Dissipation and Safety Analysis of Dimethomorph Application in Lychee by High-Performance Liquid Chromatography-Tandem Mass Spectrometry with QuEChERS.

This study presents a method for analyzing dimethomorph residues in lychee using QuEChERS extraction and HPLC-MS/MS. The validation parameters for this method, which include accuracy, precision, linearity, and recovery, indicate that it meets standard validation requirements. Following first-order kinetics, the dissipation dynamic of dimethomorph in lychee was determined to range from 6.4 to 9.2 days. Analysis of terminal residues revealed that residues in whole lychee were substantially greater than those in the pulp, indicating that dimethomorph residues are predominantly concentrated in the peel. When applied twice and thrice at two dosage levels with pre-harvest intervals (PHIs) of 5, 7, and 10 days, the terminal residues in whole lychee ranged from 0.092 to 1.99 mg/kg. The terminal residues of the pulp ranged from 0.01 to 0.18 mg/kg, with the residue ratio of whole lychee to pulp consistently exceeding one. The risk quotient (RQ) for dimethomorph, even at the recommended dosage, was less than one, indicating that the potential for damage was negligible. This study contributes to the establishment of maximum residue limits (MRLs) in China by providing essential information on the safe application of dimethomorph in lychee orchards.

Catalytic degradation of dimethomorph by nitrogen-doped rice husk biochar.

N-doped biochar is widely used for activating persulfate to degrade organic pollutants. Which type of N atom is the key factor for activation is still unclear and needs to be further explored and analyzed. In this study, four kinds of biochar were prepared using urea and rice husk as precursors, and tested for the catalytic degradation of dimethomorph. Increasing the nitrogen doping level caused the catalytic removal efficiency of dimethomorph in the presence of peroxymonosulfate increased from 16.6% to 86.8%. A correlation analysis showed that the ability of N-doped biochar to activate PMS is mainly related to the content of pyrrole N, graphite N and carbonyl and the degree of defects. In experiments on electron paramagnetic resonance and free radical suppression, the reactive species of SO4•-, 1O2,·OH and O2.- were detected, among which 1O2 was found to be the main agent in the nonradical pathway. The degradation pathways for dimethomorph were analyzed based on a total of 8 degradation products identified by high-performance liquid chromatography-time of flight mass spectrometry (HPLC-Q-TOFMS). The results of this study provide a fundamental basis for using agricultural waste to produce inexpensive and efficient nonmetal catalysts that are highly effective in reducing dimethomorph levels in agricultural lands.

Design, Synthesis, and Fungicidal Activities of Indole-Modified Cinnamamide Derivatives.

Dimethomorph is a kind of cinnamamide fungicide with high fungicidal activities for oomycete diseases. The commercially available dimethomorph is a mixture of two isomers, in which (Z)-dimethomorph possessing higher activity and (E)-dimethomorph possessing lower activity. Herein, we reported the design, synthesis and fungicidal activities of a series of novel indole-modified cinnamamide derivatives, which used the indole group to 'fix' the cis-styrene group in (Z)-dimethomorph. The modification of the molecular structure of cinnamamide compounds could be beneficial to improve its practical application performance. Tested the fungicidal activities, it was found that compounds 8j, 9a, 9e, 9i and 9j showed excellent in vivo fungicidal activities (80-100 %) against Pseudoperonospora cubensis at a concentration of 100 mg L-1 , while dimethomorph and flumorph were noneffective. Moreover, parts of synthesized indole-modified cinnamamide derivatives 8 (8a, 8c, 8d and 8j) and 9 (9c and 9j) exhibited the same in vivo fungicidal activities against Phytophthora infestans with dimethomorph or flumorph at a concentration of 50 mg L-1 with 100 % inhibition. The biological assay results indicated that indole-modified cinnamamide derivatives have promising applications in the prevention and treatment of Phytophthora infestans.

Residue behavior and dietary risk assessment of six pesticides in pak choi using QuEChERS method coupled with UPLC-MS/MS.

A reliable and simple modified QuEChERS method with UPLC-MS/MS was developed for the simultaneous determination of six pesticides (dimethomorph, imidaclothiz, lufenuron, methoxyfenozide, pyridaben, spinetoram) and their metabolites in pak choi. Method validation indicated good linearity (R2 ≥ 0.99), accuracy (recoveries of 75%-112%), sensitivity (limits of quantification, 0.002-0.01 mg kg-1), and precision (relative standard deviations ≤ 21%), and matrix effects were -36-28%. The half-lives of the six pesticides in pak choi were 2.2-12 d under open field and greenhouse conditions. Considering the short growth cycle of pak choi, the terminal residue levels (0.046-7.8 mg kg-1) and the relevant maximum residue limits (MRLs) of some countries, 5 d was recommended as the pre-harvest interval for the six pesticides on pak choi. Dietary risk assessment revealed that the risk quotients were 3.1%-58% for different gender and age groups in China, indicating none unacceptable public health risk for general population. The results showed that all the six pesticides degraded faster and the terminal residues were much lower under open field conditions than those under greenhouse conditions, which was mainly due to the influence of rainfall, sunlight and other environmental factors. This work was thus significant in assessing the dissipation fate and food safety risks of the six pesticides on pak choi and facilitated the establishment of maximum residue limits.

Fungicide Efficacy Against Pseudoperonospora humuli and Point Mutations Linked to Carboxylic Acid Amide Resistance in Michigan.

Hops have expanded as a niche crop in Michigan and other production areas in the eastern United States, but growers in these regions face annual downy mildew outbreaks incited by Pseudoperonospora humuli, exacerbated by frequent rainfall and high relative humidity. We evaluated the efficacy of foliar- and drench-applied fungicides against downy mildew and examined Michigan isolates for point mutations linked to carboxylic acid amide (CAA) resistance. Disease severity and density were assessed weekly in 2016 and 2017 in nontrellised research hop yards in Michigan. Area under the disease progress curve values for disease severity were significantly lower for plants treated with oxathiapiprolin, ametoctradin/dimethomorph, fluopicolide, cyazofamid, or mandipropamid (90.6 to 100% control) compared with those treated with fosetyl-Al (64.3 to 93.0% control) at both locations for both years. Drench treatments of fluopicolide and oxathiapiprolin/mefenoxam reduced disease density and severity at both locations but were only moderately effective (76.4 to 91.5% control). To assess CAA resistance, the cellulose synthase CesA3 gene was aligned using reference downy mildew species and primers designed to amplify the 1105 and 1109 amino acids. Point mutations conferring CAA resistance were not detected at these loci for sporangia from 42 symptomatic shoots collected from 11 commercial hop yards. These efficacy results for hop downy mildew are needed to guide disease recommendations in this expanding Michigan industry. The absence of resistant genotypes indicates that Michigan growers can continue to utilize CAA-containing commercial fungicides as part of an overall downy mildew management program.

RNA-seq analysis of the response of plant-pathogenic oomycete Phytophthora parasitica to the fungicide dimethomorph.

Phytophthora parasitica is an important oomycete that causes disease in a variety of plants, dimethomorph fungicides being specific for oomycetes. The aim of this study was to use RNA-seq to rapidly discover the mechanism by which dimethomorph acts in the treatment of P. parasitica. We found that the expression of 832 genes changed significantly after the dimethomorph treatment, including 365 up-regulated genes and 467 down-regulated genes. According to the Gene Ontology (GO) enrichment analysis, pathway enrichment and verification test results, the following conclusions are obtained: (i) the treatment of P. parasitica with dimethomorph causes changes in the expression levels of genes associated with the cell wall and cell wall synthesis; (ii) dimethomorph treatment results in reduced permeability of the cell membrane and changes in the expression of certain transport-related proteins; (iii) dimethomorph treatment increased reactive oxygen species and reduced the expression of genes related to the control of oxidative stress.

Residues, dissipation kinetics, and dietary intake risk assessment of two fungicides in grape and soil.

The residue behavior and dietary intake risk of two fungicides (dimethomorph and pyraclostrobin) in grape (Vitis vinifera L.) were investigated from field trials. A modified quick, easy, cheap, effective, rugged, and safe method for simultaneously determining dimethomorph and pyraclostrobin residues in grape and soil was established using high performance liquid chromatography-tandem mass spectrometry. The average recoveries of dimethomorph and pyraclostrobin in the grape and soil matrices varied from 76.88% to 97.05%, with relative standard deviations of 1.73%-10.38%. The degradation half-lives of dimethomorph and pyraclostrobin were 7.3-12.0 days and 3.6-7.0 days in grape and soil, respectively. The terminal residues of dimethomorph and pyraclostrobin in the two matrices were 0.05-0.87 mg/kg. For dietary exposure risk assessments, all of the hazard quotient and hazard quotient index values were below 100%, which indicated that the suspending agents of dimethomorph and pyraclostrobin were sprayed on grape at the recommended dosages with no significant potential risks for Chinese consumers. This study provides a reference for analytically evaluating residual degradation behavior and dietary intake risk of two fungicides under field conditions.

Residual dynamic and risk assessment of dimethomorph in Swiss chard grown at two different sites.

Residue analysis of dimethomorph in Swiss chard cultivated at two different locations under greenhouse conditions was conducted using high-performance liquid chromatography-ultraviolet detection and confirmed by tandem mass spectrometry. The randomly collected samples (over 14 days) were extracted with acetonitrile and purified using a Florisil solid-phase extraction cartridge. Linearity over a concentration range of 0.05-50.0 mg/L had an excellent coefficient of determination of 0.9996. Recovery rate ranged from 82.98 to 95.43% with relative standard deviations ≤5.12% and limits of detection and quantification of 0.003 and 0.01 mg/kg, respectively. The initial deposits [day 0 (2 h post-application)] were considerably lower (7.57 and 8.55 mg/kg for sites 1 and 2, respectively) than the maximum residue limit (30 mg/kg) set by the Korean Ministry of Food and Drug Safety. The dissipation half-life was approximately the same, being 5.0 and 5.1 days for sites 1 and 2, respectively. Risk assessment estimated as acceptable daily intake revealed a value of 0.084 or 0.094% (day 0) and 0.014% (10 days post-application), for sites 1 and 2, respectively. The values indicated that dimethomorph can be safely used on Swiss chard, with no hazardous effects expected for Korean consumers.

Synergistic antifungal effect and potential mechanism of Dimethomorph combined with Pyrimethanil against Phytophthora capsici.

Synergistic effect of dimethomorph (DIM) and pyrimethanil (PYM) was evaluated using the Wadley method and the molecular mechanism of the antifungal effects of the combined treatment was systematically investigated. DIM+PYM had a synergistic effect on Phytophthora capsici, with the synergistic effect being observed at 5:1, at which the synergy coefficient was 1.8536. The mycelia of the pathogen treated with DIM+PYM were branched, uneven in thickness, and swollen. Moreover, scanning electron microscopy (SEM) revealed that DIM+PYM caused mycelium breaks, swelling, and apex enlargement, while transmission electron microscopy (TEM) revealed structural damage, cavities, and cell membrane morphological abnormalities. DIM+PYM inhibited the growth of mycelia, destroyed the cell membrane, interfered with energy metabolism, reduced protein and sugar content. Additionally, the transcriptome and metabolome of fungi treated with DIM+PYM changed significantly; specifically, there were 1571 differentially expressed genes and 802 differential metabolites. DIM+PYM may mainly damage the cell membrane, energy, protein, soluble sugar pathways.

Designing a size exclusion-based hapten and the development of a quantitative and visual time-resolved fluorescence immunochromatography assay strip for detecting dimethomorph and flumorph in a group-specific manner.

Based on the size and surface properties of dimethomorph and flumorph, we used a computer simulation-assisted size exclusion hapten design strategy to develop group-specific monoclonal antibodies that can simultaneously recognize dimethomorph and flumorph. For this, we performed quantitative and visual semi-quantitative time-resolved fluorescence immunochromatography (TRFICA) to simultaneously detect dimethomorph and flumorph in potatoes and apples. In potato samples, the visual limit of detection (vLOD) for dimethomorph and flumorph was 4 ng/mL and 8 ng/mL, respectively, whereas the quantitative limit of detection (qLOD) for dimethomorph and flumorph was 0.26 and 0.33 ng/mL, respectively. The vLOD of dimethomorph and flumorph in apple samples was 8 ng/mL, whereas the qLOD of dimethomorph and flumorph was 0.17 and 0.38 ng/mL, respectively. The average recovery of potato and apple samples ranged from 77.5% to 121.7%, which indicated that the method can be used to rapidly detect dimethomorph and flumorph in food samples.

Dimethomorph degradation in vineyards examined by isomeric and isotopic fractionation.

Knowledge of the degradation extent and pathways of fungicides in the environment is scarce. Fungicides may have isomers with distinct fungal-control efficiency, toxicity and fate in the environment, requiring specific approaches to follow up the degradation of individual isomers. Here we examined the degradation of the widely used fungicide dimethomorph (DIM) in a vineyard catchment using ratios of carbon stable isotopes (δ13C) and E/Z isomer fractionation (IF(Z)). In a microcosm laboratory experiment, DIM degradation half-life in soil was 20 ± 3 days, and was associated with significant isomeric (ΔIF(Z) = +30%) and isotopic (Δδ13C up to 7‰) fractionation. This corresponds to an isomer enrichment factor of εIR = -54 ± 6%, suggesting isomer selectivity and similar carbon stable isotopic fractionation values of εDIM-(Z) = -1.6 ± 0.2‰ and εDIM-(E) = -1.5 ± 0.2‰. Isomeric and isotopic fractionation values were used to estimate DIM degradation in topsoil and transport in a vineyard catchment over two wine-growing seasons. DIM concentrations following DIM application were up to 3 µg g-1 in topsoil and 29 µg L-1 in runoff water at the catchment outlet. Accordingly, the IF(Z) and δ13C values of DIM in soil were similar to those observed in DIM commercial formulations. The gradual enrichments in DIM-(Z) and 13C of the residual DIM in soil indicated DIM biodegradation over time. DIM biodegradation estimated based on E/Z isomer and carbon stable isotope ratios in topsoil and runoff water ranged from 0% after DIM application up to 100% at the end of the wine-growing season. DIM biodegradation was overestimated compared to conventional approaches relying on DIM mass balance, field concentrations and half-lives. Altogether, our study highlights the usefulness of combining carbon stable isotopes, E/Z isomers and classical approaches to estimate fungicide degradation at the catchment scale, and uncovers difficulties in using laboratory-derived values in field studies.

Peer review of the pesticide risk assessment of the active substance dimethomorph.

The conclusions of EFSA following the peer review of the initial risk assessments carried out by the competent authorities of the rapporteur Member State The Netherlands and co-rapporteur Member State Germany for the pesticide active substance dimethomorph and the assessment of applications for maximum residue levels (MRLs) are reported. The context of the peer review was that required by Commission Implementing Regulation (EU) No 844/2012, as amended by Commission Implementing Regulation (EU) No 2018/1659. The conclusions were reached on the basis of the evaluation of the representative uses of dimethomorph as a fungicide applied via foliar spraying on field strawberry and grapevine crops and permanent greenhouse lettuce crops; via drenching on field and permanent greenhouse strawberry crops and via dripping on permanent greenhouse strawberry crops. The peer review also provided considerations on whether exposure to humans and the environment from the representative uses of dimethomorph can be considered negligible, taking into account the European Commission's draft guidance on this topic. MRLs were assessed in potatoes, other root and tuber vegetables (except radishes), stem vegetables (except celery, leeks, globe artichokes, sugar beet, cereal forage and straw). The reliable end points, appropriate for use in regulatory risk assessment and the proposed MRLs, are presented. Missing information identified as being required by the regulatory framework is listed. The concerns are reported where identified.

Additive effect of the Streptomyces albus XJC2-1 and dimethomorph control pepper blight (Capsicum annuum L.).

BACKGROUND: Pepper blight, caused by Phytophthora capsici, is a destructive soilborne disease, which poses a serious threat to pepper, Capsicum annuum L., production. Chemical fungicides, which mainly are used to control pepper blight, have a negative effect on the environment, rendering biological control as a promising alternative to maintain the balance between ecology and pest management. The purpose of this study was to screen the biocontrol bacteria, reduce the dosage of fungicides and increase the stability of biocontrol bacteria, and to find the mixing ratio of biocontrol bacteria and fungicides giving the best control effect. RESULTS: We isolated actinomycetes strains from the soil surrounding the roots of healthy pepper plants amongst field-grown plants infected with P. capsici. Of these, Streptomyces albus XJC2-1 showed a strong inhibition effect on the growth of P. capsici, with an inhibition rate of ≤85%. XJC2-1 effectively inhibited the formation of sporangium and release of zoospores of P. capsici as well as directly destroyed its hyphae, to achieve the inhibitory effect. Transcriptomic profiling of pepper leaves, postirrigation of plants with the XJC2-1 fermentation broth, revealed upregulation of genes related to the photosynthesis pathway in pepper. Furthermore, XJC2-1 treatment improved the net photosynthetic rate and intercellular CO2 concentration, thereby improving the pepper plant's resistance to pathogens. The combination of XJC2-1 with the fungicide dimethomorph (8 µg mL-1 ) displayed strong synergism in inhibition of P. capsici infection, with a control efficiency as high as 75.16%, thus providing a basis for its application in the field. CONCLUSION: Our study demonstrated that S. albus XJC2-1 inhibited Phytophthora pathogens from infecting pepper plants and enhanced plant host resistance. The combination of XJC2-1 and dimethomorph displayed a more stable and stronger control effect on pepper blight, showing potential for the future application of XJC2-1 in the field of biological control. © 2023 Society of Chemical Industry.

Comparative Transcriptome Analysis to Investigate the Immunotoxicity Mechanism Triggered by Dimethomorph on Human Jurkat T Cell Lines.

Dimethomorph (DMM) is a broad-spectrum fungicide used globally in agricultural production, but little is known regarding the immunotoxicity of DMM in humans. In this study, the immunotoxicity of DMM on human Jurkat T cells was evaluated in vitro. The results indicated that the half-effective concentration (EC50) of DMM for Jurkat cells was 126.01 mg/L (0.32 mM). To further elucidate the underlying mechanism, transcriptomics based on RNA sequencing for exposure doses of EC25 (M21) and EC10 (L4) was performed. The results indicated that compared to untreated samples (Ctr), 121 genes (81 upregulated, 40 downregulated) and 30 genes (17 upregulated, 13 downregulated) were significantly differentially regulated in the L4 and M21 samples, respectively. A gene ontology analysis indicated that the significantly differentially expressed genes (DEGs) were mostly enriched in the negative regulation of cell activities, and a KEGG pathway analysis indicated that the DEGs were mainly enriched in the immune regulation and signal transduction pathways. A quantitative real-time PCR for the selected genes showed that compared to the high-dose exposure (M21), the effect of the low-dose DMM exposure (L4) on gene expression was more significant. The results indicated that DMM has potential immunotoxicity for humans, and this toxicity cannot be ignored even at low concentrations.

Removal and fate of pesticides in a farm constructed wetland for agricultural drainage water treatment under Mediterranean conditions (Italy).

A non-waterproofed surface flow constructed wetland (SFCW), treating agricultural drainage water in Northern Italy, was investigated to gain information on the potential ability for effective pesticide abatement. A mixture of insecticide imidacloprid, fungicide dimethomorph, and herbicide glyphosate was applied, by simulating a single rain event, into 470-m-long water course of the SFCW meanders. The pesticides were monitored in the wetland water and soil for about 2 months after treatment. Even though the distribution of pesticides in the wetland was not uniform, for each of them, a mean dissipation of 50% of the applied amount was already observed at ≤7 days. The dissipation trend in the water phase of the wetland fitted (r2 ≥ 0.8166) the first-order model with calculated DT50 of 20.6, 12.0, 5.8, and 36.7 days for imidacloprid, dimethomorph, glyphosate, and the glyphosate metabolite AMPA, respectively. The pesticide behavior was interpreted based on the chemical and physical characteristics of both the substances and the water-soil system. Despite the fast abatement of glyphosate, traces were detected in the water until the end of the trial. The formation of soluble 1:1 complex between glyphosate and calcium, the most representative cation in the wetland water, was highlighted by infrared analyses. Such a soluble complex was supposed to keep traces of the herbicide in solution.

A pH- and redox-stimulated responsive hollow mesoporous silica for triggered delivery of fungicides to control downy mildew of Luffa cylindrica.

BACKGROUND: Downy mildew, a devastating disease of cucurbitaceous crops caused by Pseudoperonospora cubensis. Although a variety of fungicides are used to control downy mildew, choosing an effective product can be challenging. Environmental stimulus-responsive pesticide delivery systems have great potential to improve the effectiveness of disease and pest control and reduce the impact on environmentally beneficial organisms. RESULTS: In this work, a disulfide bond (SS)-modified and chitosan oligosaccharide (COS)-capped hollow mesoporous silica (HMS) pesticide delivery system was synthesized using a hard template method for the control of downy mildew in cucurbit crops. The synthesized nanoparticles were loaded with dimethomorph (DMM), denoted as DMM@HMS-SS-COS, and the developmental toxicity of these nanoparticles to zebrafish embryos were evaluated. The results showed that the prepared DMM@HMS-SS-COS exhibited excellent dual response properties to pH and glutathione (GSH), with an encapsulation rate of up to 24.36%. DMM@HMS-SS-COS has good ultraviolet (UV) radiation stability and adhesion properties. Compared with dimethomorph suspension concentrate (SC), DMM@HMS-SS-COS was more effective against downy mildew for up to 21 days. Toxicity tests showed that DMM@HMS-SS-COS significantly reduced the effect of DMM on the hatching rate and survival rate of zebrafish embryos. CONCLUSIONS: This work not only demonstrates that DMM@HMS-SS-COS could be used as a nanodelivery system for intelligent control of downy mildew but also emphasizes the necessity of increasing the acute toxicity of nanoformulations to non-target organisms in environmental risk assessment. © 2022 Society of Chemical Industry.

CHA-based dual signal amplification immunofluorescence biosensor for ultrasensitive detection of dimethomorph.

Dimethomorph (DMM) is a widely used high-efficiency fungicide which poses unpredictable threats to the ecological environment and public health. It is significant to establish a sensitive and robust analytical method for DMM detection. Here, we fabricated a catalytic hairpin assembly (CHA) - based immunofluorescence (IMF) biosensor by using single-strand DNA and DMM antibody co-modified gold nanoparticles (H0-Ab-Au) as anti-interference probes and DMM antigen coated 96-well plate as the immune recognition element and CHA reaction vessel. Parameters relevant to AuNP probes preparation and CHA reaction environment were optimized. After optimization, the LOD of 0.002 ng/mL was calculated, with a linear correlation in inverse proportion to DMM concentration ranging from 0.01 ng/mL to 50 ng/mL. In addition, the developed biosensor was successfully applied to a variety of complex matrix samples, with satisfactory recoveries over a range of 86.74%-118.60%. Moreover, the detection results of IMF biosensor have a good correlation with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Therefore, our proposed IMF biosensor exhibits ultra-high sensitivity and excellent specificity, as well as great potential for application to other hazards.

Human cytochrome P450 3A-mediated two-step oxidation metabolism of dimethomorph: Implications in the mechanism-based enzyme inactivation.

Dimethomorph (DMM), an effective and broad-spectrum fungicide applied in agriculture, is toxic to environments and living organisms due to the hazardous nature of its toxic residues. This study aims to investigate the human cytochrome P450 enzyme (CYP)-mediated oxidative metabolism of DMM by combining experimental and computational approaches. Dimethomorph was metabolized predominantly through a two-step oxidation process mediated by CYPs, and CYP3A was identified as the major contributor to DMM sequential oxidative metabolism. Meanwhile, DMM elicited the mechanism-based inactivation (MBI) of CYP3A in a suicide manner, and the iminium ion and epoxide reactive intermediates generated in DMM metabolism were identified as the culprits of MBI. Furthermore, three common pesticides, prochloraz (PCZ), difenoconazole (DFZ) and chlorothalonil (CTL), could significantly inhibit CYP3A-mediated DMM metabolism, and consequently trigger elevated exposure to DMM in vivo. Computational studies elucidated that the differentiation effects in charge distribution and the interaction pattern played crucial roles in DMM-induced MBI of CYP3A4 during sequential oxidative metabolism. Collectively, this study provided a global view of the two-step metabolic activation process of DMM mediated by CYP3A, which was beneficial for elucidating the environmental fate and toxicological mechanism of DMM in humans from a new perspective.

Combined Developmental Toxicity of the Pesticides Difenoconazole and Dimethomorph on Embryonic Zebrafish.

Difenoconazole (DIF) and dimethomorph (DIM) are widely used pesticides frequently detected together in environmental samples, so the deleterious effects of combined exposure warrant detailed examination. In this study, the individual and combined effects of DIM and DIF on conventional developmental parameters (hatching rate, deformity rate, lethality) and gene expression were measured in embryonic zebrafish. Both DIF and DIM interfered with normal zebrafish embryo development, and the most sensitive toxicity index for both was 96 h post-fertilization (hpf) deformity rate (BMDL10 values of 0.30 and 1.10 mg/L, respectively). The combination of DIF and DIM had mainly synergistic deleterious effects on 96 hpf deformity and mortality rates. Transcriptome analysis showed that these compounds markedly downregulated expression of mcm family genes, cdk1, and cdc20, thereby potentially disrupting DNA replication and cell cycle progression. Enhanced surveillance for this pesticide combination is recommended as simultaneous environmental exposure may be substantially more harmful than exposure to either compound alone.

Dissipation behavior, residue distribution, and dietary risk assessment of fluopimomide and dimethomorph in taro using HPLC-MS/MS.

Dimethomorph is a morpholine broad-spectrum fungicide and effectively controls taro blight, cucumber downy mildew, rice blast disease, and others. Fluopimomide is a newly developed broad-spectrum fungicide to primarily control oomycetes and rhizoctonia diseases. Taro, one of the earliest cultivated crops, is a staple food in Africa, Oceania, and Asia. Recently, a commercial suspension concentrate formulation containing 15% fluopimomide and 25% dimethomorph has been registered in China, the second largest taro producer in the world. The objective of this study was to develop a high-performance liquid chromatography tandem mass spectrometry method to detect the residues of fluopimomide and dimethomorph concurrently in taro samples. The results showed that the average recoveries of fluopimomide and dimethomorph ranged from 83 to 108%, and relative standard deviations (RSD) ranged from 1 to 11%. The limit of quantitation (LOQ) was 0.01 mg kg-1 for the two compounds. The dissipation results demonstrated that both fluopimomide and dimethomorph in taro degraded rapidly in taro fields, and the residues of the two fungicides were below the LOQ within 14 days post-application. The final residue levels of fluopimomide and dimethomorph in taro were lower than 0.066 mg kg-1 28 days post-application. For dietary risk assessments, the dietary structure of different genders and age of people in China exposure risk assessment and whole diet exposure risk assessment shows that the risk quotient (RQ) values were substantially lower than 100%, suggesting that the long-term risks of fluopimomide/dimethomorph mixed formulation in taro at the recommended dosage were negligible. In summary, our combined results from the dissipation behaviors, terminal residues, and dietary risk assessments provide the critical empirical data for the establishment of the maximum residue levels (MRLs) of the two broad-spectrum fungicides in taro, a traditional food for African, Oceanic, and South Asian cultures.