Environmental levels of azoxystrobin disturb male zebrafish behavior: Possible roles of oxidative stress, cholinergic system, and dopaminergic system.

A widely applied pesticide of azoxystrobin, is increasingly detected in the water environment. Concern has been raised against its potential detriment to aquatic ecosystems. It has been shown that exposure to azoxystrobin interfere with the locomotor behavior of zebrafish larvae. This study aims to investigate whether exposure to environmental levels of azoxystrobin (2 µg/L, 20 µg/L, and 200 µg/L) changes the behavior of male adult zebrafish. Herein, we evaluated behavioral response (locomotor, anxiety-like, and exploratory behaviors), histopathology, biochemical indicators, and gene expression in male adult zebrafish upon azoxystrobin exposure. The study showed that exposure to azoxystrobin for 42 days remarkably increased the locomotor ability of male zebrafish, resulted in anxiety-like behavior, and inhibited exploratory behavior. After treatment with 200 µg/L azoxystrobin, vasodilatation, and congestion were observed in male zebrafish brains. Exposure to 200 µg/L azoxystrobin notably elevated ROS level, MDA concentration, CAT activity, and AChE activity, while inhibiting SOD activity, GPx activity, ACh concentration, and DA concentration in male zebrafish brains. Moreover, the expression levels of genes related to the antioxidant, cholinergic, and dopaminergic systems were significantly changed. This suggests that azoxystrobin may interfere with the homeostasis of neurotransmitters by causing oxidative stress in male zebrafish brains, thus affecting the behavioral response of male zebrafish.

Comparative fitness of Monilinia fructicola isolates with multiple fungicide-resistant phenotypes.

This study characterized 52 isolates of Monilinia fructicola from peach and nectarine orchards for their multi-resistance patterns to thiophanate-methyl (TF), tebuconazole (TEB), and azoxystrobin (AZO) using in vitro sensitivity assays and molecular analysis. The radial growth of M. fructicola isolates was measured on media amended with a single discriminatory dose of 1 µg/ml for TF and AZO and 0.3 µg/ml for TEB. Cyt b, CYP51, and ß-tubulin were tested for point mutations that confer resistance to quinone outside inhibitors (QoIs), demethylation inhibitors (DMIs), and methyl benzimidazole carbamates (MBCs), respectively. Eight phenotypes were identified including isolates with single, double, and triple in vitro resistance to QoI, MBC, and DMI fungicides. All resistant phenotypes to TF and TEB presented the H6Y mutation in ß-tubulin and the G641S mutation in CYP51. None of the point mutations typically linked to QoI resistance were present in the Monilinia isolates examined. Moreover, fitness of the M. fructicola phenotypes was examined in vitro and detached fruit assays. Phenotypes with single-resistance displayed equal fitness in in vitro and fruit assays compared to the wild-type. In contrast, the dual and triple-resistance phenotypes suffered fitness penalties based on osmotic sensitivity and aggressiveness on peach fruit. In this study, multiple resistance to MBC, DMI, and QoI fungicide groups was confirmed in M. fructicola. Results suggest that Monilinia populations with multiple resistance phenotypes are likely to be less competitive in the field than those with single resistance, thereby impeding their establishment over time and facilitating disease management.

Response of Soil Microbiota, Enzymes, and Plants to the Fungicide Azoxystrobin.

The present study was aimed at assessing the impact of azoxystrobin-a fungicide commonly used in plant protection against pathogens (Amistar 250 SC)-on the soil microbiota and enzymes, as well as plant growth and development. The laboratory experiment was conducted in three analytical terms (30, 60, and 90 days) on sandy clay (pH-7.0). Azoxystrobin was applied to soil in doses of 0.00 (C), 0.110 (F) and 32.92 (P) mg kg-1 d.m. of soil. Its 0.110 mg kg-1 dose stimulated the proliferation of organotrophic bacteria and actinobacteria but inhibited that of fungi. It also contributed to an increase in the colony development index (CD) and a decrease in the ecophysiological diversity index (EP) of all analyzed groups of microorganisms. Azoxystrobin applied at 32.92 mg kg-1 reduced the number and EP of microorganisms and increased their CD. PP952051.1 Bacillus mycoides strain (P), PP952052.1 Prestia megaterium strain (P) bacteria, as well as PP952052.1 Kreatinophyton terreum isolate (P) fungi were identified in the soil contaminated with azoxystrobin, all of which may exhibit resistance to its effects. The azoxystrobin dose of 0.110 mg kg-1 stimulated the activity of all enzymes, whereas its 32.92 mg kg-1 dose inhibited activities of dehydrogenases, alkaline phosphatase, acid phosphatase, and urease and stimulated the activity of catalase. The analyzed fungicide added to the soil at both 0.110 and 32.92 mg kg-1 doses inhibited seed germination and elongation of shoots of Lepidium sativum L., Sinapsis alba L., and Sorgum saccharatum L.

The Combined Effects of Azoxystrobin and the Biosurfactant-Producing Bacillus sp. Kol B3 against the Phytopathogenic Fungus Fusarium sambucinum IM 6525.

This study aimed to evaluate how the combined presence of the synthetic fungicide azoxystrobin (AZ) and the biosurfactant-producing Bacillus sp. Kol B3 influences the growth of the phytopathogenic fungus Fusarium sambucinum IM 6525. The results showed a noticeable increase in antifungal effectiveness when biotic and abiotic agents were combined. This effect manifested across diverse parameters, including fungal growth inhibition, changes in hyphae morphology, fungal membrane permeability and levels of intracellular reactive oxygen species (ROS). In response to the presence of Fusarium and AZ in the culture, the bacteria changed the proportions of biosurfactants (surfactin and iturin) produced. The presence of both AZ and/or Fusarium resulted in an increase in iturin biosynthesis. Only in 72 h old bacterial-fungal co-culture a 20% removal of AZ was noted. In the fungal cultures (with and without the addition of the bacteria), the presence of an AZ metabolite named azoxystrobin free acid was detected in the 48th and 72nd hours of the process. The possible involvement of increased iturin and ROS content in antifungal activity of Bacillus sp. and AZ when used together are also discussed. Biosurfactants were analyzed by liquid chromatography with tandem mass spectrometry (LC-MS/MS). Microscopy techniques and biochemical assays were also used.

Recommended rates of azoxystrobin and tebuconazole seem to be environmentally safe but ineffective against target fungi.

The use of fungicides in agriculture has been playing a role in the enhancement of agricultural yields through the control of pathogens causing serious diseases in crops. Still, adverse environmental and human health effects resulting from its application have been reported. In this study, the possibility of readjusting the formulation of a commercial product combining azoxystrobin and tebuconazole (active ingredients - AIs; Custodia®) towards environmentally safer alternative(s) was investigated. Specifically, the sensitivity of non-target aquatic communities to each AI was first evaluated by applying the Species Sensitivity Distributions (SSDs) approach. Then, mixtures of these AIs were tested in a non-target organism (Raphidocelis subcapitata) denoting sensitivity to both AIs as assessed from SSDs. The resulting data supported the design of the last stage of this study, where mixtures of those AIs at equivalent vs. alternative ratios and rates as in the commercial formulation were tested against two target fungal species: Pyrenophora teres CBS 123929 and Rhynchosporium secalis CBS 110524. The comparison between the sensitivity of non-target aquatic species and the corresponding efficacy towards target fungi revealed that currently applied mixture and rates of these AIs are generally environmentally safe (antagonistic interaction; concentrations below the EC1 for R. subcapitata and generally below the HC5 for aquatic non-target communities), but ineffective against target organisms (maximum levels of inhibition of 70 and 50% in P. teres CBS 123929 and R. secalis CBS 110524, respectively). Results additionally suggest a potentiation of the effects of the AIs by the other formulants added to the commercial product at tested rates. Overall, this study corroborates that commercial products can be optimized during design stages based on a systematic ecotoxicological testing for ingredient interactions and actual efficacy against targets. This could be a valuable pathway to reduce environmental contamination during transition to a more sustainable agricultural production.

Characterization of mutants with single and combined Qi and Qo site mutations in Saccharomyces cerevisiae reveals interactions between the picolinamide fungicide CAS-649 and azoxystrobin.

Picolinamide and strobilurin fungicides bind to the Qi and Qo sites on cytochrome b, respectively, and target many of the same plant pathogens. Using Saccharomyces cerevisiae as a model system, we explore effects of amino acid changes at each site on sensitivity to a fungicide acting at the opposite site and examine the relationship between altered sensitivity and growth penalty. In addition, double mutants containing the G143A or F129L mutations responsible for strobilurin resistance in combination with Qi site mutations that confer resistance to picolinamides are characterized in terms of their sensitivity to QiI and QoI fungicides and growth rate. Mutants containing amino acid changes at the Qo site varied in their growth rate and sensitivity to the picolinamide CAS-649, and increased sensitivity was associated with a greater growth penalty. Conversely, changes at the Qi site affected sensitivity to azoxystrobin and also showed a correlation between increased sensitivity and reduced growth. There was no overall correlation between resistance to azoxystrobin and CAS-649 among mutants, however negative cross-resistance occurred in the case of mutations which conferred resistance to either compound and also carried a growth penalty. These results suggest the use of QoI fungicides to delay the emergence of pathogen resistance to QiIs, and vice versa. Double mutants containing G143A or F129L in combination with Qi site changes N31K, G37C/V or L198F that cause resistance to picolinamides generally exhibited lower resistance factors for both azoxystrobin and CAS-649 than corresponding resistant strains with a single mutation. Reduced growth was observed for all F129L-containing double mutants, whereas the growth rate of double mutants containing G143A was significantly reduced only by the Qi site mutations N31K and G37V that confer a larger growth penalty. Our results suggest that resistance to picolinamides in pathogens could emerge more readily in a strobilurin-sensitive genetic background than in a strobilurin-resistant one.

An integrated approach for evaluating the interactive effects between azoxystrobin and ochratoxin A: Pathway-based in vivo analyses.

Azoxystrobin (AZO) is a broad-spectrum strobilurin fungicide widely used in agriculture. However, its use increases the possibility of co-occurrence with mycotoxins such as ochratoxin A (OTA), which poses a significant risk to human health. Therefore, it is imperative to prioritize the evaluation of the combined toxicity of these two compounds. To assess the combined effects of AZO and OTA, the response genes and phenotypes for AZO or OTA exposure were obtained by utilizing Comparative Toxicogenomics Database, and Database for Annotation, Visualization and Integrated Discovery was used for GO and KEGG pathway enrichment analysis. In addition, we provided in-vivo evidence that AZO and OTA, in isolation and combination, could disrupt a variety of biological processes, such as oxidative stress, inflammatory response, apoptosis and thyroid hormone regulation under environmentally relevant concentrations. Notably, our findings suggest that the combined exposure group exhibited greater toxicity, as evidenced by the expression of various markers associated with the aforementioned biological processes, compared to the individual exposure group, which presents potential targets for the underlying mechanisms of induced toxicity. This study provides a novel methodological approach for exploring the mechanism of combined toxicity of a fungicide and a mycotoxin, which can shed light for conducting risk assessment of foodborne toxins.

Molecular Characterization and Fungicide Sensitivity of Jujube Pathogens Colletotrichum gloeosporioides Sensu Stricto and Colletotrichum nymphaeae in South Korea.

Jujube (Ziziphus jujuba) is cultivated across South Korea because of its medicinal and economic value. It is used as a sweetener in jam, tea, and snacks and a garnish in many cuisines. Anthracnose caused by Colletotrichum spp. accounts for huge economic losses for jujube growers. In 2019 and 2020, severe anthracnose was observed in the jujube-growing areas of South Korea. The infected fruit displayed small, water-soaked, sunken, circular spots. Infected fruit were collected from different commercial orchards of Boeungun and Gyeongsan regions of South Korea, and putative causal agents were isolated on potato dextrose agar. Based on the morphological and molecular characteristics, the fungal isolates were identified as Colletotrichum gloeosporioides sensu stricto and C. nymphaeae. The pathogenicity of these isolates was confirmed by inoculating a conidial suspension (1 × 106 conidia ml-1) on healthy fruit. The in vitro sensitivity of the fungal isolates to tebuconazole, carbendazim, and azoxystrobin was also tested. All isolates showed high sensitivity to azoxystrobin in terms of mycelial growth inhibition (half maximal effective concentration value of 0.01 to 0.6 µg/ml). To the best of our knowledge, this is also the first report of jujube anthracnose caused by C. nymphaeae in South Korea.

Determining the Distribution of QoI Fungicide-Resistant Cercospora sojina on Soybean from Indiana.

Frogeye leaf spot (FLS) is a foliar disease of soybean (Glycine max) caused by Cercospora sojina. Application of fungicide products that contain quinone outside inhibitor (QoI) active ingredients has been one of the major tools used in the management of this disease, but, since 2010, QoI-resistant C. sojina isolates have been confirmed in over 20 states in the United States, including Indiana. In summer 2019 and 2020, 406 isolates of C. sojina were collected from 32 counties across Indiana and screened for QoI resistance using a PCR-restriction fragment length polymorphism (RFLP) method. An in vitro fungicide sensitivity test was also performed on a subset of isolates to evaluate their sensitivity to three QoI fungicides: azoxystrobin, pyraclostrobin, and picoxystrobin. A discriminatory dose of picoxystrobin was established as 1 µg/ml by testing five concentrations (0.001, 0.01, 0.1, 1, and 10 µg/ml). QoI-resistant isolates were found in 29 counties, and 251 of the 406 isolates (61.8%) were confirmed to be resistant to QoI fungicides based on PCR-RFLP results. Partial nucleotide sequences of the cytochrome b gene from four resistant and four sensitive isolates corroborated the presence and absence, respectively, of the G143A mutation. Results from the sensitivity assays with discriminatory doses of azoxystrobin (1 µg/ml) and pyraclostrobin (0.1 µg/ml) also supported the findings from the PCR-RFLP assay, because all QoI-resistant isolates were inhibited less than 50% relative to a no-fungicide control when exposed to these doses. Resistant isolates harboring the G143A mutation also exhibited resistance to picoxystrobin. The effective concentrations to inhibit mycelial growth by 50% relative to the nonamended control (EC50) in QoI-sensitive isolates ranged from 0.087 to 0.243 µg/ml, with an overall mean of 0.152 µg/ml, while EC50 values in QoI-resistant isolates were established as >10 µg/ml for picoxystrobin. Results from this study indicated that QoI-resistant C. sojina isolates are spread throughout Indiana and exhibit cross-resistance to QoI fungicides.

Ternary Mixture of Azoxystrobin, Boscalid and Pyraclostrobin Disrupts the Gut Microbiota and Metabolic Balance of Honeybees (Apis cerana cerana).

There is a growing risk of pollinators being exposed to multiple fungicides due to the widespread use of fungicides for plant protection. A safety assessment of honeybees exposed to multiple commonly used fungicides is urgently required. Therefore, the acute oral toxicity of the ternary mixed fungicide of ABP (azoxystrobin: boscalid: pyraclostrobin = 1:1:1, m/m/m) was tested on honeybees (Apis cerana cerana), and its sublethal effect on foragers' guts was evaluated. The results showed that the acute oral median lethal concentration (LD50) of ABP for foragers was 12.6 µg a.i./bee. ABP caused disorder of the morphological structure of midgut tissue and affected the intestinal metabolism; the composition and structure of the intestinal microbial community was perturbed, which altered its function. Moreover, the transcripts of genes involved in detoxification and immunity were strongly upregulated with ABP treatment. The study implies that exposure to a fungicide mixture of ABP can cause a series of negative effects on the health of foragers. This work provides a comprehensive understanding of the comprehensive effects of common fungicides on non-target pollinators in the context of ecological risk assessment and the future use of fungicides in agriculture.

Response of soil phosphatase activity and soil phosphorus fractions to the application of chloropicrin and azoxystrobin in ginger cultivation.

BACKGROUND: Soil fumigation can change soil nutrient cycling processes by affecting soil beneficial microorganisms, which is a key issue for soil fertility. However, the effect of combined application of fumigant and fungicide on soil phosphorus (P) availability remains largely unclear. We investigated the effects of the fumigant chloropicrin (CP) and the fungicide azoxystrobin (AZO) on soil phosphatase activity and soil P fractions in ginger production using a 28-week pot experiment with six treatments: control (CK), a single application of AZO (AZO1), double applications of AZO (AZO2), CP-fumigated soil without AZO (CP), CP combined with AZO1 (CP + AZO1) and CP combined with AZO2 (CP + AZO2). RESULTS: AZO application alone significantly increased the soil labile P fractions (Resin-P + NaHCO3 -Pi + NaOH-Pi) at 9 weeks after planting (WAP) but decreased the soil phosphatase activity at 28 WAP. CP fumigation significantly reduced the soil phosphatase activity but increased the proportions of soil labile P fractions (Resin-P + NaHCO3 -Pi + NaHCO3 -Po) to total P (TP) by 9.0-15.5% throughout the experiment. The combined application of CP and AZO had a synergistic effect on soil phosphatase activity and soil P fractions compared with a single application. CONCLUSION: Although AZO application and CP fumigation can increase soil available P in the short term, they might negatively affect soil fertility in the long run by inhibiting soil phosphatase activity. Soil microbial activities, especially microorganisms related to P cycling, may be responsible for the variations in soil P availability, but further research is needed. © 2023 Society of Chemical Industry.

Biochemical compounds and stress markers in lettuce upon exposure to pathogenic Botrytis cinerea and fungicides inhibiting oxidative phosphorylation.

MAIN CONCLUSION: Botrytis cinerea and fungicides interacted and influenced selected biochemical compounds. DPPH and glutathione are the first line of defence against biotic/abiotic stress. Plant metabolites are correlated with fungicides level during dissipation. Botrytis cinerea is an etiological agent of gray mould in leafy vegetables and is combated by fungicides. Fluazinam and azoxystrobin are commonly used fungicides, which inhibit oxidative phosphorylation in fungi. In this study, lettuce was (i) inoculated with B. cinerea; (ii) sprayed with azoxystrobin or fluazinam; (iii) inoculated with B. cinerea and sprayed with fungicides. This investigation confirmed that B. cinerea and fungicides affected lettuce's biochemistry and stress status. B. cinerea influenced the behaviour of fungicides reflected by shortened dissipation of azoxystrobin compared to non-inoculated plants, while prolonged degradation of fluazinam. Stress caused by B. cinerea combined with fungicides reduced level of chlorophylls (53.46%) and carotenoids (75.42%), whereas increased phenolic compounds (81%), ascorbate concentrations (32.4%), and catalase activity (116.1%). Abiotic stress caused by fungicides contributed most to the induction of carotenoids (107.68 µg g-1 on dissipation day 3-1). Diphenyl picrylhydrazyl (DPPH) radical scavenging activity and glutathione concentration peaked from the first hour of fungicides dissipation. For the first time correlation between the status of plant metabolites and fungicides during their dissipation was observed. These results indicate that non-enzymatic antioxidants could be the first-line compounds against stress factors, whereas ascorbate and antioxidant enzymes tend to mitigate stress only secondarily. The findings of this study help better understand plant biochemistry under biotic/abiotic stress conditions.

Structural Elucidation of Agrochemicals and Related Derivatives Using Infrared Ion Spectroscopy.

Agrochemicals frequently undergo various chemical and metabolic transformation reactions in the environment that often result in a wide range of derivates that must be comprehensively characterized to understand their toxicity profiles and their persistence and outcome in the environment. In the development phase, this typically involves a major effort in qualitatively identifying the correct chemical isomer(s) of these derivatives from the many isomers that could potentially be formed. Liquid chromatography-mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy are often used in attempts to characterize such environment transformation products. However, challenges in confidently correlating chemical structures to detected compounds in mass spectrometry data and sensitivity/selectivity limitations of NMR frequently lead to bottlenecks in identification. In this study, we use an alternative approach, infrared ion spectroscopy, to demonstrate the identification of hydroxylated derivatives of two plant protection compounds (azoxystrobin and benzovindiflupyr) contained at low levels in tomato and spinach matrices. Infrared ion spectroscopy is an orthogonal tandem mass spectrometry technique that combines the sensitivity and selectivity of mass spectrometry with structural information obtained by infrared spectroscopy. Furthermore, IR spectra can be computationally predicted for candidate molecular structures, enabling the tentative identification of agrochemical derivatives and other unknowns in the environment without using physical reference standards.

Uptake and Translocation of a Silica Nanocarrier and an Encapsulated Organic Pesticide Following Foliar Application in Tomato Plants.

Pesticide nanoencapsulation and its foliar application are promising approaches for improving the efficiency of current pesticide application practices, whose losses can reach 99%. Here, we investigated the uptake and translocation of azoxystrobin, a systemic pesticide, encapsulated within porous hollow silica nanoparticles (PHSNs) of a mean diameter of 253 ± 73 nm, following foliar application on tomato plants. The PHSNs had 67% loading efficiency for azoxystrobin and enabled its controlled release over several days. Thus, the nanoencapsulated pesticide was taken up and distributed more slowly than the nonencapsulated pesticide. A total of 8.7 ± 1.3 µg of the azoxystrobin was quantified in different plant parts, 4 days after 20 µg of nanoencapsulated pesticide application on a single leaf of each plant. In parallel, the uptake and translocation of the PHSNs (as total Si and particulate SiO2) in the plant were characterized. The total Si translocated after 4 days was 15.5 ± 1.6 µg, and the uptake rate and translocation patterns for PHSNs were different from their pesticide load. Notably, PHSNs were translocated throughout the plant, although they were much larger than known size-exclusion limits (reportedly below 50 nm) in plant tissues, which points to knowledge gaps in the translocation mechanisms of nanoparticles in plants. The translocation patterns of azoxystrobin vary significantly following foliar uptake of the nanosilica-encapsulated and nonencapsulated pesticide formulations.

Virulence of native isolates of entomopathogenic fungi (Hypocreales) against the "sweetpotato whitefly" Bemisia tabaci (Hemiptera: Aleyrodidae), including the effects of temperature and fungicides.

Hypocrella, Moelleriella and related species in the Hypocreales (Ascomycota, Sordariomycetes) cause epizootics of whiteflies and scale insects in nature. However, studies on their host specificity, virulence, infection cycles, optimal development under laboratory conditions, and compatibility with other control methods, are unexplored for most species. Under laboratory conditions, the virulence of several isolates of field-collected hypocrealean fungi (Hypocrella, Moelleriella, Regiocrella, and Verticillium) was determined on Bemisia tabaci eggs and 4th instar nymphs. In addition to virulence, the effect of temperature and two commercial fungicides on growth rates and germination of the isolates was evaluated. None of the isolates infected the eggs, while M. libera, M. ochracea, and M. turbinata caused high nymphal mortality. Moelleriella libera was the most virulent isolate. At all temperatures, M. libera, Regiocrella sp. (P17H20), and Verticillium cf. pseudohemipterigenum had the highest germination and growth rates. The optimal growth temperature depended on the isolate, but at 23 °C and 25 °C, the probability of spore germination was higher for most isolates. Finally, the fungicides azoxystrobin and chlorothalonil inhibited growth rates and conidial germination at 24 and 48 h of exposure. This research produces vital knowledge on the virulence and infection cycles of poorly studied native species of entomopathogenic fungi. In addition, the results provide information on the optimal temperature for development in laboratory conditions and susceptibility to fungicides, which could contribute to future biological control strategies.

Effects of chloropicrin fumigation and azoxystrobin application on ginger growth and phosphorus uptake.

Soil chloropicrin (CP) fumigation helps to increase crop yields by eliminating soil-borne diseases which inhibit plant growth. However, little is known about the effect of the CP fumigation combined with fungicide application on plant growth and nutrient uptake. In this study, we conducted a mesocosm experiment with six treatments: CK (untreated soil), AZO1 (a single application of azoxystrobin (AZO)), AZO2 (double applications of AZO), CP (CP fumigation with no AZO), CP+AZO1 (CP combined with AZO1) and CP+AZO2 (CP combined with AZO2) to investigate the effects of CP fumigation and AZO application on ginger growth and phosphorus (P) uptake. Results showed that a single application of AZO had no significant effect on ginger height, biomass and P uptake whether treated with or without CP fumigation, whereas double applications of AZO combined with CP fumigation significantly improved ginger height and the total amount of P in root (P < 0.05). Meanwhile, AZO residues were similar in all treatments with the same number of applications, with less than 50% remaining in the soil after 7 days applied, indicating that CP fumigation treatment did not influence AZO degradation in ginger cultivation. In addition, although the differences in P use efficiency observed across the different treatments were not significant, they nevertheless suggest that the P budget and soil microbial activity may contribute to those differences. Therefore, further studies should be done to link P cycling with microbial communities, and how these related to fumigation and fungicide application.

Baseline Sensitivity and Control Efficacy of Two Quinone Outside Inhibitor Fungicides, Azoxystrobin and Pyraclostrobin, Against Ustilaginoidea virens.

Rice false smut caused by the filamentous fungus Ustilaginoidea virens is a devastating grain disease in rice. Fungicides have been an important measure for the control of this disease. In this study, baseline sensitivities of 179 isolates of U. virens to the quinone outside inhibitor (QoI) fungicides azoxystrobin and pyraclostrobin were established. The distribution of the 50% effective concentration (EC50) values of each fungicide was unimodal. The frequency distribution of logarithmically transformed EC50 values fit or fit closer to a normal distribution. The ranges of EC50 values for azoxystrobin and pyraclostrobin were 0.001 to 0.864 and 0.001 to 0.569 µg/ml, with means and standard errors of the mean values of 0.203 ± 0.012 and 0.079 ± 0.006 µg/ml, respectively. There was a statistically significant and moderately positive correlation (n = 100, r = 0.469, P = 0.001) in sensitivity between these two fungicides. No cross-resistance was found between azoxystrobin, pyraclostrobin, and carbendazim or sterol demethylation inhibitor fungicides. Each fungicide had a significantly higher mean preventive efficacy compared with its curative efficacy. Field assays showed that the control efficacy of pyraclostrobin against rice false smut was greater than that of azoxystrobin. Pyraclostrobin had the best control of rice false smut in three rice varieties, with the control efficacy ranging from 81.5 to 95.5%, whereas azoxystrobin decreased the disease index by 64.1 to 69.2% under the same conditions. These results provide us a reference point in the management of U. virens and future QoI fungicide resistance monitoring programs.

Phytoremediation of azoxystrobin and imidacloprid by wetland plant species Juncus effusus, Pontederia cordata and Sagittaria latifolia.

Azoxystrobin (strobilurin fungicide) and imidacloprid (neonicotinoid insecticide) have been detected in surface waters near treated agricultural, urban, and mixed landscapes. The hazards of pesticide runoff can be prevented through best management practices, including the establishment of diverse wetland plant barriers that can phytoremediate the chemicals in which they come into contact with. In this study, the wetland plant species softrush (Juncus effusus), pickerelweed (Pontederia cordata), and arrowhead (Sagittaria latifolia) were planted in sandy soil containers that were then placed in azoxystrobin or imidacloprid treated water. Every week for 2 months, water samples were collected for pesticide residue analysis using high-performance liquid chromatography (HPLC). At 14, 28, and 56 days after initiation, plants were destructively harvested and analyzed for pesticide residue in soil, above-ground vegetation, and below-ground vegetation. Results from this study report P. cordata reduced greater azoxystrobin (51.7% reduction compared to treated non-planted containers) compared to J. effusus and S. latifolia (24.9% and 28.7% reduction from non-planted containers) at 56 days. However, S. latifolia reduced greater imidacloprid (79.3% reduction compared to non-planted containers) compared to J. effusus and P. cordata (36.0% and 37.1% reduction from non-planted containers) at 56 days.Novelty statement: While research has found that wetland plants can absorb and remediate synthetic chemicals, this practice is only sustainable if used with native plants that require low maintenance and are tolerant to the applied substances. Various previous studies observe plants that are fast-growing, tolerant to environmental conditions, require low-maintenance, and are hardy. However, these plant species are not always suitable for any location and are often considered invasive and/or weed-like. The present research initiates a list of plant species which can be used within the southeastern United States and similar areas to phytoremediate commonly used pesticides azoxystrobin and imidacloprid and prevent off-target movement into sensitive water systems.

1-Tetradecanol, Diethyl Phthalate and Tween 80 Assist in the Formation of Thermo-Responsive Azoxystrobin Nanoparticles.

The occurrence of crop fungal diseases is closely related to warm environmental conditions. In order to control the release of fungicides in response to warm conditions, and enhance the efficacy, a series of thermo-responsive fungicide-loaded nanoparticles were developed. The fungicide azoxystrobin, solvent DEP, emulsifier Tween 80 and thermo-responsive component TDA were combined to create thermal-response oil phases, conditions for emulsification were then optimized. LDLS, zeta potential, FTIR, DSC, TGA, XRD, SEM and antifungal efficacy assays were carried out to investigate the characteristics and forming mechanism. The results indicated that the formula with 5 g azoxystrobin, 10 mL DEP, 6 mL Tween 80 and 2.5 g TDA constructed the proposed oil phase with the ability to transform from solid at 20 °C to softerned at 31.5 °C. Both DEP and TDA played key roles in interfering with the crystallization of azoxystrobin. The optimal T3t-c12 nanoparticles had a mean particle size of 162.1 nm, thermo-responsive morphological transformation between 20 °C and 30 °C, AZO crystal reforming after drying, the ability to attach to fungal spores and satisfied antifungal efficacy against P. nicotiana PNgz07 and A. niger A1513 at 30 °C. This report provides referable technical support for the construction of smart-release nanoparticles of other agrochemicals.

Dissipation and Residue of Metalaxyl-M and Azoxystrobin in Scallions and Cumulative Risk Assessment of Dietary Exposure to Hepatotoxicity.

Metalaxyl-M and azoxystrobin have been used to control various fungal diseases on scallion and other crops. In view of the adverse toxic effects of both on the mammalian liver, it is necessary to conduct a cumulative risk assessment of their dietary exposure to consumers. The residues of metalaxyl-M and azoxystrobin on scallion were determined by a quick, easy, cheap, effective, rugged, and safe method (QuEChERS) combined with high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS). The half-lives were about 1.15 and 3.89 days, respectively, and the final residues after a seven-day harvest interval were <0.001−0.088 mg/kg and 0.190−4.687 mg/kg, respectively. The cumulative dietary risk quotient of the two fungicides to Chinese consumers calculated by the probability model is 13.94%~41.25%. According to the results of the contribution analysis, the risk posed by azoxystrobin is much greater than that of metalaxyl-M. Although metalaxyl-M and azoxystrobin do not pose a cumulative risk to Chinese consumers, the risk to children and adolescents is significantly higher than that to adults. This suggests that in future research, more consideration should be given to the cumulative risk of compounds to vulnerable groups.