A Case of Severely Impaired Consciousness and Convulsions From Attempted Suicide by Ingesting Triflumizole Emulsion: Clinical Features and Literature Review.

Triflumizole (TFZ) is a fungicide widely used in agriculture to prevent fungal infections of fruits and vegetables. Although it is considered safe for humans and animals, its toxicity profile in humans remains largely unexplored. Here, we describe a case where an individual experienced symptoms suggestive of intoxication after ingesting TFZ emulsion. A 70-year-old man ingested TFZ emulsion (Trifumin emulsionTM) and alcohol in an attempt to commit suicide. He developed a severe disturbance of consciousness, which was not explained by the estimated blood alcohol concentration, and experienced convulsions. We managed this patient with symptomatic treatment, temporary mechanical ventilation, and antiepileptic drugs. He subsequently recovered without any sequelae. We present the first case of acute oral intoxication with TFZ emulsion. Moreover, we review the literature on TFZ-induced organ dysfunction and discuss the possible mechanisms and management of this condition.

Triflumizole Induces Developmental Toxicity, Liver Damage, Oxidative Stress, Heat Shock Response, Inflammation, and Lipid Synthesis in Zebrafish.

Triflumizole (TFZ) toxicity must be investigated in the aquatic environment to understand the potential risks to aquatic species. Accordingly, the adverse effects of TFZ exposure in zebrafish were investigated. Results demonstrate that, after TFZ exposure, the lethal concentration 50 (LC50) in 3 d post-fertilization (dpf) embryos and 6 dpf larvae were 4.872 and 2.580 mg/L, respectively. The development (including pericardium edema, yolk sac retention, and liver degeneration) was apparently affected in 3 dpf embryos. Furthermore, the alanine aminotransferase (ALT) activity, superoxide dismutase (SOD) activity, catalase (CAT) activity, and malondialdehyde (MDA) content in 6 dpf larvae were significantly increased. Additionally, the expression of heat shock response genes (including hsp70, grp78, hsp90, and grp94), inflammatory genes (including p65-nfκb, il-1ß, and cox2a), and lipid synthetic genes (including srebp1, fas, acc, and ppar-γ) in 3 dpf embryos was significantly increased, which was also partially observed in the intestinal cell line form Pampus argenteus. Taken together, TFZ could affect the development of zebrafish, accompanied by disturbances of oxidative stress, heat shock response, inflammation, and lipid synthesis. Our findings provide an original insight into the potential risks of TFZ to the aquatic ecosystem.

Fungicide Sensitivity of Colletotrichum Species Causing Bitter Rot of Apple in the Mid-Atlantic U.S.A.

Apple growers in the Mid-Atlantic region of the U.S.A. have reported increased losses to bitter rot of apple. We tested the hypothesis that this increase is because the Colletotrichum population has developed resistance to commonly used single-mode-of-action (single-MoA) fungicides. We screened 220 Colletotrichum isolates obtained from 38 apple orchards in the Mid-Atlantic region for resistance to 11 fungicides in Fungicide Resistance Action Committee (FRAC) groups 1, 7, 9, 11, 12, and 29. Eleven (5%) of these isolates were resistant to FRAC group 1 with confirmed ß-tubulin E198A mutations, and two (<1%) were also resistant to FRAC group 11 with confirmed cytochrome-b G143A mutations. Such low frequencies of resistant isolates indicate that fungicide resistance is unlikely to be the cause of any regional increase in bitter rot. A subsample of isolates was subsequently tested in vitro for sensitivity to every single-MoA fungicide registered for apple in the Mid-Atlantic U.S.A. (22 fungicides; FRAC groups 1, 3, 7, 9, 11, 12, and 29), and 13 fungicides were tested in field trials. These fungicides varied widely in efficacy both within and between FRAC groups. Comparisons of results from our in vitro tests with results from our field trials and other field trials conducted across the eastern U.S.A. suggested that EC25 values (concentrations that reduce growth by 25%) are better predictors of fungicide efficacy in normal field conditions than EC50 values. We present these results as a guideline for choosing single-MoA fungicides for bitter rot control in the Mid-Atlantic U.S.A.

Triflumizole as a Novel Lead Compound for Strigolactone Biosynthesis Inhibitor.

Strigolactones (SLs) are carotenoid-derived plant hormones involved in the development of various plants. SLs also stimulate seed germination of the root parasitic plants, Striga spp. and Orobanche spp., which reduce crop yield. Therefore, regulating SL biosynthesis may lessen the damage of root parasitic plants. Biosynthetic inhibitors effectively control biological processes by targeted regulation of biologically active compounds. In addition, biosynthetic inhibitors regulate endogenous levels in developmental stage- and tissue-specific manners. To date, although some chemicals have been found as SL biosynthesis inhibitor, these are derived from only three lead chemicals. In this study, to find a novel lead chemical for SL biosynthesis inhibitor, 27 nitrogen-containing heterocyclic derivatives were screened for inhibition of SL biosynthesis. Triflumizole most effectively reduced the levels of rice SL, 4-deoxyorobanchol (4DO), in root exudates. In addition, triflumizole inhibited endogenous 4DO biosynthesis in rice roots by inhibiting the enzymatic activity of Os900, a rice enzyme that converts the SL intermediate carlactone to 4DO. A Striga germination assay revealed that triflumizole-treated rice displayed a reduced level of germination stimulation for Striga. These results identify triflumizole as a novel lead compound for inhibition of SL biosynthesis.

Dissipation behavior, residue distribution, and risk assessment of triflumizole and FM-6-1 in greenhouse strawberries and soil.

This study aimed to develop a reliable method for the simultaneous analysis of triflumizole (TRIF) and its primary metabolite FM-6-1 (N-4-chloro-2-trifluoromethylphenyl-2-propoxy-acetamidine) in the soil and treated strawberries using solid phase extraction (SPE) coupled with gas chromatography-tandem mass spectrometry (GC-MS/MS). Using this method, TRIF and FM-6-1 degradation in strawberries and the soil under greenhouse conditions were investigated. The field trials showed that t1/2 of TRIF and total residues (the sum of TRIF and FM-6-1) were 1.6-2.2 days and 2.4-2.9 days in strawberry and 4.3-6.1 days and 5.5-6.9 days in soil, respectively. Terminal total residues were ≤ 0.39 mg/kg in strawberry and ≤ 0.42 mg/kg in soil from 5 to 10 days of harvest. The risk quotient (RQ) of TRIF was below 1.89%, showing that the dietary risk of TRIF in strawberry was low. These findings provide guidance for the use of TRIF on crops and provide reference to establish the maximum residue level (MRL) of TRIF in strawberry.

Acute toxicity of triflumizole to freshwater green algae Chlorella vulgaris.

Triflumizole is one of imidazole fungicides that works by inhibiting ergosterol biosynthesis, and is widely used for the control of powdery mildew and scabs on various fruits and crops. Triflumizole residue has been frequently detected in soil and aquatic ecosystems. While many studies have focused on its toxic effect on terrestrial and aquatic animals, little attention has been paid to aquatic algae, the primary producers of aquatic environments. Therefore, we evaluated the acute (96 h) toxicity effects of triflumizole on the freshwater algae Chlorella vulgaris, by examining growth, cell morphology, photosynthesis, and oxidative stress. The results showed that the 96 h median inhibition concentration (96 h-EC50) was 0.82 mg/L (95% confidential interval 0.70-0.98 mg/L).The growth of algal cells was conspicuously inhibited by triflumizole exposure, and the cell surfaces appeared to be shrunkThe chlorophyll content (including Chl-a, Chl-b and T-Chl) dramatically decreased at triflumizole concentrations of 0.2 and 1.0 mg/L. In addition, the transcript abundance of photosynthesis-related genes (psaB, psbC and rbcL) showed obvious decreases in above treatments after 96 h of exposure to triflumizole. Moreover, the algal growth inhibition was accompanied by an increase in intracellular reactive oxygen species and malondialdehyde content, as well as increased activity of antioxidant enzymes such as superoxide dismutase and peroxidase, indicating oxidative stress and lipid peroxidation. Our findings reveal that triflumizole has potential toxicity to the primary producers (freshwater algae) in aquatic ecosystems.

Development of triflumizole ionic liquids containing anions of natural origin for improving the utilization and minimizing the adverse impacts on aquatic ecosystems.

Triflumizole, a broad-spectrum systemic fungicide, has been widely used for the management of fungal diseases in plants. However, rapid photolysis and high risk to the aquatic environment limit its application. Ionic liquid (IL) forms of active pharmaceutical ingredients are innovative and promising agents that can optimize the application of the starting chemicals through the selection, or functionalization of the counterions (cation or anion). In this study, triflumizole was paired with various natural organic acids to develop novel ILs for improving the physicochemical properties and reducing the toxicity to fish. The results showed that the obtained ILs had low surface tension and lipophilicity and could protect triflumizole against degradation under UV irradiation as well as exhibit more excellent biological activity against Botrytis cinerea than triflumizole. The IL forms of triflumizole reduced the dosage and frequency of this fungicide, accordingly minimized the negative effect on environment. The IL contained salicylic acid as anion decreased > 4-fold toxicity to adult zebrafish over TFM. The results reported here create new application possibilities for imidazole fungicides and offer some heuristic rules for the design of active pharmaceutical ingredients-ionic liquids.

Dissipation behaviour and dietary risk assessment of boscalid, triflumizole and its metabolite (FM-6-1) in open-field cucumber based on QuEChERS using HPLC-MS/MS technique.

BACKGROUND: To resist plant diseases, boscalid and triflumizole have been applied to cucumbers frequently. However, the residue and dietary risk assessment of these fungicides in cucumber should be given attention for food safety. RESULTS: An effective and highly sensitive method based on the liquid chromatography-tandem mass spectrometry technique for simultaneous multidetermination of boscalid, triflumizole and its metabolite (FM-6-1) in a cucumber ecosystem was established and validated. Field experiments were conducted in three different locations, where boscalid and triflumizole (35% suspension concentration) were applied at 253 g of active ingredient (a.i.) per hectare (the recommended high dosage) and 379.5 g a.i. ha-1 (1.5 times the recommended high dosage) in each location. The limits of quantification and the limits of detection of the proposed method ranged from 0.01 to 0.05 mg kg-1 and 3.9 × 10-5 to 7.5 × 10-4 mg L-1 respectively. The mean recoveries and relative standard deviations of these compounds were 80-105% and 1.0-6.1% respectively. The dissipation dynamics of compounds followed pseudo-first-order kinetic models remarkably, with a half-value period of 2.3-40.8 days. The residues of boscalid and triflumizole in cucumber at harvest were below 0.66 mg kg-1 and 0.07 mg kg-1 respectively. The results of the dietary risk assessments have shown a low dietary risk of compounds in cucumber with hazard ratios <1 and hazard index <1. CONCLUSION: These results from the experiments are the most important for putting a guide on reasonable usage of these fungicides under the open-field conditions in China. © 2018 Society of Chemical Industry.

Multivariate optimization of dispersive liquid-liquid microextraction for the determination of paclobutrazol and triflumizole in water by GC-MS.

A new analytical method based on dispersive liquid-liquid microextraction with gas chromatography mass spectrometry has been optimized for the simultaneous determination of paclobutrazol and triflumizole in tap water and wastewater samples. A two-level, full-factorial design that allowed the study of main effects and factor interactions was applied to analyze the influence on microextraction process by chloroform, ethanol, potassium iodide and hand shaking period. The extraction conditions selected were 200 µL of chloroform, 3.0 mL of ethanol, 2.0 g of potassium iodide and 15 s of hand shaking. The limits of detection obtained for triflumizole and paclobutrazol under optimum conditions were 0.97 and 0.29 ng/mL, respectively. Calibration plots of both analytes were linear over a wide concentration range, and good precision was observed for replicate measurements. Applicability and accuracy of the method were determined by performing spiked recovery tests. Appreciable recovery results were obtained for municipal wastewater and matrix matching was used to obtain close to 100% recovery for tap water.

Review of the existing maximum residue levels for triflumizole according to Article 12 of Regulation (EC) No 396/2005.

According to Article 12 of Regulation (EC) No 396/2005, EFSA has reviewed the maximum residue levels (MRLs) currently established at European level for the pesticide active substance triflumizole. To assess the occurrence of triflumizole residues in plants, processed commodities, rotational crops and livestock, EFSA considered the conclusions derived in the framework of Directive 91/414/EEC, the MRLs established by the Codex Alimentarius Commission as well as the import tolerances and European authorisations reported by Member States (including the supporting residues data). Based on the assessment of the available data, MRL proposals were derived and a consumer risk assessment was carried out. Although no apparent risk to consumers was identified, some information required by the regulatory framework was missing. Hence, the consumer risk assessment is considered indicative only and all MRL proposals derived by EFSA still require further consideration by risk managers.