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.

Biochemical compounds and stress markers in lettuce upon exposure to pathogenic Botrytis cinerea and fungicides inhibiting oxidative phosphorylation.

MAIN CONCLUSION: Botrytis cinerea and fungicides interacted and influenced selected biochemical compounds. DPPH and glutathione are the first line of defence against biotic/abiotic stress. Plant metabolites are correlated with fungicides level during dissipation. Botrytis cinerea is an etiological agent of gray mould in leafy vegetables and is combated by fungicides. Fluazinam and azoxystrobin are commonly used fungicides, which inhibit oxidative phosphorylation in fungi. In this study, lettuce was (i) inoculated with B. cinerea; (ii) sprayed with azoxystrobin or fluazinam; (iii) inoculated with B. cinerea and sprayed with fungicides. This investigation confirmed that B. cinerea and fungicides affected lettuce's biochemistry and stress status. B. cinerea influenced the behaviour of fungicides reflected by shortened dissipation of azoxystrobin compared to non-inoculated plants, while prolonged degradation of fluazinam. Stress caused by B. cinerea combined with fungicides reduced level of chlorophylls (53.46%) and carotenoids (75.42%), whereas increased phenolic compounds (81%), ascorbate concentrations (32.4%), and catalase activity (116.1%). Abiotic stress caused by fungicides contributed most to the induction of carotenoids (107.68 µg g-1 on dissipation day 3-1). Diphenyl picrylhydrazyl (DPPH) radical scavenging activity and glutathione concentration peaked from the first hour of fungicides dissipation. For the first time correlation between the status of plant metabolites and fungicides during their dissipation was observed. These results indicate that non-enzymatic antioxidants could be the first-line compounds against stress factors, whereas ascorbate and antioxidant enzymes tend to mitigate stress only secondarily. The findings of this study help better understand plant biochemistry under biotic/abiotic stress conditions.

Estimating acute and chronic exposure of children and adults to chlorpyrifos in fruit and vegetables based on the new, lower toxicology data.

This paper presents, for the first time, results for chlorpyrifos (CHLP) in Polish fruits and vegetables over the course of a long period of research, 2007-2016, with toxicological aspects. The challenge of this study was to re-evaluate the impact of chlorpyrifos residues in fruit and vegetables on health risk assessed via acute and chronic exposure based on old and new, lower, established values of: Average Daily Intakes (ADIs)/Acute Reference Doses (ARfDs) and Maximum Residue Levels (MRLs). A total of 3 530 samples were collected, and CHLP in the range of 0.005-1.514 mg/kg was present in 10.2% of all samples. The MRL was exceeded in 0.7% of all samples (MRL established in 2009-2015), and recalculation yielded a much greater number of violations for the new MRL (2016), which exceeded 2.9% of all samples. Acute exposure to CHLP calculated according to the old, higher toxicological data (0.10 mg/kg bw/day), does not exceed 14% of its respective ARfDs for adults and both groups of children, but when calculated for incidental cases according to the current value (ARfD 0.005 mg/kg bw) for infants and toddlers, was above 100% of its respective ARfDs in: white cabbage (263.65% and 108.24%), broccoli (216.80% and 194.72%) and apples (153.20% and 167.70%). The chronic exposure calculated for both newly established ADI values (0.001 mg/kg bw/day and 0.100 mg/kg bw/day) appears to be relatively low for adults and children.

The Study of Anti-/Pro-Oxidant, Lipophilic, Microbial and Spectroscopic Properties of New Alkali Metal Salts of 5-O-Caffeoylquinic Acid.

Lithium, sodium, potassium, rubidium and caesium salts of 5-O-caffeoylquinic acid (chlorogenic acid, 5-CQA) were synthesized and described by FT-IR (infrared spectroscopy), FT-Raman (Raman spectroscopy), UV (UV absorption spectroscopy), ¹H (400.15 MHz), 13C (100.63 MHz) NMR (nuclear magnetic resonance spectroscopy). The quantum-chemical calculations at the B3LYP/6-311++G** level were done in order to obtain the optimal structures, IR spectra, NBO (natural bond orbital) atomic charges, HOMO (highest occupied molecular orbital) and LUMO (lowest unoccupied molecular orbital) orbitals and chemical reactivity parameters for 5-CQA and Li, Na and K 5-CQAs (chlorogenates). The DPPH (α, α-diphenyl-ß-picrylhydrazyl) and FRAP (ferric reducing antioxidant power) assays were used for the preliminary estimation of the antioxidant properties of alkali metal chlorogenates and chlorogenic acid. In the DPPH assay the EC50 parameter were equal to 7.39 µM for 5-CQA and was in the range of 4.50-5.89 µM for salts. The FRAP values for two different concentrations (5 and 2.5 µM) of the studied compounds were respectively 114.22 and 72.53 µM Fe2+ for 5-CQA, whereas for salts they were 106.92-141.13 and 78.93-132.00 µM Fe2+. The 5-CQA and its alkali metal salts possess higher antioxidant properties than commonly applied antioxidants (BHA, BHT, l-ascorbic acid). The pro-oxidant action of these compounds on trolox oxidation was studied in the range of their concentration 0.05-0.35 µM. The lipophilicity (logkw) of chlorogenates and chlorogenic acid was determined by RP-HPLC (reverse phase-high performance liquid chromatography) using five different columns (C8, PHE (phenyl), CN (cyano), C18, IAM (immobilized artificial membrane)). The compounds were screened for their in vitro antibacterial activity against E. coli, Bacillus sp., Staphylococcus sp., Streptococcus pyogenes and antifungal activity against Candida sp. The 5-CQA possessed lower antibacterial (minimal inhibitory concentration, MIC = 7.06 mM) and antifungal (MIC = 14.11 mM) properties than its alkali metal salts (MIC values: 6.46-2.63 mM and 12.91-5.27mM, respectively). The synthesized chlorogenates possessed better antioxidant, lipophilic, antimicrobial as well as lower pro-oxidant properties than the ligand alone. Moreover, a systematic change of the activity of alkali metal salts along the series Li→Cs suggests that there are correlations between the studied biological properties. The type of metal cation in the carboxylate group of chlorogenate is crucial for the activity of studied compounds.

Evolution of novel sorbents for effective clean-up of honeybee matrix in highly toxic insecticide LC/MS/MS analysis.

Highly toxic insecticides (HTIs) belonging to different chemical groups are dangerous to pollinating organisms, even in sublethal doses. An important objective of this study was to develop a method to determine over fifty HTIs at very low concentrations in the bee matrix. The novelty of this research involved obtaining extract completely free from beewaxes, lipids and proteins using EMR-lipid (enhanced matrix removal-lipid), chitin and Z-Sep+ (zirconium oxide and C18 dual-bonded to silica) as clean-up sorbents. Different parameters, such as weight of bees, extraction solvent, and freezing time were evaluated. Determinations were made using liquid chromatography-tandem mass spectrometry (LC-MS/MS). EMR-lipid allows for removing most of the fatty co-extracts and increases the overall performance of the method by reducing the matrix effects (ME) without significant analyte loss. The established modified QuEChERS method based on 1% acetic acid in acetonitrile extraction followed by EMR-lipid clean-up was validated at three different spiking levels (0.001, 0.01 and 0.1mgkg-1). Precision, calculated as relative standard deviation (RSD), was below 20%. The proposed method was used to determine sublethal doses of these insecticides in real samples of dead honeybees.

Multiclass pesticide residue analysis in fish muscle and liver on one-step extraction-cleanup strategy coupled with liquid chromatography tandem mass spectrometry.

The analysis of pesticide residues in fish samples is challenging due to the low concentrations and large number of analytes that need to be monitored and quantified in a complex matrix. This is the first report providing a novel one-step extraction-cleanup strategy for simultaneous analysis of over 340 pesticides in a fatty fish and liver matrix, coupled with liquid-chromatography tandem mass spectrometry. The samples of fish muscle and liver were prepared according to the modified QuEChERS (quick, easy, cheap, effective, rugged and safe) procedure, wherein the extraction and cleanup protocol were integrated into one step. Among the tested cleanup dispersive solid phase extraction sorbents (C18, primary-secondary amine, Z-Sep), chitin yielded the best results. Spike-in experiments were carried out at three different spiking levels in fish and liver to determine the recovery, precision and limits of detection of the method as well as the matrix effect. The method's detection limits ranged from 0.05 to 1.2µgkg-1, while recoveries of most pesticides were in the range of 70-120% with associated precision - relative standard deviations below 20%. A linear relationship was observed within the range of 0.005-1mgkg-1, and the correlation coefficient was R2>0.997. Expanded measurement uncertainty was estimated to be between 7% and 52%, on average. Matrix effects were evaluated and were not significant for the vast majority of pesticides. The validated method was employed in the analysis of 54 real fish and liver samples in which 10 different pesticides with concentrations ranging from 0.005 to 0.047mgkg-1 were detected.

Dissipation of S-metolachlor in plant and soil and effect on enzymatic activities.

The present study aimed at evaluating the dissipation of S-metolachlor (S-MET) at three doses in maize growing on diverse physico-chemical properties of soil. The effect of herbicide on dehydrogenase (DHA) and acid phosphatase (ACP) activity was estimated. A modified QuEChERS method using LC-MS/MS has been developed. The limit of quantification (0.001 mg kg-1) and detection (0.0005 mg kg-1) were very low for soil and maize samples. The mean recoveries and RSDs for the six spiked levels (0.001-0.5 mg kg-1) were 91.3 and 5.8%. The biggest differences in concentration of S-MET in maize were observed between the 28th and 63rd days. The dissipation of S-MET in the alkaline soil was the slowest between the 2nd and 7th days, and in the acidic soil between the 5th and 11th days. DT50 of S-MET calculated according to the first-order kinetics model was 11.1-14.7 days (soil) and 9.6-13.9 days (maize). The enzymatic activity of soil was higher in the acidic environment. One observed the significant positive correlation of ACP with pH of soil and contents of potassium and magnesium and negative with contents of phosphorus and organic carbon. The results indicated that at harvest time, the residues of S-MET in maize were well below the safety limit for maize. The findings of this study will foster the research on main parameters influencing the dissipation in maize ecosystems.

The influence of effective microorganisms (EM) and yeast on the degradation of strobilurins and carboxamides in leafy vegetables monitored by LC-MS/MS and health risk assessment.

The aim of this study was to determine the behaviour of strobilurin and carbocyamides commonly used in chemical protection of lettuce depending on carefully selected effective microorganisms (EM) and yeast (Y). Additionally, the assessment of the chronic health risk during a 2-week experiment was performed. The statistical method for correlation of physico-chemical parameters and time of degradation for pesticides was applied. In this study, the concentration of azoxystrobin, boscalid, pyraclostrobin and iprodione using liquid chromatography-mass spectrometry (LC-MS/MS) in the matrix of lettuce plants was performed, and there was no case of concentration above maximum residues levels. Before harvest, four fungicides and their mixture with EM (1 % and 10 %) and/or yeast 5 % were applied. In our work, the mixtures of 1%EM + Y and 10%EM + Y both stimulated and inhibited the degradation of the tested active substances. Adding 10%EM to the test substances strongly inhibited the degradation of iprodione, and its concentration decreased by 30 %, and in the case of other test substances, the degradation was approximately 60 %. Moreover, the addition of yeast stimulated the distribution of pyraclostrobin and boscalid in lettuce leaves. The risk assessment for the pesticides ranged from 0.4 to 64.8 % on day 1, but after 14 days, it ranged from 0.0 to 20.9 % for children and adults, respectively. It indicated no risk of adverse effects following exposure to individual pesticides and their mixtures with EM and yeast.

Removal of 16 pesticide residues from strawberries by washing with tap and ozone water, ultrasonic cleaning and boiling.

The effects of washing with tap and ozone water, ultrasonic cleaning and boiling on 16 pesticide (ten fungicides and six insecticides) residue levels in raw strawberries were investigated at different processing times (1, 2 and 5 min). An analysis of these pesticides was conducted using gas chromatography with nitrogen-phosphorous and electron capture detection (GC-NPD/ECD). The processing factor (PF) for each pesticide in each processing technique was determined. Washing with ozonated water was demonstrated to be more effective (reduction from 36.1 to 75.1 %) than washing with tap water (reduction from 19.8 to 68.1 %). Boiling decreased the residues of the most compounds, with reductions ranging from 42.8 to 92.9 %. Ultrasonic cleaning lowered residues for all analysed pesticides with removal of up to 91.2 %. The data indicated that ultrasonic cleaning and boiling were the most effective treatments for the reduction of 16 pesticide residues in raw strawberries, resulting in a lower health risk exposure. Calculated PFs for alpha-cypermethrin were used to perform an acute risk assessment of dietary exposure. To investigate the relationship between the levels of 16 pesticides in strawberry samples and their physicochemical properties, a principal component analysis (PCA) was performed. Graphical abstract ᅟ.

Studies of pesticide residues in tomatoes and cucumbers from Kazakhstan and the associated health risks.

The aim of the present study was to assess the level of pesticide residues in vegetables in the Almaty Region of Kazakhstan and to determine the potential health risks associated with the exposures to these pesticides. A total of 82 samples of cucumbers and tomatoes from top agro-based market and greenhouses were analysed using a gas chromatography-micro electron capture detector/nitrogen-phosphorous detector (GC-µECD/NPD), a multiresidue method to analyse 184 different pesticide types. The results indicated that more than half of samples (59 %) contained 29 pesticides, in which 10 are not registered in Kazakhstan, ranging from 0.01 to 0.88 mg kg(-1), and 28 % contained pesticide residues above maximum residue levels (MRLs). The estimated daily intakes (EDIs) ranged from 0.01 % of the acceptable daily intake (ADI) for pyrimethanil to 12.05 % of the ADI for lambda-cyhalothrin. The most critical commodity is triazophos and flusilazole in tomatoes, contributing 70.8 and 42.5 % to the acute hazard index (aHI). The results provided important information on the current pesticide contamination status of two commonly consumed vegetables and pointed an urgent need to control the use of plant protection products applied, especially potentially persistent pesticides, such as endosulfan and dicofol. These results also show that the detected pesticides may be considered a public health problem.

Dissipation, processing factors and dietary risk assessment of the bioinsecticide abamectin in herbal plants belonging to Lamiaceae family from open field to herbal tea infusion.

Abamectin, the mixture of avermectin B1a and B1b, is widely used as a bioinsecticide and is an alternative to chemical pest control from insects. To our knowledge, its behaviour is not fully recognized, especially in herbs. Thus, the objective of this study was to investigate the environmental fate of abamectin in herbal plants belonging to the Lamiaceae family, its dissipation in open field studies laboratory processing treatments and dietary risk assessment. Three medicinally and culinary important species of herbs: Melissa officinalis L., Mentha × piperita L. and Salvia L. were treated with single and double dose than recommended on the label during their cultivation (BBCH 11-29). Residues were monitored using the QuEChERS method followed by the LC-MS/MS. The dissipation pattern of the sum of avermectin B1a and B1b and their persistence were observed 14 d after spraying. Abamectin decline was very rapid in plants and followed the first-order kinetics model. The half-life (t1/2) was in the range of 0.96-1.08 d (single dose) and 0.93-1.02 d (double dose). The pre-harvest intervals (decrease to the level of 0.01 mg kg-1) were 7.29-7.92 d at single and 7.99-8.64 d at double dose application. Herbal infusion preparation in previously washed and dried mint, lemon balm and sage leaves was the key processing step in the removal of abamectin residues. The reduction of initial deposits after single dose treatment was noted up to 65% (PF = 0.35-0.67) and up to 79% after double dose application (PF = 0.21-0.72) in herbal tea. Acute risk assessment of children and adults for the highest residues in EFSA PRIMo model at single and double dose expressed as hazard quotients (HQ) were <1, indicating no risk to humans via consumption of the herbal products. The data provide a better understanding of abamectin behaviour in herbal plants and can help assure herbs' safety for consumers.

Plants, Microorganisms and Their Metabolites in Supporting Asbestos Detoxification-A Biological Perspective in Asbestos Treatment.

Many countries banned asbestos due to its toxicity, but considering its colossal use, especially in the 1960s and 1970s, disposing of waste containing asbestos is the current problem. Today, many asbestos disposal technologies are known, but they usually involve colossal investment and operating expenses, and the end- and by-products of these methods negatively impact the environment. This paper identifies a unique modern direction in detoxifying asbestos minerals, which involves using microorganisms and plants and their metabolites. The work comprehensively focuses on the interactions between asbestos and plants, bacteria and fungi, including lichens and, for the first time, yeast. Biological treatment is a prospect for in situ land reclamation and under industrial conditions, which can be a viable alternative to landfilling and an environmentally friendly substitute or supplement to thermal, mechanical, and chemical methods, often characterized by high cost intensity. Plant and microbial metabolism products are part of the green chemistry trend, a central strategic pillar of global industrial and environmental development.

The development, validation and application of a GC-dual detector (NPD-ECD) multi-pesticide residue method for monitoring bee poisoning incidents.

A simple multiresidue method based on matrix solid phase dispersion (MSPD) combined with clean-up has been developed for the simultaneous screening of 153 pesticides in honeybees suspected of suffering from pesticide poisoning during field spraying. Extraction and clean-up were carried out in a glass column containing anhydrous sulphate, 2.0g of octadecyl (C18) and a 2.0-g sample of bees (23 insects on average) macerated with 4.0g of Florisil. An additional layer of anhydrous sodium sulphate was added, and acetonitrile was used as the elution solvent. This combination of clean-up steps ensured an efficient purification. A gas chromatograph with dual selective detectors for electron capture and nitrogen-phosphorous was used. The results presented in this paper demonstrate that matrix solid-phase dispersion (MSPD) with the one-step clean-up procedure is the most effective extraction technique. MSPD method recoveries ranged from 70 to 118%, with precision values expressed as a relative standard of <20%, except for 10 pesticides that had recoveries of 50-70% and two with 120-130%. Low limits of detection (0.003-0.04µg/g) and quantification (0.005-0.05µg/g) were readily achieved with this method for all tested pesticides. A "top down" empirical model was used to estimate the expanded uncertainty at 28% on average (coverage factor k=2, confidence level 95%). The MSPD method was successfully used on real bee samples to analyse four acaricides, 55 fungicides, 16 herbicides and 78 insecticides from various regions of Poland. A total of 33 honeybee samples from suspected pesticide poisoning incidents were analysed, in which 17 different pesticides were determined (14 insecticides and three fungicides). The pesticides most often found in honeybees were cypermethrin (in 51% of the samples, 0.008-0.563µg/bee), chlorpyrifos (27%, 0.001-51.5µg/bee) and biphentin (21%, 0.002-0.012µg/bee).

Assessment of the genotoxic activity of alpha-asarone and its derivatives in the comet assay.

In our previous paper we examined mutagenic and genotoxic activity of 4 alpha-asarone isomers 2-5 exhibiting relatively high hypolipidemic activity. In the present paper, we examined genotoxic activity of alpha-asarone and its isomers as the ability to damage cellular DNA, evaluated in the comet assay. Additionally, mutagenic activity of alpha-asarone in Ames test has been examined. The Ames test for alpha-asarone was carried out in accordance with the guidelines of the PN-EN ISO 10993-3 standard. Compounds 4 and 5 were found to be devoid of any genotoxic activity while maintaining their hypolipemic potential. Because mutagenic activity of compound 4 was also minor it could be considered as a candidate for further pharmacological evaluation. Genotoxic but not mutagenic activity of alpha-asarone has been confirmed.

Effect of vermiculite feed additive on the chemical, mineral, and amino acid compositions of quail meat.

Background and Aim: Poultry farming plays a significant role in providing the population with high-quality and cheap meat products. The development and success of this industry depend on the proper use of available feed. As a rule, feed additives should compensate for missing elements in livestock diets. This study investigated the effects of vermiculite feed additives on the chemical, amino acid, and mineral compositions of quail meat. Materials and Methods: Texas Quail meat breed birds were randomly divided into three groups (35 heads each). Group I served as the control group, in which the birds were fed a standard diet without any supplements. The diet of Group II was supplemented with 3% vermiculite feed additive, whereas Group III received 5% vermiculite. The experiment was conducted over 120 days. Results: Quails in the experimental groups contained less moisture, had more ash and protein, and were richer in calories in terms of energy value. In terms of mineral composition, the calcium concentrations were 9.9% and 16.5% higher in the in the Group I and II, respectively. Furthermore, the highest percentages of nonessential and essential amino acids were found in the 5% vermiculite group. In the meat of quails that received 5% vermiculite, cadmium and lead salt levels were 20%-25% lower than those in the control variant. Conclusion: The results of our analyses confirmed that meeting the mineral requirements of quails improves animal productivity and meat product quality. Vermiculite as a feed additive improved the chemical, mineral, and amino acid compositions of quail meat when it was added to up to 5% of the feed dry matter. These results will help to improve the feed base of the poultry industry in the Republic of Kazakhstan.

Impact of technological processes on tebuconazole reduction in selected cereal species and the primary cereal product, and dietary exposure assessment.

Contamination of cereals with tebuconazole (TEB) can affect the dietary risk assessment. This study investigates, for the first time, how mechanical, thermal, physical-chemical, and biochemical processes affect the TEB level in wheat, rye, and barley. The biochemical process of malting was the most effective for tebuconazole reduction (by 86%) in cereals. Thermal processes were also effective, i.e., boiling (70%) and baking (55%). These processes considerably decreased the concentration of tebuconazole, and Procesing Factors (PFs) were from 0.10 to 0.18 (malting), 0.56 to 0.89 (boiling), and 0.44 to 0.45 (baking), respectively. The concentration of TEB was not reduced after the application of mechanical processing. The risk was estimated in dietary exposure assessment on the basis of the highest reported levels of tebuconazole residues bread. At a high level of rye bread consumption, the potential exposure to tebuconazole reached only 3.5% and 2.7% in children and adults, respectively.

Removal of triazole and pyrethroid pesticides from wheat grain by water treatment and ultrasound-supported processes.

A simple way to reduce pesticides in cereal grains is to use washing methods. The challenge of this study was to evaluate the effectiveness of reduction of 3 triazole fungicides (difenoconazole, tebuconazole, tetraconazole) and 3 pyrethroid insecticides (beta-cyfluthrin, cypermethrin, deltamethrin) commonly used in wheat protection. Four different pre-washing methods (hot and cold water washing, twice water, and ultrasound-supported washing) were evaluated. The processing factor (PF) was calculated based on the concentration of pesticides determined by LC-MS/MS in the samples of cereal grains before and after the washing process. PFs were within the range 0.01-0.97. Time, teperature and ultrasound were factors influencing the efficiency of water treatment. The study showed that ultrasound-supported washing eliminated pesticide residues to a greater extent than ordinary washing. This process significantly affected or completely reduced concentrations of triazoles in wheat grains. The highest reduction of residues (99%) was received for tebuconazole and ultrasound washing with heating temperature of 60 °C for a total of 10 min. In all washing processes, pyrethroids were removed with lower efficiency than triazoles. The lowest residue reduction was obtained for cypermethrin and washing under cold water for 5 min (3%; PF = 0.97). Beta-Cyfluthrin showed only a 6-27% reduction regardless of the process (PF: 0.73-0.95). Using static analysis, the relationship between the properties of pesticides and the reduction of their concentration in cereals was clarified and showed a strong correlation.

High and low temperature processing: Effective tool reducing pesticides in/on apple used in a risk assessment of dietary intake protocol.

Apples play an important role in everyone's diet and may contain pesticide residues that can pose a significant health problem for consumers. Various technological processes are promising methods for minimizing pesticide concentrations in fruit. Therefore, the subject of this comprehensive study was to investigate the effects of high-temperature (baking) and low-temperature (freeze-drying) processes on the change in the levels of nine fungicides in apples with skin and peeled. The investigated compounds belong to the chemical groups of benzimidazole (thiophanate methyl and carbendazim), phtalimide (captan and their metabolite tetrahydrophtalimid (THPI)), strobilurin (pyraclostrobin, trifloxystrobin) and triazole (difenoconazole, tebuconazole, tetraconazole). Processing factors (PF) were calculated for each pesticide-process-product combination. The results show that baking and freeze-drying generally reduced pesticide concentrations, with PFs ranging from 0.31 to 0.81 and 0.26 to 0.68, respectively. Apart from freeze-drying for carbendazim and baking for captan, PFs were above 1. Only for thiophanate-methyl, a complete reduction was observed, which resulted from complete degradation to carbendazim. The study also aimed to assess human risk according to the new strategy for different sub-populations with conversion using the 36 PFs obtained. The highest acute exposure (expressed as %ARfD) was obtained for tebuconazole in raw apples (initial concentration of 1.42 mg/kg; 400% ARfD) for Dutch toddlers. After food processing, this decreased to 284% (0.74 mg/kg, baking) and to 137% (0.37 mg/kg, freeze-drying), but was still above the safety limit. Similarly, for adults and the general French population for tebuconazole, the %ARfD was high as it reached the values of 104% (initial concentration of 0.89 mg/kg) in unprocessed apples, 73.9% after baking (0.73 mg/kg) and 35.6% after freeze-drying (0.35 mg/kg). The results indicate that food processing techniques can potentially be used to minimize the hazardous effects of pesticide residues on human health.

The processing factors of canning and pasteurization for the most frequently occurring fungicides and insecticides in apples and their application into dietary risk assessment.

The challenge of the present comprehensive work was to study, from apple orchards to consumer's plate, the influence of high- and low-temperature thermal treatments on the most frequently occurring fungicides (boscalid, captan, pyraclostrobin) and insecticides (acetamiprid, methoxyfenozide) in apples and processing factor (PF) application for more realistic dietary risk assessment in the new EFSA methodology. Dry pasteurization and canning combined with previous preliminary treatment gave PFs = 0.25-1.8 of the five active substances. Acute exposure (expressed as %ARfD) in the raw commodity was demonstrated to be 168.1% for acetamiprid in the worst case (input - highest residue) and 307.9% for boscalid in the most critical case (input - MRL), and after re-calculation for PF, decreased to 139.5% for acetamiprid in canned product and 203.2% for boscalid in pasteurized apples. These novel data may be helpful in estimating new threshold residue levels significant in food safety especially intended for children.

Evaluation of titanium and silicon role in mitigation of fungicides toxicity in wheat expressed at the level of biochemical and antioxidant profile.

Biostimulators are compounds that are gaining attention due to their potential to support plant growth, however, less in known about their impact on plant biochemistry. Therefore, the main goal of this comprehensive study was to evaluate the effect of two inorganic biostimulators (titanium, Ti and silicon, Si) and five fungicides (propiconazole, cyproconazole, spiroxamine, tebuconazole, triadimenol) on biochemical and antioxidant status as well as mycotoxin profile in wheat (Triticum aestivum). Moreover, the half-life of fungicides under the influence of biostimulators was evaluated. Titanium caused most efficient mitigation of fungicide-induced toxicity reflected by greater increase of protein (104.80 mg g-1), carbohydrates (43.70 mg g-1), phenolic compounds (1023.18 µg g-1), and peroxidase activity (0.151 U mg-1) compared to treatment with silicon. The application of spiroketalamine and triazole fungicides with silicon decreased the activity of acid phosphatases (35.70 µM h-1 g-1) while inducing amino acid (861.40 µg kg-1) and catalase activity (1.056 U mg-1). Moreover, silicon was most effective in the reduction of mycotoxin contamination in wheat grain (61.17 µg g-1). This study indicated a shortened dissipation of propiconazole, spiroxamine, tebuconazole and triadimenol under the influence of titanium or silicon (DT50 = 1.85-4.82), which can therefore affect the grace period of wheat. The results of this comprehensive investigation demonstrated that titanium and silicon have a beneficial influence on wheat biochemistry and mitigate fungicide toxicity, which makes them widely suitable for optimizing plant health.