Triazole Fungicides Research Articles

Benzimidazole-based bimodal periodic mesoporous organosilica for thin film microextraction of triazole fungicides in fruiting vegetables prior to gas chromatography-mass spectrometry

Benzimidazole-based bimodal periodic mesoporous organosilica for thin film microextraction of triazole fungicides in fruiting vegetables prior to gas chromatography-mass spectrometry

Antifungal activity and mechanism of mefentrifluconazole against Magnaporthe grisea causing rice blast

Rice blast, caused by Magnaporthe grisea, is one of the most destructive diseases of rice worldwide. Mefentrifluconazole, a novel isopropanol triazole fungicide, exhibits excellent bioactivity to a wide range of fungi. However, its antifungal activity against M. grisea is not well studied. In this study, the sensitivity of 103 M. oryzae strains to mefentrifluconazole was determined by mycelial growth rate method, and the effect of mefentrifluconazole on the biochemical and physiological characteristers was examined. The results indicated that mefentrifluconazole exhibited excellent inhibitory activity against M. oryzae with EC50 values of 103 M. oryzae strains tested ranging from 0.363 to 1.576 μg/mL, and the average EC50 value of 0.958 ± 0.298 μg/mL. After mefentrifluconazole treatment, the apical branches of M. oryzae mycelia increased and the mycelia became distorted, twisted and ruptured. In addition, mefentrifluconazole increased relative conductivity, disrupted cell wall and membrane integrity, and significantly reduced ergosterol content. The expression levels of demethylation inhibitor resistance-associated genes Cyp51A, ABC6 and MFS1 were all significantly upregulated, suggesting an adaption response to mefentrifluconazole of M. oryzae. Furthermore, the results of pot experiment showed that mefentrifluconazole provided >70% control efficacy against rice blast at 300 μg/mL, surpassing the performance of carbendazim, prochloraz and azoxystrobin at the same concentration. Overall, this study demonstrated the strong inhibitory activity of mefentrifluconazole against M. oryzae and the excellent efficacy in controlling rice blast, thus providing new sights into rice blast control as well as the practical application of mefentrifluconazole.

In Silico, In Vitro, and In Vivo Studies Indicate the Endocrine-Disrupting Effects of Cyproconazole Stereoisomers.

The widespread use of chiral triazole fungicide cyproconazole (CPZ) in agricultural fields has led to frequent detection of CPZ in the environment. The restriction of CPZ in the EU raised wide concerns regarding its potential endocrine-disrupting effects (EDEs). The present study was conducted to evaluate EDEs of CPZ stereoisomers in vitro, in silico, and in vivo. The reporter gene assay indicated that all CPZ stereoisomers were agonists to the human estrogenic receptor α. (2S,3S)-(+)- and (2R,3S)-(-)-CPZ exhibited stronger binding capacities to ERα compared with (2R,3R)-(-)- and (2S,3R)-(+)-CPZ. Our computational studies showed consistent results with reporter gene assay, elucidating the stereoselective binding mode of CPZ to estrogen receptor. In zebrafish embryos, the 96h-lethality of CPZ stereoisomers ordered (2R,3R)-(-)- > (2R,3S)-(-)- > (2S,3S)-(+)- > Rac- > (2S,3R)-(+)-CPZ. Stereoselective developmental toxicity of CPZ was observed while (2R,3S)-(-)-CPZ is the most toxic isomer. The estrogenic hormones were significantly decreased in (2S,3R)-(+)- and (2R,3S)-(-)-CPZ groups and enhanced in (2S,3S)-(+)- and (2R,3R)-(-)-CPZ, along with the gene expression in hypothalamic-pituitary-gonad axis altered. CPZ shows no thyroid hormone activity. These data clarified that CPZ is a new-found endocrine disruptor threatening human health and each stereoisomer of CPZ showed stereoselective EDEs by regulating the nuclear receptor-mediated gene expression.

Effects of the Fungicide Prothioconazole on Lipid Metabolism in Mice: Whitening Alterations of Brown Adipose Tissue.

With considerable concerns about the associations between metabolic disorders and agricultural biocides, there are scattered data suggesting that the triazole fungicide prothioconazole (PTC) at lower doses than the no observed adverse effect level of 5000 μg/kg/d possibly has the potential to disrupt glycolipid metabolism in mammals. Here, we investigated the effects of 50, 500, and 5000 μg/kg/d of PTC on glycolipid metabolism in mice following 8 weeks of administration via drinking water, with specific attention on brown adipose tissue (BAT) and white adipose tissue (WAT) in addition to the liver. We found that along with the increased serum triglyceride level in the 5000 μg/kg/d group, small fatty vacuoles occurred in livers in all treatment groups, indicating lipid accumulation. No change in WAT was observed, but PTC caused BAT whitening, characterized by adipocyte hypertrophy, more unilocular adipocytes with enlarged lipid droplets, reduced UCP1 levels, and down-regulated Doi2 expression, and even the dose of 50 μg/kg/d was effective. Transcriptomic analysis revealed immune inhibition and circadian rhythm disturbance in BAT from the 5000 μg/kg/d group, which are in agreement with BAT whitening and inactivation. On employing the C3H10T1/2 cells in vitro, we found that PTC treatment concentration-dependently promoted lipid accumulation in brown adipocytes, along with altered expression of thermogenesis-related and circadian genes. Taken together, our study shows that low doses of PTC caused BAT whitening, calling for much attention to the new target by pollutants.

Hydrophobic melamine sponge incorporated with carbon dots as a green sorbent for micro-solid phase extraction of triazole fungicide residues in edible fungi samples

Fabrication of hydrophobic melamine sponge incorporated with carbon dots was investigated to extract triazoles prior to HPLC-DAD quantitation. The melamine sponge was simply incorporated with carbon dots via thermal treatment under facile conditions. The optimum conditions were melamine sponge (MeS) (1 × 1 × 1 cm3), 10 mL of standard solution, 0.04 M citric acid, functionalized at 250 °C for 5 h, and the analytes was then eluted from as-prepared material using acetonitrile (200 µL). Operating the best conditions, wide linearity was gained with R2 ≥ 0.99, for all analytes. Detection limits established varied between 3–10 μg L−1. Such approached method demonstrates excellent applicability for accurately and efficiently determining triazole fungicides in edible fungi samples, and a good recovery was also obtained. In addition, the desirable method validation and cost effective, not only its potential for reuse, but also provides it a better alternative for matrix purification. The appraisal estimated by two methods, confirmed its superior environmental friendliness.

Potential Mechanisms of Hexaconazole Resistance in Fusarium graminearum.

Fusarium head blight (FHB) caused by Fusarium graminearum is a serious fungal disease that can dramatically impact wheat production. At present, disease control is mainly achieved by the use of chemical fungicides. Hexaconazole (IUPAC name: 2(2,4-dichlorophenyl)-1-(1,2,4-triazol-1-yl)hexan-2-ol) is a widely used triazole fungicide, but the sensitivity of F. graminearum to this compound has yet to be established. The current study found that the EC50 values of 83 field isolates of F. graminearum ranged between 0.06 and 4.33 μg/ml, with an average EC50 value of 0.78 μg/ml. Assessment of four hexaconazole-resistant laboratory mutants of F. graminearum revealed that their mycelial growth and pathogenicity were reduced compared with their parental isolates and that asexual reproduction was reduced by resistance to hexaconazole. Meanwhile, the mutants appeared to be more sensitive to abiotic stress associated with SDS and H2O2, while their tolerance to high concentrations of Congo red, and Na+ and K+ increased. Molecular analysis revealed numerous point mutations in the FgCYP51 target genes that resulted in amino acid substitutions, including L92P and N123S in FgCYP51A, as well as M331V, F62L, Q252R, A412V, and V488A in FgCYP51B, and S28L, S256A, V307A, D287G, and R515I in FgCYP51C, three of which (S28L, S256A, and V307A) were conserved in all of the resistant mutants. Furthermore, the expression of the FgCYP51 genes in resistant strains was found to be significantly (P < 0.05) reduced compared with their sensitive parental isolates. Positive cross-resistance was found between hexaconazole and metconazole and flutriafol, as well as with the diarylamine fungicide fluazinam, but not with propiconazole, and the phenylpyrrole fungicide fludioxonil, or with tebuconazole, which actually exhibited negative cross-resistance. These results provide valuable insight into resistant mechanisms to triazole fungicides in F. graminearum, as well as the appropriate selection of fungicide combinations for the control of FHB to ensure optimal wheat production.

Health risk assessment of triazole fungicides around a pesticide factory in China.

The environmental pollution and health effects caused by pesticide production have consistently garnered considerable research interest. In the present study, the concentrations of five triazole fungicides (TFs) in air, indoor dust, and diet were monitored around a pesticide factory in eastern China from November 2020 to May 2021. The levels of five TFs in each sample were determined via UPLC‒MS/MS. For a health risk assessment, the United States Environmental Protection Agency's deterministic method was applied. The findings revealed that the total concentrations of the five TFs around the monitoring area ranged from 0.29 to 5.85 ng/m3 in outdoor air, 287.4 to 9878.5μg/kg in indoor dust, 0.0578 to 4.948μg/kg in vegetables, and 0.447 to 3.00μg/kg in rice. Notably, tebuconazole and hexaconazole had consistently high contributions over the years. For adults and children, the average daily doses (ADDs) were 1.32 × 10-5 and 2.69 × 10-5mg/kg/day, respectively, in the monitoring area and 4.25 × 10-6 and 6.42 × 10-6mg/kg/day, respectively, in the control area. In the control area, rice and vegetables were the primary media for exposure to TFs in children and adults, collectively accounting for more than 94% of the total TF exposure. Conversely, indoor dust is identified as the main medium of TF exposure in children residing near the pesticide factory, representing approximately 40% of the total exposure. The risks of noncarcinogenic effects on children and adults in the monitoring area were significantly greater than those in the control area, being approximately ten times greater for children, warranting increased attention. The carcinogenic risk to human health is relatively safe.

Profiling the endocrine disrupting properties of triazines, triazoles and short-chain PFAS.

Persistent, mobile and toxic (PMT) compounds released to the environment are likely to pollute drinking water sources due to their slow environmental degradation (persistency) and high water solubility (mobility). The aim of the present study was to create in vitro hazard profiles for sixteen triazoles, nine triazines and eleven PFAS based on their agonistic and antagonistic effects in estrogen receptor (ER), androgen receptor (AR) and thyroid hormone receptor (TR) reporter gene assays, their ability to bind human transthyretin (TTR), and their effects on steroidogenesis. The triazole fungicides tetraconazole, bitertanol, fenbuconazole, tebuconazole, cyproconazole, difenoconazole, propiconazole, paclobutrazol and triadimenol had agonistic or antagonistic effects on the ER and AR. Difenoconazole, propiconazole and triadimenol were also found to be TR antagonists. The triazine herbicide ametryn was an ER, AR and TR antagonist. The same nine triazole fungicides and the triazines atrazine, deethyl-atrazine and ametryn affected the secretion of steroid hormones. Furthermore, PFAS compounds PFBS, PFHxS, PFHxA, PFOS, PFOA and GenX and the triazoles bitertanol, difenoconazole and 4-methyl benzotriazole were found to displace T4 from TTR. These results are in line with earlier in vitro and in vivo studies on the endocrine disrupting properties of triazines, triazoles and PFAS. The present study demonstrates that this battery of in vitro bioassays can be used to profile compounds from different classes based on their endocrine disrupting properties as a first step to prioritize them for further research, emission reduction, environmental remediation and regulatory purposes.

Synthesis of a bimetal-organic framework-cobalt oxide nanoflowers (Ni/Co-MOF@Co3O4 NFs) for sensitive tebuconazole extraction from fruit juice and water samples using micro-solid phase extraction-HPLC-DAD

Tebuconazole (TBZ) is a widely used triazole fungicide that poses potential risks to human health and the environment due to its persistence in food and water. Effective monitoring of TBZ residues is crucial for ensuring food safety and environmental protection. This study presents a novel method for synthesizing Ni/Co-MOF@Co3O4 nanoflowers (NFs) and their application in extracting TBZ from water and fruit juice samples via micro-solid phase extraction (µ-SPE). Ni/Co-MOF@Co3O4 NFs was synthesized using the hydrothermal technique followed by calcination. The synthesized nanoflowers were characterized through SEM, XRD, and FT-IR analyses. Optimal extraction conditions were established at pH 8, using 10 mg of adsorbent, with adsorption and desorption times of 3 and 2 min, respectively. TBZ analysis was conducted using HPLC with a Poroshell 120 EC-C18 column and a diode array detector (DAD). The method demonstrates excellent recovery rates (90–102 %) and RSD% of 3.7 % in spiked fruit juice and natural water samples. The analytical performance showed a wide linear range (1–2000 µg L−1) with a high correlation coefficient (R2 = 0.9998), a detection limit of 4.0 ng L−1, and a quantification limit of 13.0 ng L−1. This approach is fast, simple, and highly sensitive, offering a reliable technique for detecting trace levels of TBZ in food and water environments.

Induction of drug metabolizing enzyme and drug transporter expression by antifungal triazole pesticides in human HepaSH hepatocytes

Triazole pesticides are widely used fungicides, to which humans are rather highly exposed. They are known to activate drug-sensing receptors regulating expression of hepatic drug metabolizing enzymes and drug transporters, thus suggesting that the hepatic drug detoxification system is modified by these agrochemicals. To investigate this hypothesis, the effects of 9 triazole fungicides towards expression of drug metabolizing enzymes and transporters were characterized in cultured human HepaSH cells, that are human hepatocytes deriving from chimeric humanized liver TK-NOG mice. Most of triazoles used at 10 μM were found to act as inducers of cytochrome P-450 (CYP) 1A1, CYP1A2, CYP2B6, CYP3A4 and UDP-glucuronosyltransferase 1A1 mRNA levels and of CYP3A4 protein; some triazoles also enhanced mRNA expression of the canalicular transporters P-glycoprotein/MDR1, multidrug resistance-associated protein 2 and breast cancer resistance protein. Triazoles however concomitantly inhibited CYP2B6 and CYP3A4 activities and thus appeared as dual regulators of these CYPs, being both inducers of their expression and inhibitors of their activity. The inducing effect however predominated, at least for bromuconazole, propiconazole and tebuconazole. Bromuconazole was moreover predicted to enhance CYP2B6 and CYP3A4 expression in humans exposed to this fungicide in a chronic, acute or occupational context. These data demonstrate that key-actors of the human hepatic detoxification system are impacted by triazole pesticides, which may have to be considered for the risk assessment of these agrochemicals. They additionally highlight that the use of human HepaSH cells as surrogates to primary human hepatocytes represents an attractive and promising way for studying hepatic effects of environmental chemicals.

Design and application of magnetic deep eutectic solvent-based spontaneous liquid-liquid microextraction for the determination of triazole fungicides in water, juice, and wine

In this study, a simple, rapid, and eco-friendly technique, magnetic deep eutectic solvent-based spontaneous liquid-liquid microextraction (MDES-SLLME), was developed to detect triazole fungicides in water, juice, and wine by UHPLC. The unreported SLLME involves injecting the extractant in the whole liquid-phase microextraction, shortening the extraction and separation time without device assistance. MDES produced by heating bio-derived solvent anisole and metal chloride ferric chloride was used as the extractant to reduce the consumption of toxic solvents. Based on the properties of anisole, MDES spontaneously formed small droplets in samples to complete extraction. Based on the properties of ferric chloride, MDES achieved magnetic separation under an external magnetic field. The method was applied to three matrices (water, juice, and wine). The concentration range was 0.007–0.7 mg L−1, and the matrix effect range was 87.4 %–94.9 %. The limit of detection was 0.002 mg L−1. The spiked recoveries ranged from 85.6 % to 94.7 %. The relative standard deviation ranged from 1.7 %–6.3 %. The SLLME requires attention as a potential complement to single-drop microextraction, hollow fiber liquid-phase microextraction, dispersive liquid-liquid microextraction, homogeneous liquid-liquid microextraction, and emulsive liquid-liquid microextraction.

Prothioconazole Stress Reduces Bacterial Richness and Alters Enzyme Activity in Soybean Rhizosphere.

Prothioconazole (PTC) is currently a popular triazole fungicide. In recent years, as the use of PTC has increased, there has been growing concern about its environmental and toxicological effects. Here, we studied the effect of PTC on the growth of soybean plants and further analyzed the enzyme activity and microbial community of rhizosphere soil after PTC treatment through 16S rRNA gene high-throughput sequencing and fungal ITS. Changes in structural diversity and species richness were measured using Simpson's diversity index, Shannon's diversity index and the Chao1 and ACE algorithms. The statistical t-test was applied to test whether the index values were significantly different between the two groups. The results showed that the contents of malondialdehyde (MDA) and H2O2 increased after the recommended dose of PTC, indicating that PTC has a strong toxic effect on plant growth, thus affecting the healthy growth of plants. In the presence of PTC, the species richness of fungi and bacteria decreased in all three soil types (black soil, yellow earth and red earth), and the community structure also changed significantly (the p-values were all less than 0.05). Proteobacteria, Actinomycetota, Bacteroidota and Acidobacteriota were the main bacteria, and the abundance of Acidobacteriota and Chloroflexi increased. The dominant fungal communities were Ascomycota and Mortierellomycota. The increased abundance of potentially beneficial microorganisms, such as Sphingomonadaceae, suggested that plants may be resistant to PTC stress by recruiting beneficial microorganisms. PICRUSt analysis showed that the metabolism-related functions and membrane transport pathway of rhizosphere bacterial community were inhibited after PTC stress. Spearman correlation analysis revealed a weak correlation between key fungal taxa and rhizosphere variables in the presence of PTC. Therefore, compared with those in the fungal community, the bacterial community was more likely to help plants resist PTC stress, indicating that these key fungal groups may indirectly help soybean growth under PTC stress by affecting the bacterial community.

Engineering an Epoxide Hydrolase for Chemoenzymatic Asymmetric Synthesis of Chiral Triazole Fungicide (S)- and (R)-Flutriafol.

Flutriafol, a globally utilized triazole fungicide in agriculture, is typically applied as a racemic mixture, but its enantiomers differ in bioactivity and environmental impact. The synthesis of flutriafol enantiomers is critically dependent on chiral precursors: 2,2-bisaryl-substituted oxirane [(2-fluorophenyl)-2-(4-fluorophenyl)oxirane, 1a] and 1,2-diol [1-(2-fluorophenyl)-1-(4-fluorophenyl)ethane-1,2-diol, 1b]. Here, we engineered a Rhodotorula paludigensis epoxide hydrolase (RpEH), obtaining mutant Escherichia coli/RpehH336W/L360F with a 6.4-fold enhanced enantiomeric ratio (E) from 5.5 to 35.4. This enabled a gram-scale resolution of rac-1a by E. coli/RpehH336W/L360F, producing (S)-1a (98.2% ees) and (R)-1b (75.0% eep) with 44.3 and 55.7% analytical yields, respectively. As follows, chiral (S)-flutriafol (98.2% ee) and (R)-flutriafol (75.0% ee) were easily synthesized by a one-step chemocatalytic process from (S)-1a and a two-step chemocatalytic process from (R)-1b, respectively. This chemoenzymatic approach offers a superior alternative for the asymmetric synthesis of flutriafol enantiomers. Furthermore, molecular dynamics simulations revealed insight into the enantioselectivity improvement of RpEH toward bulky 2,2-bisaryl-substituted oxirane 1a.

Efficiency and resistance risk of flusilazole against northern corn leaf blight caused by Setosphaeria turcica

Northern corn leaf blight (NCLB) infected by Setosphaeria turcica is a devastating disease of corn worldwide. Flusilazole is a broad-spectrum triazole fungicide. However, its resistance risk and field efficiency in controlling NCLB are still unknown. The present research evaluated the antifungal activity of flusilazole against 101 S. turcica isolates, and their EC50 values ranged from 0.0013 to 0.0466 μg/mL, with a mean of 0.0157 μg/mL. Seven S. turcica mutants resistant to flusilazole were obtained from two wild-type isolates by fungicide adaptation. After 10 consecutive transfers on PDA medium without fungicide, their resistance decreased. Cross-resistance was not existed between flusilazole and fluazinam, pyraclostrobin, amobam, epoxiconazole, or fluxapyroxad. Compared to the wild-type isolates, seven flusilazole-resistant mutants showed reduced biological fitness. No point mutation was detected, however, over-expression of StCYP51 and StatrD genes were detected in the resistant mutants. In addition, in the field experiment, flusilazole exhibited over 85 % efficacy against NCLB, significantly higher than amobam. In summary, these results suggested that the resistance risk of S. turcica to flusilazole was low, and the over-expression of StCYP51 and StatrD might be related to the flusilazole resistance against S. turcica. Flusilazole showed great potential as an alternative fungicide for controlling NCLB.

Respuesta de la roya (Hemileia vastatrix) frente a condiciones climáticas y aplicaciones de fungicidas en el cultivo de café (Coffea arabica) bajo sombra

Coffee leaf rust (Hemileia vastatrix) was first reported in 1869 on a plantation on the island of Sri Lanka, Asia, and has since spread to all coffee-growing regions far from its center of origin. The coexistence between the coffee plant, primarily Coffea arabica, and the fungus Hemileia vastatrix leads to coevolution, which manifests in significant genetic variability in the plant’s resistance to the disease and the fungus’s virulence towards the plant (Avelino and Rivas, 2013). The trophic relationships between the fungus and the host are influenced by soil moisture, fruit load, genetic resistance, precipitation, relative humidity, and temperature; these are the main factors affecting plant colonization (Filho and Astorga, 2015). In this study, the behavior of H. vastatrix was recorded through sampling to describe the relationship between disease incidence and environmental conditions, such as precipitation, temperature, and relative humidity, over an 18-month period at the Santa Lucía Experimental Farm (FESL) in Heredia, Costa Rica. The main findings demonstrated that the use of triazole fungicides and copper hydroxide can maintain disease incidence below 7%, despite contributions to its development from precipitation and increased temperatures, but not from increased relative humidity. Proper management of coffee leaf rust and the presence of shade can help reduce the incidence of the disease in the coffee plantation at FESL.

A novel approach to triazole fungicides risk characterization: Bridging human biomonitoring and computational toxicology

Brazil stands as the world's leading coffee producer, where the extensive use of pesticides is economically critical yet poses health and environmental risks due to their non-selective mechanisms of action. Specifically, triazole fungicides are widely used in agriculture to manage fungal diseases and are known to disrupt mammalian CYP450 and liver microsomal enzymes. This research establishes a framework for risk characterization of human exposure to triazole fungicides by internal-dose biomonitoring, biochemical marker measurements, and integration of high-throughput screening (HTS) data via computational toxicology workflows from the Integrated Chemical Environment (ICE). Volunteers from the southern region of Minas Gerais, Brazil, were divided into two groups: farmworkers and spouses occupationally and environmentally exposed to pesticides from rural areas (n = 140) and individuals from the urban area to serve as a comparison group (n = 50). Three triazole fungicides, cyproconazole, epoxiconazole, and triadimenol, were detected in the urine samples of both men and women in the rural group. Androstenedione and testosterone hormones were significantly reduced in the farmworker group (Mann-Whitney test, p < 0.0001). The data show a significant inverse association of testosterone with cholesterol, LDL, VLDL, triglycerides, and glucose and a direct association with HDL (Spearman's correlation, p < 0.05). In the ICE workflow, active in vitro HTS assays were identified for the three measured triazoles and three other active ingredients from the pesticide formulations. The curated HTS data confirm bioactivities predominantly related to steroid hormone metabolism, cellular stress processes, and CYP450 enzymes impacted by fungicide exposure at occupationally and environmentally relevant concentrations based on the in vitro to in vivo extrapolation models. These results characterize the potentially significant human health risk, particularly from the high frequency and intensity of exposure to epoxiconazole. This study showcases the critical role of biomonitoring and utility of computational tools in evaluating pesticide exposure and minimizing the risk.

Multiple routes to fungicide resistance: Interaction of Cyp51 gene sequences, copy number and expression.

We examined the molecular basis of triazole resistance in Blumeria graminis f. sp. tritici (wheat mildew, Bgt), a model organism among powdery mildews. Four genetic models for responses to triazole fungicides were identified among US and UK isolates, involving multiple genetic mechanisms. Firstly, only two amino acid substitutions in CYP51B lanosterol demethylase, the target of triazoles, were associated with resistance, Y136F and S509T (homologous to Y137F and S524T in the reference fungus Zymoseptoria tritici). As sequence variation did not explain the wide range of resistance, we also investigated Cyp51B copy number and expression, the latter using both reverse transcription-quantitative PCR and RNA-seq. The second model for resistance involved higher copy number and expression in isolates with a resistance allele; thirdly, however, moderate resistance was associated with higher copy number of wild-type Cyp51B in some US isolates. A fourth mechanism was heteroallelism with multiple alleles of Cyp51B. UK isolates, with significantly higher mean resistance than their US counterparts, had higher mean copy number, a high frequency of the S509T substitution, which was absent from the United States, and in the most resistant isolates, heteroallelism involving both sensitivity residues Y136+S509 and resistance residues F136+T509. Some US isolates were heteroallelic for Y136+S509 and F136+S509, but this was not associated with higher resistance. The obligate biotrophy of Bgt may constrain the tertiary structure and thus the sequence of CYP51B, so other variation that increases resistance may have a selective advantage. We describe a process by which heteroallelism may be adaptive when Bgt is intermittently exposed to triazoles.

Integrating adverse effects of triazole fungicides on reproduction and physiology of farmland birds

The extensive use of pesticides has been recognized as one of the major factors negatively impacting birds in agricultural habitats. One of the pesticide groups most used worldwide are triazole fungicides due to their effectiveness in controlling phytopathogenic fungi in cereals, vineyards and orchards. In the last decades, different experimental studies have reported important negative effects on the health and fitness of birds after exposure to triazoles. Birds can be exposed throughout the year through different routes, including oral uptake, dermal contact with treated surfaces and inhalation by overspray. Yet, the ingestion of treated or sprayed material is the principal route. The most alarming effect of triazoles, which can even occur several months after cessation of the exposure, is the decreasing reproductive outputs of birds, including delay in the onset of laying dates, reduced clutch size and hatching rate, and increased mortality of chicks. In order to synthesize the data and knowledge about the toxic effects of triazoles at different levels of biological organization, here we propose an dverse outcome pathway (AOP) on the mechanisms by which triazoles can affect avian reproduction and physiology. The reported effects highlight that the current risk assessment needs some improvements to avoid undesired effects on birds, especially long‐term effects that can influence stability and viability of avian populations from agricultural habitats.

Ultrasound assisted upper critical solution temperature type switchable deep eutectic solvent based liquid-liquid microextraction for the determination of triazole in water

BackgroundThe use of pesticides to protect crops has long been an important measure to provide healthy and safe agricultural products, but excess pesticides flow into fields and rivers, causing environmental pollution. Earlier methods utilizing organic solvent liquid-liquid microextraction for pesticide residue detection were not environmentally friendly. Therefore, it is significant to find a greener and more convenient detection method to determine pesticide residues. ResultsA new method was established to detect three triazole fungicides (TFs), including myclobutanil, epoxiconazole and tebuconazole, in environmental water samples. And the determination was conducted using a high-performance liquid chromatography with the ultraviolet detector (HPLC-UV). The switchable deep eutectic solvent (SDES) can be reversibly switched between hydrophilic and hydrophobic states through temperature modulation. Additionally, the method exhibited excellent linearity for all target analytes within the concentration range of 10–2000 μg L−1, with satisfactory R2 values (≥0.9975). The limits of detection (LODs) ranged from 2.3 to 2.6 μg L−1, and the limits of quantification (LOQs) ranged from 7.8 to 8.7 μg L−1. The accuracy of the method was assessed through intra-day and inter-day precision tests, yielding relative standard deviations (RSDs) in the ranges of 2.8%–6.7% and 2.2%–7.5%, respectively. Density functional theory (DFT) results indicated that hydrogen bonding is a significant factor affecting the binding of DES with triazoles. Three different green assessment tools were used to prove that the SDES-HLLME method had good greenness and broad applicability. SignificanceThis is a homogeneous liquid-liquid microextraction (HLLME) method based on the upper critical solution temperature (UCST) type switchable deep eutectic solvent program, which can complete the extraction within a few minutes without dispersant. In terms of pesticide detection, the analytical method is simple and more conducive to environmental protection.

In silico analysis to predict the carcinogenicity and mutagenicity of a group of triazole fungicides

In silico analysis to predict the carcinogenicity and mutagenicity of a group of triazole fungicides