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In this work, a series of derivatives with disulfide bonds containing pyridine, pyrimidine, thiophene, thiazole, benzothiazole, and quinoline were designed and synthesized based on the various biological activities of allicin disulfide bond functional groups. The antimicrobial activities of the target compounds were determined, and the structure-activity relationships were discussed. Among them, compound

Four common fungicidal products were evaluated for their effect on symptoms caused by two nectriaceous black root rot fungi, Calonectria ilicicola and Dactylonectria macrodidyma, when applied as pot drenches to avocado (Persea americana) seedlings in the greenhouse. Applications of fludioxonil, thiophanate-methyl + etridiazole, prochloraz, and prochloraz MnCl at 2 and 4 weeks after inoculation with C. ilicicola significantly reduced root necrosis and improved root and aboveground plant biomass compared with water-treated controls. Fludioxonil reduced necrosis by 60% and had a significantly lower frequency of reisolation of C. ilicicola than the other three fungicide treatments. D. macrodidyma inoculation caused less severe symptoms in seedlings than C. ilicicola despite the longer duration of the trial. Pot drenches with fludioxonil, thiophanate-methyl + etridiazole, and prochloraz MnCl, but not prochloraz alone, significantly reduced root necrosis caused by D. macrodidyma. Prochloraz MnCl was the only fungicide treatment to increase root and plant biomass compared with water-treated controls. Both fludioxonil and prochloraz MnCl reduced the frequency of reisolation of D. macrodidyma from necrotic roots by about 50% compared with the other fungicides or water controls. The results indicated that drenches with these fungicides may suppress existing low to moderate black root rot infection, allowing new root growth and improved establishment in the orchard. Fungicide drenching must not replace best-practice disease management strategies in nurseries but may be a useful tool in crisis situations.

The objectives of this study were to examine cytotoxic and genotoxic damage in human BJ fibroblasts caused by three pesticides used worldwide by trypan blue dye exclusion assays and to measure the relative level of phosphorylated histone H2A.X by flow cytometry at different concentrations. Captan-based fungicide and methyl thiophanate-based fungicide (100 and 1000 µΜ) showed immediate cytotoxic effects; furthermore, after 24 h, captan-based fungicide, chlorothalonil-based fungicide and methyl thiophanate-based fungicide caused cytotoxic effects in the concentration ranges of 40-100 µM, 30-100 µM and 150-1000 µM, respectively. All fungicides generated DNA damage in the treated cells by activating ATM and H2A.X sensor proteins. The three fungicides tested generated DNA double-stranded breaks and showed cytotoxicity at concentrations 33, 34, and 5 times lower (captan, chlorothalonil and thiophanate-methyl respectively) than those used in the field, as recommended by the manufacturers.

La muerte descendente del limón ‘Persa’ (Citrus latifolia Tan.) causada por Lasiodiplodia spp., es una enfermedad de alta importancia. La intensidad de las podas en limón incrementa la susceptibilidad y la pérdida anual de hasta 35% de árboles, lo que resulta 60% de reducción de la producción. Durante los años 2016 y 2017, en el municipio de Papantla, Veracruz se evaluó la efectividad del control químico y biológico contra Lasiodiplodia spp., después de la poda. Se realizó un diseño de bloques completo al azar, se aplicaron cinco tratamientos: metil tiofanato (1 g L-1), tiabendazole (2.5 g L-1), clorotalonil (3 g L-1) y mancozeb (4 g L-1), Trichoderma harzianum + Trichoderma viridae (20 ml L-1) y un testigo (sin aplicación). Se realizaron dos aspersiones dirigidas a tronco, ramas y follaje hasta punto de goteo. Se evalúo la incidencia y severidad de la enfermedad cada siete días después de la primera aplicación. Con los datos de incidencia se realizó ajuste de modelos epidemiológicos y se calcularon los parámetros descriptivos como: intensidad inicial, tasa de incremento y área bajo la curva del progreso de la enfermedad. El fungicida metil tiofanato tuvo mayor efectividad para el control de la enfermedad, seguido de tiabendazol.

Background Thiophanate-methyl and its metabolite carbendazim are broad-spectrum fungicides used on many crops. The residues of these chemicals could result in potential environmental and human health problems. Therefore, investigations of the dissipation and residue behaviors of thiophanate-methyl and its metabolite carbendazim on cowpeas and associated dietary risk assessments are essential for the safety of agricultural products. Results A simple analytical approach using liquid chromatography with tandem mass spectrometry was developed and validated for the determination of thiophanate-methyl and carbendazim concentrations in cowpeas. Good linearity (R2 > 0.998) was obtained, and the recoveries and relative standard deviations were 80.0%-104.7% and 1.4%-5.2%, respectively. The dissipation rates of thiophanate-methyl, carbendazim and total carbendazim were high (half-lives of 1.61-2.46 days) and varied in the field cowpea samples because of the different weather conditions and planting patterns. Based on the definition of thiophanate-methyl, the terminal residues of total carbendazim in cowpea samples were below the maximum residue limits set by Japan for other legumes. The acute and chronic risk quotients of three analytes were 0.0%-27.6% in cowpea samples gathered from all terminal residue treatments, which were below 100%. Conclusion An optimized approach for detecting thiophanate-methyl and carbendazim in cowpeas was applied for the investigation of field-trial samples. The potential acute and chronic dietary risks of thiophanate-methyl, carbendazim and total carbendazim to the health of Chinese consumers were low. These results could guide the safe and proper use of thiophanate-methyl in cowpeas and offer data for the dietary risk assessment of thiophanate-methyl in cowpeas. This article is protected by copyright. All rights reserved.

The highly heterogeneous nature of

Charcoal rot disease of soybean caused by Macrophomina phaseolina is a serious problem in most of the soybean growing area of Madhya Pradesh. In this study, seven plant extracts viz., leaves of Azadirachta indica, Citrus limon, Polyalthia longifolia, Parthenium hysterophorus and Ricinus communis, bulb of Allium sativum and Allium cepa and eight fungicides viz., Captan (0.25%), Mancozeb (0.25%), Carbendazim + Mancozeb (25%), Thiophanate Methyl (0.1%), Pyraclostrobin (0.2%), Carbendazim (0.1%) and Blue copper (0.3% ) were evaluated for an effective management of charcoal rot of soybean caused by M. phaseolina (Tassi) Goid under in vitro and in vivo condition. Among plant extracts, garlic clove extract was found most effective showing 77.3 % growth inhibition and poor microslerotia formation of M. phaseolina by 77.3 % followed by parthenium leaf extract (75.2% inhibition) at 15 % concentration. Two soil drenching of garlic clove extracts @ 15.0 % concentration also found most effective for the management of disease under field condition recorded minimum disease incidence (13.5%) and highest yield (14.6q/ha). Among fungicides, Carbendazim (0.1%) and Thiophanate Methyl (0.1%) showed 100 % inhibition of radial growth and microsclerotia production of M phaseolina under in vitro condition. Two soil drenching of Carbendazim @ 0.1% found to be most effective for the management of charcoal rot of soybean under field condition showing minimum disease incidence (5.36%) and producing highest yield (16.0 q/ha) followed by Thiophanate Methyl. These results suggested that the toxic effect of Carbendazim and Thiophanate Methyl and A. sativum inhibited maximum mycelium growth in vitro and provide management of charcoal rot disease under field conditions.

A dual-emission fluorescent (FL) probe was constructed by coordinating Cu

The impact of fungicides on the head blight (FHB) development disease index, percent spike harvest index (SHI), and deoxynivalenol (DON) accumulation in wheat kernels under field conditions was evaluated after artificial spike inoculation with

Chemical seed treatment is an established practice in agriculture to protect crops from soil-borne pathogens and pests. Arbuscular mycorrhizal fungi (AMF) benefit plants by extending soil exploration as well as water and nutrient uptake. The objective of this work was to analyze the effects of combinations of seed treatments with doses of inoculant containing Rhizoglomus intraradices on vegetative development, root colonization and nutrition of Phaseolus vulgaris plants and soil microbiota.Seed treatment benefited the vegetative development and nutrition of beans, with the treatments metalaxyl + fludioxonil + tiabendazole and pyraclostrobin + thiophanate methyl + fipronil standing out regarding the contents of nitrogen (N), phosphorus (P), iron (Fe) and zinc (Zn) of the aerial parts. Mycorrhizal inoculation linearly increased dehydrogenase activity, root biomass and total plant biomass, with increments reaching 27%. There was an interaction between seed treatment and inoculation dose for aboveground biomass and the contents of potassium (K), calcium (Ca), magnesium (Mg), sulfur (S), manganese (Mn) and root colonization, with expressive results for the combination of the two highest doses of inoculant with metalaxyl + fludioxonil + tiabendazole or pyraclostrobin + methylthiophanate + fipronil in the seeds.Chemical seed treatment and mycorrhizal inoculation benefited bean plants and their nutritional status. The best combinations for the bean crop were metalaxyl + fludioxonil + tiabendazole with 41.4 mg of the inoculant per 100 seeds and pyraclostrobin + thiophanate methyl + fipronil with 62.1 mg of the inoculant per 100 seeds. © 2022 Society of Chemical Industry.

Exposure to pesticides through eyes, skin, ingestion and inhalation may affects human health by interfering with immune cells, such as macrophages. We evaluated, in vitro, the effect of six pesticides widely used in apple arboriculture on the functions of human monocyte-derived macrophages (hMDMs). hMDMs were cultured for 4 or 24 h with or without pesticides (0.01, 0.1, 1, 10 μmol.L

Outbreaks of bitter rot were observed in three commercial apple orchards in Illinois despite best management efforts during the 2018 production season. Three isolates from symptomatic fruit from these orchards and two isolates from an orchard in South Carolina were identified to the species level using morphological tools and calmodulin, glyceraldehyde-3-phosphate dehydrogenase, and beta-tubulin gene sequences. The isolates from Illinois were identified as Colletotrichum siamense of the Colletotrichum gloeosporioides species complex and the ones from South Carolina as Colletotrichum fioriniae and Colletotrichum fructicola of the Colletotrichum acutatum and the C. gloeosporioides species complex, respectively. Two of the three C. siamense isolates from Illinois were resistant to azoxystrobin and thiophanate-methyl as determined in mycelial growth tests in vitro. EC50 values were >100 μg/ml for both fungicides. One isolate was only resistant to azoxystrobin. None of the isolates from South Carolina was resistant to either of the two compounds. All five isolates were sensitive to fludioxonil (EC50 values

Sclerotinia homoeocarpa causes dollar spot disease on turfgrass and is a serious problem on many species worldwide. Fludioxonil, a phenylpyrrole fungicide, is not currently registered for dollar spot control in China. In this study, the baseline sensitivity to fludioxonil was established using an in vitro assay for 105 isolates of S. homoeocarpa collected from 10 locations in different regions of China. Results indicate that the frequency distribution of effective concentration for 50% inhibition of mycelial growth (EC50) values of the S. homoeocarpa isolates was unimodal (W = 0.9847, P = .2730). The mean EC50 value was 0.0020 ± 0.0006 μg/ml with a range from 0.0003 to 0.0035 μg/ml. A total of 7 fludioxonil-resistant mutants were obtained in laboratory, the mutants were stable in fludioxonil sensitivity after the 10th transfer, with resistance factor (RF) ranging from 4.320 to >13,901.4. The mutants showed a positive cross-resistance between fludioxonil and the dicarboximide fungicide iprodione, but not propiconazole, fluazinam, and thiophanate-methyl. When mycelial growth rate, pathogenicity and osmotic sensitivity were assessed, the mutants decreased in the fitness compared with their parental isolates. Sequence alignment of the histidine kinase gene Shos1 revealed a 13-bp fragment deletion only in one mutant, no mutations were observed on Shos1 in the rest resistant mutants.

Brown rot disease broke out in stone fruit orchards of Fujian, China in 2019, despite pre-harvest application of methyl benzimidazole carbamate (MBC). To determine the reason, a total of 44 Monilinia fructicola strains were collected from nectarine, plum and peach fruits in this study, among which 79.5% strains were resistant to thiophanate-methyl, indicated by discriminatory dose of 5 μg/mL. The resistance of these strains was confirmed by treating detached peach fruit with label rates of formulated thiophanate-methyl which only completely inhibit infection of the sensitive strains, but not the resistant strains. Further analysis of the mechanism of MBC resistance revealed that all resistant strains carry a H6Y mutation in β-tubulin protein Tub2, which was only reported previously in the M. fructicola strains from California, USA, and do not display obvious fitness penalties, as no significant defects in mycelial growth rate, sporulation, conidia germination, aggressiveness on detached peach fruit and temperature sensitivity was detected. In addition, we found that diethofencarb, the agent for managing MBC-resistance strains, was unable to inhibit growth of the H6Y strains. Taken together, our study, for the first time, identified a mutation form of H6Y in the β-tubulin protein of M. fructicola in China, rendering the strains wide resistance to thiophanate-methyl. This mechanism of M. fructicola gaining resistance to MBC fungicides needs to be fully considered, when designing management strategies to control brown rot disease in stone fruit orchards.

Multiple

Though pesticide mixtures can reflect the real-life situation in the water ecosystem, the quantification of their toxicity is still not fully understood. Combined effects of thiophanate-methyl (THM) and fenvalerate (FEN) on embryonic zebrafish (Danio rerio) and underlying mechanism were conducted in this study. Results showed that the 96-h LC50 values of THM to D. rerio at different growth periods ranged from 12.1 to 26.1 mg L−1, which were lower in comparison with those of FVR ranging from 0.025 to 2.8 mg L−1. Mixture of THM and FVR exhibited a synergetic response to zebrafish embryos. Activities of Cu/Zn-SOD, POD, caspase 3 and caspase 9 were significantly different in most of single and mixture administrations compared with the control group. In addition, five genes (P53, Cu/Zn-sod, crh, ERα and IL-8) associated with oxidative stress, cellular apoptosis, immune system and endocrine system showed greater variations of expressions when administrated to pesticide mixtures compared with single chemicals. Our experimental results exhibited that mixtures of thiophanate-methyl and fenvalerate produced higher toxicity towards aqueous vertebrates than when determined singly. Collectively, upcoming environmental risk assessments established according to single administrations might not be enough to protect the water ecosystem.

An 18-ha commercial pecan orchard was sampled over 3 years to study the spatial and temporal variation in fungicide sensitivity of

Twenty-eight isolates of Sclerotinia homoeocarpa, causal agent of dollar spot disease in turf, were assessed for fungicide hormesis at sublethal concentrations of thiophanate-methyl (T-methyl). Each isolate was grown in corn meal agar amended with 11 concentrations of T-methyl (30,500 to 0.047 µg/liter), and the area of mycelial growth was determined relative to the control. Three replicates were used per concentration, and the experiment was repeated three to five times for each isolate. Reference isolates (EC50 > 20 µg/liter), with no prior history of T-methyl exposure, were highly sensitive and not stimulated by low doses. Likewise, no stimulation was observed in two highly sensitive isolates (EC50 > 30 µg/liter) that had been preconditioned by exposure to T-methyl, or in four T-methyl-tolerant isolates. Seventeen (81%) preconditioned T-methyl-tolerant isolates (EC50 = 294 to1,550 µg/liter) had statistically significant growth stimulation, in the range of 2.8 to 19.7% relative to the control. These results support that hormesis (low-dose stimulation, high-dose inhibition) is a common dose response in preconditioned S. homoeocarpa, particularly in response to subtoxic doses of T-methyl.

Sclerotinia homoeocarpa causes dollar spot disease on many turfgrass species and is a significant problem worldwide. Thiophanate-methyl (TM), a methyl benzimidazole carbamate (MBC) fungicide, has been used for over forty years to manage dollar spot. Here we describe genetic mutations linked to three distinct TM fungicide resistance phenotypes: sensitive (S), moderately resistant (MR) and highly resistant (HR). These were established using multiple doses of TM, compared to previous studies using single discriminatory doses. In total, 19 S, 3 MR and 22 HR isolates were detected. Analysis of the β-tubulin gene revealed the MR isolates had a point mutation from T to A at codon 200 changing phenylalanine (TTC) to tyrosine (TAC). Twenty HR isolates had a mutation at codon 198 changing glutamic acid (GAG) to alanine (GCG) and two HR isolates had a mutation at codon 198 changing glutamic acid (GAG) to lysine (AAG). Allele-specific PCR assays were developed for rapid detection of these mutations in isolates of S. homoeocarpa. In addition, our results suggest a two-dose system for in vitro screening provides useful information for monitoring the development of resistance.

Incidence of blossom blight and Botrytis fruit rot (BFR), caused by Botrytis cinerea, on two southern highbush blueberry cultivars was evaluated in several blueberry fields grown in the vicinity (BB-Str(+)) or not (BB-Str(−)) of strawberry fields in central Florida. Blossom blight and BFR incidence were higher in BB-Str(+) fields in 2014 and significantly higher in 2015 compared to BB-Str(−) fields. In total, 613 B. cinerea isolates (i.e., 181 and 432 isolates from BB-Str(−) and BB-Str(+) fields, respectively) were collected. The isolates were evaluated for sensitivity to eight single-site and one multisite fungicides using a spore germination and a germ tube elongation assay. Overall, 5, 15, 24, 28, 54, and 93% of isolates collected from BB-Str(−) were resistant to penthiopyrad, cyprodinil, boscalid, fenhexamid, pyraclostrobin, and thiophanate-methyl, respectively. Respective resistance frequencies in BB-Str(+) isolates were 10, 30, 65, 66, 89, and 99%. Resistance frequencies for all fungicides were always higher in BB-Str(+) fields compared to BB-Str(−) fields. Isolates exhibiting resistance to six or five fungicides simultaneously were predominant (50 to 70%) in blueberry fields regardless if they were grown in the vicinity of strawberry fields or not. Among 308 and 305 B. cinerea isolates tested in 2014 and 2015, 41.8 and 47.1%, respectively, showed reduced sensitivity to the multisite fungicide captan. The lower label rate of captan applied preventively did not control isolates with reduced sensitivity on detached blueberry fruit. These findings suggest a potential population flow between strawberry and blueberry fields that may impact blossom blight and gray mold development in blueberry fields. The relatively lower fungicide input applied to blueberry fields compared with strawberry fields seems to be sufficient to select for resistance and multiple-resistant phenotypes in B. cinerea populations in blueberry.

The emerging infectious disease chytridiomycosis, caused by the fungus Batrachochytrium dendrobatidis, is a major driver pushing many amphibian species to the brink of extinction. Substantial efforts to develop effective protocols that use antifungal drugs have had notable success. Here, we used the antifungal agents itraconazole and thiophanate-methyl, singly and in combination, in an attempt to treat common midwife toad Alytes obstetricans larvae naturally infected with the globalized hypervirulent lineage of B. dendrobatidis. Despite the successful use of itraconazole in a closely related species (A. muletensis), our results show that these antifungal treatments are not always effective and that full clearance of animals cannot be assumed following treatment., This research was conducted under permits of the Consejería de Medio Ambiente of Aragón, Castilla y León and Madrid, and supported by the Fundación BBVA. T.W.J.G. was supported by NERC Standard Grant NE/N009967/1.

An efficient method was developed and validated for simultaneous determination of tebuconazole, thiophanate-methyl and its metabolite carbendazim in grape and soil using QuEChERS procedure and liquid chromatography-tandem mass spectrometry. The average recoveries of the method were 83.2%–105.4%, the limit of detection (LOD) ranged from 2.1 × 10−5 to 8.7 × 10−4 mg L−1, and the limit of quantitation (LOQ) was 0.1 mg kg−1. The field results showed that tebuconazole and thiophanate-methyl in soil and grape were rapidly dissipated with half-lives less than 24.7 days. The terminal residues of all analytes in grape were lower than the maximum residue limit (MRL) set by China (2 mg kg−1 for tebuconazole; 3 mg kg−1 for both thiophanate-methyl and carbendazim). Risk assessment showed that tebuconazole, thiophanate-methyl and its metabolite would be unlikely to cause health problems. However, carbendazim, the higher active metabolites of thiophanate-methyl, should receive more concerns.

In 2014 and 2015, a total of 151 tobacco brown spot (Alternaria alternata) isolates were collected from Guizhou Province in China to evaluate their resistance to the benzimidazole thiophanate-methyl, the carbamate diethofencarb, and the dicarboximide procymidone. Resistance to thiophanate-methyl and diethofencarb was observed in all isolates. Resistance to all the three fungicides, thiophanate-methyl, diethofencarb, and procymidone was detected at a frequency of 6.0%. The F167Y single mutation in the β-tubulin gene was found to be associated with resistance to thiophanate-methyl,but no mutation was found in the coiled-coil region of the histidine kinase-encoding gene OS1, a fungal gene for dicarboximide resistance. Procymidone applied at the rate of 20 mg l−1 inhibited spot lesion formation on tobacco leaves with an efficacy of 51.7% for the low resistance (LR) isolates and 74.2% for the procymidone-sensitive isolates. Thiophanate-methyl applied at 100 mg l−1, however, slightly promoted the expansion of disease lesions with an efficacy of −7.7%. Azoxystrobin applied at 10 and 20 mg l−1 provided efficacies of 91.1 and 100%, respectively, regardless of whether the isolates were thiophanate-methyl resistant or procymidone-LR. Further studies suggested that azoxystrobin exhibited excellent protective activity and good curative activity against A. alternata in plants. The baseline sensitivity to azoxystrobin was then determined. In the presence SHAM, the mean EC50 values for conidial germination inhibition were 0.49 ± 0.22 (Mean ± SD) mg l−1. Interestingly, no resistance was recovered through UV irradiation or Agrobacterium tumefaciens-mediated mutagenesis. This research indicated widespread resistance to thiophanate-methyl and diethofencarb, low frequency of (6.0%) resistance to procymidone in A. alternata populations from tobacco, and suggested that azoxystrobin could potentially constitute a good alternative for the management of tobacco brown spot disease.

Effect of pre-storage fungicidal treatment on prevalence of seed-borne fungi and quality of onion (Allium cepa L.) seeds was evaluated under laboratory condition. Seeds were treated separately with five fungicides namely Bavistin 50 WP (Carbendazim) and Dithane M-45 (Mancozeb) @ 2.2 g/kg, Homai (Thiophanate) and Rovral (Iprodione) @ 2.0 g/kg and Provax-200 (Carboxin) @ 3.0 g/kg of seeds before storage. Prevalence of seed-borne fungi, percent seeds germination and vigor index were evaluated just before storage, 75 and 150 days after storage. Fungicides produced significant negative impact on seed-borne fungi and positive impact in maintaining quality during storage. Provax-200, Rovral, Dithane M-45 and Bavistin effectively reduced seed-borne fungi and performed better for germination and vigor of seeds. Seed treatment with Provax-200, Rovral, Bavistin or Dithane M-45 would therefore be suggested as effective fungicides to control seed-borne fungal infection and maintain better germination and vigor of onion seed in storage.The Agriculturists 2017; 15(2) 52-58

The causal agents of apple bull?s-eye rot in southern Brazil have recently been described as Neofabraea actinidiae and N. brasiliensis. Isolates of both species were evaluated for response of mycelial growth index (MGI) to different temperatures, enzyme production, mycelial growth inhibition and effective concentrations (EC 50 and EC 100 ) of the fungicides tri fl umizole, pyrimethanil and thiophanate methyl, as well as aggres- siveness on fruits of "Fuji " hybrid and " Pink Lady " . There was signi ficantly lower mycelium growth in N. brasiliensis compared with N. actinidiae at all temperatures tested. Neither species grew at 3 and 32°C. There were minor differences in production of enzymes in the two species, with all N. brasiliensis isolates showing no production of pectolyase at pH 7. The lowest EC 50 and EC 100 values were observed with thiophanate methyl. In general, " Fuji " fruits were more susceptible to Neofabraea infection and had larger lesions, while N. brasiliensis isolates showed greater aggressiveness on " Fuji " hybrid and " Pink lady " fruits compared with N. actinidiae. Made available in DSpace on 2018-03-03T00:34:48Z (GMT). No. of bitstreams: 1 CharacterizationofNeofabraeaactinidiaeandNbrasiliensisascausalagentsofapplebullseyerotinsouthernBrazil.pdf: 563520 bytes, checksum: a0be3950124fd2c673c066146382a841 (MD5) Previous issue date: 2018-03-02

Anthracnose, caused by Colletotrichum musae, is the most important postharvest disease of banana and is widely distributed among the banana production regions in Brazil. Although thiophanate-methyl is a fungicide frequently used in Brazilian banana orchards to control Sigatoka leaf spot, Collettotrichum populations are also exposed, resulting in the evolution of fungicide resistance and the inability to manage banana anthracnose. We investigated 139 Brazilian isolates of C. musae for thiophanate-methyl sensitivity in vitro. The 50% mycelial growth inhibition (EC50) values varied between 0.003 and 48.73 μg/ml. One-hundred and thirty isolates were classified as sensitive, with EC50 values ranging from 0.003 to 4.84 μg/ml, while the remaining nine isolates were considered moderately resistant, with EC50 values ranging between 10.43 and 48.73 μg/ml. Resistant or highly resistant isolates (EC50 > 100 μg/ml) were not found. A substitution of TAC for TTC at codon 200 in a coding region of the β-tubulin gene was associated with the moderately resistant phenotype. Applications of thiophanate-methyl formulation to detached banana fruit at the label rate (500 μg/ml) showed low efficacy in controlling the moderately resistant isolates on banana fruits. However, there is no indication of a reduction in fitness associated with fungicide resistance as sensitive and moderately resistant isolates do not differ with respect to mycelial growth rate (P = 0.098), spore production (P = 0.066), spore germination (P = 0.366), osmotic sensitivity (P = 0.051), and virulence (P = 0.057). Our results revealed absence of adaptability cost for the moderately resistant isolates, suggesting that they can be dominant in population if the fungicide continue to be applied.

This research aims at determining residues of thiophanate methyl and propiconazole in grape and mango fruits as an indication for their persistence in this environmental compartment. Fruit extracts were analyzed for thiophanate methyl using High Performance Liquid Chromatography and using Gas Chromatography Electron Capture Detector (GC/ECD), respectively. The results indicated that propiconazole had a less environmental impact since propiconazole had shorter residue half-lives which were 1.24 and 1.19 days in grape and mango fruits, respectively, while thiophanate methyl had half-lives of 2.49 and 2.64 days in mango and grape, respectively. The degradation rates of propiconazole in grape and mango fruits did not change significantly and neither did those of thiophanate methyl. According to the maximum residue level, the pre-harvest intervals of propiconazole were set to be 3 and 7 days for grape and mango fruits, respectively, and the pre-harvest intervals for thiophanate methyl were 15 days for both grape and mango fruits. Propiconazole was generally considered to be less hazardous to humans and will leave the environment less altered because of its faster degradation than that of thiophanate methyl.

Surface-enhanced Raman scattering (SERS) imaging coupling with multivariate analysis in spectral region of 200 to 1800cm-1 was developed to quantify and visualize thiophanate-methyl (TM) and its metabolite carbendazim residues in red bell pepper (Capsicum annuum L.). Least squares support vector machines (LS-SVM) and support vector machines (SVM) models based on seven optimized characteristic peaks that showed SERS effects of TM and its metabolite carbendazim residues were employed to establish prediction models. SERS spectra with first derivative (1st) and second derivative (2nd) method were subsequently compared and the optimized model of 1st-LS-SVM acquired showed the best performance (RPD=6.08, R2P=0.986 and RMSEP=0.473). The results demonstrated that SERS imaging with multivariate analysis had the potential for rapid determination and visualization of the trace TM and its metabolite carbendazim residues in complex food matrices.

Thiophanate methyl is a broad-spectrum benzimidazole fungicide extensively applied in pre- and post-harvest, for the control of a wide range of fruit and vegetable pathogens. In the current work, the residue behavior of thiophanate methyl after application on strawberries and an estimation of the consumer dietary exposure was performed. Supervised field trials were conducted in Egypt (Qaluobiya Governorate) as to investigate the residue dynamics and terminal residues at different PHIs of thiophanate methyl and its metabolite carbendazim in strawberries under Egyptian conditions. For the measurement of residues in fruits, a QuEChERS-based protocol coupled with LC–MS/MS was optimized and successfully validated at 0.01 mg kg−1. The half-life (t1/2) of thiophanate methyl in strawberries was estimated, and a dietary risk assessment was performed employing both FAO/WHO and EFSA approaches.

In this study, a mycelial growth assay was used to evaluate the sensitivity to thiophanate-methyl of 144

Chlorothalonil and thiophanate-methyl are fungicides widely used in agriculture. The aim of this study was to assess maternal toxicity and embryotoxic potential of exposure to chlorothalonil and thiophanate-methyl during organogenesis period in rats. Pregnant rats were divided into four groups: control and exposed to 400 (CT400), 800 (CT800) and 1200 mg-1kg bw-1 day (CT1200) of commercial formulation constituted of 200 g of thiophanate-methyl kg−1 and 500 g of chlorothalonil kg−1 by gavage, from 6th to 15th gestational day. Maternal toxicity, liver, kidney and placenta histology, reproductive performance, and external, skeletal and visceral malformations of fetuses were evaluated. Maternal liver weight was decreased in CT1200 group and focal necrosis and microvesicular steatosis, inflammatory infiltrate and hepatocytes with pyknotic nucleus were observed in CT800 and CT1200 groups. Reproductive performance was similar among groups. The percentage of fetuses small for pregnancy age was increase in CT400 and CT800 groups. Moreover, incidence of skeletal anomalies was increased in the three groups exposed to fungicides. Chlorothalonil and thiophanate-methyl exposure showed affect the prenatal development and induce maternal toxicity.

Winter is the key period for the control of apple diseases, and fungicides are needed to protect the trunk or main branches. Fungicide residue in apple tree bark is an important basis for the action of the pesticide, but there are no reports on analytical methods or dissipation patterns. In this work, thiophanate-methyl, carbendazim, tebuconazole and pyraclostrobin were selected as typical fungicides and a new QuEChERS-HPLC-VWD(QuEChERS extraction followed by high-performance liquid chromatography detection with a variable wavelength detector) analytical method was developed to estimate their residue kinetics in apple tree bark during the winter months. In the pretreatment step, the sorbent for the clean-up of extracts was optimized as 60 mg/ml primary secondary amine and a gradient-elution model followed by a variable wavelength detection was developed for instrumental analysis. Then this method was validated and applied to the analysis of apple tree bark samples with the linearity range of 0.010-50.00 mg/L, quantification limit range of 0.028-0.080 mg/kg and recovery range of 86.1-101.4%. The dissipation kinetics of thiophanate-methyl and pyraclostrobin could be described by the first-order and two-phase kinetics models, respectively. For carbendazim and tebuconazole, two new models were developed to describe their residue kinetics.

A multi-residue method for the determination of 107 pesticide residues in wolfberry has been developed and validated. Similar pretreatment approaches were compared, and the linearity, matrix effect, analysis limits, precision, stability and accuracy were validated, which verifies the satisfactory performance of this new method. The LODs and LOQs were in the range of 0.14&ndash, 1.91 &micro, g/kg and 0.46&ndash, 6.37 &micro, g/kg, respectively. The recovery of analytes at three fortification levels (10 &micro, g/kg, 50 &micro, g/kg, 100 &micro, g/kg) ranged from 63.3&ndash, 123.0%, 72.0&ndash, 118.6% and 67.0&ndash, 118.3%, respectively, with relative standard deviations (RSDs) below 15.0%. The proposed method was applied to the analysis of fifty wolfberry samples collected from supermarkets, pharmacies and farmers&rsquo, markets in different cities of Shandong Province. One hundred percent of the samples analyzed included at least one pesticide, and a total of 26 pesticide residues was detected in fifty samples, which mainly were insecticides and bactericide. Several pesticides with higher detection rates were 96% for acetamiprid, 82% for imidacloprid, 54% for thiophanate-methyl, 50% for blasticidin-S, 42% for carbendazim, 42% for tebuconazole and 36% for difenoconazole in wolfberry samples. This study proved the adaptability of the developed method to the detection of multiple pesticide residues in wolfberry and provided basis for the research on the risks to wolfberry health.

Stem-end rot caused by Lasiodiplodia theobromae is one of the most devastating diseases of papaya in northeastern Brazil. It is most effectively controlled by applications of fungicides, including site-specific fungicides at risk for resistance development. This study investigated the molecular mechanisms of reduced sensitivity to the QoI fungicide azoxystrobin and resistance to the MBC fungicide thiophanate-methyl in L. theobromae from Brazilian orchards. The EC50 values for azoxystrobin in sixty-four isolates ranged from 0.36 μg/ml to 364.24 μg/ml and the frequency distribution of EC50 values formed a multimodal curve, indicating reduced sensitivity to azoxystrobin. In detached fruit assays reduced sensitive isolates were not controlled as effectively as sensitive isolates at lowest label rate. Partial fragments were obtained from target genes β-tubulin (751 bp) and Cytb (687 bp) of isolates resistant to thiophanate-methyl and reduced sensitivity to azoxystrobin. Sequence analysis of the β-tubulin fragment revealed a mutation corresponding to E198K in all thiophanate-methyl-resistant isolates, while reduced sensitivity to axoxystrobin was not attributable to Cytb gene alterations. The target gene-based mechanism conferring resistance to thiophanate-methyl will likely be stable even if selection pressure subsides. However, the mechanism conferring reduced sensitivity to azoxystrobin is not based on target gene modifications and thus may not be as stable as other genotypes with mutations in Cytb gene.

Residue analysis to detect thiophanate-methyl and its primary metabolite (carbendazim) during oyster mushroom (Pleurotus ostreatus var. florida) cultivation was done for two consecutive years 2017 and 2018. Wheat straw substrate was chemically treated with different treatments of thiophate-methyl, viz, thiophanate-methyl 30 ppm + formalin 500 ppm (T1), thiophanate-methyl 40 ppm + formalin 500 ppm (T2), thiophanate-methyl 50 ppm + formalin 500 ppm (T3), thiophanate-methyl 60 ppm + formalin 500 ppm (T4), and formalin 500 ppm (T5 as control and recommended concentration), and utilized for cultivation of oyster mushroom. Treatments T3 and T4 exhibited significant difference in pH levels during both the trials. Minimum spawn run, pinhead formation, and fruit body formation time were recorded in treatments T3 and T4. Significantly higher biological efficiency (%) was recorded in treatments T3 and T4 as compared with all other treatments. No incidence of competitor molds was recorded in T3 and T4. Pesticide residue analysis for detection of thiophanate-methyl and its metabolite (carbendazim) was done in the fruit body produced in T3 and T4 treatments using liquid chromatography with tandem mass spectrometry method. No residue of thiophanate-methyl and carbendazim was detected at 50 ppm concentration of thiophanate-methyl during both the trials. However, in trial II, residue of carbendazim (5.39 μg/kg) was detected at 60 ppm. Based on the findings of the trials I and II, T3 (thiophanate-methyl 50 ppm + formalin 500 ppm) may be utilized for substrate sterilization for oyster mushroom cultivation and Pleurotus ostreatus var. florida could be recognized as microorganism which could play a role in degradation of thiophanate-methyl.

Mancozeb (MZ), chlorothalonil (CT), and thiophanate methyl (TM) are pesticides commonly used in agriculture due to their efficacy, low acute toxicity to mammals, and short environmental persistence. Although the toxic effects of these pesticides have been previously reported, studies regarding their influence on the immune system are limited. As such, this study focused on the immunomodulatory effect of MZ, CT, and TM pesticides on macrophage cells. RAW 264.7 cells were exposed to a range of concentrations (0.1–100 μg/mL) of these pesticides. CT exposure promoted an increase in reactive oxygen species (ROS) and nitric oxide (NO) levels. The MTT and ds-DNA assay results demonstrated that MZ, CT, and TM exposure induced macrophage proliferation. Moreover, MZ, CT, and TM promoted cell cycle arrest at S phase, strongly suggesting macrophage proliferation. The levels of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, and IFN-γ) and caspases (caspase 1, 3, and 8) in macrophages exposed to MZ, CT, and TM pesticides increased, whereas the anti-inflammatory cytokine levels decreased. These results suggest that MZ, CT, and TM exert an immunomodulatory effect on the immune system, inducing macrophage activation and enhancing the inflammatory response.