Concentrations Of Vinclozolin Research Articles

Effects of Subtoxic Concentrations of Atrazine, Cypermethrin, and Vinclozolin on microRNA-Mediated PI3K/Akt/mTOR Signaling in SH-SY5Y Cells.

Endocrine-disrupting chemicals (EDCs) are different natural and synthetic chemicals that may interfere with several mechanisms of the endocrine system producing adverse developmental, metabolic, reproductive, and neurological effects in both human beings and wildlife. Among pesticides, numerous chemicals have been identified as EDCs. MicroRNAs (miRNAs) can regulate gene expression, making fine adjustments in mRNA abundance and regulating proteostasis. We hypothesized that exposure to low doses of atrazine, cypermethrin, and vinclozolin may lead to effects on miRNA expression in SH-SY5Y cells. In particular, the exposure of SH-SY5Y cells to subtoxic concentrations of vinclozolin is able to downregulate miR-29b-3p expression leading to the increase in the related gene expression of ADAM12 and CDK6, which may promote a pro-oncogenic response through the activation of the PI3K/Akt/mTOR pathway and counteracting p53 activity. A better understanding of the molecular mechanisms of EDCs could provide important insight into their role in human disease.

Effects of chronic exposure to the fungicide vinclozolin on gut microbiota community in an aquatic turtle

Environmental issues associated with the widespread use of agricultural chemicals are being seriously concerned. Of them, toxicological impacts of fungicides in aquatic organisms are often overlooked. Here, soft-shelled turtle (Pelodiscus sinensis) hatchlings were exposed to different concentrations of vinclozolin (0, 5, 50, 500 and 5000 μg/L) for 60 days to investigate the impact of fungicide exposure on their gut microbial composition and diversity. Vinclozolin exposure significantly affected the composition of the gut microbiota in hatchling turtles. Unexpectedly, gut bacterial diversity and richness of vinclozolin-exposed turtles (but not for the 5000 μg/L-exposed group) were relatively higher than control ones. At the phylum level, the abundance of Firmicutes was decreased, while that of Proteobacteria was increased in high-concentration groups. At the genus level, some bacterial genera including Cellulosilyticum, Romboutsia and Clostridium_sensu_stricto, were significantly changed after vinclozolin exposure; and some uniquely observed in high-concentration groups. Gene function predictions showed that genes related to amino acid metabolism were less abundant, while those related to energy metabolism more abundant in high-concentration groups. The prevalence of some pathogens inevitably affected gut health status of vinclozolin-exposed turtles. Such gut microbiota dysbiosis might be potentially linked with hepatic metabolite changes induced by vinclozolin exposure.

Functional and hepatic metabolite changes in aquatic turtle hatchlings exposed to the anti-androgenic fungicide vinclozolin

Many man-made chemicals that are released into water bodies in agricultural landscapes have been identified as endocrine disruptors and can cause serious impacts on the growth and survival of aquatic species living in these environments. However, very little attention has been paid to their toxicological effects in cultured non-fish species, such as aquatic turtles. We exposed hatchlings of the Chinese soft-shelled turtle (Pelodiscus sinensis) to different concentrations of vinclozolin (0, 5, 50 and 500 μg/L) for 60 days to assess physiological and metabolic impacts of this fungicide. Despite no death occurrence, hatchling turtles exposed to the highest concentration of vinclozolin consumed less food, grew more slowly (resulting in smaller body size after exposure) and performed more poorly in behavioral swimming tests than controls and turtles exposed to lower concentrations. Hepatic metabolite profiles acquired via liquid chromatography-mass spectrometry (LC-MS) revealed multiple metabolic perturbations related to amino acid, lipid, and fatty acid metabolism in animals exposed to environmentally relevant concentrations. Specifically, many critical metabolites involved in energy-related metabolic pathways (such as some intermediates in the tricarboxylic acid cycle, lactate, and some amino acids) were present in livers of hatchling turtles exposed vinclozolin, though at lower concentrations, reflecting energy metabolism dysregulation induced by exposure to this fungicide. Overall, our results suggest that the changes in growth and behavioral performances caused by chronic vinclozolin exposure may be associated with internal physiological and metabolic disorders mediated at the biochemical level.

Electrochemical Reduction Bahaviour of Vinclozolin Fungicide at Platinum Electrode

The electrochemical behavior of vinclozolin (3-(3,5-dichlorophenyl)-5-methyl-5-vinyl-1,3 oxazolidine-2,4-dion) in dimethylsulfoxide was performed by cyclic voltammetry (CV) and linear sweep voltammetry (LSV). The main decomposition pathway included in the cleavage of chlorine atoms at platinum electrode. The general reduction mechanism of this compound has been suggested based on the results obtained. The peak potential and peak current were found to be depending on pH of the buffer solution. The measuring system response was linear over the vinclozolin concentration range from 1.0 ×10 -3 to 5.0 ×10 -7 mol L -1 and the detection limit (LOD) achieved was 1.1 ×10 -8 mol L -1 (2.5 µg L -1 ) at pH 4.0. Vinclozolin was determined in spiked serum sample.

Influence of environmentally relevant concentrations of vinclozolin on quality, DNA integrity, and antioxidant responses of sterlet Acipenser ruthenus spermatozoa

The effects of vinclozolin (VIN), an anti-androgenic fungicide, on quality, oxidative stress, DNA integrity, and ATP level of sterlet (Acipenser ruthenus) spermatozoa were investigated in vitro. Fish spermatozoa were incubated with different concentrations of vinclozolin (0.5, 2, 10, 15, 20 and 50μg/l) for 2h. A dose-dependent reduction in spermatozoa motility and velocity was observed at concentrations of 2–50μg/l. A dramatic increase in DNA fragmentation was recorded at concentrations 10μg/l and above. After 2h exposure at higher test concentrations (10–50μg/l), oxidative stress was apparent, as reflected by significantly higher levels of protein and lipid oxidation and significantly greater superoxide dismutase activity. Intracellular ATP content of spermatozoa decreased with increasing concentrations of VIN. The results demonstrated that VIN can induce reactive oxygen species stress in fish spermatozoa, which could impair the sperm quality, DNA integrity, ATP content, and the antioxidant defense system.

Development of partial life-cycle experiments to assess the effects of endocrine disruptors on the freshwater gastropod Lymnaea stagnalis: a case-study with vinclozolin

Long-term effects of endocrine disruptors (EDs) on aquatic invertebrates remain difficult to assess, mainly due to the lack of appropriate sensitive toxicity test methods and relevant data analysis procedures. This study aimed at identifying windows of sensitivity to EDs along the life-cycle of the freshwater snail Lymnaea stagnalis, a candidate species for the development of forthcoming test guidelines. Juveniles, sub-adults, young adults and adults were exposed for 21 days to the fungicide vinclozolin (VZ). Survival, growth, onset of reproduction, fertility and fecundity were monitored weekly. Data were analyzed using standard statistical analysis procedures and mixed-effect models. No deleterious effect on survival and growth occurred in snails exposed to VZ at environmentally relevant concentrations. A significant impairment of the male function occurred in young adults, leading to infertility at concentrations exceeding 0.025 μg/L. Furthermore, fecundity was impaired in adults exposed to concentrations exceeding 25 μg/L. Biological responses depended on VZ concentration, exposure duration and on their interaction, leading to complex response patterns. The use of a standard statistical approach to analyze those data led to underestimation of VZ effects on reproduction, whereas effects could reliably be analyzed by mixed-effect models. L. stagnalis may be among the most sensitive invertebrate species to VZ, a 21-day reproduction test allowing the detection of deleterious effects at environmentally relevant concentrations of the fungicide. These results thus reinforce the relevance of L. stagnalis as a good candidate species for the development of guidelines devoted to the risk assessment of EDs.

An environmentally relevant endocrine-disrupting antiandrogen, vinclozolin, affects calling behavior of male Xenopus laevis

Vinclozolin (VIN) is an antiandrogenic model substance as well as a common fungicide that can affect the endocrine system of vertebrates. The objective of this study was to investigate how VIN affects mate calling behavior of South African clawed frogs ( Xenopus laevis) and whether it is effective at environmentally relevant concentrations. Male X. laevis were injected with human chorionic gonadotropin (hCG) to stimulate their androgen-controlled mate calling behavior and were treated with VIN at concentrations of 10 − 6, 10 − 8 and 10 − 10 M. VIN at 10 − 6 M reduced calling activity. Furthermore, the vocalization composition of VIN-treated X. laevis was altered. The call types advertisement calls and chirping are uttered by reproductively active males, whereas the call types growling, ticking, and rasping indicate a sexually unaroused state of a male. VIN at any of the tested concentrations led to a decrease in utterance of calls, which indicate a sexually aroused state of the males, and an increase in relative proportions of calls, indicating a sexually unaroused state of the males. Additionally, the mean duration of clicks and the number of accentuated clicks during the advertisement calls decreased at all concentrations of VIN. No significant differences were observed in any other temporal or spectral calling parameters between the treatments. This study illustrates that exposure to the antiandrogen VIN might result in a reduced reproductive success by altering mate calling behavior of X. laevis. Moreover, it suggests that the behavioral parameters examined in this study can be used as sensitive biomarkers for detecting antiandrogenic endocrine disrupting compounds in amphibians.

Fate of vinclozolin, thiabendazole and dimethomorph during storage, handling and forcing of chicory.

As part of ongoing research for a sustainable production of Belgian endives, the fate of three fungicides during storage, handling and forcing of witloof chicory roots was investigated. Storage roots are protected against Sclerotinia sp. Fuckel and Phoma exigua var. exigua Desm. by means of vinclozolin and thiabendazole respectively. During hydroponic forcing, the most imminent pathogen is Phytophthora cryptogea Pethybr. & Laff., which is controlled by the use of dimethomorph. Vinclozolin and thiabendazole concentrations on roots remained constant during storage at -1 degrees C. Dermal exposure of the workers in hydroponics was exceeded. Vinclozolin and thiabendazole residues were not detected 2 weeks after hydroponic forcing; dimethomorph was still detected at harvest. At harvest, the vinclozolin concentration in the chicory heads was below the maximum residue limit, but the chicory roots contained residues much above the thiabendazole and dimethomorph maximum residue level. Vinclozolin and thiabendazole residues applied before storage are still present on the roots at the start of the forcing cycle. During the set-up of chicory roots, preventive measures are recommended, as effects of repeated human exposure to low doses of applied fungicides cannot be excluded. Dimethomorph applied at the start of the hydroponic forcing is the only pesticide detected in the drainage water at harvest. The chicory heads were safe for human consumption. However, more attention should be paid to the residues of fungicides in the roots used for cattle feeding.

Vinclozolin, a widely used fungizide, enhanced BaP-induced micronucleus formation in human derived hepatoma cells by increasing CYP1A1 expression

Vinclozolin, a widely used fungicide, can be identified as a residue in numerous vegetable and fruit samples. To get insight in its genetic toxicity, we investigated the genotoxic effect of vinclozolin in the human derived hepatoma cell line HepG2 using the micronucleus (MN) assay. Additionally, to evaluate the co- or anti-mutagenic potency of vinclozolin, we treated HepG2 cells with different concentrations of vinclozolin for 24 h. Subsequently, the cells were exposed to benzo[ a]pyrene (BaP) for 1 h. Exposure of HepG2 cells to 50–400 μM vinclozolin alone did not cause any induction of micronuclei. However, a pronounced co-mutagenic effect was observed. MN frequencies caused by BaP increased by 30.6%, 52.8% and 65.3% after pretreatment of the cell cultures with 50, 100 and 200 μM vinclozolin, respectively. The highest concentration (400 μM) of vinclozolin tested caused cytotoxicity. Therefore, micronuclei were not considered for that concentration. To clarify the mechanism of cogenotoxicity, we assayed cytochrome P450 1A1 (CYP1A1), which plays a pivotal role in activation of BaP. Cells exposed to vinclozolin led to significant increase of CYP1A1 expression in Western blot. The result suggested that induction of CYP1A1 by vinclozolin account for its enhancing effect on genotoxicity caused by BaP.

Effect of Pathogen Aggressiveness and Vinclozolin on Efficacy of Rhodotorula glutinis PM4 Against Botrytis cinerea on Geranium Leaf Disks and Seedlings

Efficacy of the yeast Rhodotorula glutinis isolate PM4 as a biological control agent against 29 isolates of Botrytis cinerea obtained from greenhouse-grown ornamentals was assessed in vitro on geranium leaf disks. Isolates of B. cinerea varied in aggressiveness in the absence of either biological or chemical controls; diameters of lesions produced on leaf disks ranged from 0.8 to 12.3 mm. Efficacy of R. glutinis PM4 against the different isolates of B. cinerea varied greatly; lesion diameters ranged from 0.2 to 10.3 mm when the yeast was present. The yeast significantly reduced lesion development by 16 B. cinerea isolates in each of two replicate trials and by 9 isolates in one of the trials; however, 3 isolates were not inhibited by the yeast on geranium leaf disks. The yeast significantly reduced lesion development by B. cinerea isolate 01, used as a standard for comparison, in four of six trials. Fourteen of the B. cinerea isolates were inoculated onto geranium seedlings and produced a range of lesion sizes (2.9 to 16.4 mm), similar to that produced on leaf disks. Efficacy of the yeast in combination with a reduced rate (0.1×) of the fungicide vinclozolin (50 μg of active ingredient ml-1) was evaluated on geranium seedlings against 10 isolates of B. cinerea that were resistant to vinclozolin. Addition of vinclozolin to the yeast significantly reduced lesion diameter by five of the isolates compared with diameters of lesions produced in the presence of the yeast alone. Lesions produced by nine of the resistant isolates were 2.6 mm or smaller in both trials on plants treated with the mixture of yeast and vinclozolin. The effect of vinclozolin concentration (0 to 500 μg a.i. ml-1) on biocontrol efficacy of R. glutinis PM4 was evaluated using three resistant isolates of B. cinerea and geranium seedlings. Disease control was significantly better at higher concentrations of fungicide for two of the isolates. Linear regression of lesion diameter against vinclozolin concentration showed a significant effect on yeast biocontrol efficacy with B. cinerea isolate FL-2-b (y = 6.20 - 0.63x; r2= 0.74) and isolate BR-1 (y = 4.10 - 0.32x; r2 =0.28) but there was no significant effect with isolate GG-2-b. Overall, PM4 exhibited biocontrol activity on both geranium leaf disks and seedlings against a number of isolates of B. cinerea that varied in aggressiveness. Variability in biocontrol efficacy against isolates resistant to vinclozolin usually was reduced by the addition of vinclozolin.

Transfer of the fungicide vinclozolin from treated to untreated plants via volatilization

Head lettuce plantlets ( Lactuca sativa L. var. capitata) were potted, treated with vinclozolin at the six-leaf stage according to application standards and allowed to dry for 24 h. The potted plantlets were then placed in either growth chambers with controlled temperature (20 and 25 °C, respectively) or in a greenhouse (approximately 12 °C), together with untreated spinach ( Spinacia oleracea L.) and standardized grass cultures ( Lolium multiflorum Lam. ssp.) While the treated lettuce pots remained in the respective growing compartments until the end of the experiments, spinach and grass were exposed to the compartment air for 24 h and their shoot material was analyzed for vinclozolin by GC-ECD and GC-high resolution mass spectrometry. Exposure and analysis of untreated spinach and grass were carried out at two- or three-day intervals during the course of the experiments. Also, air samples were taken from the compartments at intervals and analyzed for vinclozolin. Maximum vinclozolin concentration in the growth chamber air was about 330 ng m −3 while vinclozolin contamination of the untreated plants ranged from 50 to 200 μg kg −1 FW (fresh weight). In the greenhouse atmospheric vinclozolin concentration reached approximately 15 ng m −3 and maximum contamination of spinach and grass were 30–40 μg kg −1 FW. Our data clearly show that unintended contamination of plants growing in the vicinity of vinclozolin-treated plants can occur even if the fungicide layer is completely dry. Implications for safety testing and food plants are discussed.

Effects of the mammalian antiandrogen vinclozolin on development and reproduction of the fathead minnow ( Pimephales promelas)

Previous work with the chlorinated fungicide vinclozolin and its metabolites, 2-{[(3,5-dichlorophenyl)-carbamoyl]oxy}-2-methyl-3-butenoic acid (M1) and 3′,5′-dichloro-2-hydroxy-2-methylbut-3-enanilide (M2), indicated antiandrogenic properties expressed in vivo as abnormalities in sexual differentiation of male rats after maternal exposures. In this study, we attempted to determine whether vinclozolin might also exhibit antiandrogenic properties in a model fish species, the fathead minnow, Pimephales promelas. In one study, embryonic (<6 h old) fathead minnows were exposed for approximately 34 days to five toxicant concentrations, ranging from 90 to 1200 μg l −1, delivered via a flow-through diluter. The embryos were periodically sampled to determine survival, growth and gross pathology, and then placed in clean water for 4–6 months to assess long-term effects on sexual differentiation and subsequent reproductive success. Except for slightly reduced growth after 34 days in the highest vinclozolin concentration, no adverse effects were noted with respect to any of these endpoints. In a second experiment, adult fathead minnows were exposed to vinclozolin concentrations of approximately 200 or 700 μg l −1 for 21 days, following which, gonadal morphology was assessed and serum sex steroid concentrations determined. Tissue samples from the exposed adults were assayed for vinclozolin and its metabolites. There was a slight increase in the serum β-estradiol concentration of the male fathead minnows exposed to 700 μg vinclozolin l −1, and a marked reduction in gonadal condition of female fish from this treatment. The possibility that vinclozolin and its metabolites would bind to androgen receptors in the fathead minnow was investigated through competitive radioligand binding studies. Vinclozolin, M1 and M2 failed to compete for high-affinity, low-capacity testosterone binding sites in fathead minnow brain and ovary cytosolic fractions, suggesting that these chemicals might not act as antiandrogens in the fathead minnow. More experimentation is necessary to determine whether responses observed in vivo might be due to the effects of vinclozolin (or its metabolites) on some other aspect of endocrine function.

Cytogenetic damage and induction of pro‐oxidant state in human lymphocytes exposed in vitro to gliphosate, vinclozolin, atrazine, and DPX‐E9636

We analyzed chromosome aberrations (CAs), sister chromatid exchanges (SCEs), mitotic index (MI), and glucose 6-phosphate dehydrogenase (G6PD) enzyme activity in human peripheral lymphocytes from three healthy donors exposed in vitro to different concentrations of gliphosate, vinclozolin, atrazine, and DPX-E9636. The pesticides gliphosate, vinclozolin, and atrazine have been studied in a broad range of genetic tests with predominantly conflicting or negative results, whereas little is known about the genotoxicity of DPX-E9636. In our experimental conditions, each chemical compound tested produced a dose-related increase in the percent of aberrant cells and an increase of SCE/cell. Furthermore, at the highest concentrations of vinclozolin, atrazine, and DPX-E9636, we observed a significant reduction of the mitotic index. The increase of G6PD activity in exposed lymphocyte cultures strongly indicated an induction of a pro-oxidant state of the cells as an initial response to pesticide exposure. Environ. Mol. Mutagen. 32:39–46, 1998 © 1998 Wiley-Liss, Inc.

Cytogenetic damage and induction of pro-oxidant state in human lymphocytes exposed in vitro to gliphosate, vinclozolin, atrazine, and DPX-E9636

We analyzed chromosome aberrations (CAs), sister chromatid exchanges (SCEs), mitotic index (MI), and glucose 6-phosphate dehydrogenase (G6PD) enzyme activity in human peripheral lymphocytes from three healthy donors exposed in vitro to different concentrations of gliphosate, vinclozolin, atrazine, and DPX-E9636. The pesticides gliphosate, vinclozolin, and atrazine have been studied in a broad range of genetic tests with predominantly conflicting or negative results, whereas little is known about the genotoxicity of DPX-E9636. In our experimental conditions, each chemical compound tested produced a dose-related increase in the percent of aberrant cells and an increase of SCE/cell. Furthermore, at the highest concentrations of vinclozolin, atrazine, and DPX-E9636, we observed a significant reduction of the mitotic index. The increase of G6PD activity in exposed lymphocyte cultures strongly indicated an induction of a pro-oxidant state of the cells as an initial response to pesticide exposure.

Environmental fate of pesticides used in Australian viticulture: Behaviour of dithianon and vinclozolin in the soils of the South Australian Riverland

The red calcareous earth soils of the South Australian Riverland produce more than one-third of the grapes used in Australian winemaking. As part of on-going investigations into pesticide transport in Australian vineyard soils, the movement of the fungicides dithianon and vinclozolin through such strongly alkaline soils was investigated. Small, undisturbed soil cores were extracted from the inter-row topsoil of a vineyard adjacent to the River Murray, approximately 10 km S.W. of Overland Corner, South Australia. The vines were grown in a deep (1 – 4 m) reddish brown, strongly alkaline, sandy loam with a low organic carbon content (1 – 2 %). Surface fluxes of pesticide were applied at the maximum recommended application rates to the surface of the cores, which were then irrigated, and pesticide residues in the leachate determined by HPLC. No leaching of either dithianon or vinclozolin occurred. Dithianon was immobilised in the top 2 cm of the soil. Dithianon concentrations were low ( 0 – 37 % applied dose) suggesting that rapid degradation of this compound occurs in these soils (63 – 100 % degradation in 10 days). Extremely low concentrations of vinclozolin were found throughout the soil core profiles (0.05 – 1.4 % applied dose) suggesting that this fungicide was somewhat mobile, but also that it too was unstable in such alkaline soils (> 98 % degradation in 10 days). These results suggest that the irrigated vineyard soils of this region are unlikely to be prone to leaching of dithianon or vinclozolin, and therefore that groundwater supplies in this area are unlikely to be at any significant risk of contamination through viticultural use of these compounds.

Long-Term Studies of Fungicide Concentrations in Greenhouses. 2. Fungicide Concentrations in Air and on Leaves after Different Exposure Times and under Different Climate Conditions

The decline of the two fungicides vinclozolin (formulation Ronilan FL) and triadimefon (formulation Bayleton Special) was studied in greenhouses. The decline of vinclozolin was also studied in climate chambers. In greenhouse experiments the fungicides were applied with either a low-volume sprayer or a high-volume sprayer, while in the climate chamber experiments the fungicide was applied with a pipet. Concentrations on leaves 1 day after spraying in the greenhouses with Ronilan FL were 0.87 μg of vinclozolin/cm2 and after spraying with Bayleton Special close to the detection limit (0.002 μg of triadimefon/cm2). Concentrations were higher on floors than on leaves. Air concentrations were higher immediately after low-volume spraying than after high-volume spraying but decreased rapidly. Air concentrations of vinclozolin during the first harvest, the third day after application, were below the detection limit (0.4 μg/m3). Climate chamber experiments showed no significant differences in residue decline rate between different climate conditions, with temperatures in the range of 18−26 °C and a vapor pressure deficit between 0.26 and 0.79 kPa. Keywords: Spraying; greenhouse; fungicide; pesticide; residues; decline; vinclozolin

The fungistatic and fungicidal activity of vinclozolin against Botrytis cinerea

The germination of Botrytis cinerea conidia in nutrient medium was completely inhibited by 20 μ m vinclozolin. Probits of the percentage inhibition of spore germination were linearly correlated with log vinclozolin concentration indicating that vinclozolin acted as a single compound with one mechanism of action. Treatment of resting (ungerminated) spores suspended in buffer with vinclozolin concentrations as high as 750–1000 μ m resulted in no significant release of cellular potassium and inorganic phosphate or decrease in cell viability. On the contrary, for germinated B. cinerea conidia growing in liquid medium, vinclozolin showed a slow but marked fungicidal activity. After 3 and 20 h of treatment with 2 μ m vinclozolin, 63 and 4% of the cells survived, respectively. However, if vinclozolin was added to germinated spores in non-growing conditions, no significant reduction in cell viability was observed after 20 h of treatment. The results indicated that vinclozolin has a strong fungistatic activity, but is fungicidal only in actively growing cells, and that it causes no drastic alteration in the permeability of the cytoplasmic membrane of resting conidial cells. The implications of these results for the chemical control of plant mycoses are briefly discussed.