RESUMEN

INTRODUCTION: Some studies have reported the occurrence of microorganisms isolated from water. Considering these microorganisms, fungi are known to occur ubiquitously in the environment, including water, and some are pathogenic and may cause health problems, especially in immunocompromised individuals. The aim of this study was to identify fungi in hospital water samples and to correlate their presence with the concentration of free residual chlorine. METHODS: Water samples (100 mL) were collected from taps (n = 74) and water purifiers (n = 14) in different locations in a university hospital. Samples were filtered through a nitrocellulose membrane and placed on Sabouraud dextrose agar and incubated for 24 hours at 30°C. Fungi were identified according to established methods based on macroscopic and microscopic characteristics (filamentous) and physiological tests (yeasts). Free chlorine residual content was measured at the time of sample collection. RESULTS: Seventy species of fungi were identified in the water samples and about 56% of the water samples contained culturable fungi. Cladosporium oxysporum, Penicillium spinulosum, and Aspergillus fumigatus were the most common filamentous fungi. Aureobasidium pullulans and Candida parapsilosis were the most common yeasts. Chemical analyses revealed that free residual chlorine was present in 81.8% of the samples within recommended concentrations. Among samples from water purifiers, 92.9% showed low levels of free residual chlorine (<0.2 mg/L). There was no significant association between chlorine concentrations (either within or outside the recommended range) and the presence of filamentous fungi and yeasts. CONCLUSIONS: This study showed that hospital water can be a reservoir for fungi, some of which are potentially harmful to immunocompromised patients. Free residual chlorine was ineffective in some samples.

Asunto(s)

Biodiversidad , Hongos/aislamiento & purificación , Hospitales Universitarios , Microbiología del Agua , Abastecimiento de Agua , Aspergillus fumigatus/aislamiento & purificación , Aspergillus fumigatus/fisiología , Aureobasidium/aislamiento & purificación , Aureobasidium/fisiología , Biopelículas/crecimiento & desarrollo , Brasil , Candida parapsilosis/aislamiento & purificación , Candida parapsilosis/fisiología , Cloro/análisis , Cladosporium/aislamiento & purificación , Cladosporium/fisiología , Hongos/clasificación , Hongos/fisiología , Humanos , Micosis/microbiología , Penicillium/aislamiento & purificación , Penicillium/fisiología , Agua/análisis , Agua/química

RESUMEN

Fungal endophyte associations have been suggested as a possible strategy of Antarctic vascular plants for surviving the extreme environmental conditions of Antarctica. However, the mechanisms by which this occurs are still poorly understood. The role of root fungal endophytes in nitrogen mineralization and nutrient uptake, as well as their impact on the performance of Antarctic plants, were studied. We tested root endophytes, isolated from Colobanthus quitensis and Deschampsia antarctica, for lignocellulolytic enzyme production, nitrogen mineralization, and growth enhancement of their host plants. Penicillium chrysogenum and Penicillium brevicompactum were identified using a molecular approach as the main root endophytes inhabiting C. quitensis and D. antarctica, respectively. Both root endophytes were characterized as psychrophilic fungi displaying amylase, esterase, protease, cellulase, hemicellulase, phosphatase and urease enzymatic activities, mainly at 4 °C. Moreover, the rates and percentages of nitrogen mineralization, as well as the final total biomass, were significantly higher in symbiotic C. quitensis and D. antarctica individuals. Our findings suggest that root endophytes exert a pivotal ecological role based not only to breakdown different nutrient sources but also on accelerating nitrogen mineralization, improving nutrient acquisition, and therefore promoting plant growth in Antarctic terrestrial ecosystems.

Asunto(s)

Nitrógeno , Penicillium , Desarrollo de la Planta , Regiones Antárticas , Ecosistema , Endófitos , Humanos , Penicillium/fisiología , Plantas

RESUMEN

Introduction: Infectious diseases represent one of the leading causes of death worldwide. Considering the growing global challenge of antimicrobial resistance, research into new sources of potentially effective antimicrobial agents from natural origins is of great importance for world health. Objective: To evaluate the antimicrobial activity of endophytic fungi from Mammea americana and Moringa oleifera upon Staphylococcus aureus (ATCC 29213), S. aureus (resistant strain USb003), Escherichia coli (ATCC 25922), and E. coli (resistant strain USb007). Materials and methods: We isolated endophytic fungi from the leaves, seeds, and stems of the two plants under study. We evaluated their antimicrobial activity through the formation of sensitivity haloes in dual tests in vitro, as well as in trials using crude ethanolic extracts from the endophytes. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and cytotoxicity o the substances were analyzed. Results: Three ethanolic extracts of Penicillium sp., Cladosporium (001), and Cladosporium (002) exhibited the greatest inhibition halos in sensitive and resistant strains of E. coli and S. aureus. The MIC and CBM found were statistically significant (p≤0.05) compared with the gentamicin control. Furthermore, the cytotoxicity test results of CC50>1,000 demonstrated that the endophytic fungi studied exhibit bactericidal characteristics without causing unintended damage. Conclusion: The endophytic fungi M. oleifera and M. americana represent a source of active secondary metabolites with antimicrobial and non-toxic properties. In light of these findings, further research should proceed with chemical identification of the compounds and the study of their mechanisms of action, especially given the paucity of current scientific knowledge concerning the isolation of endophytes in these plants.

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Introducción. Las enfermedades infecciosas son una causa importante de muertes en el mundo. La resistencia antimicrobiana es un problema global, por lo que es conveniente la investigación de nuevas fuentes de agentes antimicrobianos de origen natural potencialmente efectivos. Objetivo. Evaluar la actividad antimicrobiana de hongos endófitos de Mammea americana y Moringa oleifera en la cepa sensible (ATCC 29213) y en la cepa resistente (USb003) de Staphylococcus aureus, así como en la cepa sensible (ATCC 25922) y la cepa resistente (USb007) de Escherichia coli. Materiales y métodos. Se aislaron 14 hongos endófitos de las hojas, semillas y tallos de las dos plantas en estudio. Se evaluó su actividad antimicrobiana mediante la formación de halos de sensibilidad por ensayo dual in vitro y pruebas con extractos etanólicos crudos provenientes de los endófitos a los que se les evaluó la concentración mínima inhibitoria (CMI), la concentración bactericida mínima (CBM) y la citotoxicidad. Resultados. Tres extractos etanólicos de Penicillium sp., Cladosporium sp. (001) y Cladosporium sp. (002) presentaron mayores halos de inhibición en cepas sensibles y resistentes de E. coli y S. aureus. La CMI y la CBM halladas fueron estadísticamente significativas (p≤0,05), comparadas con el control de gentamicina. Las pruebas de citotoxicidad (concentración citotóxica, CC50>1.000) demostraron que los hongos endófitos poseen características bactericidas y no ocasionan daño alguno. Conclusión. Se halló una fuente de metabolitos secundarios activos con propiedades antimicrobianas y no tóxicas en los hongos endófitos de M. oleifera y M. americana; estos hallazgos son importantes para continuar con la identificación química de los compuestos y el estudio de sus mecanismos de acción en estas plantas en las que el aislamiento de endófitos ha sido escaso.

Asunto(s)

Antibacterianos/aislamiento & purificación , Endófitos/fisiología , Hongos/fisiología , Mammea/microbiología , Moringa oleifera/microbiología , Plantas Medicinales/microbiología , Animales , Antibacterianos/farmacología , Chlorocebus aethiops , Cladosporium/química , Cladosporium/aislamiento & purificación , Cladosporium/fisiología , Evaluación Preclínica de Medicamentos , Farmacorresistencia Bacteriana Múltiple , Endófitos/aislamiento & purificación , Escherichia coli/efectos de los fármacos , Etanol , Hongos/aislamiento & purificación , Pruebas de Sensibilidad Microbiana , Penicillium/química , Penicillium/aislamiento & purificación , Penicillium/fisiología , Hojas de la Planta/microbiología , Tallos de la Planta/microbiología , Semillas/microbiología , Staphylococcus aureus/efectos de los fármacos , Células Vero

RESUMEN

Penicillium digitatum is the major source of postharvest decay in citrus fruits worldwide. This fungus shows a limited host range, being able to infect mainly mature fruit belonging to the Rutaceae family. This highly specific host interaction has attracted the interest of the scientific community. Researchers have investigated the chemical interactions and specialized virulence strategies that facilitate this fungus's fruit colonization, thereby leading to a successful citrus infection. There are several factors that mediate and affect the interaction between P. digitatum and its host citrus, including hydrogen peroxide modulation, secretion of organic acids and consequently pH control, and other strategies described here. The recently achieved sequencing of the complete P. digitatum genome opened up new possibilities for exploration of the virulence factors related to the host-pathogen interaction. Through such techniques as RNAseq, RT-PCR and targeted gene knockout mediated by Agrobacterium tumefaciens, important genes involved in the fungal infection process in citrus have been reported, helping to elucidate the molecular mechanisms, metabolites and genetic components that are involved in the pathogenicity of P. digitatum. Understanding the infection process and fungal strategies represents an important step in developing ways to protect citrus from P. digitatum infection, possibly leading to more productive citriculture.

Asunto(s)

Citrus/microbiología , Penicillium/fisiología , Enfermedades de las Plantas/microbiología , Citrus/fisiología , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulación Fúngica de la Expresión Génica , Especificidad del Huésped , Interacciones Huésped-Patógeno , Penicillium/genética

RESUMEN

Ochratoxin A (OTA) produced by mycotoxigenic fungi (Aspergillus and Penicillium spp.) is an extremely toxic and carcinogenic metabolite. The use of cold plasma to inhibit toxin-producing microorganisms in coffee could be an important alternative to avoid proliferation of mycotoxigenic fungi. Roasted coffee samples were artificially inoculated with A. westerdijikiae, A. steynii, A. versicolor, and A. niger, and incubated at 27 °C over 21 days for OTA production. Samples were cold plasma treated at 30 W input power and 850 V output voltage with helium at 1.5 L/min flow. OTA production in coffee was analyzed by high performance liquid chromatography coupled to a mass spectrometer (HPLC-MS). After 6 min of treatment with cold plasma, fungi were completely inhibited (4 log reduction). Cold plasma reduces 50% of OTA content after 30 min of treatment. Toxicity was estimated for extracts of artificially contaminated roasted coffee samples using the brine shrimp (Artemia salina) lethality assay. Toxicity for untreated roasted coffee was shown to be "toxic", while toxicity for cold plasma treated coffee was reduced to "slightly toxic". These results suggested that cold plasma may be considered as an alternative method for the degradation and reduction of toxin production by mycotoxigenic fungi in the processing of foods and feedstuffs.

Asunto(s)

Aspergillus/efectos de los fármacos , Café/microbiología , Contaminación de Alimentos/prevención & control , Ocratoxinas/análisis , Penicillium/efectos de los fármacos , Gases em Plasma/farmacología , Animales , Artemia , Aspergillus/fisiología , Penicillium/fisiología

RESUMEN

Penicillium italicum (Blue mold) is a major postharvest disease of citrus. An alternative to controlling the disease is through the use of yeasts. The purpose of the present study was to screen effective yeast antagonists against P. italicum, isolated from soil, leaves, flowers, and citrus fruits, to assess the action mechanisms of the yeast isolates that were demonstrated to be effective for biocontrol, and to identify the most effective yeast isolates for the biocontrol of blue mold. The in vitro assays showed that six yeast strains inhibited up to 90% of the pathogen's mycelial growth. In vivo assays, evaluating the incidence of blue mold on sweet oranges, the strains ACBL-04, ACBL-05, ACBL-10 and ACBL-11 were effective, demonstrating the potential for the blue mold control when preventively applied, whereas the ACBL-08 strain showed a high potential to preventive and curative applications. Additional studies on the modes of action of these yeast strains showed that most of the evaluated yeast strains did not produce antifungal substances, in sufficient quantities to inhibit the pathogen growth. Competition for nutrients was not a biocontrol strategy used by the yeast strains. The 'killer' activity might be the main action mechanism involved in P. italicum biocontrol. This study indicated that the multiple modes of action against the pathogen presented by yeasts may explain why these strains provided P. italicum control under in vitro and in vivo conditions. However, further studies in future might be able to elucidate the 'killer' activity and its interaction with pathogen cells and the bioproduct production using Candida stellimalicola strains for control postharvest diseases.

Asunto(s)

Antibiosis/fisiología , Citrus/microbiología , Microbiología de Alimentos , Penicillium/fisiología , Levaduras/fisiología , Enfermedades de las Plantas/microbiología

RESUMEN

A comparative study on the lipase-producing fungus Penicillium simplissicimum, grown on a tray type solid-state fermentation (SSF) bioreactor, was performed using stereoscopy, focus Z-stacking stereoscopic images, field emission scanning electron microscopy (FE-SEM) and environmental scanning electron microscopy (ESEM) to better characterize the morphology of filamentous fungi in SSF and their distribution over the solid matrix. The imaging of live fungal samples using a stereomicroscope with focus Z-stacking showed differences in colonization between a static SSF and an intermittent agitated SSF. A comparison of the stereomicroscopy, hi-vacuum and environmental scanning electron microscopy results obtained using different protocols for sample processing showed that fixation with osmium tetroxide vapor and subsequent hydrated imaging is the best combination of sample preparation and imaging conditions for keeping the arrangement of the aerial hyphae and conidia morphology closer to the natural state. These combined methodologies can be applied in the SSF of fungal growth to characterize the formation of conidiophores over time, the conidia morphology and the spatial organization after their release from conidiophores. Mycelium colonization over the matrix, which is an important characteristic related to the production of different biotechnological products, could be observed and provide more knowledge about fungal physiology behavior during SSF.

Asunto(s)

Fermentación , Hongos/crecimiento & desarrollo , Hongos/ultraestructura , Penicillium/crecimiento & desarrollo , Penicillium/ultraestructura , Hongos/citología , Hongos/fisiología , Hifa/ultraestructura , Microscopía/instrumentación , Microscopía/métodos , Microscopía Electrónica de Rastreo/métodos , Micelio/ultraestructura , Penicillium/citología , Penicillium/fisiología , Esporas Fúngicas/ultraestructura , Propiedades de Superficie

RESUMEN

A total of 172 Brazil nut samples (114 in shell and 58 shelled) from the Amazon rainforest region and São Paulo state, Brazil was collected at different stages of the Brazil nut production chain: rainforest, street markets, processing plants and supermarkets. The mycobiota of the Brazil nut samples were evaluated and also compared in relation to water activity. A huge diversity of Aspergillus and Penicillium species were found, besides Eurotium spp., Zygomycetes and dematiaceous fungi. A polyphasic approach using morphological and physiological characteristics, as well as molecular and extrolite profiles, were studied to distinguish species among the more important toxigenic ones in Aspergillus section Flavi and A. section Nigri. Several metabolites and toxins were found in these two sections. Ochratoxin A (OTA) was found in 3% of A. niger and 100% of A. carbonarius. Production of aflatoxins B and G were found in all isolates of A. arachidicola, A. bombycis, A. nomius, A. pseudocaelatus and A. pseudonomius, while aflatoxin B was found in 38% of A. flavus and all isolates of A. pseudotamarii. Cyclopiazonic acid (CPA) was found in A. bertholletius (94%), A. tamarii (100%), A. caelatus (54%) and A. flavus (41%). Tenuazonic acid, a toxin commonly found in Alternaria species was produced by A. bertholletius (47%), A. caelatus (77%), A. nomius (55%), A. pseudonomius (75%), A. arachidicola (50%) and A. bombycis (100%). This work shows the changes of Brazil nut mycobiota and the potential of mycotoxin production from rainforest to consumer, considering the different environments which exist until the nuts are consumed.

Asunto(s)

Biodiversidad , Abastecimiento de Alimentos , Hongos/aislamiento & purificación , Micobioma , Nueces/microbiología , Aflatoxinas/análisis , Aspergillus/aislamiento & purificación , Aspergillus/fisiología , Aspergillus flavus/aislamiento & purificación , Aspergillus flavus/fisiología , Brasil , Seguridad de Productos para el Consumidor , Microbiología de Alimentos , Hongos/fisiología , Micobioma/fisiología , Penicillium/aislamiento & purificación , Penicillium/fisiología , Bosque Lluvioso , Ácido Tenuazónico/análisis

RESUMEN

Economic losses caused by postharvest diseases represent one of the main problems of the citrus industry worldwide. The major diseases affecting citrus are the "green mold" and "blue mold", caused by Penicillium digitatum and P. italicum, respectively. To control them, synthetic fungicides are the most commonly used method. However, often the emergence of resistant strains occurs and their use is becoming more restricted because of toxic effects and environmental pollution they generate, combined with trade barriers to international markets. The aim of this work was to isolate indigenous killer yeasts with antagonistic activity against fungal postharvest diseases in lemons, and to determine their control efficiency in in vitro and in vivo assays. Among 437 yeast isolates, 8.5% show to have a killer phenotype. According to molecular identification, based on the 26S rDNA D1/D2 domain sequences analysis, strains were identified belonging to the genera Saccharomyces, Wickerhamomyces, Kazachstania, Pichia, Candida and Clavispora. Killers were challenged with pathogenic molds and strains that caused the maximum in vitro inhibition of P. digitatum were selected for in vivo assays. Two strains of Pichia and one strain of Wickerhamomyces depicted a significant protection (p <0.05) from decay by P. digitatum in assays using wounded lemons. Thus, the native killer yeasts studied in this work showed to be an effective alternative for the biocontrol of postharvest fungal infections of lemons and could be promising agents for the development of commercial products for the biological control industry.

Asunto(s)

Citrus/microbiología , Penicillium/fisiología , Control Biológico de Vectores/métodos , Enfermedades de las Plantas/microbiología , Levaduras/fisiología , Citrus/crecimiento & desarrollo , Fenotipo , Plásmidos/genética , Levaduras/genética

RESUMEN

This study aimed at evaluating if essential oils misted in broiler houses reduce environmental fungi counts. The investigation was conducted in three experimental rooms, where broiler chickens were reared between 1 to 42 d of age. Every three days, the rooms were fogged with pure water (control) or with aqueous solutions of peppermint or thyme oils. On the next day, fogging samples from the air, flat surfaces, and litter were collected and quantitatively and qualitatively analysed for fungal contamination. The treatment with essential oils showed promising results. In the room fogged with thyme oil, aerial fungi growth was not as evident as in the control room, and presented the lowest average fungi count. Thyme oil was also the most effective in reducing fungi colonization on drinker surfaces and litter. The use of peppermint oil also reduced the population of air, wall, surface and litter fungi, although some exceptions were noted. Aspergillus, Penicillium, Fusarium and Saccharomyces genera were identified most frequently. The effect of essential oils was noticeable in the last two weeks, when the counts of Aspergillus sp. were 75% (thyme oil) and 46% (peppermint oil) lower in comparison with the control group. The results show that fogging broiler houses with essential oils may be an effective prevention method against fungal aerosol in broiler houses. However, further investigations to determine the synergistic effect of different oils and their compounds, and the best possible doses and methods of application in the field are needed.

Asunto(s)

Animales , Antifúngicos/análisis , Hongos/fisiología , Pollos/inmunología , Mentha piperita/fisiología , Thymus serpyllum/análisis , Aceites Volátiles/análisis , Aspergillus/fisiología , Fusarium/fisiología , Indicadores de Contaminación/análisis , Penicillium/fisiología , Contaminación Ambiental/análisis , Saccharomyces/fisiología

RESUMEN

This study aimed at evaluating if essential oils misted in broiler houses reduce environmental fungi counts. The investigation was conducted in three experimental rooms, where broiler chickens were reared between 1 to 42 d of age. Every three days, the rooms were fogged with pure water (control) or with aqueous solutions of peppermint or thyme oils. On the next day, fogging samples from the air, flat surfaces, and litter were collected and quantitatively and qualitatively analysed for fungal contamination. The treatment with essential oils showed promising results. In the room fogged with thyme oil, aerial fungi growth was not as evident as in the control room, and presented the lowest average fungi count. Thyme oil was also the most effective in reducing fungi colonization on drinker surfaces and litter. The use of peppermint oil also reduced the population of air, wall, surface and litter fungi, although some exceptions were noted. Aspergillus, Penicillium, Fusarium and Saccharomyces genera were identified most frequently. The effect of essential oils was noticeable in the last two weeks, when the counts of Aspergillus sp. were 75% (thyme oil) and 46% (peppermint oil) lower in comparison with the control group. The results show that fogging broiler houses with essential oils may be an effective prevention method against fungal aerosol in broiler houses. However, further investigations to determine the synergistic effect of different oils and their compounds, and the best possible doses and methods of application in the field are needed.(AU)

Asunto(s)

Animales , Antifúngicos/análisis , Mentha piperita/fisiología , Aceites Volátiles/análisis , Thymus serpyllum/análisis , Pollos/inmunología , Hongos/fisiología , Contaminación Ambiental/análisis , Indicadores de Contaminación/análisis , Aspergillus/fisiología , Penicillium/fisiología , Fusarium/fisiología , Saccharomyces/fisiología

RESUMEN

The goal of the present research work was to assess the management of sewage sludge (SS) by composting using fermented water hyacinth (WHferm) as an amendment. The water hyacinth was fermented, and a higher production of volatile fatty acids (VFAs) (782.67 mg L(-1)) and soluble organic carbon (CSOL) (4788.34 mg L(-1)) was obtained using a particle size of 7 mm compared to 50 mm. For composting, four treatments (10 kg fresh weight each) were evaluated: treatment A (100 % SS + 0 % WHferm), treatment B (75 % SS + 25 % WHferm), treatment C (50 % SS + 50 % WHferm), and treatment D (25 % SS + 75 % WHferm). The WHferm added to SS, especially in treatments C (50 %) and D (75 %), increased the initial contents of organic matter (OM), organic carbon (CORG), CSOL, the C/N ratio, and the germination index (GI). The heavy metal content (HMC) (As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn) at the beginning was below the maximum allowed by USEPA regulations. All of the samples were free of Salmonella sp. from the beginning. The reduction of the CORG, CSOL, total Kjeldahl nitrogen (TKN), and C/N ratio indicated the degradation of the OM by day 198. The treatments with WHferm (B, C, and D) yielded higher values of electrical conductivity, cation exchange capacity, and GI than SS at day 198. No significant differences were observed in GI among the treatments with WHferm. The fecal coliforms were eliminated (<3 MPN g(-1)) and the helminths were reduced to ≤5 eggs/2 g during the process. The competition for nutrients and the presence of suppressive fungi of the genera Penicillium, Rhizopus, Paecilomyces (penicillin producers), and Fusariella isolated from the compost may have promoted the elimination of pathogens since no thermophile temperatures were obtained. WHferm as an amendment in the composting of SS improved the characteristics of the final product, especially when it was used in proportions of 25 and 50 %. An excellent product was obtained in terms of HMC, and the product was B class in terms of pathogens.

Asunto(s)

Eichhornia/química , Contaminantes Ambientales/análisis , Aguas del Alcantarillado/química , Suelo/química , Biodegradación Ambiental , Ácidos Grasos Volátiles/química , Fermentación , Metales Pesados/análisis , Nitrógeno , Paecilomyces/fisiología , Penicillium/fisiología , Rhizopus/fisiología , Microbiología del Suelo , Temperatura

RESUMEN

Proteins containing Zn(II)(2)Cys(6) domains are exclusively found in fungi and yeasts. Genes encoding this class of proteins are broadly distributed in fungi, but few of them have been functionally characterized. In this work, we have characterized a gene from the filamentous fungus Penicillium roqueforti that encodes a Zn(II)(2)Cys(6) protein, whose function to date remains unknown. We have named this gene pcz1. We showed that the expression of pcz1 is negatively regulated in a P. roqueforti strain containing a dominant active Gαi protein, suggesting that pcz1 encodes a downstream effector that is negatively controlled by Gαi. More interestingly, the silencing of pcz1 in P. roqueforti using RNAi-silencing technology resulted in decreased apical growth, the promotion of conidial germination (even in the absence of a carbon source), and the strong repression of conidiation, concomitant with the downregulation of the genes of the central conidiation pathway brlA, abaA and wetA. A model for the participation of pcz1 in these physiological processes in P. roqueforti is proposed.

Asunto(s)

Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Penicillium/fisiología , Carbono/metabolismo , Proteínas Fúngicas/química , Regulación Fúngica de la Expresión Génica , Silenciador del Gen , Fenotipo , Subunidades de Proteína , Interferencia de ARN , ARN Interferente Pequeño/genética

RESUMEN

Putative mechanisms of action associated with the biocontrol capacity of four yeast strains (Cryptoccocus albidus NPCC 1248, Pichia membranifaciens NPCC 1250, Cryptoccocus victoriae NPCC 1263 and NPCC 1259) against Penicillium expansum and Botrytis cinerea were studied by means of in vitro and in situ assays. C. albidus(YP), a commercial yeast was also evaluated for comparative purposes. The yeast strains exhibited a variety of different mechanisms including: wound colonization, germination inhibition, biofilm formation, secretion of killer toxins, competition for nutrient and secretion of hydrolytic enzymes (protease, chitinase and glucanase). The relationship between strains (and their associated antagonist mechanisms) and in situ antagonist activity was also evaluated. Results indicate that mechanisms such as production of hydrolytic enzymes, the ability for colonization of wounds, production of killer toxin and inhibition of germination are the most important for biocontrol activity. Our study indicate that multiple modes of action may explain why P. membranifaciens NPCC 1250 and C. victoriae NPCC 1263 provided excellent control of postharvest pears disease.

Asunto(s)

Agentes de Control Biológico , Botrytis/fisiología , Penicillium/fisiología , Enfermedades de las Plantas/prevención & control , Pyrus/microbiología , Levaduras/fisiología , Antibiosis , Recuento de Colonia Microbiana , Cryptococcus/fisiología , Frutas/microbiología , Hidrolasas , Factores Asesinos de Levadura/metabolismo , Pichia/fisiología , Enfermedades de las Plantas/microbiología

RESUMEN

Trichomonas vaginalis is the causative agent of trichomonosis, the most common non-viral sexually transmitted disease. Infection with this protozoan may have serious consequences, especially for women. Currently, 5-nitroimidazole drugs are the treatment of choice for trichomonosis, but the emergence of resistance has limited the effectiveness of this therapy. In this context, this study aimed to evaluate the anti-T. vaginalis activity of marine-associated fungi found in the South Brazilian Coast. A total of 42 marine-associated fungal species (126 filtrate samples) isolated from 39 different marine organisms, mainly sponges, were selected to be screened against T. vaginalis. Of these, two filtrate samples from Hypocrea lixii F02 and Penicillium citrinum F40 showed significant growth-inhibitory activity (up to 100%) against ATCC 30236 and fresh clinical isolates, including a metronidazole-resistant isolate. Minimum inhibitory concentration (MIC) values of H. lixii F02 and P. citrinum F40 samples for all isolates tested, including the metronidazole-resistant isolate, were 2.5 mg/mL. The kinetic growth curve showed that the filtrate samples were able to reduce the density of parasites to zero within 24 h of incubation, which was confirmed by microscopy. Both fungal filtrate samples exhibited no hemolytic activity, and the P. citrinum F40 filtrate sample showed low cytotoxicity against Vero cells. These data suggest that marine-associated fungi from the South Brazilian Coast may produce potential candidates for further investigation and possible use in the treatment of metronidazole-resistant trichomonosis.

Asunto(s)

Hongos/fisiología , Trichomonas vaginalis/microbiología , Animales , Antiprotozoarios/farmacología , Brasil , Chlorocebus aethiops , Resistencia a Medicamentos , Eritrocitos/efectos de los fármacos , Eritrocitos/microbiología , Hongos/química , Hemólisis/efectos de los fármacos , Humanos , Hypocrea/química , Hypocrea/fisiología , Metronidazol/farmacología , Datos de Secuencia Molecular , Pruebas de Sensibilidad Parasitaria , Penicillium/química , Penicillium/fisiología , Poríferos/microbiología , Agua de Mar , Trichomonas vaginalis/efectos de los fármacos , Células Vero

RESUMEN

AIM: To investigate the cellular damage on Penicillium digitatum produced by a sequential oxidative treatment (SOT), previously standardized in our laboratory, to prevent the conidia growth. Lethal SOT consists of 2-min preincubation with 10 ppm NaClO followed by 2-min incubation with 6 mmol l(-1) CuSO(4) and 100 mmol l(-1) H(2)O(2) at 25°C. METHODS AND RESULTS: After the application of lethal SOT or sublethal SOT (decreasing only the H(2)O(2) concentration), we analysed several conidia features such as germination, oxygen consumption, ultrastructure and integrity of the cellular wall and membrane. Also, we measured the production of reactive oxygen species (ROS) and the content of thiobarbituric acid-reactive species (TBARS). With the increase of H(2)O(2) concentration in the SOT, germination and oxygen consumption of conidia became inhibited, while the membrane permeability, ROS production and TBARS content of conidia increased. Several studies revealed ultrastructural disorganization in P. digitatum conidia after lethal SOT, showing severe cellular damage without apparent damage to the cell wall. In addition, mycelium of P. digitatum was more sensitive than conidia to the oxidative treatment, because growth ceased and permeability of the membranes increased after exposure of the mycelium to a SOT with only 50 mmol l(-1) H(2)O(2) compared to a SOT of 100 mmol l(-1) for these effects to occur on conidia. CONCLUSION: Our insights into cellular changes produced by the lethal SOT are consistent with the mode of action of the oxidant compounds, by producing both alteration of membrane integrity and intracellular damage. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results allow the understanding of SOT effects on P. digitatum, which will be useful to develop a reliable treatment to control postharvest diseases in view of its future application in packing houses.

Asunto(s)

Oxidantes/toxicidad , Penicillium/efectos de los fármacos , Permeabilidad de la Membrana Celular/efectos de los fármacos , Pared Celular/efectos de los fármacos , Micelio/efectos de los fármacos , Micelio/crecimiento & desarrollo , Estrés Oxidativo , Penicillium/fisiología , Penicillium/ultraestructura , Esporas Fúngicas/efectos de los fármacos , Esporas Fúngicas/crecimiento & desarrollo , Esporas Fúngicas/ultraestructura

RESUMEN

AIMS: To investigate the presence of fungi during three human decomposition stages: bloated, putrefaction and skeletonization. METHODS AND RESULTS: The samples were gathered in the city of Fortaleza, Ceará, Brazil, from the public morgue and cemeteries. The material was submitted to conventional mycological procedures by direct examination and macro/micro morphological and biochemical analyses. The main fungi isolated were Aspergillus spp., Penicillium spp. and Candida spp. in the bloated stage (n = 34 cadavers) and in the putrefaction stage (n = 6 cadavers), while in the skeletonization stage (n = 20 cadavers), the main fungi were Aspergillus spp., Penicillium spp. and Mucor sp. CONCLUSIONS: Aspergillus, Penicillium and Candida species were associated with decomposed human cadavers. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings enable tracing out the profile of fungal communities of human cadavers for the first time. However, much more research will be necessary to develop this new segment of mycology and to enable its routine use in forensic science.

Asunto(s)

Ecosistema , Ciencias Forenses , Hongos Mitospóricos/fisiología , Cambios Post Mortem , Aspergillus/aislamiento & purificación , Aspergillus/fisiología , Biodiversidad , Brasil , Candida/aislamiento & purificación , Candida/fisiología , Humanos , Hongos Mitospóricos/aislamiento & purificación , Penicillium/aislamiento & purificación , Penicillium/fisiología

RESUMEN

Apples and apple products are excellent substrates for Penicillium expansum to produce patulin. In an attempt to avoid excessive levels of patulin, limiting or reducing P. expansum contamination levels on apples designated for storage in packinghouses and/or during apple juice processing is critical. The aim of this work was (i) to determine the thermal resistance of P. expansum spores in apple juice, comparing the abilities of the Bigelow and Weibull models to describe the survival curves and (ii) to determine the inactivation of P. expansum spores in aqueous chlorine solutions at varying concentrations of chlorine solutions, comparing the abilities of the biphasic and Weibull models to fit the survival curves. The results showed that the Bigelow and Weibull models were similar for describing the heat inactivation data, because the survival curves were almost linear. In this case, the concept of D- and z-values could be used, and the D-values obtained were 10.68, 6.64, 3.32, 1.14, and 0.61 min at 50, 52, 54, 56, and 60 degrees C, respectively, while the z-value was determined to be 7.57 degrees C. For the chlorine treatments, although the biphasic model gave a slightly superior performance, the Weibull model was selected, considering the parsimony principle, because it has fewer parameters than the biphasic model has. In conclusion, the typical pasteurization regimen used for refrigerated apple juice (71 degrees C for 6 s) is capable of achieving a 6-log reduction of P. expansum spores.

Asunto(s)

Cloro/farmacología , Desinfectantes/farmacología , Calor , Penicillium/efectos de los fármacos , Penicillium/fisiología , Malus , Penicillium/metabolismo

RESUMEN

The use of fungal starter strains in the casing of dry fermented sausages allows standardization of the manufacturing process and ensures consumer safety. Penicillium nalgiovense is normally used for this purpose. Even though this species is reported as safe with respect to the production of the most common mycotoxins, its safety may be strain specific. The aim of the present work was to assess the toxicogenic potential of nine P. nalgiovense strains isolated from dry fermented sausages that were previously suitable as starters. The strains were tested for toxicity on brine shrimp larvae and the human cell line MCF7, for mutagenicity in the Ames test, and for antibacterial activity against gram-positive and gram-negative bacteria. According to our results, several P. nalgiovense strains were positive in more than one bioassay. Therefore, it is important to use different toxicological assays when characterizing strains intended for food use. Strains S1-2 and S14-4, which belong to biotypes 6 and 5, respectively, were nontoxigenic under the conditions tested. Overall, strain S1-2 of P. nalgiovense proved to be best suited as a starter in dry fermented sausage manufacture because in addition of being nontoxicogenic it produces white conidia, which is a desirable feature.

Asunto(s)

Seguridad de Productos para el Consumidor , Manipulación de Alimentos/métodos , Productos de la Carne/microbiología , Penicillium/fisiología , Animales , Línea Celular/microbiología , Recuento de Colonia Microbiana , Fermentación , Microbiología de Alimentos , Tecnología de Alimentos , Humanos , Microbiología Industrial , Penaeidae/microbiología , Especificidad de la Especie , Porcinos

RESUMEN

Tissues from Coffea arabica, C. congensis, C. dewevrei and C. liberica collected in Colombia, Hawaii and at a local plant nursery in Maryland were sampled for the presence of fungal endophytes. Surface sterilized tissues including roots, leaves, stems and various berry parts were plated on yeast-malt agar. DNA was extracted from a set of isolates visually recognized as Penicillium, and the internal transcribed spacer region and partial LSU-rDNA was amplified and sequenced. Comparison of DNA sequences with GenBank and unpublished sequences revealed the presence of 11 known Penicillium species: P. brevicompactum, P. brocae, P. cecidicola, P. citrinum, P. coffeae, P. crustosum, P. janthinellum, P. olsonii, P. oxalicum, P. sclerotiorum and P. steckii as well as two possibly undescribed species near P. diversum and P. roseopurpureum. Ochratoxin A was produced by only four isolates, one isolate each of P. brevicompactum, P. crustosum, P. olsonii and P. oxalicum. The role these endophytes play in the biology of the coffee plant remains enigmatic.

Asunto(s)

Coffea/microbiología , Ocratoxinas/biosíntesis , Penicillium/aislamiento & purificación , Penicillium/fisiología , Colombia , ADN de Hongos/química , ADN de Hongos/genética , ADN de Hongos/aislamiento & purificación , ADN Ribosómico/química , ADN Ribosómico/genética , ADN Ribosómico/aislamiento & purificación , ADN Espaciador Ribosómico/química , ADN Espaciador Ribosómico/genética , ADN Espaciador Ribosómico/aislamiento & purificación , Hawaii , Maryland , Datos de Secuencia Molecular , Penicillium/clasificación , Filogenia , Estructuras de las Plantas/microbiología , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido Nucleico