Resistance risk assessment of mefentrifluconazole in Corynespora cassiicola and the control of cucumber target spot by a two-way mixture of mefentrifluconazole and prochloraz.

The cucumber target spot, caused by Corynespora cassiicola, is a major cucumber disease in China. Mefentrifluconazole, a new triazole fungicide, exhibits remarkable efficacy in controlling cucumber target spot. However, the resistance risk and mechanism remain unclear. In this study, the inhibitory activity of mefentrifluconazole against 101 C. cassiicola isolates was determined, and the results indicated that the EC50 values ranged between 0.15 and 12.85 µg/mL, with a mean of 4.76 µg/mL. Fourteen mefentrifluconazole-resistant mutants of C. cassiicola were generated from six parental isolates in the laboratory through fungicide adaptation or UV irradiation. The resistance was relatively stable after ten consecutive transfers on a fungicide-free medium. No cross-resistance was observed between mefentrifluconazole and pyraclostrobin, fluopyram, prochloraz, mancozeb, or difenoconazole. Investigations into the biological characteristics of the resistant mutants revealed that six resistant mutants exhibited an enhanced compound fitness index (CFI) compared to the parental isolates, while others displayed a reduced or comparable CFI. The overexpression of CcCYP51A and CcCYP51B was detected in the resistant mutants, regardless of the presence or absence of mefentrifluconazole. Additionally, a two-way mixture of mefentrifluconazole and prochloraz at a concentration of 7:3 demonstrated superior control efficacy against the cucumber target spot, achieving a protection rate of 80%. In conclusion, this study suggests that the risk of C. cassiicola developing resistance to mefentrifluconazole is medium, and the overexpression of CcCYP51A and CcCYP51B might be associated with mefentrifluconazole resistance in C. cassiicola. The mefentrifluconazole and prochloraz two-way mixture presented promising control efficacy against the cucumber target spot.

Expanding the insight of ecological risk on the novel chiral pesticide mefentrifluconazole: Mechanism of enantioselective toxicity to earthworms (Eisenia fetida).

Continued application of new chiral fungicide mefentrifluconazole (MFZ) increases its risk to soil ecosystem. However, the toxicity of MFZ enantiomers to soil fauna and whether stereoselectivity exists remains poorly elucidated. Based on multilevel toxicity endpoints and transcriptomics, we investigated the negative effects of racemic, R-(-)-, and S-(+)-MFZ on Eisenia fetida. After exposure to S-(+) configuration at 4 mg/kg for 28 day, its reactive oxygen species levels were elevated by 15.4% compared to R-(-) configuration, inducing enantiospecific oxidative stress and transcriptional aberrations. The S-(+) isomer induced more severe cell membrane damage and apoptosis than the R-(-) isomer, and notably, the selectivity of apoptosis is probably dominated by the mitochondrial pathway. Mechanistically, differential mitochondrial stress lies in: S-(+) isomer specifically up-regulated mitochondrial cellular component compared to R-(-) isomer and identified more serious mitochondrial fission. Furthermore, S-(+) conformation down-regulated biological processes associated with ATP synthesis and metabolism, with specific inhibition of mitochondrial respiratory electron transport chain complex I and IV activity resulting in more severe electron flow disturbances. These ultimately mediated enantioselective ontogenetic process disorders, which were supported at phenotypic (weight loss), genetic, and protein (reverse modulate TCTP and Sox2 expression) levels. Our findings offer an important reference for elucidating the enantioselective toxicological mechanism of MFZ in soil fauna.

Mefentrifluconazole-Resistant Risk and Resistance-Related Point Mutation in FpCYP51B of Fusarium pseudograminearum.

Mefentrifluconazole, a triazole fungicide, exhibits remarkable efficacy in combating Fusarium spp. The mean EC50 value of mefentrifluconazole against 124 isolates of Fusarium pseudograminearum was determined to be 1.06 µg/mL in this study. Fungicide taming produced five mefentrifluconazole-resistant mutants with resistance factors ranging from 19.21 to 111.34. Compared to the original parental isolates, the fitness of three resistant mutants was much lower, while the remaining two mutants displayed enhanced survival fitness. There was evidence of positive cross-resistance between tebuconazole and mefentrifluconazole. Mefentrifluconazole resistance in F. pseudograminearum can be conferred by FpCYP51BL144F, which was identified in four mutants according to molecular docking and site-directed transformation experiments. Overexpression of FpCYP51s was also detected in the resistant mutants. In conclusion, mefentrifluconazole has a low-to-medium resistance risk in F. pseudograminearum, and the L144F mutation in FpCYP51B and the increased expression level of FpCYP51s may be responsible for mefentrifluconazole resistance in F. pseudograminearum.

Mefentrifluconazole sensitivity amongst European Zymoseptoria tritici populations and potential implications for its field efficacy.

BACKGROUND: Septoria tritici blotch caused by Zymoseptoria tritici continues to be one of the most economically destructive diseases of winter wheat in north-western Europe. Control is heavily reliant on the application of fungicides, in particular those belonging to the azole group. Here we describe the sensitivity of European Z. tritici populations to the novel azole mefentrifluconazole and the analysis of associated mechanisms of resistance. RESULTS: A wide range of sensitivity to mefentrifluconazole was observed amongst the Z. tritici collections examined, with strong cross-resistances also observed between mefentrifluconazole, difenoconazole and tebuconazole. Overall, the Irish population displayed the lowest sensitivity to all azoles tested. Further detailed analysis of the Irish population in 2021 demonstrated differences in sensitivity occurred between sampling sites, with these differences associated with the frequencies of key resistance mechanisms (CYP51 alterations and MFS1 promoter inserts linked to overexpression). Under glasshouse conditions reductions in the efficacy of mefentrifluconazole were observed towards those strains exhibiting the lowest in vitro sensitivities. CONCLUSIONS: This study demonstrates that a large range of sensitivity to mefentrifluconazole exists in European Z. tritici populations. Those strains exhibiting the lowest sensitivity to the azoles tested had the most complex CYP51 haplotypes in combination with the 519 bp insert, associated with enhanced activity of MFS1. The future use of mefentrifluconazole should take these findings into consideration to minimise the selection of these strains. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Resistance to the DMI fungicide mefentrifluconazole in Monilinia fructicola: risk assessment and resistance basis analysis.

BACKGROUND: Brown rot disease, caused by Monilinia fructicola, poses a significant challenge to peach production in China. The efficacy of mefentrifluconazole, a new triazole fungicide, in controlling brown rot in peaches has been remarkable. However, the resistance risk and mechanism associated with this fungicide remain unclear. This study was designed to assess the resistance risk of M. fructicola to mefentrifluconazole and reveal the potential resistance mechanism. RESULTS: The mean median effective concentration (EC50 ) of 101 M. fructicola isolates to mefentrifluconazole was 0.003 µg mL-1 , and the sensitivity exhibited a unimodal distribution. Seven mefentrifluconazole-resistant mutants were generated from three parental isolates in the laboratory through fungicide adaption. The biological characteristics of the resistant mutants revealed that three of them exhibited enhanced survival fitness compared to the parental isolates, whereas the remaining four mutants displayed reduced survival fitness. Mefentrifluconazole showed strong positive cross-resistance with fenbuconazole, whereas no cross-resistance was observed with pyrimethanil, procymidone or pydiflumetofen. No overexpression of MfCYP51 gene was detected in the resistant mutants. Multiple sequence alignment revealed that three resistant mutants (MXSB2-2, Mf12-1 and Mf12-2) had a point mutation (G461S) in MfCYP51 protein. Molecular docking techniques confirmed the contribution of this point mutation to mefentrifluconazole resistance. CONCLUSION: The risk of M. fructicola developing resistance to mefentrifluconazole is relatively low-to-medium and point mutation G461S in MfCYP51 could confer mefentrifluconazole resistance in M. fructicola. This study provided essential data for monitoring the emergence of resistance and developing resistance management strategies for mefentrifluconazole. © 2023 Society of Chemical Industry.

Bioactivity of mefentrifluconazole against different Fusarium spp.

Emergence and development of resistance to 14α-demethylase inhibitors (DMIs) have become a critical issue in both agriculture and medical fields. Mefentrifluconazole, the first isopropanol triazole fungicide belonging to a new subclass of DMIs, has been proposed to show high activity, minimal adverse side effects, and inconsistent cross resistance with other DMIs due to its high structural flexibility. In this study, mefentrifluconazole showed disparate inhibitory activity against the mycelium growth of seven tested Fusarium species. The most sensitive species included F. oxysporum, F. proliferatum, F. commuae, and F. fujikuroi, followed by F. equiseti and F. graminearum, while F. solani was most insensitive. Consistently, mefentrifluconazole presented the strongest inhibiting effects on conidium germination, cell membrane integrity, and ergosterol biosynthesis in F. fujikuroi, followed by F. graminearum, while F. solani ranked last. Further results indicated that all F. fujikuroi isolates causing rice bakanae disease (RBD) were sensitive to mefentrifluconazole regardless of their resistance to prochloraz, tebuconazole, carbendazim, and phenamacril. Additionally, the inoculation tests found that mefentrifluconazole presented a better protective efficacy on rice seedlings when applied 12 h before the F. fujikuroi inoculation, compared to applied 12 h post the inoculation. Overall, this study demonstrated the various bioactivity of mefentrifluconazole combating Fusarium spp. and put new insights into RBD management as well as the applications of DMIs.

Magnetic deep eutectic solvent-based dispersive liquid-liquid microextraction for enantioselectively determining chiral mefentrifluconazole in cereal samples via ultra-high-performance liquid chromatography.

A simple, rapid, and efficient pretreatment method of mefentrifluconazole enantiomers in cereal samples was established by dispersive liquid-liquid microextraction coupled with ultra-high-performance liquid chromatography-diode array detector (UHPLC-DAD). In this study, a novel ternary magnetic deep eutectic solvent (MDES) [octyltrimethylammonium bromide][cobalt chloride][acetic acid] was synthesized as the extractant. Acetic acid was used as the dispersant to promote the in situ dispersion of binary MDES [octyltrimethylammonium bromide][cobalt chloride]. The microdroplets of binary MDES containing mefentrifluconazole were separated by an external magnet. Device-assisted dispersion and time-consuming centrifugation steps were eliminated to ensure simplicity and rapidity of the pretreatment. Good linearity ranging from 0.01 µg g-1 to 2 µg g-1 was obtained. The extraction recovery varied from 82.9 to 95.0%. The limit of detection was 0.003 µg g-1. Finally, this established approach has been applied for the enantioselective detection of chiral mefentrifluconazole in corn, rice, wheat, millet, and sorghum samples.

Fluconazole Population Pharmacokinetics after Fosfluconazole Administration and Dosing Optimization in Extremely Low-Birth-Weight Infants.

A prospective single-center study was conducted to characterize the pharmacokinetics (PK) of fluconazole (FLCZ) in extremely low-birth-weight infants (ELBWIs) who received fosfluconazole (F-FLCZ). Intravenous F-FLCZ was administered at a dose of 3 mg/kg of body weight every 72 h during the first 2 weeks of life, every 48 h during the third and fourth weeks of life, and every 24 h after 5 weeks of life. Blood samples from ELBWIs treated with F-FLCZ were collected using scavenged samples. The concentration of FLCZ was determined using liquid chromatography-tandem mass spectrometry. The population pharmacokinetic model was established using Phenix NLME 8.2 software. In total, 18 ELBWIs were included in this analysis. Individual PK parameters were determined by a one-compartment analysis with first-order conversion. Postmenstrual age (PMA), serum creatinine (SCr), and alkaline phosphatase were considered covariates for clearance (CL). The mean population CL and the volume of distribution were 0.011 L/h/kg0.75 and 0.95 L/kg, respectively. Simulation assessments with the final model revealed that the current regimen (3 mg/kg every 72 h) could achieve the proposed target FLCZ trough concentration (>2 µg/mL) in 43.3% and 72.2% of infants with a PMA of ≥37 and 30 to 36 weeks, respectively, and an SCr level of <0.5 mg/dL. Shortened dosing intervals (every 48 or 24 h) might improve the probability of target attainment. This study was the first to assess the PK of F-FLCZ in ELBWI, as well as the first to provide fundamental information about FLCZ exposure after F-FLCZ administration, with the goal of facilitating dose optimization in the ELBWI population. IMPORTANCE Invasive fungal infection is an important cause of mortality and morbidity in very preterm or very-low-birth-weight infants. In order to limit the risk of invasive fungal infections in this population, the administration of fluconazole is generally recommended for extremely low-birth-weight infants admitted to a neonatal intensive care unit with a Candida species colonization prevalence rate of >10%, under the guidelines of the Infectious Diseases Society of America. Fosfluconazole can reduce the volume of solution required for intravenous therapy compared to fluconazole because it has increased solubility, which is a major advantage for infants undergoing strict fluid management. To date, no study has demonstrated the fluconazole pharmacokinetics after fosfluconazole administration in neonates and infants, and this needs to be clarified. Here, we characterized the pharmacokinetics of fluconazole in extremely low-birth-weight infants who received F-FLCZ and explored the appropriate dosage in this patient population.

Switchable deep eutectic solvent-based homogenous liquid-liquid microextraction combined with high-performance liquid chromatography-diode-array detection for the determination of the chiral fungicide mefentrifluconazole in water, fruit juice, and fermented liquor.

A switchable deep eutectic solvent-based homogeneous liquid-liquid microextraction (SDES-HLLME) technique was developed and combined with high-performance liquid chromatography-diode-array detection for the determination of the chiral fungicide mefentrifluconazole. A (green) SDES was synthesized from 4-methoxyphenyl and 3-amino-1-propanol and used as an extraction solvent (thus avoiding the use of toxic extraction solvents). To improve the efficiency of the extraction process, a hydrophobic extraction solvent was subsequently generated in situ by adjusting the pH. The detection process is linear in the range of 0.01 to 1 µg ml-1 . The limit of detection and limit of quantification were determined to be 0.003 and 0.01 µg ml-1 , respectively. Recovery rates of 79.2% to 104.6% were acquired with relative standard deviations of 0.6% to 2.5%. The method is fast, simple, and environmentally friendly. Moreover, it was successfully used to enantioselectively determine the concentrations of mefentrifluconazole residues in water, fruit juice, and fermented liquor samples.

Enantioselective toxic effects of mefentrifluconazole in the early life stage of zebrafish (Danio rerio).

The research on the enantioselective toxic effects of chiral pesticides on non-target aquatic organisms has attracted more and more attention. This study investigated the enantioselective toxic effects of mefentrifluconazole (MFZ) on acute toxicity, developmental toxicity, locomotor behaviors, and the mRNA relative expression levels of genes related to neurodevelopment and cardiac development in zebrafish embryos or larvae. The 96-h lethal concentration 50 (LC50 ) values (exposed to racemate and enantiomers of MFZ, that is, rac-MFZ/(-)-MFZ/(+)-MFZ) were 1.010, 1.552, and 0.753 mg/L for embryo, and 0.753, 1.187, and 0.553 mg/L for larvae. The rac-MFZ/(-)-MFZ/(+)-MFZ can affect the heart development of zebrafish embryos, accompanied by heart rate inhibition, yolk sac deformities, pericardial deformities, and down-regulation of genes related to cardiotoxicity in larvae in an enantioselective manner. Moreover, the rac-MFZ/(-)-MFZ/(+)-MFZ also can affect the neural development of zebrafish embryos, accompanied by autonomic movement inhibition, swimming speed and swimming distance abnormalities, and down-regulation of genes related to neurotoxicity in larvae in an enantioselective manner. For all toxicity endpoints, the effect of the (+)-MFZ to early-staged zebrafish were significantly greater than that of (-)-MFZ. These results will help distinguishing the difference of MFZ enantiomers to zebrafish, and provide scientific reference for improving the risk assessment of chiral pesticides MFZ.

Enantioselectivity of new chiral triazole fungicide mefentrifluconazole: Bioactivity against phytopathogen, and acute toxicity and bioaccumulation in earthworm (Eisenia fetida).

Elaborating the environmental behavior of mefentrifluconazole, a novel triazole fungicide, in stereoselective level is of paramount importance for the application of the pesticide in agriculture. In this study, the enantioselective bioactivity, acute toxicity and stereoselective bioaccumulation of mefentrifluconazole in earthworm (Eisenia fetida) were investigated. Bioactivity tests against four pathogens revealed that R-(-)-mefentrifluconazole exhibited approximately 11-113 times higher bioactivity than its S-(+)-mefentrifluconazole. However, the LC50 of S-(+)-, rac- and R-(-)-mefentrifluconazole to earthworm was measured to be 4.1, 11.4 and 7.3 µg/cm2, respectively, indicating active ingredient R-(-)-mefentrifluconazole is less toxic than its racemate and S-form. Accumulation of mefentrifluconazole in earthworms was non-enantioselective and negatively related to its adsorption onto soils. The concentration of mefentrifluconazole in in situ pore water (CIPW) and CaCl2 extraction (CCaCl2) was closely related to its accumulation in earthworms, suggesting that CIPW and CCaCl2 could be appropriate indicators for estimation of the bioavailability of mefentrifluconazole in soil. Conclusively, our study provides necessary information for the risk assessment of mefentrifluconazole in agriculture.

Enantioselective toxic effects of mefentrifluconazole in the liver of adult zebrafish (Danio rerio) based on transcription level and metabolomic profile.

Mefentrifluconazole, a new type of chiral triazole fungicide, is widely applied to control a variety of fungal diseases in crops. However, the toxicological effects of mefentrifluconazole on aquatic organisms are unknown, especially at the enantiomer level. In the present study, zebrafish were selected as a typical model for mefentrifluconazole enantiomer exposure. Metabolomic and transcription analyses were performed with 0.01 and 0.10 mg/L mefentrifluconazole and its enantiomers (i.e., rac-mfz/(-)-mfz/(+)-mfz) at 28 days. The 1H nuclear magnetic resonance (NMR)-based metabolomics analysis showed that 9, 10 and 4 metabolites were changed significantly in the rac-mfz, (+)-mfz and (-)-mfz treatment groups compared with the control group, respectively. The differential metabolites were related to energy metabolism, lipid metabolism and amino acid metabolism. The qRT-PCR analysis revealed that the expression of lipid metabolism-, apoptosis- and CYP-related genes in the livers of female zebrafish in rac-mfz and (+)-mfz was 1.61-108.92 times and 2.37-551.34 times higher than that in (-)-mfz, respectively. The results above indicate that exposure to mefentrifluconazole induced enantioselective liver toxicity in zebrafish. Our study underlined the importance of distinguishing different enantiomers, which will contribute to environmental protection.

Preparation, spectroscopic, characterizations and biological studies of new charge transfer complexes formed between fluconazole drug with various acceptors.

Charge transfer complexes developed during the interaction of Fluconazole drug (FLU) as an electron donor with different types of electron acceptors, including σ-type as iodine (I2), and π-types as 2,3-dinitrosalsylic acid (HDNS), Tetracyanoethylene (TCNE) and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ). The formed complexes were characterized using various techniques as UV-Vis spectra, Thermal analyses, spectrophotometric measurements, 1H NMR and FTIR Spectroscopy. It was found that the stoichiometry of all developed complexes was a 1:1 M ratio between fluconazole and acceptors (I2, HDNS, TCNE and DDQ). The characteristic physical parameters data such as ionization potential (ID), The oscillator strength (ƒ), formation constant (KCT), transition dipole moment (µ), free energy (ΔG), and energy of interaction (ECT) of the formed CT-complexes have also been reported. Eventually, the synthesized complexes were screened for their microbial and antioxidant activities.

Enantiomeric profiling of mefentrifluconazole in watermelon across China: Enantiochemistry, environmental fate, storage stability, and comparative dietary risk assessment.

Elucidating the enantiomeric chemistry and enantioselective fate of the novel chiral triazole fungicide mefentrifluconazole is of vital importance for agroecosystem safety and human health. The absolute configuration of mefentrifluconazole was identified firstly as S-(+)-mefentrifluconazole and R-(-)-mefentrifluconazole on a cellulose tris(3-chloro-4-methylphenylcarbamate) chiral phase. A baseline resolution (Rs, 2.51), favorable retention (RT ≤ 2.24 min), and high sensitivity (LOQ, 0.5 µg/kg) of enantiomer pair were achieved by reversed-phase liquid chromatography tandem mass spectrometry combined with a 3D response surface strategy. Nationwide field trials were undertaken to clarify the enantiomer occurrence, enantioselective dissipation, terminal concentrations, and storage stability of S-mefentrifluconazole and R-mefentrifluconazole in watermelon across China. The original deposition of the sum of enantiomer pair was estimated to be 14.4-163.7 µg/kg, and terminally decreased to < LOQ-59.3 µg/kg 10 days after foliage application. S-mefentrifluconazole preferentially degraded (T1/2, 3.3-6.0 days), resulting in the relative enrichment of R-mefentrifluconazole (T1/2, 3.9-6.6 days) in watermelon. A probabilistic model is recommended for the dietary risk assessment, although both acute (%ARfD, 0.435-22.188%) and chronic (%ADI, 1.697-9.658%) risks are acceptable for associated population. The long-term exposures should be continuously emphasized given the increasing applications and persistent fate of mefentrifluconazole, especially for urban children.

Absolute Configuration, Enantioselective Bioactivity, and Degradation of the Novel Chiral Triazole Fungicide Mefentrifluconazole.

Mefentrifluconazole is a new chiral triazole fungicide with a pair of enantiomers. However, the enantioselective differences in the biological effects and environmental behaviors of mefentrifluconazole are unclear. In the present work, a new simultaneous determination method of mefentrifluconazole enantiomers was established using ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The absolute configuration of the two mefentrifluconazole enantiomers was confirmed by comparing the experimental and calculated ECD spectra. The enantioselective bioactivity to target fungi and degradation in cucumber samples were also assessed. The absolute configurations of the two enantiomers eluted on the Superchiral IG-3 column were confirmed as R-(-)-mefentrifluconazole and S-(+)-mefentrifluconazole. The R-(-)-mefentrifluconazole possessed 5-473 times higher bioactivity than S-(+)-mefentrifluconazole toward six kinds of target pathogenic fungi. In addition, R-(-)-mefentrifluconazole exhibited stronger efficacy of suppression of ergosterol biosynthesis. The molecular docking results indicated that R-(-)-mefentrifluconazole had shorter binding distances and lower energies with the target protein than S-(+)-mefentrifluconazole, which may result in the enantioselective bioactivity. The high-efficiency enantiomer of R-(-)-mefentrifluconazole has longer duration in cucumber samples due to the relatively long half-life of 4.0 days. This research has clarified the bioactivity differences and mechanism between mefentrifluconazole enantiomers against target fungi and laid the foundation for an in-depth study of mefentrifluconazole at the chiral level.

Evaluation of the antifungal and biochemical activities of mefentrifluconazole against Botrytis cinerea.

Mefentrifluconazole is the first product of a new sub-class of triazoles fungicides, i.e., the isopropanol triazoles, with the broad spectrum and high activity. In this study, the potential and biochemical activities of mefentrifluconazole against Botrytis cinerea were investigated. The frequency distribution of all EC50 values of mefentrifluconazole against mycelial growth and germ tube elongation of 106 isolates formed unimodal curve, with the mean EC50 values of 0.124 ± 0.025 and 0.015 ± 0.008 µg/mL, respectively. The effect of mefentrifluconazole against gray mold was determined on detached leaves of cucumber in vivo, the treatment of mefentrifluconazole at 200 µg/mL provided 100% preventative efficacy and 72.7% curative efficacy. No evident correlation was detected between the sensitivity of B. cinerea to mefentrifluconazole and that to tebuconazole, difenoconazole, myclobutanil, hexaconazole, triadimefon, flusilazole and pyrisoxazole (P > 0.05). Mefentrifluconazole treatment resulted in the increase of mycelium branch, the decrease of ergosterol content and the changes of the permeability of cell membrane. These studies evaluated the potential of mefentrifluconazole to control gray mold and helped us to understand the possible biochemical activity of mefentrifluconazole against B.cinerea.

Cross-resistance to the new fungicide mefentrifluconazole in DMI-resistant fungal pathogens.

In the European Union (EU), regulation of sterol demethylation inhibiting (DMI) fungicides is tightened due to their suspected endocrine disrupting properties. However, the new DMI fungicide mefentrifluconazole was reported to have high fungicidal activity with minimal adverse side effects. In addition, some evidence suggests inconsistent cross resistance between mefentrifluconazole and other azoles. In this study, mefentrifluconazole and other triazoles were examined for activity to select pathogens sensitive or resistant to DMIs using mycelial growth tests on fungicide-treated culture medium or spray trials using cucumber plants. Cross-resistance was confirmed for all of the fungal species tested but activity levels varied. The sensitivity of Monilinia fructicola from peach to mefentrifluconazole was higher compared to other DMIs. In contrast, the inhibitory activity of mefentrifluconazole was equal or slightly inferior compared to difenoconazole, tebuconazole, propiconazole in Colletotrichum spp., Alternaria alternaria sp. complex and Cercospora beticola isolated from peach and sugar beet, respectively. Similar tendencies (i.e. equal or slightly inferior activity and cross-resistance) were observed for cucumber powdery mildew (Podosphaera xanthii) resistant to triflumizole, myclobutanil, and difenoconazole. Despite cross-resistance to other DMIs, mefentrifluconazole is a promising fungicide for fungal disease control on peach and other crops, with a reportedly more favorable toxicity profile.

Activity of the Novel Fungicide Mefentrifluconazole Against Colletotrichum scovillei.

The prevalence and destructiveness of anthracnose, caused by Colletotrichum scovillei, in pepper production regions seriously affects pepper yield and quality. Mefentrifluconazole, the first of the isopropanol-azole subgroup of triazole fungicides, was introduced for the control of pepper anthracnose. However, the growth characteristics of pepper fruit and rapid spread of anthracnose suggest that the fungicide application method must be optimized to enhance fungicide efficacy. The sensitivity of C. scovillei to mefentrifluconazole was determined by mycelial growth and germ tube elongation assays using 157 single-spore isolates with mean 50% effective concentration values of 0.462 ± 0.138 and 0.359 ± 0.263 mg/liter, respectively. The in vivo data also showed that mefentrifluconazole had favorable protective and curative effects against pepper anthracnose. Mefentrifluconazole significantly affected C. scovillei infection on pepper by reducing appressorium formation and sporulation, shriveling spores and germ tubes, and causing the abnormal development of appressoria and conidiophores. Mefentrifluconazole could move acropetally, horizontally, and basipetally in pepper plants. Compared with a knapsack sprayer, mefentrifluconazole applied by mist sprayer exhibited significantly better activity against pepper anthracnose. Additionally, as the spray volume increased from 45 to 150 liters/ha, the control efficacy of mefentrifluconazole first increased and then tended to be steady, with an optimal spray volume of 90 liters/ha. The difference in disease control efficacy was related to the deposition and droplet distribution of mefentrifluconazole on the pepper fruit. These results provide scientific guidance for the application of mefentrifluconazole in pepper fields and improved fungicide utilization.

A fast and sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry method for determining mefentrifluconazole in plant- and animal-derived foods.

A rapid and sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry method, in conjunction with multiwalled carbon nanotube purification, was developed to determine the mefentrifluconazole levels in grapes, cucumbers, tomatoes, peppers, wheat, maize, eggs, milk, pork, chicken, and fish. After purification, tandem mass spectrometry of mefentrifluconazole required <3.0 min. Matrix-matched external standard curves were used to quantify the residual mefentrifluconazole. The method meets the requirements of the European Union Document SANTE/11813/2017. Quantification was linear between 5 and 500 µg/kg (R2 ≥ 0.9988), and both the intra- and interday relative standard deviations were ≤13.7%. Analyte recovery ranged from 81.5% to 107.6%. The limit of mefentrifluconazole quantification was 5 µg/kg for all matrices. The method successfully detected and quantified mefentrifluconazole that had been applied to cucumbers and tomatoes grown in a test field. These results imply that the proposed method is effective and reliable for detecting mefentrifluconazole residues in plant- and animal-derived foods.

Innovative selection approach for a new antifungal agent mefentrifluconazole (Revysol®) and the impact upon its toxicity profile.

Mefentrifluconazole (trade name: Revysol®) is an agrochemical active ingredient from the new sub-class of isopropanol-triazole fungicides, with high selective fungicide activity. A full program of toxicity testing conducted according to OECD guidelines has shown mefentrifluconazole (MFZ) to be non-genotoxic and non-carcinogenic. Repeated dose studies in rats, mice and dogs identified the liver as the main target organ. Prenatal developmental toxicity studies in rats and rabbits did not indicate treatment-related embryofetal toxicity or teratogenicity up to the highest dose levels tested. In a two-generation dietary study in rats, the high dose level resulted in reduced food consumption and body weight gain throughout the dosing-period. Mating performance and fertility, estrous cycles, gestation length and pre-and post-natal survival of offspring were essentially unaffected and there was no evidence of masculinization of female pups or feminization of male pups. The screening strategy that led to the selection of MFZ was aimed to identify candidates with both high fungicidal activity and minimal likelihood of adverse side effects thought to arise from aromatase inhibition. The success of the selection strategy has been illustrated for MFZ by the absence in toxicity studies of effects that would indicate an endocrine disrupting potential.