Evaluation of acetamiprid and azoxystrobin residues and their hormonal disrupting effects on male rats using liquid chromatography-tandem mass spectrometry.

Endocrine-disrupting compounds as pesticides affect the hormonal balance, and this can result in several diseases. Therefore, the analysis of representative hormones with acetamiprid (AC) and azoxystrobin (AZ) was a good strategy for the investigation of the endocrine-disrupting activity of pesticides. Hence, a sensitive and rapid analytical method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed. The method was validated for the analysis of AC, AZ, estriol, estrone, progesterone, and testosterone in the serum, testis, and liver of rats. The correlation between the residues of pesticides and the disturbance of the endocrine system was evaluated. The different mass parameters, mobile phase types, analytical columns, injection volumes, and extraction solvents were compared to get the lowest limit of detection of the studied compounds. The detection limits of AC, AZ, estriol, estrone, progesterone, and testosterone were 0.05, 0.05, 1.0, 10, and 1.0 ng/ml, respectively. The method developed was applied to evaluate the changes in these hormones induced by the duration of exposure to AC and AZ in rat testis and serum. The hormones level in rat serum and testis had a significant decrease as they were oral gavage treated with different high concentrations of studied pesticides. Both pesticides were distributed in the body of rats by the multi-compartment model (liver, testis, and serum).

Resistance mechanisms and fitness of pyraclostrobin-resistant isolates of Lasiodiplodia theobromae from mango orchards.

BACKGROUND: Stem-end rot, caused by Lasiodiplodia theobromae (Pat.) Griffon & Maubl is a serious postharvest disease in mango. In China, a high prevalence of the QoI fungicides resistance has been reported in the last decade. The study aimed to discuss factors determining rapid development of pyraclostrobin-resistance and its resistance mechanisms. METHODS: To determine the resistance stability and fitness of pyraclostrobin resistance in L. theobromae, three phenotypes of pyraclostrobin resistance were compared and analyzed for the EC50 values, mycelial growth, virulence and temperature sensitivity and osmotic stress sensitivity. The relative conductivity and enzyme activities of different phenotypes were compared under fungicide stress to explore possible biochemical mechanisms of pyraclostrobin resistance in L. theobromae. The Cytb gene sequences of different phenotypes were analysed. RESULTS: All isolates retained their original resistance phenotypes during the 10 subcultures on a fungicide-free PDA, factor of sensitivity change (FSC) was approximately equal to 1. The resistance-pyraclostrobin of the field isolates should be relatively stable. Two pyraclostrobin-resistant phenotypes shared similar mycelial growth, virulence and temperature sensitivity with pyraclostrobin-sensitive phenotype. After treated by pyraclostrobin, the relative conductivity of the sensitive phenotype was significantly increased. The time of Pyr-R and Pyr-HR reached the most conductivity was about 8-10 times than that of Pyr-S, the time for the maximum value appearance showed significant differences between sensitive and resistant phenotypes. The activities of Glutathione S-transferase (GST), catalase (CAT) and peroxidase (POD) of Pyr-HR were 1.78, 5.45 and 1.65 times respectively, significantly higher than that of Pyr-S after treated by 200 mg/l pyraclostrobin. CONCLUSION: The results showed that the pyraclostrobin-resistant phenotypes displayed high fitness and high-risk. The nucleotide sequences were identical among all pyraclostrobin-resistant and -sensitive isolates. The pyraclostrobin resistance was not attributable to Cytb gene alterations, there may be some of other resistance mechanisms. Differential response of enzyme activity and cell membrane permeability were observed in resistant- and sensitive-isolates suggesting a mechanism of metabolic resistance.

Evaluation of the acute toxic effect of azoxystrobin on non-target crayfish (Astacus leptodactylus Eschscholtz, 1823) by using oxidative stress enzymes, ATPases and cholinesterase as biomarkers.

Azoxystrobin is a broad-spectrum fungicide used worldwide. Since azoxystrobin spreads to large areas, its toxic effects on non-target organisms have aroused interest. In this study, the acute toxicity (96 h) of azoxystrobin on the crayfish (Astacus leptodactylus) was examined by using various biomarkers. The 96 h-LC50 dose (1656 mg L-) and its three sub-doses (828, 414, 207 mg L-1) were applied to crayfish. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities were increased significantly compared to the control in hepatopancreas, gill and muscle tissues. The activities of acetylcholinesterase (AChE) and glutathione S-transferase (GST) increased, and glutathione reductase (GR) activity decreased significantly in hepatopancreas. Level of reduced glutathione (GSH) decreased significantly. The content of malondialdehyde (MDA) increased in a dose-dependent manner in all azoxystrobin treatments with the exception of the lowest dose (207 mg L-1)treatment. ATPases (Na+/K+ -ATPase, Mg2+ -ATPase, Ca2+ -ATPase, total ATPase) were significantly inhibited in gill and muscle tissues. The results of the present study indicate that azoxystrobin induces oxidative stress, and has adverse effects on activities of AChE and ATPases in crayfish.

Choice of vehicle affects pyraclostrobin toxicity in mice.

Pyraclostrobin is a strobilurin fungicide that inhibits mitochondrial complex III of fungal and mammalian cells. In toxicity studies that were used to estimate the safety factor, pyraclostrobin was added to animal feed or to aqueous vehicles. However, foods containing residues of pyraclostrobin and other strobilurin fungicides (azoxystrobin, trifloxystrobin, fluoxastrobin) are frequently prepared in vegetable oil prior to human consumption. The primary objective of this study was to determine if pyraclostrobin dissolved in an oil-based vehicle had adverse health outcomes in mice when compared to aqueous-based vehicles. We found that pyraclostrobin does not fully dissolve in aqueous methyl cellulose (MC) or carboxymethyl cellulose (CMC), two vehicles used in industry-sponsored toxicity studies, but does fully dissolve in corn oil. Moreover, C57BL/6 mice receiving pyraclostrobin in corn oil displayed adverse health outcomes, including loss of body weight, hypothermia and diarrhea at lower doses than when added to feed or to aqueous vehicles. Our data suggest that previous studies underestimated the true toxicity of pyraclostrobin in mammals. Additional toxicity tests using oil-based vehicles are recommended to verify current safety recommendations for strobilurin fungicides.

Compound pesticide controlled release system based on the mixture of poly(butylene succinate) and PLA.

Controlled release system has been widely developed and utilised in agriculture to optimise efficacy and minimise environmental pollution. Here, the azoxystrobin and difenoconazole compound pesticide microsphere was constructed by the solvent evaporation method. Blending poly (butylene succinate) (PBS) and poly (lactic acid) (PLA) as shells, in order to increase its entrapment efficiency and match the goal of complex synergism. The resulting microsphere processes high entrapment efficiency with a mean diameter of 7.2 µm. Acute toxicity suggested microsphere on zebrafish was moderate toxicity. Comparative studies revealed that the microsphere had a longer period of sustained release than difenoconazole-azoxystrobin (5:8) 32.5% w/v suspension concentrate (SC). In addition, the concentration ratio of active ingredients closed to the optimum compound stoichiometry, which can achieve the objective of improving the efficacy of the pesticides. These results demonstrated that such a compound pesticide microsphere delivery system may be a considerable potential for further exploration.

Evaluation of the TOM-CAST Forecasting Model in Asparagus for Management of Stemphylium Leaf Spot in Ontario, Canada.

In the last 5 years, asparagus acreage in Canada has increased by over 25%. Stemphylium leaf spot, caused by Stemphylium vesicarium, has emerged as the predominant foliar pathogen of asparagus. Typically, contact fungicides are applied every 14 days; however, regardless of the number of applications, growers are not achieving adequate control of the disease. The TOM-CAST forecasting model is used widely in Michigan asparagus fields, but it has never been assessed for suitability in Ontario or in the popular cultivar, Guelph Millennium. Six field trials were conducted in 2012 and 2013 to evaluate the TOM-CAST forecasting model in two asparagus cultivars. The fungicides chlorothalonil or azoxystrobin/difenoconazole were applied according to the forecasting model or on a 14-day interval. The effectiveness of the forecasting model differed between sites and cultivars. Even though TOM-CAST is used in all cultivars in Michigan, TOM-CAST was not effective on Guelph Millennium. In the cultivar Jersey Giant, however, TOM-CAST with a 20 disease severity value spray interval improved control of Stemphylium leaf spot without increasing the number of sprays, compared with a 14-day treatment. The results in Guelph Millennium differed between sites. At one site, TOM-CAST maintained similar levels of Stemphylium leaf spot, but increased the number of applications, compared with a 14-day application interval. Of more concern, none of the fungicide treatments differed greatly from the untreated control at the other site. Our results show that forecasting models need to be validated locally in asparagus cultivars relevant to production today.

Evaluation of Self-Propelled High-Energy Ultrasonic Atomizer on Azoxystrobin and Tebuconazole Application in Sunlit Greenhouse Tomatoes.

In this study, a self-propelled high-energy ultrasonic atomizer was evaluated in terms of deposition on the canopy, the loss to the ground, and fungicide residues in cherry tomato and tomato. Artificial collectors fixed to the upper side and underside of the leaves at different depths and heights were used to collect the depositions. A reliable analytical method for determination of azoxystrobin and tebuconazole in artificial collectors and residue samples was developed by using liquid chromatography triple-quadrupole mass spectrometry. The results showed that the atomizer distributed the droplets evenly throughout the greenhouse with good uniformity (CVs below 39%). The ratio of depositions on the internal and external sides was 66⁻83%, and the ratio of depositions on the underside and upper side was 39⁻50%. There were no significant differences in depositions between two different height crops. The residues of azoxystrobin and tebuconazole in tomato and cherry tomato fruits were far below the maximum residue limits at harvest time. In general, self-propelled high-energy ultrasonic atomizer used in a greenhouse could increase the depositions, especially on the underside and internal side of the canopies, and lead to a reduction of operator exposure risk.

Differences in Small Plot and On-Farm Trials for Yield Response to Foliar Fungicide in Soybean.

Soybean yield response variability to foliar fungicide applications was evaluated in on-farm replicated strip trials (OFTs) and small-plot trials (SPTs) from 2008 through 2015 in Iowa. A total of 230 OFTs and 49 SPTs were compared for yield response to pyraclostrobin, pyraclostrobin + fluxapyroxad, or trifloxystrobin + prothioconazole fungicides. OFTs (18 to 55 m wide and 200 to 800 m long strips) were harvested with farmers' combines equipped with yield monitors and GPS, while SPTs (3.0 to 4.6 m wide and 10.7 to 15.3 m long plots) were harvested by small research plot combines. Variance component and power analyses were conducted with a subset of data consisting of 12 OFTs and SPTs, each with pyraclostrobin and evaluated in 2008 and 2009. While average yield responses were similar, the residual random yield variation was smaller in OFTs than SPTs. Power analysis showed that SPTs need more replications than OFTs to detect the same overall treatment differences. To detect a yield response of 134 kg/ha, it would require at least three treatment replications with 12 locations in OFTs and seven replications with 12 locations in SPTs. Researchers need to acknowledge the differences in statistical power of detecting yield responses to foliar fungicide on soybean in different types of field experiments, especially with smaller plot sizes in situations with less foliar disease.

First study of hormesis effect on mushroom cultivation.

The use of fungicides is common in mushroom cultivation, but no study was carried out applying reduced doses of fungicides in order to increase yield, taking account the hormesis effect. The aim of this manuscript was to verify the effects of different concentrations of fungicides to stimulate the productivity of different strains of Agaricus bisporus. Two stages were developed, an in vitro study to define the best concentration to be applied in the second experiment an agronomic study, which consisted of the application of the selected fungicides, in their respective concentrations, in an experiment carried out in the mushroom chamber. Clearly, the result of the hormesis effect on mushroom cultivation can be verified. The results obtained in the 1st stage of the study (in vitro) were not always reproduced in the 2nd stage of the study (in vivo). The kresoxim methyl active ingredient may be an important chemical agent, while strain ABI 15/01 may be an extremely important biological agent to increase yield in the study of hormesis effects.