Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 788
Filtrar
1.
BMC Microbiol ; 23(1): 71, 2023 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-36922770

RESUMO

Microfungal isolates were routinely identified depending on both macro and micro morphological characteristics, sometimes, some fungal isolates appeared to be similar and such cases caused severe confusion for mycologists during the preliminary identification. During our previous studies dealing with isolation of fungi for some biotechnological applications; two mystifying species Aspergillus flavus and Aspergillus oryzae showed similar cultural and macroscopic features. Therefore, the current study aimed to easily distinguish between these two species depending on simple approaches which are routinely followed by a large segment of researchers. Investigation of the macroscopic features was performed to check the fungal growth on four different media (PDA, MEA, YES, and CYA) followed by microscopic examination using an ordinary light microscope, and scanning electron microscope SEM. Also, screening of secondary metabolites for both strains was preliminarily identified to find out the difference between their metabolic profiles. Finally, ITS rDNA was involved to clarify the molecular differences along their partial sequence. Conclusively, the BLAST strategy confirmed the similarity of ITS rDNA segments of both fungal strains that supported our hypothesis. The color of the fungal growth is a very critical factor whereas it is extensively influenced by the type of cultivation media. Accordingly, the YES medium was an inspiring tool assisting in prompt differentiation during the culture investigation step whereas A. oryzae and A. flavus appeared significant mustard yellow and olive green respectively. During the microscopic examination, the CYA medium also had a robust effect on the formation of the conidial chain whereas the knit long chain was observed in A. oryzae while the conidia appeared scattered and not in a chain in the case of A. flavus. Likewise, both two strains possessed different metabolic profiles where A. oryzae is not an Afla toxin producer, unlike A. flavus.


Assuntos
Aflatoxinas , Aspergillus oryzae , Aspergillus flavus , Aspergillus oryzae/genética , Aspergillus oryzae/metabolismo , Aflatoxinas/genética
2.
Plant Dis ; 2023 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-36947838

RESUMO

Optimizing synthetic antimicrobial peptides for safe and enhanced activity against fungal and bacterial pathogens is useful for genetic engineering of plants for resistance to plant pathogens and their associated mycotoxins. Nine synthetic peptides modeled after lytic peptides tachyplesin 1, D4E1 from cecropin A and protegrin 1 were added to germinated spores of fungal species Aspergillus flavus, Rhizopus stolonifer, Fusarium oxysporum f. sp. vasinfectum, F. verticillioides, F. graminearum, Claviceps purpurea, Verticillium dahliae, Thielaviopsis basicola and bacterial cultures of Psuedomonas syringae p.v. tabaci and Xanthomonas campestris p.v. campestris at different doses and inhibitory dose response curves were modeled to assess antimicrobial activity. Peptides GV185 and GV187, modified from tachyplesin 1, had superior abilities to inhibit fungal and bacterial growth (50% inhibitory concentrations or IC50 ranging from 0.1 to 8.7 µM). Rhizopus stolonifer (IC50 = 8.1 µM), A. flavus (IC50 = 3.1 µM) and F. graminearum (IC50 = 2.2 µM) were less inhibited by GV185 and GV187 than all the remaining fungi (IC50 = 1.4 µM) and bacteria (IC50 = 0.1 µM). Of the remaining peptides, GV193, GV195 and GV196 (IC50 range 0.9 to 6.6 µM) inhibited fungal growth of A. flavus, F. verticillioides and F. graminearum less than GV185 and GV187 (IC50 range 0.8 to 3.9 µM), followed by GV197 (IC50 range 0.8 - 9.1 µM) whereas GV190 and GV192 inhibited poorly (IC50 range 28.2 to 36.6 µM and 15.5 to 19.4 µM, respectively) and GV198 stimulated growth. GV185 and GV187 had slightly weaker hydrophobic and cationic residues than other tachyplesin 1 modified peptides, but still had unexpectedly high lytic activity. Germinated fungal spores of R. stolonifer and F. graminearum exposed to these two peptides and D4E1 and AGM182 appeared wrinkled with perforations near potential cytoplasmic leakage, which provided evidence of plasma membrane and cell wall lysis. We conclude that peptides GV185 and GV187 are promising candidates for genetic engineering of crops for resistance to plant pathogenic bacteria and fungi including A. flavus and aflatoxin contamination.

3.
Int J Food Microbiol ; 394: 110178, 2023 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-36947915

RESUMO

The objective of this study was to evaluate the chemical composition and antifungal activity of free and encapsulated Cinnamomum cassia essential oil (EO) against Penicillium crustosum, Alternaria alternata, and Aspergillus flavus, and the aroma persistence in maize flour. Trans-cinnamaldehyde (TC) was identified as the major compound (86 %) in the C. cassia EO. The EO was encapsulated by spray-dryer with 45.26 % efficiency using gum arabic (GA) and maltodextrin (MD) in a ratio of 1:1 (m/m). C. cassia EO showed antifungal activity against A. alternata, A. flavus, and P. crustosum, with a minimum inhibitory concentration (MIC) of 0.5 % for both free and standard TC, and 5 % for the encapsulated EO. Fungal growth inhibition was evaluated under exposition to vapors at different concentrations of C. cassia EO and TC standard, with MIC of 6 % and 8 % against P. crustosum, 4 % and 1 % A. alternata, and 4 % A. flavus, respectively. The sensory analysis results of the free and encapsulated C. cassia EO in maize flour showed a significant difference between the treated samples in relation to the standard sample (p < 0.05). The sample with free EO has high aroma intensity persistence, while the samples treated with encapsulated EO were evaluated as being closer to the standard sample. The results suggest that the encapsulated C. cassia EOs can be used as natural alternatives to control fungi in maize flour.

4.
J Hazard Mater ; 449: 131030, 2023 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-36827728

RESUMO

Mold contamination in foodstuffs causes huge economic losses, quality deterioration and mycotoxin production. Thus, non-destructive and accurate monitoring of mold occurrence in foodstuffs is highly required. We proposed a novel whole-cell biosensor array to monitor pre-mold events in foodstuffs. Firstly, 3 volatile markers ethyl propionate, 1-methyl-1 H-pyrrole and 2,3-butanediol were identified from pre-mold peanuts using gas chromatography-mass spectrometry. Together with other 3 frequently-reported volatiles from Aspergillus flavus infection, the volatiles at subinhibitory concentrations induced significant but differential response patterns from 14 stress-responsive Escherichia coli promoters. Subsequently, a whole-cell biosensor array based on the 14 promoters was constructed after whole-cell immobilization in calcium alginate. To discriminate the response patterns of the whole-cell biosensor array to mold-contaminated foodstuffs, optimal classifiers were determined by comparing 6 machine-learning algorithms. 100 % accuracy was achieved to discriminate healthy from moldy peanuts and maize, and 95 % and 98 % accuracy in discriminating pre-mold stages for infected peanuts and maize, based on random forest classifiers. 83 % accuracy was obtained to separate moldy peanuts from moldy maize by sparse partial least square determination analysis. The results demonstrated high accuracy and practicality of our method based on a whole-cell biosensor array coupling with machine-learning classifiers for mold monitoring in foodstuffs.


Assuntos
Técnicas Biossensoriais , Fungos , Fungos/química , Algoritmos , Cromatografia Gasosa-Espectrometria de Massas , Arachis , Aprendizado de Máquina
5.
Int J Biol Macromol ; 234: 123684, 2023 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-36791939

RESUMO

Aflatoxin B1 (AFB1), a potent natural group 1 carcinogen produced by Aspergillus flavus is considered an unavoidable toxic contaminant of herbal raw materials, which often deteriorates their active ingredients making them less effective and hazardous during their formulation in herbal drugs. The present investigation reports the antifungal (0.5 µl/ml) and AFB1 inhibitory (0.4 µl/ml) effects of the developed formulation CIM based on a mixture of essential oils (Carum carvi, and Illicium verum), and methyl anthranilate using mathematical modeling. The insight into the mechanism of action has also been explored using biochemical, molecular docking, and RT-PCR. Further, the nanoencapsulation of CIM (Ne-CIM) was prepared using a green facile synthesis of chitosan-based nanomatrix and characterized by Dynamic light scattering (DLS), Fourier transform-infrared, (FTIR), and X-ray diffraction analysis (XRD). The in-situ results showed that at MIC doses Ne-CIM effectively controls the A. flavus (81.25-89.57 %), AFB1 contamination (100 %), and protects the active ingredients deterioration of Piper nigrum, P. longum, Andrographis paniculata, Silybum marianum, and Withania somnifera caused by toxigenic species of A. flavus without affecting their sensory properties. Hence, Ne-CIM could be used as a green chemical agent to protect the biodeterioration of active ingredients of herbal raw materials caused by toxigenic species of A. flavus.


Assuntos
Quitosana , Óleos Voláteis , Antifúngicos/farmacologia , Antifúngicos/química , Quitosana/farmacologia , Quitosana/química , Simulação de Acoplamento Molecular , Óleos Voláteis/química , Aspergillus flavus
6.
Food Res Int ; 163: 112300, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36596201

RESUMO

The antimicrobial effects of continuous treatment with essential oils (EOs) in both liquid and gaseous phases have been intensively studied. Due to their rapid volatility, the effects of EOs on microorganisms after transient treatment are also worth exploring. In this work, the persistent effects of cinnamaldehyde (CA) vapor on Aspergillus flavus were detected by a series of biochemical analyses. Transcriptome analysis was also conducted to study the gene expression changes between recovered and normal A. flavus. When CA vapor was removed, biochemical analyses showed that the oxidative stress induced by the antimicrobial atmosphere was alleviated, and almost all the damaged functions were restored apart from mitochondrial function. Remarkably, the suppressed aflatoxin production intensified, which was confirmed by the up-regulation of most genes in the aflatoxin synthetic gene cluster, the velvet-related gene FluG and the aflatoxin precursor acetyl-CoA. Transcriptomic analysis also demonstrated significant changes in secondary metabolism, energy metabolism, oxidative stress, and amino acid metabolism in the recovery group. Taken together, these findings provide new insights into the mechanisms underlying the response of A. flavus to CA vapor treatment and will guide the rational application of EOs.


Assuntos
Aflatoxinas , Aspergillus flavus , Aflatoxinas/metabolismo , Acroleína/farmacologia , Acroleína/metabolismo , Perfilação da Expressão Gênica
7.
Plants (Basel) ; 12(2)2023 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-36678949

RESUMO

Aspergillus flavus is an important pathogenic fungus affecting many crops and is one of the main sources of their aflatoxin contamination. The primary method of limiting this pathogen is using chemical fungicides, but researchers focus on searching for other effective agents for its control due to many disadvantages and limitations of these agrochemicals. The results obtained in the present study indicate the high potential of two yeast strains, Aureobasidium pullulans PP4 and A. pullulans ZD1, in the biological control of A. flavus. Under in vitro conditions, mycelial growth was reduced by 53.61% and 63.05%, and spore germination was inhibited by 68.97% and 79.66% by ZD1 and PP4 strains, respectively. Both strains produced the lytic enzymes chitinase and ß-1,3-glucanase after 5 days of cultivation with cell wall preparations (CWP) of A. flavus in the medium as a carbon source. In addition, the tested yeasts showed the ability to grow over a wide range of temperatures (4-30 °C), pH (4-11), and salinity (0-12%) and showed tolerance to fungicides at concentrations corresponding to field conditions. Both isolates tested were highly tolerant to cupric oxychloride, showing biomass gains of 85.84% (ZD1) and 87.25% (PP4). Biomass growth in the presence of fungicides azoxystrobin was 78.71% (ZD1) and 82.65% (PP4), while in the presence of difenoconazole, it was 70.09% (ZD1) and 75.25% (PP4). The yeast strains were also tested for antagonistic effects against A. flavus directly on tomato fruit. Both isolates acted effectively by reducing lesion diameter from 29.13 mm (control) to 8.04 mm (PP4) and 8.83 mm (ZD1).

8.
J Fungi (Basel) ; 9(1)2023 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-36675939

RESUMO

Aspergillus flavus is an opportunistic pathogen responsible for millions of dollars in crop losses annually and negative health impacts on crop consumers globally. A. flavus strains have the potential to produce aflatoxin and other toxic secondary metabolites, which often increase during plant colonization. To mitigate the impacts of this international issue, we employ a range of strategies to directly impact fungal physiology, growth and development, thus requiring knowledge on the underlying molecular mechanisms driving these processes. Here we utilize RNA-sequencing data that are obtained from in situ assays, whereby Zea mays kernels are inoculated with A. flavus strains, to select transcription factors putatively driving virulence-related gene networks. We demonstrate, through growth, sporulation, oxidative stress-response and aflatoxin/CPA analysis, that three A. flavus strains with knockout mutations for the putative transcription factors AFLA_089270, AFLA_112760, and AFLA_031450 demonstrate characteristics such as reduced growth capacity and decreased aflatoxin/CPA accumulation in kernels consistent with decreased fungal pathogenicity. Furthermore, AFLA_089270, also known as HacA, eliminates CPA production and impacts the fungus's capacity to respond to highly oxidative conditions, indicating an impact on plant colonization. Taken together, these data provide a sound foundation for elucidating the downstream molecular pathways potentially contributing to fungal virulence.

9.
J Fungi (Basel) ; 9(1)2023 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-36675946

RESUMO

Di-n-butyl phthalate (DBP) is one of the most extensively used plasticizers for providing elasticity to plastics. Being potentially harmful to humans, investigating eco-benign options for its rapid degradation is imperative. Microbe-mediated DBP mineralization is well-recorded, but studies on the pollutant's fungal catabolism remain scarce. Thus, the present investigation was undertaken to exploit the fungal strains from toxic sanitary landfill soil for the degradation of DBP. The most efficient isolate, SDBP4, identified on a molecular basis as Aspergillus flavus, was able to mineralize 99.34% dibutyl phthalate (100 mg L-1) within 15 days of incubation. It was found that the high production of esterases by the fungal strain was responsible for the degradation. The strain also exhibited the highest biomass (1615.33 mg L-1) and total soluble protein (261.73 µg mL-1) production amongst other isolates. The DBP degradation pathway scheme was elucidated with the help of GC-MS-based characterizations that revealed the formation of intermediate metabolites such as benzyl-butyl phthalate (BBP), dimethyl-phthalate (DMP), di-iso-butyl-phthalate (DIBP) and phthalic acid (PA). This is the first report of DBP mineralization assisted with A. flavus, using it as a sole carbon source. SDBP4 will be further formulated to develop an eco-benign product for the bioremediation of DBP-contaminated toxic sanitary landfill soils.

10.
Molecules ; 28(2)2023 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-36677769

RESUMO

The risk of resistance development and adverse effects on human health and the environment has increased in the last decade. Furthermore, many antifungal agents fail to inhibit the pathogenesis of azole-resistant Aspergillus flavus. In this report, we isolated and identified azole-resistant A. flavus isolates from two sources of maize (white and yellow maize). The susceptibilities of Aspergillus flavus isolates were investigated by conventional antifungals such as Terbinfine, Fluconazole, Ketoconazole, Voricazole, Amphotericin, and Nystatin. Then zinc oxide nanoparticles associated with Chlorella vulgaris, which are synthesized by using the precipitation method, were examined against isolated fungi. The results showed that twelve species of white corn were isolated out of fifty isolates, while the number of isolates from the yellow corn source was only four. Interestingly, the following antifungals have an impact effect against azole-resistant A. flavus isolates: the inhibition zones of ketoconazole, voricazole, and terbinafine were 40 mm, 20 mm, and 12 mm, respectively, while the remaining antifungal agents have no effect. Similarly, the inhibition zones of the following antifungal agents were as follows: 41 mm for Terbinfine, 13 mm for Voricazole, and 11 mm for Ketoconazole against Aspergillus flavus that was isolated from yellow corn. The physiochemical characterization of zinc oxide nanoparticles provides evidence that ZnO-NPs associate with Chlorella vulgaris and have been fabricated by the precipitation method with a diameter of 25 nm. The zinc oxide nanoparticle was then used to isolate azole-resistant A. flavus, and the results show that ZnO-NPs have an effect on azole-resistant A. flavus isolation. The inhibition zone of zinc oxide nanoparticles against A. flavus (that was isolated from white corn) was 50 mm with an MIC of 50 mg/mL, while the inhibition zone of zinc oxide nanoparticles against Azole-resistant A. flavus isolated from yellow corn was 14 nm with an MIC of 25 mg/mL, which indicated that zinc oxide nanoparticles gave a better result against Azole-resistant A. flavus isolated from maize.


Assuntos
Chlorella vulgaris , Óxido de Zinco , Humanos , Antifúngicos/farmacologia , Aspergillus flavus , Zea mays , Óxido de Zinco/farmacologia , Azóis/farmacologia , Cetoconazol/farmacologia , Testes de Sensibilidade Microbiana
11.
J Food Sci Technol ; 60(2): 752-760, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36712224

RESUMO

In this study, the possibilities of using cold plasma technology in solving the mold problem, which is one of the most important problems in Kashar cheese, were investigated. For this purpose Kashar cheeses were exposed to cold plasma with different gas compositions. As a result of the study 3-4 log reduction was achieved for both Aspergillus flavus and Penicillium crysogenum. The pH and aw values of samples were decreased with cold plasma application. The b* values of samples increased while L* and a* values decreased. When all the results obtained are considered as a whole, it can be said that cold plasma technology improves the physicochemical properties of Kashar cheese and provides significant decrease in mold count of the product.

12.
Toxins (Basel) ; 15(1)2023 01 14.
Artigo em Inglês | MEDLINE | ID: mdl-36668896

RESUMO

Semen coicis is not only a traditional Chinese medicine (TCM), but also a typical food in China, with significant medical and healthcare value. Because semen coicis is rich in starch and oil, it can be easily contaminated with Aspergillus flavus and its aflatoxins (AFs). Preventing and controlling the contamination of semen coicis with Aspergillus flavus and its aflatoxins is vital to ensuring its safety as a drug and as a food. In this study, the endosphere bacteria Pseudomonas palleroniana strain B-BH16-1 produced volatiles that strongly inhibited the mycelial growth and spore formation activity of A. flavus. Gas chromatography-mass spectrometry profiling revealed three volatiles emitted from B-BH16-1, of which 1-undecene was the most abundant. We obtained authentic reference standards for these three volatiles; these significantly reduced mycelial growth and sporulation in Aspergillus, with dimethyl disulfide showing the most robust inhibitory activity. Strain B-BH16-1 was able to completely inhibit the biosynthesis of aflatoxins in semen coicis samples during storage by emitting volatile bioactive components. The microscope revealed severely damaged mycelia and a complete lack of sporulation. This newly identified plant endophyte bacterium was able to strongly inhibit the sporulation and growth of Aspergillus and the synthesis of associated mycotoxins, thus not only providing valuable information regarding an efficient potential strategy for the prevention of A. flavus contamination in TCM and food, but potentially also serving as a reference in the control of toxic fungi.


Assuntos
Aflatoxinas , Coix , Aspergillus flavus , Aflatoxinas/análise , Pseudomonas , Aspergillus
13.
Braz. j. biol ; 83: e246389, 2023. graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1285638

RESUMO

Abstract Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.


Resumo A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.


Assuntos
Animais , Galinhas , Plumas , Fermentação , Fungos , Resíduos Industriais , Queratinas/metabolismo
14.
Braz. J. Biol. ; 83: 1-8, 2023. graf
Artigo em Inglês | VETINDEX | ID: vti-765419

RESUMO

Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.(AU)


A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.(AU)


Assuntos
Aspergillus flavus/isolamento & purificação , Queratinas/análise , Queratinas/toxicidade , Biotransformação
15.
Mol Cell Proteomics ; 22(2): 100490, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36566904

RESUMO

Aspergillus flavus is a common saprophytic and pathogenic fungus, and its secondary metabolic pathways are one of the most highly characterized owing to its aflatoxin (AF) metabolite affecting global economic crops and human health. Different natural environments can cause significant variations in AF synthesis. Succinylation was recently identified as one of the most critical regulatory post-translational modifications affecting metabolic pathways. It is primarily reported in human cells and bacteria with few studies on fungi. Proteomic quantification of lysine succinylation (Ksuc) exploring its potential involvement in secondary metabolism regulation (including AF production) has not been performed under natural conditions in A. flavus. In this study, a quantification method was performed based on tandem mass tag labeling and antibody-based affinity enrichment of succinylated peptides via high accuracy nano-liquid chromatography with tandem mass spectrometry to explore the succinylation mechanism affecting the pathogenicity of naturally isolated A. flavus strains with varying toxin production. Altogether, 1240 Ksuc sites in 768 proteins were identified with 1103 sites in 685 proteins quantified. Comparing succinylated protein levels between high and low AF-producing A. flavus strains, bioinformatics analysis indicated that most succinylated proteins located in the AF biosynthetic pathway were downregulated, which directly affected AF synthesis. Versicolorin B synthase is a key catalytic enzyme for heterochrome B synthesis during AF synthesis. Site-directed mutagenesis and biochemical studies revealed that versicolorin B synthase succinylation is an important regulatory mechanism affecting sclerotia development and AF biosynthesis in A. flavus. In summary, our quantitative study of the lysine succinylome in high/low AF-producing strains revealed the role of Ksuc in regulating AF biosynthesis. We revealed novel insights into the metabolism of AF biosynthesis using naturally isolated A. flavus strains and identified a rich source of metabolism-related enzymes regulated by succinylation.


Assuntos
Aflatoxinas , Aspergillus flavus , Humanos , Aspergillus flavus/metabolismo , Lisina/metabolismo , Proteômica , Aflatoxinas/metabolismo , Processamento de Proteína Pós-Traducional
16.
Microbiol Spectr ; 11(1): e0421222, 2023 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-36541770

RESUMO

The filamentous fungus Aspergillus flavus causes devastating diseases not only to cash crops but also to humans by secreting a series of secondary metabolites called aflatoxins. In the cotranslational or posttranslational process, N-myristoyltransferase (Nmt) is a crucial enzyme that catalyzes the myristate group from myristoyl-coenzyme A (myristoyl-CoA) to the N terminus or internal glycine residue of a protein by forming a covalent bond. Members of the Nmt family execute a diverse range of biological functions across a broad range of fungi. However, the underlying mechanism of AflNmt action in the A. flavus life cycle is unclear, particularly during the growth, development, and secondary metabolic synthesis stages. In the present study, AlfNmt was found to be essential for the development of spore and sclerotia, based on the regulation of the xylose-inducible promoter. AflNmt, located in the cytoplasm of A. flavus, is also involved in modulating aflatoxin (AFB1) in A. flavus, which has not previously been reported in Aspergillus spp. In addition, we purified, characterized, and crystallized the recombinant AflNmt protein (rAflNmt) from the Escherichia coli expression system. Interestingly, the crystal structure of rAlfNmt is moderately different from the models predicted by AlphaFold2 in the N-terminal region, indicating the limitations of machine-learning prediction. In conclusion, these results provide a molecular basis for the functional role of AflNmt in A. flavus and structural insights concerning protein prediction. IMPORTANCE As an opportunistic pathogen, A. flavus causes crop loss due to fungal growth and mycotoxin contamination. Investigating the role of virulence factors during infection and searching for novel drug targets have been popular scientific topics in the field of fungal control. Nmt has become a potential target in some organisms. However, whether Nmt is involved in the developmental stages of A. flavus and aflatoxin synthesis, and whether AlfNmt is an ideal target for structure-based drug design, remains unclear. This study systematically explored and identified the role of AlfNmt in the development of spore and sclerotia, especially in aflatoxin biosynthesis. Moreover, although there is not much difference between the AflNmt model predicted using the AlphaFold2 technique and the structure determined using the X-ray method, current AI prediction models may not be suitable for structure-based drug development. There is still room for further improvements in protein prediction.


Assuntos
Aflatoxinas , Aspergillus flavus , Humanos , Antifúngicos/metabolismo , Aciltransferases/genética , Aciltransferases/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo
17.
Appl Microbiol Biotechnol ; 107(1): 341-354, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36477927

RESUMO

The prevention of fungal proliferation in postharvest grains is critical for maintaining grain quality and reducing mycotoxin contamination. Fumigation with natural gaseous fungicides is a promising and sustainable approach to protect grains from fungal spoilage. In this study, the antifungal activities of (E)-2-alkenals (C5-C10) on Aspergillus flavus were tested in the vapor phase, and (E)-2-heptenal showed the highest antifungal activity against A. flavus. (E)-2-Heptenal completely inhibited A. flavus growth at 0.0125 µL/mL and 0.2 µL/mL in the vapor phase and liquid contact, respectively. (E)-2-Heptenal can disrupt the plasma membrane integrity of A. flavus via leakage of intracellular electrolytes. Scanning electron microscopy indicated that the mycelial morphology of A. flavus was remarkably affected by (E)-2-heptenal. Metabolomic analyses indicated that 49 metabolites were significantly differentially expressed in A. flavus mycelia exposed to 0.2 µL/mL (E)-2-heptenal; these metabolites were mainly involved in galactose metabolism, starch and sucrose metabolism, the phosphotransferase system, and ATP-binding cassette transporters. ATP production was reduced in (E)-2-heptenal-treated A. flavus, and Janus Green B staining showed reduced cytochrome c oxidase activity. (E)-2-Heptenal treatment induced oxidative stress in A. flavus mycelia with an accumulation of superoxide anions and hydrogen peroxide and increased activities of superoxide dismutase and catalase. Simulated storage experiments showed that fumigation with 400 µL/L of (E)-2-heptenal vapor could completely inhibit A. flavus growth in wheat grains with 20% moisture; this demonstrates its potential use in preventing grain spoilage. This study provides valuable insights into understanding the antifungal effects of (E)-2-heptenal on A. flavus. KEY POINTS : • (E)-2-Heptenal vapor showed the highest antifungal activity against A. flavus among (C5-C10) (E)-2-alkenals. • The antifungal effects of (E)-2-heptenal against A. flavus were determined. • The antifungal actions of (E)-2-heptenal on A. flavus were revealed by metabolomics and biochemical analyses.


Assuntos
Antifúngicos , Aspergillus flavus , Antifúngicos/farmacologia , Antifúngicos/metabolismo , Aldeídos/metabolismo , Metabolômica
18.
Environ Res ; 216(Pt 1): 114407, 2023 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-36216116

RESUMO

Fungal abetted processes are among the finest approaches for the transformation or degradation and decolorization of dyes in effluents. In this piece of research; biodegradation and metabolic pathways of two toxic dyes Congo Red (CR) and Reactive black 5 (RB5) by two strains of Aspergillus sp. fungus in batch experiments has been investigated. Morphological characteristics of the isolates were observed with both light and electron microscopies. Based on molecular characterization the isolates were identified as Aspergillus flavus and Aspergillus niger. The degradation was also optimized via. operational parameters such as pH, temperature, incubation time, inoculums size, dye concentration, carbon sources and nitrogen sources. Degradation measurements revealed that the isolates effectively degraded 90% and 96% of CR and RB5 respectively. Metabolites were identified with Liquid chromatography-mass spectrometry (LCMS) and degradation pathways of the dyes were proposed. Toxicity assay Phaseolus mungo seeds showed that pure CR and RB5 dyes exhibits significant toxicity whereas fungal treated dye solution resulted in an abatement of the toxicity and cell viability was increased. The results stipulated in this article clearly showed the effectiveness of the isolates on detoxification of CR and RB5 dyes.


Assuntos
Corantes , Águas Residuárias , Corantes/química , Cinética , Biodegradação Ambiental , Vermelho Congo/metabolismo , Aspergillus niger/metabolismo , Compostos Azo/toxicidade , Compostos Azo/metabolismo
19.
Pol J Microbiol ; 71(4): 589-599, 2022 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-36537059

RESUMO

Aflatoxin (AF)-producing fungi such as Aspergillus flavus commonly contaminate animal feeds, causing high economic losses. A. flavus is the most prevalent and produces AFB1, a potent mutagen, and carcinogen threatening human and animal health. Aspergillaceae is a large group of closely related fungi sharing number of morphological and genetic similarities that complicate the diagnosis of highly pathogenic strains. We used here morphological and molecular assays to characterize fungal isolates from animal feeds in Southwestern Algeria. These tools helped to identify 20 out of 30 Aspergillus strains, and 15 of them belonged to the Aspergillus section Flavi. Further analyses detected four out of 15 as belonging to Aspergillus flavus-parasiticus group. PCR targeting the AF genes' aflR-aflS(J) intergenic region amplified a single 674 bp amplicon in all four isolates. The amplicons were digested with a BglII endonuclease, and three specific fragments were observed for A. flavus but A. parasitucus lacked two typical fragments. Sequencing data of four amplicons confirmed the presence of the two BglII restriction sites yielding the three fragments, confirming that all four strains were A. flavus. In addition, this analysis illustrated the genetic variability within the A. flavus strains.


Assuntos
Aflatoxinas , Aspergillus flavus , Animais , Humanos , Aspergillus flavus/genética , Aspergillus , Aflatoxinas/análise , Aflatoxinas/genética , Reação em Cadeia da Polimerase , Ração Animal
20.
J Agric Food Chem ; 70(50): 15928-15944, 2022 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-36508213

RESUMO

Oxylipins play important signaling roles in aflatoxin (AF) biosynthesis in Aspergillus flavus. We previously showed that exogenous supply of autoxidated linolenic acid (AL) inhibited AF biosynthesis in A. flavus via oxylipins, but the molecular mechanism is still unknown. Here, we performed multiomics analyses of A. flavus grown in media with or without AL. Targeted metabolite analyses and quantitative reverse transcription (qRT)-polymerase chain reaction (PCR) showed that the imizoquin (IMQ) biosynthetic pathway was distinctly upregulated in the presence of AL. 13C-glucose labeling confirmed in parallel that the tricarboxylic acid cycle was also enhanced by AL, consistent with observed increases in mycelial growth. Moreover, we integrated thermal proteome profiling and molecular dynamics simulations to identify a potential receptor of AL; AL was found to interact with a transporter (ImqJ) located in the IMQ gene cluster, primarily through hydrophobic interactions. Further analyses of strains with an IMQ pathway transcription factor overexpressed or knocked out confirmed that this pathway was critical for AL-mediated inhibition of AF biosynthesis. Comparison of 22 assembled A. flavus and Aspergillus oryzae genomes showed that genes involved in the IMQ pathway were positively selected in A. oryzae. Taken together, the results of our study provide novel insights into oxylipin-mediated regulation of AF biosynthesis and suggest potential methods for preventing AF contamination of crops.


Assuntos
Aflatoxinas , Aspergillus flavus , Aspergillus flavus/metabolismo , Oxilipinas/metabolismo , Ácido alfa-Linolênico , Fatores de Transcrição/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA