Selected Fungicides as Potential EDC Estrogenic Micropollutants in the Environment.

An increasing level of pesticide exposition is being observed as a result of the consumption of large amounts of fruits, vegetables and grain products, which are key components of the vegetarian diet. Fungicides have been classified as endocrine-disrupting compounds, but their mechanisms of action have not yet been clarified. The effect of boscalid (B), cyprodinil (C) and iprodione (I) combined with Tamoxifen (T) and 17ß-estradiol (E2) on cell viability, cell proliferation, reporter gene expression, ROS content, the cell membrane's function, cell morphology and antioxidant enzymes gene expression in MCF-7 and T47D-KBluc cell lines were investigated. The cell lines were chosen due to their response to 17ß -estradiol. The selected fungicides are commonly used in Poland to protect crops against fungi. Our results revealed that the studied fungicides caused significant increases in cell viability and proliferation, and estrogenic activity was present in all studied compounds depending on their concentrations. Oxidative stress activated uncontrolled cancer cell proliferation by inducing ROS production and by inhibiting antioxidant defense. Our findings verify that the studied fungicides could possibly exhibit endocrine-disrupting properties and exposure should be avoided.

Fungicide Sensitivity of Colletotrichum Species Causing Bitter Rot of Apple in the Mid-Atlantic U.S.A.

Apple growers in the Mid-Atlantic region of the U.S.A. have reported increased losses to bitter rot of apple. We tested the hypothesis that this increase is because the Colletotrichum population has developed resistance to commonly used single-mode-of-action (single-MoA) fungicides. We screened 220 Colletotrichum isolates obtained from 38 apple orchards in the Mid-Atlantic region for resistance to 11 fungicides in Fungicide Resistance Action Committee (FRAC) groups 1, 7, 9, 11, 12, and 29. Eleven (5%) of these isolates were resistant to FRAC group 1 with confirmed ß-tubulin E198A mutations, and two (<1%) were also resistant to FRAC group 11 with confirmed cytochrome-b G143A mutations. Such low frequencies of resistant isolates indicate that fungicide resistance is unlikely to be the cause of any regional increase in bitter rot. A subsample of isolates was subsequently tested in vitro for sensitivity to every single-MoA fungicide registered for apple in the Mid-Atlantic U.S.A. (22 fungicides; FRAC groups 1, 3, 7, 9, 11, 12, and 29), and 13 fungicides were tested in field trials. These fungicides varied widely in efficacy both within and between FRAC groups. Comparisons of results from our in vitro tests with results from our field trials and other field trials conducted across the eastern U.S.A. suggested that EC25 values (concentrations that reduce growth by 25%) are better predictors of fungicide efficacy in normal field conditions than EC50 values. We present these results as a guideline for choosing single-MoA fungicides for bitter rot control in the Mid-Atlantic U.S.A.

Exposure to the AhR agonist cyprodinil impacts the cardiac development and function of zebrafish larvae.

Cyprodinil is a broad-spectrum pyrimidine amine fungicide that has been reportedly used worldwide. However, toxicity studies of cyprodinil on aquatic organisms, specifically zebrafish (Danio rerio), are lacking. In our present study, we predicted cyprodinil binding to the aryl hydrocarbon receptor (AhR) by using molecular docking simulation. Then, we used recombinant HepG2 cells and Tg(cyp1a1-12DRE:egfp) transgenic zebrafish to further assess the AhR agonistic activity of cyprodinil. Besides, the significant upregulation of cyp1a1 further verified that statement. Moreover, we found that zebrafish exposure to cyprodinil induced developmental toxicity in the larvae, particularly during cardiac development. The expression levels of cardiac development-related genes, namely tbx5, nkx2.5, gata4, and tnnt2, were markedly altered, which might cause the adverse effects of cyprodinil on cardiac function and development. In summary, we found that cyprodinil, as an AhR agonist, induced development toxicity in zebrafish larvae, especially on cardiac. Data here can assess the potential effects on organisms in the aquatic environment and promote the regulation and safe use of cyprodinil.

Complete genome sequence of the cyprodinil-degrading bacterium Acinetobacter johnsonii LXL_C1.

Acinetobacter johnsonii LXL_C1, a cyprodinil degrader, was isolated and purified from cyprodinil-contaminated agricultural soil. Here, we report the complete genome sequence of LXL_C1. The genome comprises one 3,398,706 bp circular chromosome with 41.2% G + C content and one 44,866 bp plasmid. Annotation based on COG and KEGG database analyses revealed genes encoding a cytochrome P450 monooxygenase and hydrolase, which can effectively degrade cyprodinil. The complete genome sequence of LXL_C1 can facilitate genetic engineering of a recombinant cyprodinil degrader.

Biobegradation and metabolic mechanism of cyprodinil by strain Acinetobacter sp. from a contaminated-agricultural soil in China.

Using sequential soil and liquid culture enrichments with cyprodinil as the sole carbon source, a Gram-negative cyprodinil-degrader from cyprodinil-polluted agricultural soil was isolated. The sequencing analysis of 16 S rRNA indicated that the strain showed 99% homology to Acinetobacter sp. The strain could effectively degrade cyprodinil at the neutral condition. At the initial concentrations of 10, 20, 50, 100, 150 and 200 mg L-1 in minimal medium, cyprodinil was degraded by 10, 20, 49.3, 64.2, 57 and 24 mg L-1 within 14 days, respectively. Two metabolites (4-cyclopropyl-6-methyl-2-pyrimidpyridine amine and monohydroxylated para-substitution) were identified using high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (HPLC-QTOF-MS/MS). A biodegradation pathway involving imines hydrolysis and monohydroxyl substitution on benzene ring was proposed on basis of the identified metabolites. Acinetobacter sp. would have a potential application in bioremediation of cyprodinil-contaminated soil, and the strain might have important implications in detoxification and bioremediation of pyrimidine analogues.

Molecular and biological characterization of Sclerotinia sclerotiorum resistant to the anilinopyrimidine fungicide cyprodinil.

Cyprodinil belongs to the chemical class of anilinopyrimidines fungicides. In this study, baseline sensitivity of Sclerotinia sclerotiorum (Lib.) de Bary to cyprodinil was determined using 100 strains collected from the fields in Jiangsu Province of China. The EC50 (50% effective concentration) values ranged from 0.0636-0.8163 µg/ml with a mean value of 0.1869 (±0.1118) ug/ml for mycelial growth. Nine cyprodinil-resistant mutants (Range of resistance factor: 20.22-271.59) were obtained from sensitive strains exposed on PDA medium amended with cyprodinil and the resistance was stable after their ten transfers on PDA without the fungicide or stored at 4 °C for two months. There was positive cross-resistance between cyprodinil and pyrimethanil but not to fludioxonil, dimetachlone, procymidone, carbendazim and boscalid in S. sclerotiorum. Compared with the parental strains, all of the nine cyprodinil-resistant mutants decreased in sclerotial production. The dry weight of mycelia, pathogenicity and cell membrane permeability of most resistant mutants decreased. The mycelial growth, oxalic acid content, and the response to various stress for resistant mutants were almost the same as the sensitive parental strains. Sequencing alignment results showed that there was no alteration of amino acid in cystathionine γ-synthase (MetB) and cystathionine ß-lyase (MetC) between cyprodinil-resistant mutants and their sensitive parental strains, which indicated that MetB or MetC was not the molecular target of cyprodinil in S. sclerotiorum. The addition of amino acids L-methionine, L-cystine or L-cysteine decreased the inhibition of cyprodinil against mycelial growth of S. sclerotiorum, which indicated that cyprodinil could not only inhibited methionine biosynthesis but also suppressed cystine and cysteine biosynthesis. These results will contribute to evaluating the resistance risk of cyprodinil for management of the plant diseases of Sclerotinia stem rot caused by S. sclerotiorum and further increase our understanding about the mode of action of cyprodinil.

3,3'-Diindolylmethane Suppressed Cyprodinil-Induced Epithelial-Mesenchymal Transition and Metastatic-Related Behaviors of Human Endometrial Ishikawa Cells via an Estrogen Receptor-Dependent Pathway.

Cyprodinil (CYP) is a pyrimidine amine fungicide that has been extensively used in agricultural areas. 3,3'-Diindolylmethane (DIM) is a derivative of the dietary phytoestrogen, indole-3-carbinol (I3C), which is derived from cruciferous vegetables and considered to be a cancer-preventive phytonutrient agent. In this study, the effects of CYP and DIM were examined on the cell viability, invasion, and metastasis of human endometrial cancer cells, Ishikawa, via epithelial mesenchymal transition (EMT). CYP increased the level of cell viability of Ishikawa cells compared to DMSO as a control, as did E2. Ishikawa cells lost cell-to-cell contact and obtained a spindle-shaped or fibroblast-like morphology in response to the application of E2 or CYP by the cell morphology assay. In the cell migration and invasion assay, CYP enhanced the ability of migration and invasion of Ishikawa cells, as did E2. E2 and CYP increased the expressions of N-cadherin and Snail proteins, while decreasing the expression of E-cadherin protein as EMT-related markers. In addition, E2 and CYP increased the protein expressions of cathepsin D and MMP-9, metastasis-related markers. Conversely, CYP-induced EMT, cell migration, and invasion were reversed by fulvestrant (ICI 182,780) as an estrogen receptor (ER) antagonist, indicating that CYP exerts estrogenic activity by mediating these processes via an ER-dependent pathway. Similar to ICI 182,780, DIM significantly suppressed E2 and CYP-induced proliferation, EMT, migration, and invasion of Ishikawa cancer cells. Overall, the present study revealed that DIM has an antiestrogenic chemopreventive effect to withdraw the cancer-enhancing effect of E2 and CYP, while CYP has the capacity to enhance the metastatic potential of estrogen-responsive endometrial cancer.

Evaluation of nitrate effects in the photodegradability of cyprodinil. Kinetics study and transformation products elucidation.

The effects of nitrate in the kinetics and the transformation routes of the fungicide cyprodinil (CYP) were investigated using aqueous solutions, ultrapure water and river water samples, spiked with the precursor compound and containing different levels of nitrate. Samples were exposed either to 254 nm radiation or to solar light, depending on the experiment. Time course of CYP and formation of transformation products (TPs) were simultaneously assessed by direct injection of different irradiation time aliquots in a liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) system. Empirical formulae and structures of TPs were inferred from their accurate MS and MS/MS scan spectra, respectively. Under all investigated situations, nitrate anions enhanced the degradability of CYP with a noticeable reduction of its half-life (t 1/2). TPs arising from aqueous photodegradation of CYP were formed through three different routes: (1) hydroxylation of the benzenic ring; (2) hydroxylation of the pyrimidine cycle, followed by ring opening and further dealkylation; and (3) nitration of the benzenic ring. The latter group of TPs display higher estimated acute toxicities than CYP and remained stable for long irradiation times. Graphical Abstract Photonitration of cyprodinil.

Effect of fenhexamid and cyprodinil on the expression of cell cycle- and metastasis-related genes via an estrogen receptor-dependent pathway in cellular and xenografted ovarian cancer models.

Fenhexamid and cyprodinil are antifungal agents (pesticides) used for agriculture, and are present at measurable amounts in fruits and vegetables. In the current study, the effects of fenhexamid and cyprodinil on cancer cell proliferation and metastasis were examined. Additionally, the protein expression levels of cyclin D1 and cyclin E as well as cathepsin D were analyzed in BG-1 ovarian cancer cells that express estrogen receptors (ERs). The cells were cultured with 0.1% dimethyl sulfoxide (DMSO; control), 17ß-estradiol (E2; 10(-9)M), and fenhexamid or cyprodinil (10(-5)-10(-7)M). Results of a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that fenhexamid and cyprodinil increased BG-1 cell proliferation about 1.5 to 2 times similar to E2 (5 times) compared to the control. When the cells were co-treated with ICI 182,780 (10(-8)M), an ER antagonist, the proliferation of pesticide-treated BG-1 cells was decreased to the level of the control. A wound healing assay revealed that the pesticides reduced the disrupted area in the BG-1 cell monolayer similar to E2. Protein levels of cyclin D1 and E as well as cathepsin D were increased by fenhexamid and cyprodinil. This effect was reversed by co-treatment with ICI 182,780. In a xenograft mouse model with transplanted BG-1 cells, cyprodinil significantly increased tumor mass formation about 2 times as did E2 (6 times) compared to the vehicle (0.1% DMSO) over an 80-day period. In contrast, fenhexamid did not promote ovarian tumor formation in this mouse model. Cyprodinil also induced cell proliferation along with the expression of proliferating cell nuclear antigen (PCNA) and cathepsin D in tumor tissues similar to E2. Taken together, these results imply that fenhexamid and cyprodinil may have disruptive effects on ER-expressing cancer by altering the cell cycle- and metastasis-related gene expression via an ER-dependent pathway.

Effect of cyprodinil and fludioxonil pesticides on bovine liver catalase activity.

This study investigated if the use of the pesticides cyprodinil and fludioxonil produced an inhibitory effect on the bovine liver catalase (CAT) activity. It was documented that the activity of the enzyme decreased with increasing concentrations of cyprodinil and fludioxonil from 0 to 500 ppm. At pesticide concentrations of 250 and 500 ppm, the activity of CAT remained unchanged and passed to a steady state. The exposure to cyprodinil in concentrations of 10, 50, 100, 250 and 500 ppm, led to a decrease in the per cent of the CAT enzyme activity calculated as 45.4, 68.0, 73.0, 77.8 and 77.4, respectively. Similarly, the exposure to fludioxonil in concentrations of 10, 50, 100, 250 and 500 ppm, produced the following percentage decrease in the CAT enzyme activity: 20.0, 30.8, 42.8, 46.3 and 45.9, respectively. Cyprodinil inhibited CAT competitively, whereas the mechanism of fludioxonil inhibition over the enzyme was non-competitive.

Assessing solvent derivatization and carbon dioxide supercritical fluid simultaneous extraction/derivatization of cyprodinil.

Derivatization of cyprodinil with different reagents and solvents has been evaluated to improve the GC/MS characterization of this fungicide. After assessing some preliminary acylation and silylation reactions, derivatization with anhydrous heptafluorobutyric anhydride (HFBA) was selected as the best derivatization option for cyprodinil. The HFBA-cyprodinil derivative was clearly identified and characterized by GC/MS (ion-trap). The spectrum of the HFBA derivative of cyprodinil was characterized by the base peak, 252 m/z ion, and two other ions with relative abundances of 5% (224 m/z ion) and 4% (420 m/z molecular ion). Conversion rates in the range of 83-92% were obtained when 0.1-1 µg cyprodinil were derivatized in vial without solvent at 25ºC temperature for 120 min, with 5 µL HFBA and 5 µL pyridine. Simultaneous extraction-derivatization of cyprodinil in supercritical carbon dioxide was only achieved when no modifier was present, but conversion/recovery rates obtained in the replicate experiments carried out with 15 mL supercritical carbon dioxide at 50°C and 200 atm (n = 5), 300 atm (n = 7), and 400 atm (n = 5) were no reproducible (RSD > 50%) and ranged between 10% and 45% (related to the signal obtained for derivatization in vial).

Resveratrol ameliorates cyprodinil-induced zebrafish cardiac developmental defects as an aryl hydrocarbon receptor antagonist.

Cyprodinil, a globally utilized broad-spectrum pyrimidine amine fungicide, has been observed to elicit cardiac abnormality. Resveratrol (RSV), a naturally occurring polyphenolic compound, showcases remarkable defensive properties in nurturing cardiac development. To investigate whether RSV could protect against cyprodinil-induced cardiac defects, we exposed zebrafish embryos to cyprodinil (500 µg/L) in the presence or absence of RSV (1 µM). Our results showed that RSV significantly mitigated the decrease of survival rate and embryo movement and the hatching delay induced by cyprodinil. In addition, RSV also improved cyprodinil-induced zebrafish cardiac developmental toxicity, including pericardial edema and cardiac function impairment. In mechanism, RSV attenuated the cyprodinil-induced changes in mRNA expression involved in cardiac development, such as myh6, myl7, tbx5, and gata4, and calcium ion channels, such as ncx1h, slc8a4a, and atp2a2b. We further showed that RSV might inhibit the activity of aryl hydrocarbon receptor (AhR) signaling pathways induced by cyprodinil. In summary, our findings establish that the protective effects of RSV against the cardiac developmental toxicity are induced by cyprodinil due to its remarkable ability to inhibit AhR activity. Our findings not only shed light on a new avenue for regulating and ensuring the safe utilization of cyprodinil but also presents a novel concept to promote its responsible use.

(-)-Epicatechin gallate ameliorates cyprodinil-induced cardiac developmental defects through inhibiting aryl hydrocarbon receptor in zebrafish.

BACKGROUND: Cyprodinil is a widely used fungicide with broad-spectrum activity, but it has been associated with cardiac abnormalities. (-)-Epicatechin gallate (ECG), a natural polyphenolic compound, has been shown to possess protective properties in cardiac development. METHODS: In this study, we investigated whether ECG could mitigate cyprodinil-induced heart defects using zebrafish embryos as a model. Zebrafish embryos were exposed to cyprodinil with or without ECG. RESULTS: Our results demonstrated that ECG significantly improved the survival rate, embryo movement, and hatching delay induced by cyprodinil. Furthermore, ECG effectively ameliorated cyprodinil-induced cardiac developmental toxicity, including pericardial anomaly and impairment of cardiac function. Mechanistically, ECG attenuated the cyprodinil-induced alterations in mRNA expression related to cardiac development, such as amhc, vmhc, tbx5, and gata4, as well as calcium ion channels, such as ncx1h, atp2a2a, and cdh2. Additionally, ECG was found to inhibit the activity of the aryl hydrocarbon receptor (AhR) signaling pathways induced by cyprodinil. CONCLUSIONS: In conclusion, our findings provide evidence for the protective effects of ECG against cyprodinil-induced cardiac developmental toxicity, mediated through the inhibition of AhR activity. These findings contribute to a better understanding of the regulatory mechanisms and safe utilization of pesticide, such as cyprodinil.

Conversion of fungicide cyprodinil to salts with organic acids: preparation, characterization, advantages.

BACKGROUND: As an effective strategy to improve the basic properties of drugs, salt formation was less used in the field of pesticides than the medicine field. It is worth trying to improve the inherent shortcomings of cyprodinil (high Kow values; polymorphism) in this way to enhance its practicality. RESULTS: Eight cyprodinil salts (CYP-Salts) were prepared. The properties of CYP-Salts, including solubility in various solvents, polymorphic behavior, soil absorption, photolysis in aquatic water, in vitro fungicidal activity and curative activity, were assessed. It was observed that compared with those of cyprodinil, CYP-Salts had lower soil adsorption, while also having lower log Kow values and could be more easily photodegraded in water. That is, CYP-Salts have lower impacts on water bodies and aquatic organisms than cyprodinil. Three CYP-Salts showed higher in vitro antifungal activities and curative activity. CYP-Salts have enhanced practicality, as they could avoid possible agglomeration caused by recrystallization. CONCLUSION: Salt forming enhanced the properties of Cyprodinil in many aspects. CYP-Salts may potentially become a better substitute for cyprodinil. This study offers a more economical and effective strategy to prepare better alternatives to existing fungicides. © 2022 Society of Chemical Industry.

Reproductive toxicity of long-term exposure to environmental relevant concentrations of cyprodinil in female zebrafish (Danio rerio).

In recent years, the widespread use of the pesticide cyprodinil has attracted attention due to its harmful effects on aquatic organisms. The purpose of this study was to evaluate the adverse effects of long-term exposure to cyprodinil on the reproductive system of female zebrafish. After the embryos had been treated with 0.1, 1 and 10 µg/L cyprodinil for 180 days, we observed that female fish treated with 1 and 10 µg/L cyprodinil showed decreased sexual attractiveness, a decreased proportion of primordial follicles in the ovary, an increased proportion of mature follicles, and increased egg production. Moreover, exposed females that mated with normal males produced offspring with increased rates of mortality and deformity (the F1 generation). In addition, the levels of gonadotropin and testosterone (T) were increased in females after cyprodinil exposure, especially in the 10 µg/L treated group. After cyprodinil treatment, some key genes in the hypothalamic-pituitary-gonad axis underwent significant changes. For example, gene expression of brain gonadotropin-releasing hormone receptors (gnrhr1, gnrhr2 and gnrhr4) was significantly downregulated after cyprodinil treatment. The study found that expression of the aromatase (cytochrome P450 family 19 subfamily A polypeptide 1a, cyp19a1a) responsible for converting T into estradiol was significantly downregulated after cyprodinil treatment, consistent with elevated T levels in the ovaries and muscles. In summary, these data provide a more comprehensive understanding of the toxicity of cyprodinil and may inform evaluation of the ecotoxicity of cyprodinil to female reproduction at environmentally relevant concentrations.

Detection of Venturia inaequalis Isolates with Multiple Resistance in Greece.

The excessive use of fungicides against Venturia inaequalis, the causal agent of apple scab, has led to the emergence of resistant populations to multiple fungicides over the years. In Greece, there is no available information on fungicide resistance, despite the fact that control failures have been reported on certain areas. An amount of 418 single-spore isolates were collected from three major apple production areas and tested for their sensitivity to eight commonly used fungicides from unrelated chemical groups. The isolates were tested on malt extract agar media enriched with the discriminatory dose of each fungicide using the point inoculation method. To define the discriminatory dose for assessing the levels of resistance, EC50 values on both spore germination and mycelial growth assays were previously determined. Isolates exhibiting high resistance to trifloxystrobin (92% in total) and difenoconazole (3%); and moderate resistance to cyprodinil (75%), dodine (28%), difenoconazole (36%), boscalid (5%), and fludioxonil (7%) were found for the first time in Greece. A small percentage of the isolates were also found less sensitive to captan (8%) and dithianon (6%). Two isolates showed various levels of resistance to all eight fungicides. Despite the occurrence of strains with multiple resistances to many fungicides, we concluded that this practical resistance in the field arose mainly due to the poor control of apple scab with trifloxystrobin and difenoconazole.

Agricultural mulching and fungicides-impacts on fungal biomass, mycotoxin occurrence, and soil organic matter decomposition.

Plastic and straw coverage (PC and SC) are often combined with fungicide application but their influence on fungicide entry into soil and the resulting consequences for soil quality are still unknown. The objective of this study was to investigate the impact of PC and SC, combined with fungicide application, on soil residual concentrations of fungicides (fenhexamid, cyprodinil, and fludioxonil), soil fungal biomass, mycotoxin occurrence, and soil organic matter (SOM) decomposition, depending on soil depth (0-10, 10-30, 30-60 cm) and time (1 month prior to fungicide application and respectively 1 week, 5 weeks, and 4 months afterwards). Soil analyses comprised fungicides, fusarium mycotoxins (deoxynivalenol, 15-acetyldeoxynivalenol, nivalenol, and zearalenone), ergosterol, soil microbial carbon and nitrogen, soil organic carbon, dissolved organic carbon, and pH. Fludioxonil and cyprodinil concentrations were higher under SC than under PC 1 week and 5 weeks after fungicide application (up to three times in the topsoil) but no differences were observed anymore after 4 months. Fenhexamid was not detected, presumably because of its fast dissipation in soil. The higher fludioxonil and cyprodinil concentrations under SC strongly reduced the fungal biomass and shifted microbial community towards larger bacterial fraction in the topsoil and enhanced the abundance and concentration of deoxynivalenol and 15-acetyldeoxynivalenol 5 weeks after fungicide application. Independent from the different fungicide concentrations, the decomposition of SOM was temporarily reduced after fungicide application under both coverage types. However, although PC and SC caused different concentrations of fungicide residues in soil, their impact on the investigated soil parameters was minor and transient (< 4 months) and hence not critical for soil quality.

DNA damage effect of cyprodinil and thiacloprid in adult zebrafish gills.

Cyprodinil and thiacloprid are two of the most commonly used pesticides in Turkey. It is more likely to reach humans or animals due to their widespread use. This study aims to investigate whether there is a DNA damage risk due to cyprodinil and thiacloprid exposure. Zebrafish, which is used as a model organism in health and environmental research, and comet assay were chosen to demonstrate this damage. Ten zebrafish per group were exposed to 2 different concentrations for each pesticides (0.31 and 0.155 mg/L for cyprodinil and 1.64 and 0.82 mg/L for thiacloprid) for 21 days. After, gills were excised and comet assay was performed. Photos of an average of 50 cells per slide were taken and were analyzed with visual evaluation program. DNA damage was found to be increased in the 0.31 mg/L cyprodinil, 0.82 mg/L thiacloprid, and 1.64 mg/L thiacloprid treatment groups when compared to the control group (p < 0.001). Average tail DNA percentage parameter values were 9.45 ± 0.51, 10.30 ± 0.34, 11.17 ± 0.33, and 2.47 ± 0.06 respectively. Cyprodinil and thiacloprid were identified as genotoxic agents that should be investigated further.

Long-term exposure to cyprodinil causes abnormal zebrafish aggressive and antipredator behavior through the hypothalamic-pituitary-interrenal axis.

Cyprodinil, one of the main pyrimidinamine fungicides, has been used to control fungal diseases in plants and vegetables worldwide. Previous studies have investigated the influences of cyprodinil on the developmental and reproductive toxicity of fish. However, it remains unknown whether it affects fish behaviors and the underlying mechanisms. In our current study, zebrafish, an ideal model animal for behavioral studies, were exposed to cyprodinil from fertilization to 240 days postfertilization at 0.1 µg/L (environmentally relevant concentration) and 1, 10 µg/L. Firstly, we observed that aggressive behavior of zebrafish was significantly enhanced after exposure to 0.1-10 µg/L cyprodinil and antipredator behavior was decreased after exposure. Cyprodinil exposure altered the adrenocorticotropic hormone and cortisol levels, which regulate cortisol homeostasis and were significantly reduced in all exposure groups (0.1-10 µg/L). In addition, most of the key genes in the hypothalamic-pituitary-interrenal gland axis, such as corticotropin-releasing hormone and melanocortin 2 receptor, were downregulated significantly in all exposure groups, which was consistent with the hormone levels. In addition, in the hypothalamus, the number of apoptotic cells increased in a dose-dependent manner in the cyprodinil exposure groups. Moreover, these changes were potentially responsible for the increased aggression of zebrafish during the mirror-like aggressive test and for the reduced antipredator behavior during the predator avoidance test. Overall, the data provided herein further our understanding of cyprodinil toxicity and can be used to assess the ecological effects of cyprodinil on the induction of abnormal behaviors at the environmental level.

Modification of the existing maximum residue levels for cyprodinil in blueberries, cranberries, currants and gooseberries.

In accordance with Article 6 of Regulation (EC) No 396/2005, the applicant Syngenta Agro GmbH submitted a request to the competent national authority in Germany to modify the existing maximum residue levels (MRL) for the active substance cyprodinil in blueberries, cranberries, currants and gooseberries. The data submitted in support of the request were found to be sufficient to derive MRL proposals for blueberries, cranberries, currants and gooseberries. Adequate analytical methods for enforcement are available to control the residues of cyprodinil on the commodities under consideration at the validated limit of quantification (LOQ) of 0.01 mg/kg. Based on the risk assessment results, EFSA concluded that the uses of cyprodinil on blueberries, cranberries, currants and gooseberries according to the reported agricultural practices will not result in a consumer exposure exceeding the toxicological reference value and therefore is unlikely to present a risk to consumer health.