Microbial degradation mechanism and pathway of the insecticide thiamethoxam by isolated Bacillus Cereus from activated sludge.

The high water solubility and ecotoxicity of thiamethoxam (TMX) is a potential hazard to ecosystems and human health. Here, a strain of Bacillus cereus with high TMX degradation activity was isolated from the sediment of the A2O process in the wastewater treatment plant and was able to utilize TMX as its sole carbon source. Under different environmental conditions, the degradation efficiency of TMX by Bacillus cereus-S1 (strain S1) ranged from 41.0% to 68.9% after 216 h. The optimum degradation conditions were DO = 3.5 mg/L and pH 9.0. The addition of an appropriate carbon-to-nitrogen ratio could accelerate the degradation of TMX. A plausible biodegradation pathway has been proposed based on the identified metabolites and their corresponding degradation pathways. TMX can be directly converted into Clothianidin (CLO), TMX-dm-hydroxyl and TMX-Urea by a series of reactions such as demethylation, oxadiazine ring cleavage and C=N substitution by hydroxy group. The main products were TMX-dm-hydroxyl and TMX-Urea, the amount of CLO production is relatively small. This study aims to provide a new approach for efficient degradation of TMX; furthermore, strain S1 is a promising biological source for in situ remediation of TMX contamination.

Thiamethoxam dynamics in pepper plants: Deciphering deposition and dissipation pattern across diverse planting modes and regions.

To reduce the application dosage of thiamethoxam (TMX), we investigated the deposition and dissipation patterns in a pepper-planted ecosystem under different planting modes across four regions in China, namely Hainan (HN), Zhejiang (ZJ), Anhui (AH) and Hebei (HB). This study focused on the deposition and dissipation of TMX at concentrations of 63.00, 47.25, 31.50, 23.63 and 15.75 g a.i.hm-2. As the application dose increased, the deposition amount of TMX initially increased in the plants and cultivated soil, showing obvious geographic differences in four cultivation areas. Surprisingly, the initial amount of TMX deposited the pepper-cultivated greenhouse of ZJ and AH was 1.1-2.1-fold and 1.0-3.6-fold higher than that in the open field system at the same application dose, respectively. In pepper leaves, stems, fruits and soil, the dissipation exhibited rapid growth and then slowed. However, the residual concentration showed an increasing trend, followed by a subsequent decrease in the pepper roots. In different planting regions, the dissipation rate of TMX followed the order HN > ZJ > AH > HB in pepper plants and cultivated soil. In comparison to the open field, the total TMX retention rate in greenhouse was higher, indicating overall greater persistence in the greenhouse conditions. These findings reveal the deposition and dissipation characteristics of TMX within the pepper-field ecosystem, offering a significant contribution to the risk assessment of pesticides.

Hormetic effects of thiamethoxam on Schizaphis graminum: demographics and feeding behavior.

In agroecosystems, insects contend with chemical insecticides often encountered at sublethal concentrations. Insects' exposure to these mild stresses may induce hormetic effects, which has consequences for managing insect pests. In this study, we used an electrical penetration graph (EPG) technique to investigate the feeding behavior and an age-stage, two-sex life table approach to estimate the sublethal effects of thiamethoxam on greenbug, Schizaphis graminum. The LC5 and LC10 of thiamethoxam significantly decreased longevity and fecundity of directly exposed adult aphids (F0). However, the adult longevity, fecundity, and reproductive days (RPd)-indicating the number of days in which the females produce offspring - in the progeny generation (F1) exhibited significant increase when parental aphids (F0) were treated with LC5 of the active ingredient. Subsequently, key demographic parameters such as intrinsic rate of increase (r) and net reproductive rate (R0) significantly increased at LC5 treatment. EPG recordings showed that total durations of non-probing (Np), intercellular stylet pathway (C), and salivary secretion into the sieve element (E1) were significantly increased, while mean duration of probing (Pr) and total duration of phloem sap ingestion and concurrent salivation (E2) were decreased in F0 adults exposed to LC5 and LC10. Interestingly, in the F1 generation, total duration of Np was significantly decreased while total duration of E2 was increased in LC5 treatment. Taken together, our results showed that an LC5 of thiamethoxam induces intergenerational hormetic effects on the demographic parameters and feeding behavior of F1 individuals of S. graminum. These findings have important implications on chemical control against S. graminum and highlight the need for a deeper understanding of the ecological consequences of such exposures within pest management strategies across the agricultural landscapes.

Identification of the cytochrome P450 gene AccCYP6A13 in Apis cerana cerana and its response to environmental stress.

Cytochrome P450 plays a crucial role in regulating insect growth, development, and resisting a variety of stresses. Insect metamorphosis and response to external stress are altered by deleting CYP450 genes. In this study, we identified and analyzed a novel gene of CYP450 family, AccCYP6A13, from Apis cerana cerana, and explored its role in the response of Apis cerana cerana to adverse external stressors. It was found that the expression of AccCYP6A13 was spatiotemporal specificity. The expression level increased with age and reached its highest value in the adult stage. The primarily expressiong location were legs, brain, and epidermis of honeybees. Stress conditions can affect the expression of AccCYP6A13 depending on treatment times. RNA interference experiments have shown that knocking down AccCYP6A13 reduces antioxidant activity and deactivates detoxification enzymes, resulting in oxidative damage accumulation and a decline in detoxification capability in bees, as well as inhibiting numerous antioxidant genes. Additionally, knockdown of the AccCYP6A13 gene in Apis cerana cerana resulted in increased sensitivity to pesticides and increased mortality when treated with neonicotinoid pesticides such as thiamethoxam. AccCYP6A13 overexpression in a prokaryotic system further confirmed its role in resistance to oxidative stress. To summarize, AccCYP6A13 may play an essential role in the normal development and response to environmental stress in Apis cerana cerana. Furthermore, this study contributed to the theoretical understanding of bee resistance biology.

Chronic Paternal/Maternal Exposure to Environmental Concentrations of Imidacloprid and Thiamethoxam Causes Intergenerational Toxicity in Zebrafish Offspring.

Imidacloprid (IMI) and thiamethoxam (THM) are ubiquitous in aquatic ecosystems. Their negative effects on parental fish are investigated while intergenerational effects at environmentally relevant concentrations remain unclear. In this study, F0 zebrafish exposed to IMI and THM (0, 50, and 500 ng L-1) for 144 days post-fertilization (dpf) was allowed to spawn with two modes (internal mating and cross-mating), resulting in four types of F1 generations to investigate the intergenerational effects. IMI and THM affected F0 zebrafish fecundity, gonadal development, sex hormone and VTG levels, with accumulations found in F0 muscles and ovaries. In F1 generation, paternal or maternal exposure to IMI and THM also influenced sex hormones levels and elevated the heart rate and spontaneous movement rate. LncRNA-mRNA network analysis revealed that cell cycle and oocyte meiosis-related pathways in IMI groups and steroid biosynthesis related pathways in THM groups were significantly enriched in F1 offspring. Similar transcriptional alterations of dmrt1, insl3, cdc20, ccnb1, dnd1, ddx4, cox4i1l, and cox5b2 were observed in gonads of F0 and F1 generations. The findings indicated that prolonged paternal or maternal exposure to IMI and THM could severely cause intergenerational toxicity, resulting in developmental toxicity and endocrine-disrupting effects in zebrafish offspring.

Thiamethoxam adsorption by ZnCl2 modified cow manure biochar: Mechanism and quantitative prediction.

The overuse of thiamethoxam (THM) has threatened the survival of living organisms and it is necessary to find an environmentally friendly material to remove THM frequently detected in water. Biochar prepared from cow manure modified with ZnCl2 (Zn-CBC) was used to remove THM. Compared to the unmodified cow manure biochar (CBC), the removal ratio of THM by Zn-CBC was enhanced 35 times. In the mechanistic analysis, SEM and BET showed that Zn-CBC had a good pore structure and its specific surface area (166.502 m2 g-1) increased to 17 times that of CBC, indicating that Zn-CBC had good pore adsorption properties. The adsorption kinetic and isotherm implied that the main mechanism was chemisorption including π-π interaction and H-bonding. Furthermore, the stable graphitized structure of Zn-CBC allowed for efficient adsorption and reusability. In addition, this study constructed an intelligent prediction model using batch experiment data, and the high R2 (0.978) and low RMSE (0.057) implied that the model could accurately and quantitatively predict the adsorption efficiency. This paper provides a novel perspective to simultaneously remove the neonicotinoid insecticides and realize the resource utilization of cow manure.

What does not kill it makes it stronger! The tolerance of the F1 larvae of Chironomus xanthus to a neonicotinoid insecticide formulation.

Thiamethoxam (TMX) is a systemic neonicotinoid that acts as a partial agonist of the nicotinic acetylcholine receptors (nAChRs). However, target species have shown resistance to formulations based on such neonicotinoids, which can also be expected for non-target insects. This research aimed to study the effects of a formulation based on TMX [Cruiser® 350 FS (CRZ)] on the life traits of Chironomus xanthus filial generation (F1) and compare it with the parental generation (P). Environmentally relevant concentrations of CRZ significantly decreased larvae growth P generation , also slowing and decreasing their emergence. Larvae of the F1 generation were less sensitive than their parents, suggesting that the progeny were able to thrive and perform basic physiological functions better than the parental generation. Our results highlight that insect resistance to neonicotinoids may be associated with the better performance of the filial generation, which is related to the change in affinities of the active ingredient for the sub-units constituting the nAChRs subtypes of F1 organisms, inherited from P organisms that were able to survive and reproduce. Moreover, further studies using biochemical and omics tools should be performed to disentangle the specific changes occurring at the nAChRs throughout insect development.

MOF@Au NPs/aptamer fluorescent probe for the selective and sensitive detection of thiamethoxam.

The luminescence performance of fluorescent reagents plays a crucial role in fluorescence analysis. Therefore, in this study, a novel bi-ligand Zn-based metal-organic framework, Au nanoparticle (NP) fluorescent material was synthesized using a hydrothermal method with Zn as the metal source. Simultaneously, a DNA aptamer was introduced as a molecular recognition element to develop a Zn-based MOF@Au NPs/DNA aptamer fluorescent probe for the ultra-trace detection of thiamethoxam residues in agricultural products. The probe captured different concentrations of the target molecule, thiamethoxam, through the DNA aptamer, causing a conformational change in the DNA aptamer and bursting the fluorescence of the probe, therefore establishing a fluorometric method for thiamethoxam detection. This method is highly sensitive due to the excellent luminescence properties of the Zn-based MOF@Au NPs, and the DNA aptamer can specifically recognize thiamethoxam, offering high selectivity. The linear range of the method was 2.5-6000 × 10-11  mol L-1 , with a detection limit of 8.33 × 10-12  mol L-1 . This method was applied to the determination of actual samples, such as bananas, and the spiked recovery rate was found to be in the range 84.05-109.07%. Overall, the proposed probe has high sensitivity, high selectivity, and easy operation for the detection of thiamethoxam residues in actual samples.

Chemosensory proteins are associated with thiamethoxam tolerance in bird cherry-oat aphid Rhopalosiphum padi.

Rhopalosiphum padi (L.) is an important cosmopolitan pest of cereal crops. Thiamethoxam is widely used for control R. padi in some regions. Chemosensory proteins (CSPs) are a class of transporter proteins in arthropods which play a key role in various physiological processes including response to insecticide exposure. However, the role of R. padi CSPs (RpCSPs) in insecticide binding and susceptibility has not been well clarified. In this study, we found that the expression levels of RpCSP1, RpCSP4, RpCSP5, RpCSP7, RpCSP10 were dramatically upregulated after exposure to thiamethoxam. Suppression of RpCSP4 and RpCSP5 transcription by RNA interference significantly enhanced the susceptibility of R. padi to thiamethoxam. Molecular docking and fluorescence competitive binding showed that RpCSP4 and RpCSP5 had high binding affinity with thiamethoxam. The present results prove that RpCSP4 and RpCSP5 are related to insecticide resistance through high binding affinity to reduce the toxicity of insecticide.

The interaction of nicotinic acetylcholine receptor subunits Ldα3, Ldα8 and Ldß1 with neonicotinoids in Colorado potato beetle, Leptinotarsa decemlineata.

The Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), is an extremely destructive notifiable quarantine pest. Over the last two decades, neonicotinoid insecticides, particularly thiamethoxam and imidacloprid, have been used to control it in Xinjiang, and local field populations have developed different levels of resistance in consequence. However, the contributions of nicotinic acetylcholine receptors (nAChRs) to neonicotinoid resistance are currently poorly understood in CPB. Previous studies have shown that nAChRα1, α3, α8 and ß1 are major target subunits for neonicotinoids in some model and important agricultural insects including nAChRα1 subunit of L. decemlineata (Ldα1). In this study, the expression levels of Ldα3, Ldα8 and Ldß1 following 72 h of treatments with median lethal doses of thiamethoxam and imidacloprid were compared using real-time quantitative PCR. These genes were then individually and simultaneously knocked down with Ldα1 by RNA interference (RNAi) using a double-stranded RNA (dsRNA) feeding method for six days to explore their roles in CPB susceptibility to imidacloprid and thiamethoxam. The results showed that the expressions of Ldα3, Ldα8 and Ldß1 were significantly decreased by 36.99-74.89% after thiamethoxam and imidacloprid treatments, compared with the control. The significant downregulation of the target genes resulting from RNAi significantly reduced the mortality of adults exposed to thiamethoxam and imidacloprid by 34.53% -56.44% and 28.78%-43.93%, respectively. Furthermore, the adult survival rates were not affected by every dsRNA-feeding treatment, while the body weight of the test adults significantly deceased after four and six days of individual gene RNAi. This study showed that Ldα3, Ldα8 and Ldß1 are down-regulated by thiamethoxam and imidacloprid and play important roles in the tolerance of CPB to neonicotinoids.

Effects of Pesticides on Red Rot of Planted Sugarcane.

Red rot, caused by Colletotrichum falcatum, is an important constraint to sugarcane production. In Louisiana, red rot primarily affects planted seed-cane and is more severe when billets (stalk sections) are planted rather than whole stalks. At planting, application of seed-treatment pesticides, particularly a combination of a fungicide and the insecticide thiamethoxam, has improved stand establishment and increased yields in billet plantings in Louisiana. However, information on the effect of chemicals on disease development is lacking. Greenhouse experiments were conducted to evaluate stalk rot symptom severity and initial plant growth for billets dip-treated with a combination of the fungicides azoxystrobin and propiconazole, thiamethoxam, a combination of both fungicides and the insecticide, and, as a control, untreated billets. Reductions in disease severity recorded for different treatments were similar for billets inoculated with the fungus or exposed to natural inoculum. Disease severity was consistently reduced by the combination treatment, while reductions resulting from treatment with fungicides and insecticide alone were variable. Reductions occurred for both internode and node rot severity. The effects of pesticide treatments on plant growth after 6 weeks were minor; however, there was evidence of disease adversely affecting germination, particularly for nontreated billets exposed to natural inoculum, where germination was reduced by one third. The treatments that reduced disease severity prevented this reduction. The results provide evidence that reduction in disease severity is an important contributor to the stand establishment and yield improvements observed for treated billets in field experiments.

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 500 µM 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 100 µM, and thiamethoxam exposures at 200 µM and 500 µM (>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.

Efficient removal of thiamethoxam by freshwater microalgae Scenedesmus sp. TXH: Removal mechanism, metabolic degradation and application.

Neonicotinoids, as the most widely used pesticides in the world, help improve the production of crops. Meanwhile, it also brings potential threats to surrounding environments and other organisms because of its wide use and even abuse. In this study, Scenedesmus sp. TXH isolated from a wastewater treatment plant was used to remove the neonicotinoid pesticide thiamethoxam (THIA). The removal efficiency, degradation pathway, metabolite fate of THIA and physicochemical effects on microalgae cells were studied. Meanwhile, the feasibility of using microalgal technology to remove THIA from municipal wastewater was also explored. The results showed that 5-40 mg/L of THIA slightly promoted the growth of microalgae, while 60 mg/L THIA severely inhibited microalgal growth. It was observed that malondialdehyde content and superoxide dismutase activity in 60 mg/L THIA group increased significantly (p < 0.05) in the early stage of the experiment, indicating that THIA caused oxidative damage to microalgae. Scenedesmus sp. TXH showed high-efficient degradation ability and high resistance to THIA, with 100% removal of THIA at 5, 20 and 40 mg/L groups and 97.5% removal of THIA at 60 mg/L group on day 12. THIA was mainly removed by biodegradation, accounting for 78.18%, 93.50%, 96.81% and 91.35% under 5, 20, 40 and 60 mg/L on day 12, respectively. Six degradation products were identified, and four potential degradation pathways were proposed. In practical wastewater, the removal efficiency of total dissolved nitrogen, total dissolved phosphorus, ammonia nitrogen and THIA reached 85.68%, 90.00%, 98.43% and 100%, respectively, indicating that Scenedesmus sp. TXH was well adapted to the wastewater and effectively removed THIA and conventional pollutants.

Residual Pattern of Chlorantraniliprole, Thiamethoxam, Flubendiamide and Deltamethrin in Tomato Fruit and Soil.

Tomato, Lycopersicon esculentum L. is grown widely as an important day-to-day demand vegetable. The crop is attacked by various polyphagous insect pests like tomato fruit borer, stink bug, cabbage looper, flea beetle, aphids, whitefly, two-spotted spider mite, etc., and oligophagous insects like leaf-miner, five-spotted hawkmoth, etc. To combat the damage and yield loss, various chemical insecticides were sprayed on tomatoes under field conditions. The residual pattern of insecticides like chlorantraniliprole, thiamethoxam, flubendiamide, and deltamethrin residues was studied following applications of chlorantraniliprole 18.5% SC (Coragen) @ 30 g a.i./ha, thiamethoxam 25% WG (Actara) @ 50 g a.i./ha, flubendiamide 39.35 M/M SC (Fame) @ 48 g a.i./ha and deltamethrin 2.8% EC (Decis 100) @ 12.5 g a.i./ha using Reverse Phase High-Performance Liquid Chromatography (RP-HPLC). Fruit samples were collected at 0 (1 h after application), 1, 2, 3, 5, 7 days and at harvest time. All the residues of insecticides such as chlorantraniliprole (0.09 mg kg- 1), thiamethoxam (0.03 mg kg- 1), flubendiamide (0.02 mg kg- 1), and deltamethrin (0.01 mg kg- 1) were persisted up to 5th day. There were no residues found at harvest time. The residues of chlorantraniliprole and deltamethrin persisted up to 3rd day of spraying whereas the residues of flubendiamide and thiamethoxam were not detected on the same day in the soil.

An amino acid transporter-like protein (OsATL15) facilitates the systematic distribution of thiamethoxam in rice for controlling the brown planthopper.

Characterization and genetic engineering of plant transporters involved in the pesticide uptake and translocation facilitate pesticide relocation to the tissue where the pests feed, thus improving the bioavailability of the agrichemicals. We aimed to identify thiamethoxam (THX) transporters in rice and modify their expression for better brown planthopper (BPH) control with less pesticide application. A yeast library expressing 1385 rice transporters was screened, leading to the identification of an amino acid transporter-like (ATL) gene, namely OsATL15, which facilitates THX uptake in both yeast cells and rice seedlings. In contrast to a decrease in THX content in osatl15 knockout mutants, ectopic expression of OsATL15 under the control of the CaMV 35S promoter or a vascular-bundle-specific promoter gdcsPpro significantly increased THX accumulation in rice plants, thus further enhancing the THX efficacy against BPH. OsATL15 was localized in rice cell membrane and abundant in the root transverse sections, vascular bundles of leaf blade, and stem longitudinal sections, but not in hull and brown rice at filling stages. Our study shows that OsATL15 plays an essential role in THX uptake and its systemic distribution in rice. OsATL15 could be valuable in achieving precise pest control by biotechnology approaches.

Multi-biomarkers approach to assess the toxicity of novel insecticide (Voliam flexi®) on Clarias gariepinus: From behavior to immunotoxicity.

This study was conducted to determine for the first time the immunological, histopathological, histochemical, and ultrastructural changes; hematological and biochemical alterations; and poikilocytosis induced in Clarias gariepinus by Voliam flexi® 40% WG (thiamethoxam + chlorantraniliprole). Beside control fish, juvenile C. gariepinus were subjected to three sublethal concentrations of Voliam flexi® (43.5, 87.5, and 175 mg/L) for 15 days. Voliam flexi® induced immunotoxic impairments in C. gariepinus, such as a decrease in some immunity variables (lysozyme and phagocyte activity, immunoglobulin concentration, and nitro blue tetrazolium level). It also caused an extreme increase in the levels of primary cytokines (interleukin-1ß and IL-6), compared with the control. The toxic effects of Voliam flexi® increased gradually with the increasing concentrations tested. Histological examination of the liver demonstrated necrosis, vacuolated hepatocytes (fatty deposition), melanomacrophage centers, foci of inflammatory cells, congested and dilated blood sinusoids, hepatic degeneration, fibrosis increment (Sirius Red stain), and glycogen depletion, as well as cytopathological alterations. We conclude that the toxic effects of Voliam flexi® must be restricted or prevented by using control mechanisms in aquatic systems.

Impurity identification in thiamethoxam by high resolution mass spectrometry and computer assisted elucidation.

Thiamethoxam (TMX) is a widely used neonicotinoid insecticide in pest control. Identification of structurally related impurities is very important during certified reference material development and pesticide registration, thus it needs to be carefully characterized. In this study, a combined strategy with liquid chromatography-high resolution mass spectrometry and computer assisted elucidation (SIRIUS) has been developed for the impurity elucidation in TMX material. MS and MS/MS spectra were used to score the impurity candidates by isotope score and fragment tree in SIRIUS. TMX, the main component, worked as an anchor for formula identification and structure elucidation of impurity. With this strategy, four impurities were identified, including two byproducts (TMX-OCH3 and TMX-Cl) and two metabolites (clothianidin and TMX-urea). Their fragmentation pathways were concluded, and mechanism of impurity formation was also proposed. This result showed successful application of combining human intelligence with machine learning in impurity identification from chemicals.

Uptake, translocation, and metabolism of thiamethoxam in soil by leek plants.

Thiamethoxam (TMX) is commonly applied on leek plants by root irrigation. It might be taken up by leek plants and thus has lasting dietary risk. In this study, the uptake, translocation, and metabolism of TMX in leek plants were investigated. The results obtained from both the hydroponic and soil experiments indicated that TMX could be easily translocated upward and accumulated in leek shoots after being absorbed by roots. The total absorbed TMX amount (Mtotal) in leek plants from the tested soils varied greatly with its adsorption governed by soil characteristics. Interestingly, Mtotal was closely correlated with the concentration of TMX in in situ pore water, indicating that TMX in in situ pore water could be a useful approach to predict uptake of this chemical by leek plants from various soils. Profoundly, clothianidin (CLO) was detected with concentration of 0.07-1.54 mg/kg in roots and 0.27-4.12 mg/kg in shoots at 14 d, respectively, suggesting that TMX is easily converted into CLO in leek plants. The results showed that TMX used in soil is easily absorbed by leek and accumulated in edible parts accompanying with formation of CLO.

The ubiquity of neonicotinoid contamination: Residues in seabirds with different trophic habits.

Neonicotinoids are one of the most diffusely used classes of pesticides whose level of danger toward non-target invertebrate and vertebrate species has raised increasing concern in the last decades. Among vertebrates, birds are particularly susceptible to unintentional neonicotinoid poisoning since they can be exposed through different pathways, including ingestion of dressed seeds, sucking of contaminated pollen, ingestion of sprayed insects, predation on contaminated aquatic and terrestrial preys. In the present study, we investigated the possible exposure of seabirds by measuring the residues of five neonicotinoids (acetamiprid, clothianidin, imidacloprid, thiacloprid and thiamethoxam) in samples of pooled feathers collected from fledglings of the strictly piscivorous Sandwich tern (Thalasseus sandvicensis) and the mixotrophic species Mediterranean gull (Ichthyaetus melanocephalus). At least one neonicotinoid was quantified in all the Mediterranean gull samples (n = 11) and 89% of the analysed Sandwich tern samples (n = 36). The active principles with the highest quantification rates were imidacloprid (100% in Mediterranean gulls and 58% in Sandwich terns) and clothianidin (100% in Mediterranean gulls and 61% in Sandwich terns), while thiacloprid was the less frequently detected pesticide (<20% of samples in both species). Mean concentrations ± standard error for imidacloprid, clothianidin and thiamethoxam were 8.8 ± 1.4, 4.5 ± 0.19 and 0.16 ± 0.02 ng g-1 for the Mediterranean gull, and 5.8 ± 0.55, 0.60 ± 0.08 and 0.36 ± 0.03 ng g-1for the Sandwich tern, respectively. Our data evidenced the exposure of seabirds to neonicotinoids and the further need to investigate the extent of neonicotinoid contamination in non-agricultural ecosystems.

Potency matters: Impacts of embryonic exposure to nAChR agonists thiamethoxam and nicotine on hatching success, growth, and neurobehavior in larval zebrafish.

Thiamethoxam (TM) is a neonicotinoid insecticide that acts as a nicotinic acetylcholine receptor (nAChR) agonist. While designed to specifically target invertebrate nAChRs, recent studies have reported adverse effects of neonicotinoid exposure in early life-stage fish. This study examined the health and neurobehavioral impacts of chronic exposure to various concentrations of TM or nicotine (NIC) in early life zebrafish (Danio rerio) in conjunction with in-silico molecular docking to compare their ligand-receptor interactions with vertebrate nAChR. Chronic exposure to both reduced survival by approximately 20% (163 µg TM/l) and 25-100% (≥0.49 µg NIC/l). Hatching and growth were impaired following exposure to ≥0.21 µg TM/l or 4.9 µg NIC/l. Both TM and NIC produced morphological and behavioral indicators of neurotoxicity, with more potent effects following NIC exposure. NIC impaired embryonic motor activity by 40% (49 µg NIC/l), while both TM and NIC significantly altered predator escape response in larvae, specifically the latency and the initial burst movement of the response were impacted. Molecular docking predicted variations in the type and strength of interactions that occur between NIC or TM and vertebrate nAChR. These findings demonstrate that chronic exposure to TM might impact general health and neurobehavior of early-stage zebrafish. Our data support hypotheses that TM presents low affinity for vertebrate nAChR but may still pose an adverse risk to larval fish growth and neurobehavior.