Molecular Docking Analysis at the Human α7-nAChR and Proliferative and Evoked-Calcium Changes in SH-SY5Y Cells by Imidacloprid and Acetamiprid Insecticides.

Acetamiprid (ACE) and Imidacloprid (IMI) are widely-used neonicotinoid insecticides (NNIs) with functional activity at human acetylcholine nicotinic receptors and, therefore, with putative toxic effects. The objective of this study was the evaluation of the interactions between NNIs and α7-nAChR, as this receptor keeps intracellular Ca2+ ([Ca2+]i) to an optimum for an adequate neuronal functioning. Possible interactions between NNIs and the cryo-EM structure of the human α-7 nAChR were identified by molecular docking. Additionally, NNI effects were analyzed in neuroblastoma SH-SY5Y cells, as they naturally express α-7 nAChRs. Functional studies included proliferative/cytotoxic effects (MTT test) in undifferentiated SH-SY-5Y cells and indirect measurements of [Ca2+]i transients in retinoic acid-differentiated SH-SY-5Y cells loaded with Fluo-4 AM. Docking analysis showed that the binding of IMI and ACE occurred at the same aromatic cage that the specific α-7 nAChR agonist EVP-6124. IMI showed a better docking strength than ACE. According to the MTT assays, low doses (10-50 µM) of IMI better than ACE stimulated neuroblastoma cell proliferation. At higher doses (250-500 µM), IMI also prevailed over ACE and dose-dependently triggered more abrupt fluorescence changes due to [Ca2+]i mobilization in differentiated SH-SY5Y neurons. Indeed, only IMI blunted nicotine-evoked intracellular fluorescence stimulation (i.e., nicotine cross-desensitization). Summarizing, IMI demonstrated a superior docking strength and more robust cellular responses compared to ACE, which were likely associated with a stronger activity at α-7nAChRs. Through the interaction with α-7nAChRs, IMI would demonstrate its high neurotoxic potential for humans. More research is needed for investigating the proliferative effects of IMI in neuroblastoma cells.

Evaluation of the protective effect of melatonin on oocyte, embryo and ovarian tissue parameters in female mice exposed to acetamiprid.

OBJECTIVE: Acetamiprid (ACP) causes infertility and its effect appears to occur via oxidative stress. Melatonin has antioxidative properties. Thus, in this experimental study, we examined the protective effect of melatonin against toxic pathologic changes from ACP on reproductive system parameters of female mice. METHODS: The study included 30 female mice divided into 5 groups (6 mice in each group), as follows: Saline (control group); ACP (10, 20 mg/kg); ACP (10mg/kg) + melatonin (10mg/kg); and ACP (20mg/kg) + melatonin (10mg/kg). All mice were given intraperitoneal injections daily for one month. The groups were evaluated for ovarian histopathological changes and oocyte quality based on in vitro fertilization (IVF) parameters. RESULTS: ACP induced histological damages in the ovaries of mice and caused increases in the number of atretic follicles and decreases in the quality of oocytes based on IVF parameters. These alterations were significantly reduced by melatonin. CONCLUSIONS: Melatonin can decrease the toxic effects of ACP in the female reproductive system of mice. Further studies are needed.

Chronic effects of a binary insecticide Acer 35 EC on Nile tilapia Oreochromis niloticus through a multi-biomarker approach.

Acer 35 EC is a widely used insecticide (a binary mixture of lambda-cyhalothrin and acetamiprid) in pest control in many West African countries, particularly in the cotton culture in north Benin. The aim of this study was to investigate the chronic effects of Acer 35 EC on Nile tilapia Oreochromis niloticus juveniles using a multi-biomarker approach under laboratory conditions. For this purpose, fish were exposed to sublethal concentrations of Acer 35 EC (0, 1 and 10% of LC50- 96 h value). After 28 and 56 days of exposure, several biomarkers were measured in males and females including enzymatic activities related to detoxification and oxidative stress, neurotoxicity and immune responses, sex steroid hormones (testosterone, 17ß-estradiol and 11-keto-testosterone) and histological alterations of liver, kidney and gonads. An Integrated Biomarker Response (IBR) was then calculated. The results showed a reduction of cholinesterase activity in muscles, and intercellular superoxide anion production in both sexes. Female steroidogenesis and gametogenesis were affected, especially testosterone levels and oocyte growth. More alterations were observed in liver after exposure to Acer 35 EC. In both sexes, IBR values were higher after 56 days than after 28 days of exposure. In conclusion, based on a large set of biomarkers and IBR values, the chronic exposure to low doses of insecticide Acer 35 EC seems to impair different physiological functions in Nile tilapia juveniles on a time-dependent manner, with a stronger impact on females than on males.

Foliar Spraying of Tomato Plants with Systemic Insecticides: Effects on Feeding Behavior, Mortality and Oviposition of Bemisia tabaci (Hemiptera: Aleyrodidae) and Inoculation Efficiency of Tomato Chlorosis Virus.

Tomato chlorosis virus (ToCV) is a phloem-limited crinivirus transmitted by whiteflies and seriously affects tomato crops worldwide. As with most vector-borne viral diseases, no cure is available, and the virus is managed primarily by the control of the vector. This study determined the effects of the foliar spraying with the insecticides, acetamiprid, flupyradifurone and cyantraniliprole, on the feeding behavior, mortality, oviposition and transmission efficiency of ToCV by B. tabaci MEAM1 in tomato plants. To evaluate mortality, oviposition and ToCV transmission in greenhouse conditions, viruliferous whiteflies were released on insecticide-treated plants at different time points (3, 24 and 72 h; 7 and 14 days) after spraying. Insect mortality was higher on plants treated with insecticides; however, only cyantraniliprole and flupyradifurone differed from them in all time points. The electrical penetration graph (DC-EPG) technique was used to monitor stylet activities of viruliferous B. tabaci in tomato plants 72 h after insecticide application. Only flupyradifurone affected the stylet activities of B. tabaci, reducing the number and duration of intracellular punctures (pd) and ingestion of phloem sap (E2), a behavior that possibly resulted in the lower percentage of ToCV transmission in this treatment (0-60%) in relation to the control treatment (60-90%) over the periods evaluated. Our results indicate that flupyradifurone may contribute to management of this pest and ToCV in tomato crops.

Computational study on acetamiprid-molecular imprinted polymer.

In the present work we investigate, through DFT calculations, the mechanism of formation of a molecular imprinted polymer for the acetamiprid (ACT) insecticide, using four different functional monomers, four molar ratios attempts, and considering eight distinct solvents. As the main result we obtain the following theoretical protocol for the MIP synthesis: methacrylic acid (MMA) as functional monomer, 1:4 M ratio, i.e., one ACT to four MMAs, and chloroform as solvent. This DFT calculated condition shows more favorable energies for the formed complexes. We consider this work quite relevant since it can be used by experimentalists in order to reach an efficient MIP synthesis for ACT, avoiding wasted time and laboratory resources. Graphical abstract Best MIP Synthesis Protocol for Acetamiprid.

Neonicotinoids transference from the field to the hive by honey bees: Towards a pesticide residues biomonitor.

The beehive as a quantitative monitor of pesticide residues applied over a soybean crop was studied through a semi field experiment of controlled exposure of honey bees to pesticides in macro tunnels. The distribution within exposed beehives of pesticides commonly used in soybean plantation, was assessed. Residue levels of insecticides in soybean leaves, honey bees, wax, honey and pollen were analyzed. The transference from pesticides present in the environment into the beehive was evidenced. The obtained results allow relating pesticide concentrations present in the environment with traces found in foraging bees. Therefore, pesticide transference ratios could be calculated for each detected compound (acetamiprid, imidacloprid and thiamethoxam) which showed a linear inverse trend with their 1-octanol/water partition coefficient (Kow). The least transferred pesticide to the hive (acetamiprid) has the highest vapor pressure (Vp). This study gives new insights on the usefulness of monitoring the environment through beehives aiming to evaluate if agroecosystems remain sustainable. It also contributes to generate valuable information for model building aiming to predict environmental quality through beehive's analysis.

Status of Resistance of Bemisia tabaci (Hemiptera: Aleyrodidae) to Neonicotinoids in Iran and Detoxification by Cytochrome P450-Dependent Monooxygenases.

Nine Bemisia tabaci (Gennadius) populations were collected from different regions of Iran. In all nine populations, only one biotype (B biotype) was detected. Susceptibilities of these populations to imidacloprid and acetamiprid were assayed. The lethal concentration 50 values (LC50) for different populations showed a significant discrepancy in the susceptibility of B. tabaci to imidacloprid (3.76 to 772.06 mg l-1) and acetamiprid (4.96 to 865 mg l-1). The resistance ratio of the populations ranged from 9.72 to 205.20 for imidacloprid and 6.38 to 174.57 for acetamiprid. The synergistic effects of piperonylbutoxide (PBO) and S,S,S-tributylphosphorotrithioate (DEF) were evaluated for the susceptible (RF) and resistant (JR) populations for the determination of the involvement of cytochrome P450-dependent monooxygenase and carboxylesterase, respectively, in their resistance mechanisms. The results showed that PBO overcame the resistance of the JR population to both imidacloprid and acetamiprid, with synergistic ratios of 72.7 and 106.9, respectively. Carboxylesterase, glutathione S-transferase and cytochrome P450-dependent monooxygenase were studied biochemically, for the purpose of measuring the activity of the metabolizing enzymes in order to determine which enzymes are directly involved in neonicotinoid resistance. There was an increase in the activity of cytochrome P450-dependent monooxygenase up to 17-fold in the resistant JR population (RR = 205.20). The most plausible activity of cytochrome P450-dependent monooxygenase correlated with the resistances of imidacloprid and acetamiprid, and this suggests that cytochrome P450-dependent monooxygenase is the only enzyme system responsible for neonicotinoid resistance in the nine populations of B. tabaci.

Eficiência de Inseticidas no Controle de Pragas em Sementes e Mudas de Pau-rosa (Aniba rosaeodora Ducke), em Viveiros, Manaus, Amazonas

The rosewood tree (Aniba rosaeodora Ducke) is known for its linalool which provides a strong, fragrant smell in all parts of the plant. Due to its exploitation in the Amazon for its linalool in the use of perfumes, the rosewood is in danger of extinction. Its irregular fruiting increases the difficulty in obtaining seeds for producing seedlings and, in addition, part of its production is lost due to pests attacking the seeds. Due to these factors, the object of this study was to achieve the application of insecticides on seeds and seedlings in nursery to avoid or minimize the attack of pests, thus improving seedling production. 300 seeds were used, distributed into 2 treatments and 1 witness. Each treatment (n = 100) was set for 4 repetitions (trays) with 25 seeds. The insecticide used was acetamiprid and fipronil in concentrations of 0.09 mg / plant and 0.18 mg / plant, respectively for treatment 1 (T1) and the treatment 2 (T2), distributed in the form of "sachês". The T1 received the application from 1 "sachê" and T2 the implementation of two "sachês", buried with washed sand in seaders. The results were evaluated by analysis of Variance (ANOVA) using the Systat 9 program and the averages compared by the Tukey test at 5% of significance. The rates of germination were: witness with 62%; T1 with 73% and T2 with 79%. There was an attack of insects only in the witness treatment with 23% of the total (n = 300), and the results were statistically significant (F = 31.263 p 0.001). There were significant differences only between the height of the seedlings in treatments T1 and T2 and witness treatment (F = 15.090 p 0.001).The main pest identified was Heilipus odoratus (Coleoptera: Curculionidae). We, therefore, recommended the concentration of 0.09 mg of acetamiprid and fipronil distributed in "sachês" in sowing and transplanted to reduce the attack of pests in seeds and seedlings of rosewood in nurseries.

O pau-rosa (Aniba rosaeodora) é conhecido pela presença do álcool linalol, que fornece um odor forte e perfumado em todas as partes da planta. A sua exploração na Amazônia ao longo dos últimos anos, para utilização do linalol como fixador de perfumes, coloca o pau-rosa em risco de extinção, a sua frutificação irregular aumenta a dificuldade em se obter sementes para a produção de mudas e, além disso, parte de sua produção é perdida face ao ataque de pragas nas sementes. Em razão desses fatores o presente estudo teve como objetivo realizar a aplicação de inseticidas nas sementes e mudas em viveiro a fim de evitar ou minimizar o ataque de pragas, aumentando, assim, a produção de mudas. Utilizou-se um total de 300 sementes, distribuídas em 2 tratamentos e 1 testemunha. Cada tratamento (n = 100) foi constituído por 4 repetições (bandejas) com 25 sementes. O inseticida utilizado nos tratamentos foi acetamiprid e fipronil nas concentrações de 0,09 mg/planta e 0,18 mg/planta, respectivamente, para o tratamento 1 (T1) e o tratamento 2 (T2), distribuídos na forma de sachês. O T1 recebeu a aplicação de 1 sachê e o T2 a aplicação de dois sachês, enterrados com areia lavada nas sementeiras. Para avaliar os resultados foi feita uma análise de Variância (ANOVA), usando o programa Systat 9 e as médias comparadas pelo teste de Tukey em nível de 5%. As taxas de germinação foram: testemunha com 62%; T1 com 73% e T2 com 79%. Houve ataque de insetos em sementes apenas na testemunha com 23% do total (n = 100) e os resultados foram estatisticamente significativos (F = 31,263; p 0,001). Houve diferenças significativas apenas entre a altura das plântulas nos tratamentos T1 e T2 e a testemunha (F = 15,090; p 0,001). A principal praga identificada atacando as sementes foi Heilipus odoratus (Coleoptera: Curculionidae). Recomenda-se, portanto, a concentração de 0,09 mg de acetamiprid e fipronil distribuídos em sachês nas sementeiras e no transplantio para diminuir o ataque de pragas em sementes e mudas de pau-rosa em viveiros.

Eficiência de Inseticidas no Controle de Pragas em Sementes e Mudas de Pau-rosa (Aniba rosaeodora Ducke), em Viveiros, Manaus, Amazonas

The rosewood tree (Aniba rosaeodora Ducke) is known for its linalool which provides a strong, fragrant smell in all parts of the plant. Due to its exploitation in the Amazon for its linalool in the use of perfumes, the rosewood is in danger of extinction. Its irregular fruiting increases the difficulty in obtaining seeds for producing seedlings and, in addition, part of its production is lost due to pests attacking the seeds. Due to these factors, the object of this study was to achieve the application of insecticides on seeds and seedlings in nursery to avoid or minimize the attack of pests, thus improving seedling production. 300 seeds were used, distributed into 2 treatments and 1 witness. Each treatment (n = 100) was set for 4 repetitions (trays) with 25 seeds. The insecticide used was acetamiprid and fipronil in concentrations of 0.09 mg / plant and 0.18 mg / plant, respectively for treatment 1 (T1) and the treatment 2 (T2), distributed in the form of "sachês". The T1 received the application from 1 "sachê" and T2 the implementation of two "sachês", buried with washed sand in seaders. The results were evaluated by analysis of Variance (ANOVA) using the Systat 9 program and the averages compared by the Tukey test at 5% of significance. The rates of germination were: witness with 62%; T1 with 73% and T2 with 79%. There was an attack of insects only in the witness treatment with 23% of the total (n = 300), and the results were statistically significant (F = 31.263 p 0.001). There were significant differences only between the height of the seedlings in treatments T1 and T2 and witness treatment (F = 15.090 p 0.001).The main pest identified was Heilipus odoratus (Coleoptera: Curculionidae). We, therefore, recommended the concentration of 0.09 mg of acetamiprid and fipronil distributed in "sachês" in sowing and transplanted to reduce the attack of pests in seeds and seedlings of rosewood in nurseries.

O pau-rosa (Aniba rosaeodora) é conhecido pela presença do álcool linalol, que fornece um odor forte e perfumado em todas as partes da planta. A sua exploração na Amazônia ao longo dos últimos anos, para utilização do linalol como fixador de perfumes, coloca o pau-rosa em risco de extinção, a sua frutificação irregular aumenta a dificuldade em se obter sementes para a produção de mudas e, além disso, parte de sua produção é perdida face ao ataque de pragas nas sementes. Em razão desses fatores o presente estudo teve como objetivo realizar a aplicação de inseticidas nas sementes e mudas em viveiro a fim de evitar ou minimizar o ataque de pragas, aumentando, assim, a produção de mudas. Utilizou-se um total de 300 sementes, distribuídas em 2 tratamentos e 1 testemunha. Cada tratamento (n = 100) foi constituído por 4 repetições (bandejas) com 25 sementes. O inseticida utilizado nos tratamentos foi acetamiprid e fipronil nas concentrações de 0,09 mg/planta e 0,18 mg/planta, respectivamente, para o tratamento 1 (T1) e o tratamento 2 (T2), distribuídos na forma de sachês. O T1 recebeu a aplicação de 1 sachê e o T2 a aplicação de dois sachês, enterrados com areia lavada nas sementeiras. Para avaliar os resultados foi feita uma análise de Variância (ANOVA), usando o programa Systat 9 e as médias comparadas pelo teste de Tukey em nível de 5%. As taxas de germinação foram: testemunha com 62%; T1 com 73% e T2 com 79%. Houve ataque de insetos em sementes apenas na testemunha com 23% do total (n = 100) e os resultados foram estatisticamente significativos (F = 31,263; p 0,001). Houve diferenças significativas apenas entre a altura das plântulas nos tratamentos T1 e T2 e a testemunha (F = 15,090; p 0,001). A principal praga identificada atacando as sementes foi Heilipus odoratus (Coleoptera: Curculionidae). Recomenda-se, portanto, a concentração de 0,09 mg de acetamiprid e fipronil distribuídos em sachês nas sementeiras e no transplantio para diminuir o ataque de pragas em sementes e mudas de pau-rosa em viveiros.