RESUMO
In the present study, halophilic bacteria communities were explored in saline soils of Howze-Soltan playa in Iran with special attention to their biological activity against an aflatoxigenic Aspergillus parasiticus NRRL 2999. Halophilic bacteria were isolated from a total of 20 saline soils using specific culture media and identified by 16S rRNA sequencing in neighbor-joining tree analysis. Antifungal and antiaflatoxigenic activities of the bacteria were screened by a nor-mutant A. parasiticus NRRL 2999 using visual agar plate assay and confirmed by high-performance liquid chromatography. Among a total of 177 halophilic bacteria belonging to 11 genera, 121 isolates (68.3%) inhibited A. parasiticus growth and/or aflatoxin production. The most potent inhibitory bacteria of the genera Bacillus, Paenibacillus and Staphylococcus were distributed in three main phylogenetic clusters as evidenced by 16S rRNA sequence analysis. A. parasiticus growth was inhibited by 0.7-92.7%, while AFB1 and AFG1 productions were suppressed by 15.1-98.9 and 57.0-99.6%, respectively. Taken together, halophilic bacteria identified in this study may be considered as potential sources of novel bioactive metabolites as well as promising candidates to develop new biocontrol agents for managing toxigenic fungi growth and subsequent aflatoxin contamination of food and feed in practice.
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Antibiose , Antifúngicos/metabolismo , Bacillus/metabolismo , Halobacteriales/metabolismo , Microbiologia do Solo , Staphylococcus/metabolismo , Aflatoxinas/antagonistas & inibidores , Antifúngicos/química , Antifúngicos/farmacologia , Aspergillus/efeitos dos fármacos , Bacillus/química , Halobacteriales/química , Microbiota , Staphylococcus/químicaRESUMO
Finding new compounds with antifungal properties is an important task due to the side effects of common antifungal drugs and emerging antifungal resistance in fungal strains. ∆24-sterol methyltransferase (24-SMT) is a crucial enzyme that plays important roles in fungal ergosterol biosynthesis pathway and is not found in humans. In the present study, the effects of α-bisabolol on Aspergillus fumigatus Af239 growth and ergosterol synthesis on the base of 24-SMT enzyme activity were studied; in addition, the expression of erg6, the gene encoded 24-SMT, was considered. To our knowledge, this is the first report demonstrating that α-bisabolol inhibits A. fumigatus growth specifically via suppressing fungal 24-SMT. Since this enzyme is a specific fungal enzyme not reported to exist in mammalian cells, α-bisabolol may serve as a lead compound in the development of new antifungal drugs. Fungi were cultured in presence of serial concentrations of α-bisabolol (0.281-9 mM) for 3 days at 35 °C. Mycelia dry weight was determined as an index of fungal growth and ergosterol content was assessed. Microsomal 24-SMT activity was assayed in presence of α-bisabolol as an inhibitor, lanosterol as a substrate and [methyl-H3] AdoMet (S-Adenosyl methionin). In addition, the expression of erg 6 gene (24-SMT encoding gene) was determined after treatments with various concentrations of α-bisabolol. Our results demonstrated that α-bisabolol strongly inhibited A. fumigatus growth (35.53-77.17%) and ergosterol synthesis (26.31-73.77%) dose-dependently and suppressed the expression of erg 6 gene by 76.14% at the highest concentration of 9 mM. α-bisabolol inhibited the activity of 24-SMT by 99% at the concentration of 5 mM. Taken together, these results provides an evidence for the first time that α-bisabolol inhibits A. fumigatus Af239 growth via affecting microsomal ∆24-sterol methyltransferase as a crucial enzyme in ergosterol biosynthetic pathway.
Assuntos
Antifúngicos/farmacologia , Aspergillus fumigatus/crescimento & desenvolvimento , Metiltransferases/antagonistas & inibidores , Sesquiterpenos/farmacologia , Aspergillus fumigatus/efeitos dos fármacos , Aspergillus fumigatus/enzimologia , Vias Biossintéticas/efeitos dos fármacos , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Ergosterol/biossíntese , Proteínas Fúngicas/antagonistas & inibidores , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Sesquiterpenos MonocíclicosRESUMO
BACKGROUND: Dermatophytosis is the most important superficial fungal infection which affects nearly 20% of human population worldwide. Recurrence of disease and emerging resistance of Trichophyton rubrum to synthetic antifungals are the main problems in control of dermatophytosis. The purpose of this study was to evaluate the effect of cold atmospheric plasma (CAP) on T. rubrum growth, ergosterol biosynthesis and keratinase activity. METHODS: A CAP system, comprised of helium 98% - oxygen 2% (He/O2), was used. Trichophyton rubrum conidia suspensions were treated with CAP in time periods of 90, 120, 150 and 180 s in 96-well microplates. Fungal growth was evaluated by counting the colony forming unit (CFU). Fungal dry weight, ergosterol biosynthesis and keratinase activity were evaluated in CAP-treated T. rubrum and untreated controls. RESULTS: T. rubrum growth was significantly inhibited by 62%-91%. CAP strongly suppressed fungal ergosterol biosynthesis by 27%-54%. The keratinase activity was increased by 7.30%-21.88% up to 120 s CAP exposure. CONCLUSION: Our results demonstrated for the first time that CAP inhibits T. rubrum growth, suppresses ergosterol biosynthesis and increases moderately keratinase activity in a dose-dependent manner. Overall, CAP exposure could be a potentially useful method for treatment of clinical cases of human and animal dermatophytoses.
Assuntos
Ergosterol/biossíntese , Peptídeo Hidrolases/química , Gases em Plasma/farmacologia , Trichophyton/enzimologia , Animais , Antifúngicos/farmacologia , Atmosfera , Temperatura Baixa , Hélio/química , Humanos , Oxigênio/química , Tinha/tratamento farmacológicoRESUMO
Aflatoxin contamination of grains and agro-products is a serious food safety issue and a significant economic concern worldwide. In the present study, the effects of eugenol on Aspergillus parasiticus growth and aflatoxin production were studied in relation to the expression of some essential genes involved in aflatoxin biosynthetic pathway. The fungus was cultured in presence of serial two-fold concentrations of eugenol (15.62-500 µg mL(-1)) for 3 days at 28 °C. Mycelia dry weight was determined as an index of fungal growth, while aflatoxin production was assessed by high performance liquid chromatography. The expression of aflatoxin biosynthetic genes including ver-1, nor-1, pksA, omtA and aflR were evaluated by real-time PCR. Eugenol strongly inhibited A. parasiticus growth in the range of 19.16-95.83 % in a dose-dependent manner. Aflatoxin B1 production was also inhibited by the compound in the range of 15.07-98.0 %. The expressions of ver-1, nor-1, pksA, omtA and aflR genes were significantly suppressed by eugenol at concentrations of 62.5 and 125 µg mL(-1). These results indicate that eugenol may be considered as a good candidate to control toxigenic fungal growth and the subsequent contamination of food, feed and agricultural commodities by carcinogenic aflatoxins.
Assuntos
Aflatoxina B1/biossíntese , Antifúngicos/farmacologia , Aspergillus/efeitos dos fármacos , Vias Biossintéticas/efeitos dos fármacos , Eugenol/farmacologia , Aspergillus/crescimento & desenvolvimento , Relação Dose-Resposta a Droga , Regulação para Baixo , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Genes Fúngicos/efeitos dos fármacosRESUMO
Background and Objectives: Candida tropicalis is one of the major non-albicans species causing nosocomial infection. There is limited data about mechanisms of azole-resistance and virulence factors of Candida tropicalis. This study was designed to investigate molecular mechanism of azole-resistance and major virulence factors of C. tropicalis isolated from oropharyngeal candidiasis in head and neck cancer patients. Materials and Methods: After collecting 38 C. tropicalis clinical isolates, antifungal susceptibility pattern and the expression levels of ERG11, CDR1, CDR2 and MDR1 were evaluated. Moreover, proteinase and phospholipase activity and biofilm formation of the isolates were investigated as virulence factors. Results: We detected fluconazole resistance in 7 C. tropicalis isolates. The expression levels of CDR1, ERG11 and MDR1 were increased respectively. Protease activity and biofilm formation were seen in all isolates. Five isolates did not exhibit phospholipase activity. Conclusion: Taken together, the overexpressions of ERG11, CDR1 and MDR1 genes were found in fluconazole resistant C. tropicalis, isolated from oropharyngeal candidiasis patients. Also, voriconazole was an effective antifungal against C. tropicalis isolates. The observed high protease enzyme activity and biofilm formation suggested strong pathogenicity of these isolates.
RESUMO
OBJECTIVE: The aim of the current work was to assess the molecular mechanisms of fluconazole-resistant Candida glabrata strains isolated from oropharyngeal candidiasis (OPC) in head and neck patients, as well as evaluation of virulence factors. DESIGN: Antifungal susceptibility pattern of sixty six clinical isolates of C. glabrata were evaluated by broth-microdilution method. The expression of ERG11, CDR1, CDR2, PDR1 genes as well as ERG11 gene capable of possible mutations was also detected in 21 fluconazol-resistant C. glabrata isolates. Phospholipase and proteinase activity of these isolates was estimated, too. The correlation between the virulence factors, antifungal susceptibility patterns and cancer type was also analyzed. RESULTS: Seven synonymous and four non-synonymous mutations were found in 21 fluconazole-resistant C. glabrata isolates; subsequently, four amino acid substitutions including H257P, Q47H, S487Y and I285N were then reported for the first time. High expression of CDR1 and PDR1 in related to other gene findings were tested in these isolates. Additionally, there was no significant difference between stage of cancer and MIC of all antimicrobial drugs. Significant differences between MIC of fluconazole, voriconazole and cancer types were also, found. The proteinase activity (92.4%) was higher than phospholipase activity in the isolates. Further, no significant difference between proteinase (rs: 0.003), phospholipase (rs: -0.107) activity and fluconazole MICs was observed. CONCLUSION: C. glabrata isolated from OPC in head and neck patients represented high capacities for proteolytic enzymes activity and high mRNA level of CDR1 and PDR1 gene and ERG11 mutations play an important role in azole drug resistance.
Assuntos
Candidíase Bucal , Neoplasias de Cabeça e Pescoço , Humanos , Antifúngicos/farmacologia , Azóis/farmacologia , Fluconazol/farmacologia , Candida glabrata/genética , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/farmacologia , Farmacorresistência Fúngica/genética , Fatores de Virulência , Testes de Sensibilidade MicrobianaRESUMO
Background: The Candida albicans is one of the most important global opportunistic pathogens, and the incidence of candidiasis has increased over the past few decades. Despite the established role of skin in defense against fungal invasion, little has been documented about the pathogenesis of Candida species when changing from normal flora to pathogens of vaginal and gastrointestinal epithelia. This study was carried out to determine the in vivo and in vitro pathogenesis of clinical C. albicans strains isolated from skin lesions. Methods: In this study, association of in vivo and in vitro pathogenesis of C. albicans isolates with different evolutionary origins was investigated. Oral and systemic experimental candidiasis was established in BALB/C mice. The expression levels of secreted aspartyl proteinases (SAP1-3 genes), morphological transformation, and biofilm-forming ability of C. albicans were evaluated. Results: All the strains showed in vitro and in vivo pathogenicity by various extents. The SAP1, SAP2, and SAP3 genes were expressed in 50%, 100%, and 75% of the strains, respectively. The biofilm formation ability was negative in 12% of the strains, while it was considerable in 38% of the strains. Fifty percent of the strains had no phospholipase activity, and no one demonstrated high level of this pathogenesis factor. Relatively all the strains had very low potency to form pseudohyphae. Conclusion: Our findings demonstrated that Candida albicans strains isolated from cutaneous candidiasis were able to cause oral and systemic infections in mice, so they could be considered as the potential agents of life-threatening nosocomial candidiasis in susceptible populations.
Assuntos
Candida albicans/isolamento & purificação , Candida albicans/patogenicidade , Candidíase/microbiologia , Fatores de Virulência/metabolismo , Animais , Candidíase/patologia , Feminino , Rim/patologia , Camundongos Endogâmicos BALB C , Filogenia , Língua/patologia , VirulênciaRESUMO
BACKGROUND AND OBJECTIVES: Cutaneous candidiasis is a multipicture fungal infection caused by members of the genus Candida which is considered as a public health problem all over the world with urgency of effective treatment and control. This study was performed to analyze the clinical epidemiology and molecular aspects of cutaneous candidiasis in Tehran-Iran in relation to antifungal susceptibility and virulence factors of etiologic Candida species. MATERIALS AND METHODS: Candida species were isolated from skin (27.3%) and nail scrapings (72.7%) of suspected patients and identified by ITS sequencing. Phylogeny of the isolates was evaluated using multilocus sequence typing (MLST) and antifungal susceptibility and virulence factors of the isolates were determined in relation to clinical presentation. RESULTS: Candida albicans was the most prevalent species (39.8%), followed by C. parapsilosis (32.9%), C. orthopsilosis (10.4%), C. tropicalis (7.9%), C. glabrata and C. guilliermondii, each (4.5%). Molecular typing of 35 C. albicans isolates by MLST revealed 28 novel sequence types with 11 singletons with 80.0% new diploid sequence types (DSTs). Majority of the isolates were susceptible to amphotericin B (91.5%), followed by posaconazole (90.3%), fluconazole (84.3%), itraconazole (74.1%), caspofungin (53.6%), and voriconazole (26.8%). Biofilm formation, yeast-to-hyphae transformation and phospholipase activity were reported species-dependent. CONCLUSION: Our results demonstrated clinical epidemiology of various Candida species from cutaneous candidiasis distributed in new molecular types with increasing importance of drug resistant of non-albicans Candida species. Our results showed that drug susceptibility and genetic variability of Candida species may be attributed to their clinical features and source of isolation.
RESUMO
The purpose of the current study is to evaluate the effect of silibinin on human hepatocellular carcinoma HepG-2 cells. Microculture tetrazolium test (MTT assay), Lactate dehydrogenase (LDH) release, Gelatin zymography, Griess reaction, Cell-based the extracelluar signal-regulated kinase (ERK) 1/2 phosphorylation assay and quantitative real-time RT-PCR were employed to appraise the effect of silibinin on cell proliferation, cytotoxicity, metastatic potential, nitric oxide (NO) production, ERK 1/2 phosphorylation and activation in HepG-2 cells. Silibinin inhibited cell proliferation, matrix metalloproteinase 2 enzymatic activity, NO production and ERK 1/2 phosphorylation in a dose-dependent manner without exerting any cytotoxicity effect. In addition, an expressive increase in mRNA levels of Raf kinase inhibitor protein (RKIP), sprouty-related protein 1 with EVH-1 domain (Spred-1), sprouty-related protein with EVH-1 domain 2 (Spred-2) coupled with a significant reduction in transcriptional levels of highly expressed in cancer (Hec1) and MMP-2 were observed. Altogether, these issues show for the first time that silibinin treatment could inhibit cell proliferation and invasive potential of HepG-2 cells through inhibition of ERK 1/2 cascade both directly (through suppression of ERK 1/2 phosphorylation) and indirectly (through up-regulation of RKIP, Spred-1 and Spred-2). In addition, cell growth and proliferation may be inhibited by silibinin through down-regulation of Hec1.
Assuntos
Antioxidantes/farmacologia , Carcinoma Hepatocelular/metabolismo , Proteínas Adaptadoras de Transdução de Sinal , Antioxidantes/administração & dosagem , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/efeitos dos fármacos , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Neoplasias Hepáticas/metabolismo , Proteínas de Membrana/efeitos dos fármacos , Proteínas de Membrana/metabolismo , Proteína Quinase 1 Ativada por Mitógeno/efeitos dos fármacos , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/efeitos dos fármacos , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Óxido Nítrico/biossíntese , Proteína de Ligação a Fosfatidiletanolamina/efeitos dos fármacos , Proteína de Ligação a Fosfatidiletanolamina/metabolismo , Fosforilação/efeitos dos fármacos , RNA Mensageiro/metabolismo , Proteínas Repressoras/efeitos dos fármacos , Proteínas Repressoras/metabolismo , Silibina , Silimarina/administração & dosagem , Silimarina/farmacologiaRESUMO
AIM: N-acetylcysteine can inhibit the formation of intracellular reactive oxygen intermediates. Cellular redox state plays a role in regulating the secretion of matrix metalloproteinase-2. We investigated the effects of N-acetylcysteine on the expression of matrix metalloproteinase-2 and tissue inhibitor of matrix metalloproteinase-2. METHODS: Bile duct ligated rats were used as a model of hepatic fibrosis. We compared the level of gene expression (using real-time reverse transcription polymerase chain reaction [RT-PCR]), liver function parameters, hepatic reactive oxygen production, lipid peroxidation and glutathione state in experimental groups. RESULTS: N-acetylcysteine treatment significantly improved liver function parameters including the plasma levels of aspartate aminotransferase, alkaline phosphatase, gamma-glutamyl transpeptidase and bilirubin. In addition, significant improvement of glutathione state and reactive oxygen production were observed. Hepatic lipid peroxidation was reversed by N-acetylcysteine treatment. Although N-acetylcysteine treatment did not completely normalize the increased matrix metalloproteinase-2 expression, it significantly decreased its level by 65%. N-acetylcysteine treatment also significantly decreased matrix metalloproteinase-2 activity and normalized tissue inhibitor of matrix metalloproteinase-2 expression. CONCLUSION: Collectively, N-acetylcysteine showed inhibition of matrix metalloproteinase-2 expression and activity. In addition, administration of N-acetylcysteine was associated with downregulation of the expression of tissue inhibitor of matrix metalloproteinase-2 and amelioration of oxidative stress in the liver of bile duct ligated rats.
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BACKGROUND: Although pyrimethamine (Tindurin) appears to be effective in the prevention and treatment of some infectious diseases, very little information exists on its unpredictable properties. We design this study to evaluate its anti-tumoral effect on a model of cell line. METHODS: The cytotoxic influence of Pyrimethamine on prostate cell line was investigated using an in vitro colometric assay. The potential modulatory effects on metastasis, apoptosis, and immortality characteristics of cells were assessed with gelatin zymography, terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay and telomeric repeat amplification protocol, respectively. RESULTS: Cytotoxicity analysis of pyrimethamine revealed a dose-dependent fashion. An apoptotic influence of pyrimethamine was also confirmed by data obtained from TUNEL assay. Dose-dependent inhibitory effect on matrix metalloproteinases (MMP) was seen in pyrimethamine. A potent inhibitory effect of pyrimethamine was also established by data achieved from TRAPeze telomerase detection kit. CONCLUSIONS: Collectively, as induction of apoptosis together with MMP and telomerase inhibition could be indicative of cancer treatment, pyrimethamine might be considered as a chemopreventative agent in cancer.
Assuntos
Inibidores Enzimáticos/farmacologia , Pirimetamina/farmacologia , Telomerase/antagonistas & inibidores , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Humanos , Masculino , Metaloproteinase 2 da Matriz/metabolismoRESUMO
In spite of developing new drugs and modern formulations, the treatments of chronic fungal infections are still challenging. Fibrous wound dressings are new suggestions for the treatment of chronic superficial infections. In the present study, we formulated an antifungal agent, terbinafine hydrochloride (TFH), which is a hydrophobic drug, in wound dressings prepared by electrospun polycaprolactone, polycaprolactone/gelatin (50:50 w/w) and gelatin. To obtain more water-stable meshes, the preparations were treated by glutaraldehyde and their properties were determined before and after treatment. The morphology of fibrous meshes was observed by scanning electron microscopy. Drug loading efficiency and release rate were measured by high performance liquid chromatography (HPLC) and the release rate was monitored for 144h. Antifungal tests were performed on Trichophyton mentagrophytes, Aspergillus fumigatus and Candida albicans cultured on Muller-Hinton agar. The toxicity of the meshes was measured after 24h and 14days by MTT assay. Terbinafine loading of polycaprolactone/gelatin (50:50) was 100% and it released the highest amount of TFH too. In antifungal tests, all samples were able to hinderT. mentagrophytes and A. fumigatus but not C. albicans growth among them, polycaprolactone fibers made the largest inhibition zone. In MTT assay, none of prepared samples showed toxicity against L929 cells. Teken together, the prepared TFH-loaded PCL/gelatin electrospun meshes were able to release TFH slowly and in a steady state in time. With respect to no obvious cytotoxicity in MTT assay and stong antifungal activity toward T. mentagrophytesin vitro, these TFH-based meshes could be considered as potential candidates in clinical application as wound dressing for treatment of chronic dermatophytosis.
Assuntos
Antifúngicos/uso terapêutico , Bandagens/microbiologia , Micoses/tratamento farmacológico , Naftalenos/uso terapêutico , Ferimentos e Lesões/tratamento farmacológico , Ferimentos e Lesões/microbiologia , Animais , Antifúngicos/farmacologia , Linhagem Celular , Doença Crônica , Liberação Controlada de Fármacos , Gelatina/química , Camundongos , Micoses/microbiologia , Nanofibras/química , Nanofibras/ultraestrutura , Naftalenos/farmacologia , Poliésteres/química , TerbinafinaRESUMO
BACKGROUND: The pathogenic yeast Candida albicans is the most common opportunistic fungal pathogen that is responsible for a wide array of infections in susceptible individuals. Despite recent progress in developing novel antifungal drugs which combat Candida-related disorders, this fungus is still a major cause of life-threatening infections all over the world. In the present study, the effect of cold atmospheric plasma (CAP) was evaluated on the growth of C. albicans with special attention to the ability of the CAP-treated fungus for biofilm formation, ergosterol biosynthesis and phospholipase and proteinase secretory production. METHODS: C. albicans cell suspensions were irradiated over time-scales ranging of 90, 120, 150 and 180s under cold atmospheric plasma contained He/O2 (2%). Treated and untreated yeast cells were analyzed for the growth, biofilm formation, ergosterol content, and activities of phospholipase and proteinase. RESULTS: Our results showed that CAP remarkably suppressed the growth of C. albicans by 31-82% at the given times. Likewise, CAP strongly inhibited the ergosterol biosynthesis by the fungus in the range of 40-91%, biofilm formation by 43-57% and the activities of phospholipase and proteinase enzymes by 4-45%, dose-dependently. CONCLUSION: CAP strongly inhibits the growth and virulence factors of C. albicans and thus, it could be a potential candidate to treat Candida-related superficial and cutaneous infections in practice.
Assuntos
Biofilmes/crescimento & desenvolvimento , Candida albicans/fisiologia , Ergosterol/biossíntese , Gases em Plasma/administração & dosagem , Fatores de Virulência/metabolismo , Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/fisiologia , Regulação para Baixo/efeitos dos fármacos , Ergosterol/metabolismoRESUMO
Fusarium culmorum, a fungal pathogen of small grain cereals, produces 4-deoxynivalenol and its acetylated derivatives that may cause toxicoses on humans or animals consuming contaminated food or feed. Natural and natural-like compounds belonging to phenol and hydroxylated biphenyl structural classes were tested in vitro to determine their activity on vegetative growth and trichothecene biosynthesis by F. culmorum. Most of the compounds tested at 1.5 or 1.0 mM reduced 3-acetyl-4-deoxynivalenol production by over 70% compared to the control, without affecting fungal growth significantly. Furthermore, several compounds retained their ability to inhibit toxin in vitro production at the lowest concentrations of 0.5 and 0.25 mM. Magnolol 27 showed fungicidal activity even at 0.1 mM. No linear correlation was observed between antioxidant properties of the compounds and their ability to inhibit fungal growth and mycotoxigenic capacity. A guaiacyl unit in the structure may play a key role in trichothecene inhibition.
Assuntos
Fungicidas Industriais/farmacologia , Fusarium/efeitos dos fármacos , Fusarium/metabolismo , Micotoxinas/metabolismo , Fenóis/farmacologia , Doenças das Plantas/microbiologia , Tricotecenos/metabolismo , Triticum/microbiologiaRESUMO
In the present study, genetic diversity and mycotoxin profiles of Aspergillus flavus isolated from air (indoors and outdoors), levels (surfaces), and soils of five hospitals in Southwest Iran were examined. From a total of 146 Aspergillus colonies, 63 isolates were finally identified as A. flavus by a combination of colony morphology, microscopic criteria, and mycotoxin profiles. No Aspergillus parasiticus was isolated from examined samples. Chromatographic analyses of A. flavus isolates cultured on yeast extract-sucrose broth by tip culture method showed that approximately 10% and 45% of the isolates were able to produce aflatoxin B(1) (AFB(1)) and cyclopiazonic acid (CPA), respectively. Around 40% of the isolates produced sclerotia on Czapek-Dox agar. The isolates were classified into four chemotypes based on the ability to produce AF and CPA that majority of them (55.5%) belonged to chemotype IV comprising non-mycotoxigenic isolates. Random amplified polymorphic DNA (RAPD) profiles generated by a combination of four selected primers were used to assess genetic relatedness of 16 selected toxigenic and non-toxigenic isolates. The resulting dendrogram demonstrated the formation of two separate clusters for the A. flavus comprised both mycotoxigenic and non-toxigenic isolates in a random distribution. The obtained results in this study showed that RAPD profiling is a promising and efficient tool to determine intra-specific genetic variation among A. flavus populations from hospital environments. A. flavus isolates, either toxigenic or non-toxigenic, should be considered as potential threats for hospitalized patients due to their obvious role in the etiology of nosocomial aspergillosis.