR subunits (α2, 4, 5, 6, 7 and ß1, 2, 4) expressed in neonatal ovaries and antral follicles. Chrnb1 was expressed equally in neonatal ovaries and antral follicles. Chrna2 and Chrnb2 expression was higher in antral follicles compared to neonatal ovaries and Chrna4, Chrna5, Chrna6, Chrna7 and Chrnb4 expression was higher in neonatal ovaries compared to antral follicles. The α subunits were detected throughout the ovary, especially in oocytes and granulosa cells. IMI and DNI dysregulated expression of multiple nAChR subunits in neonatal ovaries, but only dysregulated one subunit in antral follicles. These data indicate that mammalian ovaries contain nAChRs, and their susceptibility to IMI and DNI exposure varies with the stage of follicle maturity.

Non-acute exposure of neonicotinoids, health risk assessment, and evidence integration: a systematic review.

Neonicotinoid pesticides are utilized against an extensive range of insects. A growing body of evidence supports that these neuro-active insecticides are classified as toxicants in invertebrates. However, there is limited published data regarding their toxicity in vertebrates and mammals. the current systematic review is focused on the up-to-date knowledge available for several neonicotinoid pesticides and their non-acute toxicity on rodents and human physiology. Oral lethal dose 50 (LD50) of seven neonicotinoids (i.e. imidacloprid, acetamiprid, clothianidin, dinotefuran, thiamethoxam, thiacloprid, and nitenpyram) was initially identified. Subsequently, a screening of the literature was conducted to collect information about non-acute exposure to these insecticides. 99 studies were included and assessed for their risk of bias and level of evidence according to the Office of Health and Translation (OHAT) framework. All the 99 included papers indicate evidence of reproductive toxicity, hepatotoxicity, nephrotoxicity, neurotoxicity, immunotoxicity, and oxidative stress induction with a high level of evidence in the health effect of rodents and a moderate level of evidence for human health. The most studied type of these insecticides among 99 papers was imidacloprid (55 papers), followed by acetamiprid (22 papers), clothianidin (21 papers), and thiacloprid (11 papers). While 10 of 99 papers assessed the relationship between clothianidin, thiamethoxam, dinotefuran, and nitenpyram, showing evidence of liver injury, dysfunctions of oxidative stress markers in the reproductive system, and intestinal toxicity. This systematic review provides a comprehensive overview of the potential risks caused by neonicotinoid insecticides to humans and rodents with salient health effects. However, further research is needed to better emphasize and understand the patho-physiological mechanisms of these insecticides, taking into account various factors that can influence their toxicity.

Adult mosquitoes of the sibling species Anopheles gambiae and Anopheles coluzzii exhibit contrasting patterns of susceptibility to four neonicotinoid insecticides along an urban-to-rural gradient in Yaoundé, Cameroon.

BACKGROUND: Neonicotinoids are potential alternatives for controlling pyrethroid-resistant mosquitoes, but their efficacy against malaria vector populations of sub-Saharan Africa has yet to be investigated. The aim of the present study was to test the efficacy of four neonicotinoids against adult populations of the sibling species Anopheles gambiae and Anopheles coluzzii sampled along an urban-to-rural gradient. METHODS: The lethal toxicity of three active ingredients for adults of two susceptible Anopheles strains was assessed using concentration-response assays, and their discriminating concentrations were calculated. The discriminating concentrations were then used to test the susceptibility of An. gambiae and An. coluzzii mosquitoes collected from urban, suburban and rural areas of Yaoundé, Cameroon, to acetamiprid, imidacloprid, clothianidin and thiamethoxam. RESULTS: Lethal concentrations of neonicotinoids were relatively high suggesting that this class of insecticides has low toxicity against Anopheles mosquitoes. Reduced susceptibility to the four neonicotinoids tested was detected in An. gambiae populations collected from rural and suburban areas. By contrast, adults of An. coluzzii that occurred in urbanized settings were susceptible to neonicotinoids except acetamiprid for which 80% mortality was obtained within 72 h of insecticide exposure. The cytochrome inhibitor, piperonyl butoxide (PBO), significantly enhanced the activity of clothianidin and acetamiprid against An. gambiae mosquitoes. CONCLUSIONS: These findings corroborate susceptibility profiles observed in larvae and highlight a significant variation in tolerance to neonicotinoids between An. gambiae and An. coluzzii populations from Yaoundé. Further studies are needed to disentangle the role of exposure to agricultural pesticides and of cross-resistance mechanisms in the development of neonicotinoid resistance in some Anopheles species.

From Water to Water: Insight into the Translocation of Pesticides from Plant Rhizosphere Solution to Leaf Guttation and the Associated Ecological Risks.

Plant guttation is an important source of water/nutrients for many beneficial insects, while the presence of pesticides in guttation has been considered as a new exposure route for nontarget insects. This study aimed to elucidate how 15 diverse pesticides are translocated from growth media to guttation by maize plants through a hydroponic experiment. All pesticides were effectively translocated from the growth solution to maize guttation and reached a steady state within 5 days. The strong positive correlation (R2 = 0.43-0.84) between the concentrations of pesticides in guttation and in xylem sap demonstrated that xylem sap was a major source of pesticides in guttation. The relationship between the bioaccumulation of pesticides in guttation (BCFguttation) and the chemical Kow was split into two distinct patterns: for pesticides with log Kow > 3, we identified a good negative linear correlation between log BCFguttation and log Kow (R2 = 0.71); however, for pesticides with log Kow < 3, all data fall close to a horizontal line of BCFguttation ≅ 1, indicating that hydrophilic pesticides can easily pass through the plants from rhizosphere solution to leaf guttation and reach saturation status. Besides, after feeding with pesticide-contaminated guttation, the mortality of honeybees was significantly impacted, even at very low levels (e.g., ∑600 µg/L with a mortality of 93%). Our results provide essential information for predicting the contamination of plant guttation with pesticides and associated ecological risks.

Clothianidin-resistant Anopheles gambiae adult mosquitoes from Yaoundé, Cameroon, display reduced susceptibility to SumiShield® 50WG, a neonicotinoid formulation for indoor residual spraying.

BACKGROUND: Chronic exposure of mosquito larvae to pesticide residues and cross-resistance mechanisms are major drivers of tolerance to insecticides used for vector control. This presents a concern for the efficacy of clothianidin, an agricultural neonicotinoid prequalified for Indoor Residual Spraying (IRS). METHODS: Using standard bioassays, we tested if reduced susceptibility to clothianidin can affect the efficacy of SumiShield® 50WG, one of four new IRS formulations containing clothianidin. We simultaneously monitored susceptibility to clothianidin and to SumiShield 50WG, testing adults of Anopheles gambiae, An. coluzzii and Culex sp sampled from urban, suburban and agricultural areas of Yaoundé, Cameroon. RESULTS: We found that in this geographic area, the level of susceptibility to the active ingredient predicted the efficacy of SumiShield 50WG. This formulation was very potent against populations that reached 100% mortality within 72 h of exposure to a discriminating concentration of clothianidin. By contrast, mortality leveled off at 75.4 ± 3.5% within 7 days of exposure to SumiShield 50WG in An. gambiae adults collected from a farm where the spraying of the two neonicotinoids acetamiprid and imidacloprid for crop protection is likely driving resistance to clothianidin. CONCLUSIONS: Despite the relatively small geographic extend of the study, the findings suggest that cross-resistance may impact the efficacy of some new IRS formulations and that alternative compounds could be prioritized in areas where neonicotinoid resistance is emerging.

Navigating the complexities of docking tools with nicotinic receptors and acetylcholine binding proteins in the realm of neonicotinoids.

Molecular docking, pivotal in predicting small-molecule ligand binding modes, struggles with accurately identifying binding conformations and affinities. This is particularly true for neonicotinoids, insecticides whose impacts on ecosystems require precise molecular interaction modeling. This study scrutinizes the effectiveness of prominent docking software (Ledock, ADFR, Autodock Vina, CDOCKER) in simulating interactions of environmental chemicals, especially neonicotinoid-like molecules with nicotinic acetylcholine receptors (nAChRs) and acetylcholine binding proteins (AChBPs). We aimed to assess the accuracy and reliability of these tools in reproducing crystallographic data, focusing on semi-flexible and flexible docking approaches. Our analysis identified Ledock as the most accurate in semi-flexible docking, while Autodock Vina with Vinardo scoring function proved most reliable. However, no software consistently excelled in both accuracy and reliability. Additionally, our evaluation revealed that none of the tools could establish a clear correlation between docking scores and experimental dissociation constants (Kd) for neonicotinoid-like compounds. In contrast, a strong correlation was found with drug-like compounds, bringing to light a bias in considered software towards pharmaceuticals, thus limiting their applicability to environmental chemicals. The comparison between semi-flexible and flexible docking revealed that the increased computational complexity of the latter did not result in enhanced accuracy. In fact, the higher computational cost of flexible docking with its lack of enhanced predictive accuracy, rendered this approach useless for this class of compounds. Conclusively, our findings emphasize the need for continued development of docking methodologies, particularly for environmental chemicals. This study not only illuminates current software capabilities but also underscores the urgency for advancements in computational molecular docking as it is a relevant tool to environmental sciences.

Interactive effects of dinotefuran and Nosema ceranae on the survival status and gut microbial community of honey bees.

Growing evidences have shown that the decline in honey bee populations is mainly caused by the combination of multiple stressors. However, the impacts of parasitic Nosema ceranae to host fitness during long-term pesticide exposure-induced stress is largely unknown. In this study, the effects of chronic exposure to a sublethal dose of dinotefuran, in the presence or absence of N. ceranae, was examined in terms of survival, food consumption, detoxification enzyme activities and gut microbial community. The interaction between dinotefuran and Nosema ceranae on the survival of honey bee was synergistic. Co-exposure to dinotefuran and N. ceranae led to less food consumption and greater changes of enzyme activities involved in defenses against oxidative stress. Particularly, N. ceranae and dinotefuran-N. ceranae co-exposure significantly impacted the gut microbiota structure and richness in adult honey bees, while dinotefuran alone did not show significant alternation of core gut microbiota compared to the control group. We herein demonstrated that chronical exposure to dinotefuran decreases honey bee's survival but is not steadily associated with the gut microbiota dysbiosis; by contrast, N. ceranae parasitism plays a dominant role in the combination in influencing the gut microbial community of the host honey bee. Our findings provide a comprehensive understanding of combinatorial effects between biotic and abiotic stressors on one of the most important pollinators, honey bees.

Decreases in the number of microglia and neural circuit dysfunction elicited by developmental exposure to neonicotinoid pesticides in mice.

Neonicotinoids are insecticides widely used in the world. Although neonicotinoids are believed to be toxic only to insects, their developmental neurotoxicity in mammals is a concern. Therefore, we examined the effects of developmental exposure to neonicotinoids on immune system in the brain and post-developmental behaviors in this study. Imidacloprid or clothianidin was orally administered to dams at a dosage of 0.1 mg/kg/day from embryonic day 11 to postnatal day 21. Imidacloprid decreased sociability, and both imidacloprid and clothianidin decreased locomotor activity and induced anxiety, depression and abnormal repetitive behaviors after the developmental period. There was no change in the number of neurons in the hippocampus of mice exposed to imidacloprid. However, the number and activity of microglia during development were significantly decreased by imidacloprid exposure. Imidacloprid also induced neural circuit dysfunction in the CA1 and CA3 regions of the hippocampus during the early postnatal period. Exposure to imidacloprid suppressed the expression of csf1r during development. Collectively, these results suggest that developmental exposure to imidacloprid decreases the number and activity of microglia, which can cause neural circuit dysfunction and abnormal behaviors after the developmental period. Care must be taken to avoid exposure to neonicotinoids, especially during development.

Amino-functional magnetic covalent organic framework as an effective adsorbent for the determination of neonicotinoids in food samples.

Recently, covalent organic frameworks have gained popularity in sample pretreatment. However, the application of covalent organic frameworks in the enrichment of hydrophilic compounds remains a challenge. Thus, a functionalized magnetic covalent organic framework equipped with amino groups was constructed using a bottom-up functionalization strategy. Considering the advantages of this novel adsorbent such as high porosity, large adsorption capacity, and hydrophilic surface, a sensitive magnetic solid-phase extraction coupled with high-performance liquid chromatography-tandem mass spectrometry method was established for the effective determination of neonicotinoids. This method exhibited good linearities with correlation coefficients ranging from 0.9983 to 0.9995, low detection limits in the range 0.003-0.009 ng g-1 and 0.001-0.013 ng mL-1, and limits of quantification in the range 0.010-0.031 ng g-1 and 0.004-0.044 ng mL-1. Furthermore, satisfactory repeatability with relative standard deviations ≤ 6.7% and spiked recoveries between 82.3 and 99.8% were obtained. This work not only provided a promising adsorbent for the sensitive determination of trace-level neonicotinoids but also represented a unique insight for effective enrichment of super hydrophilic hazards.

Characterization of near-field temporal and spatial variations of pesticide residues using honeybee specimens as bio-sensing matrices.

This study was prompted by recent reports of the ubiquity of neonicotinoids (neonics) in environment and the likelihood of exposures and health hazards to non-target organisms. We aimed to quantify neonics levels in time- and location-match pollen and nectar samples foraged by honeybees (Apis mellifera) and characterized the temporal and spatial variations using a relative potency factor method to determine the total neonic levels, expressed as the imidacloprid-adjusted total neonics, IMIRPF (ng/g). Six pairs of pollen and nectar samples, a total of twelve samples, were collected from each of the thirty-two experimental hives during the active foraging months of March, April, and June and analyzed for eight neonics. We found 59% and 64% of pollen and nectar contained at least one neonic, respectively. Among those neonic-detected pollen and nectar samples, 45% and 77% of them contained more than one neonic, respectively. Imidacloprid and acetamiprid in pollen and clothianidin and thiamethoxam in nectar accounted for 60% and 83% detection, respectively. The highest 3-month average of IMIRPF in pollen (6.56 ng/g) and nectar (11.19 ng/g) were detected in a location with the predominant production of citrus fruit. The temporal and spatial variations of IMIRPF levels demonstrated the robustness of using paired pollen and nectar data as the bio-sensing matrices to facilitate the assessment of near-field exposure to total neonics and the delineation of risks.

Fabrication of Nitrogen Based Magnetic Conjugated Microporous Polymer for Efficient Extraction of Neonicotinoids in Water Samples.

Facile and sensitive methods for detecting neonicotinoids (NEOs) in aquatic environments are crucial because they are found in extremely low concentrations in complex matrices. Herein, nitrogen-based magnetic conjugated microporous polymers (Fe3O4@N-CMP) with quaternary ammonium groups were synthesized for efficient magnetic solid-phase extraction (MSPE) of NEOs from tap water, rainwater, and lake water. Fe3O4@N-CMP possessed a suitable specific surface area, extended π-conjugated system, and numerous cationic groups. These properties endow Fe3O4@N-CMP with superior extraction efficiency toward NEOs. The excellent adsorption capacity of Fe3O4@N-CMP toward NEOs was attributed to its π-π stacking, Lewis acid-base, and electrostatic interactions. The proposed MSPE-HPLC-DAD approach based on Fe3O4@N-CMP exhibited a wide linear range (0.1-200 µg/L), low detection limits (0.3-0.5 µg/L), satisfactory precision, and acceptable reproducibility under optimal conditions. In addition, the established method was effectively utilized for the analysis of NEOs in tap water, rainwater, and lake water. Excellent recoveries of NEOs at three spiked levels were in the range of 70.4 to 122.7%, with RSDs less than 10%. This study provides a reliable pretreatment method for monitoring NEOs in environmental water samples.

Effects of imidacloprid on the survival and biomarker responses of Eristalis tenax larvae (Diptera: Syrphidae): a comparative study between indoor and outdoor exposures.

Imidacloprid is a widely used pesticide in agriculture. It is being found in aquatic ecosystems in agricultural regions. This study aimed to evaluate its effects on the survival rates, acetylcholinesterase (AChE) and catalase (CAT) responses of larval Eristalis tenax hoverflies. The larvae were exposed for 3, 7 and 14 days to increasing concentrations of imidacloprid (0, 0.1, 0.5 and 2 mg L-1) both indoors at a constant temperature of 20 °C and outdoors under varying environmental conditions. The results revealed that indoors and outdoors, the mortality of E. tenax significantly increased with increasing imidacloprid concentration and duration of exposure. Median lethal concentrations (LC50) varied from 0.03 to 0.17 mg L-1 depending on the duration and conditions of exposure. Indoors, AChE activity decreased in all the treatments for all three exposure durations, whereas outdoors the decrease was observed after the short (3-day) and long (14-day) exposure durations. AChE inhibition ranged from 6% to 62% (indoors) and 12% to 62% (outdoors). Variations in CAT activity were observed for both experimental setups, with a decrease outdoors in larvae exposed to 0.5 mg L-1 for 7 days and a gradual dose-dependent increase indoors for exposure lasting 3 and 7 days. This study sheds light on the potential ecological implications of imidacloprid contamination which may cause the decline of aquatic insect populations and pollination rates, leading to disruptions of the food chain and the overall decline of aquatic and terrestrial ecosystem health.

Neonicotinoid Insecticides: Molecular Targets, Resistance, and Toxicity.

Neonicotinoids have been used to protect crops and animals from insect pests since the 1990s, but there are concerns regarding their adverse effects on nontarget organisms, notably on bees. Enhanced resistance to neonicotinoids in pests is becoming well documented. We address the current understanding of neonicotinoid target site interactions, selectivity, and metabolism not only in pests but also in beneficial insects such as bees. The findings are relevant to the management of both neonicotinoids and the new generation of pesticides targeting insect nicotinic acetylcholine receptors.

Insights into the ubiquity, persistence and microbial intervention of imidacloprid.

Imidacloprid, a neonicotinoid pesticide, is employed to increase crop productivity. Meanwhile, its indiscriminate application severely affects the non-target organisms and the environment. As an eco-friendly and economically workable option, the microbial intervention has garnered much attention. This review concisely outlines the toxicity, long-term environmental repercussions, degradation kinetics, biochemical pathways, and interplay of genes implicated in imidacloprid remediation. The studies have highlighted imidacloprid residue persistence in the environment for up to 3000 days. In view of high persistence, effective intervention is highly required. Bacteria-mediated degradation has been established as a viable approach with Bacillus spp. being among the most efficient at 30 â„ƒ and pH 7. Further, a comparative metagenomic investigation reveals dominant neonicotinoid degradation genes in agriculture compared to forest soils with distinctive microbial communities. Functional metabolism of carbohydrates, amino acids, fatty acids, and lipids demonstrated a significantly superior relative abundance in forest soil, implying its quality and fertility. The CPM, CYP4C71v2, CYP4C72, and CYP6AY3v2 genes that synthesize cyt p450 monooxygenase enzyme play a leading role in imidacloprid degradation. In the future, a systems biology approach incorporating integrated kinetics should be utilized to come up with innovative strategies for moderating the adverse effects of imidacloprid on the environment.

Developments in food neonicotinoids detection: novel recognition strategies, advanced chemical sensing techniques, and recent applications.

Neonicotinoid insecticides (NEOs) are a new class of neurotoxic pesticides primarily used for pest control on fruits and vegetables, cereals, and other crops after organophosphorus pesticides (OPPs), carbamate pesticides (CBPs), and pyrethroid pesticides. However, chronic abuse and illegal use have led to the contamination of food and water sources as well as damage to ecological and environmental systems. Long-term exposure to NEOs may pose potential risks to animals (especially bees) and even human health. Consequently, it is necessary to develop effective, robust, and rapid methods for NEOs detection. Specific recognition-based chemical sensing has been regarded as one of the most promising detection tools for NEOs due to their excellent selectivity, sensitivity, and robust interference resistance. In this review, we introduce the novel recognition strategies-enabled chemical sensing in food neonicotinoids detection in the past years (2017-2023). The properties and advantages of molecular imprinting recognition (MIR), host-guest recognition (HGR), electron-catalyzed recognition (ECR), immune recognition (IR), aptamer recognition (AR), and enzyme inhibition recognition (EIR) in the development of NEOs sensing platforms are discussed in detail. Recent applications of chemical sensing platforms in various food products, including fruits and vegetables, cereals, teas, honey, aquatic products, and others are highlighted. In addition, the future trends of applying chemical sensing with specific recognition strategies for NEOs analysis are discussed.

Infantile Internal and External Exposure to Neonicotinoid Insecticides: A Comparison of Levels across Various Sources.

Little is known about exposure of infants to neonicotinoid insecticides (NEOs). In this study, concentrations of six parent NEOs (p-NEOs) and N-desmethyl-acetamiprid (N-dm-ACE) were measured in urine and whole blood samples from infants, in addition to breast milk, infant formula, and tap water collected in South China. The p-NEO with the highest median concentration in urine (0.25 ng/mL) and blood (1.30) samples was dinotefuran (DIN), while imidacloprid (IMI) was abundant in breast milk (median: 0.27 ng/mL), infant formula (0.22), and tap water (0.028). The older infants (181-360 days) might face higher NEO and N-dm-ACE exposure than younger infants (0-180 days). Blood samples contained a significantly (p < 0.01) higher median concentration of ∑6p-NEOs (2.03 ng/mL) than that of urine samples (0.41), similar to acetamiprid (ACE), IMI, thiacloprid (THD), DIN, and N-dm-ACE, suggesting that NEOs readily partition into blood. Furthermore, breast-fed infants tend to have higher exposure levels than formula-fed infants. Infant formula prepared with tap water augmented the daily intake of ∑NEOs. The external sources contributed 80% of the total dose to IMI and clothianidin (CLO) exposure, while other unknown sources contributed to ACE, THD, and DIN exposure in infants. To the best of our knowledge, this is the first study to assess levels and sources of infantile exposure to NEOs through internal and external exposure assessment.

MtDNA Copy Number in Oral Epithelial Cells Serves as a Potential Biomarker of Mitochondrial Damage by Neonicotinoid Exposure: A Cross-Sectional Study.

As the mitochondrial DNA copy number (mtDNAcn) has been reported to be a biomarker for mtDNA damage in honeybees when exposed to sublethal neonicotinoids, the feasibility of using human mitochondria as a predictor upon neonicotinoid exposure remains elusive. This study investigated the association between the urinary neonicotinoid and the relative mtDNAcn (RmtDNAcn) of oral epithelial cells collected in a cross-sectional study with repeated measurements over 6 weeks. The molecular mechanism underlying neonicotinoid-caused mitochondrial damage was also examined by in vitro assay. Herein, the average integrated urinary neonicotinoid (IMIRPF) concentration ranged from 8.01 to 13.70 µg/L (specific gravity-adjusted) during the sampling period. Concomitantly, with an increase in the urinary IMIRPF, the RmtDNAcn significantly increased from 1.20 (low group) to 1.93 (high group), indicating potential dose-dependent mitochondrial damage. Furthermore, the linear regression analysis confirmed the significant correlation between the IMIRPF and RmtDNAcn. Results from in vitro assays demonstrated that neonicotinoid exposure led to the inhibition of the genes encoding mitochondrial oxidative phosphorylation (OXPHOS) complexes I and III (e.g., ND2, ND6, CytB, and CYC1), accompanied by increased reactive oxygen species production in SH-SY5Y cells. Conjointly, neonicotinoid exposure led to mitochondrial dysfunction and a resulting increase in the RmtDNAcn, which may serve as a plausible biomarker in humans.

Connecting the Pipes: Agricultural Tile Drains and Elevated Imidacloprid Brain Concentrations in Juvenile Northern Leopard Frogs (Rana pipiens).

Neonicotinoids are neurotoxic insecticides and are often released into nearby wetlands via subsurface tile drains and can negatively impact nontarget organisms, such as amphibians. Previous studies have indicated that imidacloprid, a commonly used neonicotinoid, can cross the amphibian blood-brain barrier under laboratory conditions; however, little is known about the impact of low concentrations in a field-based setting. Here, we report aqueous pesticide concentrations at wetland production areas that were either connected or not connected to agricultural tile drains, quantified imidacloprid and its break down products in juvenile amphibian brains and livers, and investigated the relationship between imidacloprid brain concentration and brain size. Imidacloprid concentrations in brain and water samples were nearly 2.5 and 5 times higher at tile wetlands (brain = 4.12 ± 1.92 pg/mg protein; water = 0.032 ± 0.045 µg/L) compared to reference wetlands, respectively. Tile wetland amphibians also had shorter cerebellums (0.013 ± 0.001 mm), depicting a negative relationship between imidacloprid brain concentration and cerebellum length. The metabolite, desnitro-imidacloprid, had liver concentrations that were 2 times higher at tile wetlands (2 ± 0.3 µg/g). Our results demonstrate that imidacloprid can cross the amphibian blood-brain barrier under ecological conditions and may alter brain dimensions and provide insight into the metabolism of imidacloprid in amphibians.