Diversity of fungi obtained from bats captured in urban forest fragments in Sinop, Mato Grosso, Brazil / Diversidade de fungos em morcegos de fragmentos florestais urbanos de Sinop, Mato Grosso, Brasil

Bats are important for the homeostasis of ecosystems and serve as hosts of various microorganisms including bacteria, viruses, and fungi with pathogenic potential. This study aimed to isolate fungi from biological samples obtained from bats captured in the city of Sinop (state of Mato Grosso, Brazil), where large areas of deforestation exist due to urbanization and agriculture. On the basis of the flow of people and domestic animals, 48 bats were captured in eleven urban forest fragments. The samples were processed and submitted to microbiological cultures, to isolate and to identify the fungal genera. Thirty-four (70.83%) of the captured bats were positive for fungi; 18 (37.5%) and 16 (33.33%) of these bats were female and male, respectively. Penicillium sp., Scopulariopsis sp., Fusarium sp., Aspergillus sp., Alternaria sp., Cryptococcus sp., Trichosporon sp., and Candida sp., which may cause opportunistic infections, were isolated. The bat species with the highest number of fungal isolates was Molossus molossus: 21 isolates (43.8%). According to our results, bats captured in urban forest fragments in Sinop harbor pathogenic fungi, increasing the risk of opportunistic fungal infections in humans and domestic animals.

Os morcegos apresentam grande importância na homeostasia dos ecossistemas e são hospedeiros de uma rica diversidade de micro-organismos como bactérias, vírus e fungos com potencial patogênico. Portanto, este estudo visou isolar fungos presentes em amostras biológicas de morcegos na cidade de Sinop - MT, que possui grandes áreas de desmatamento devido à urbanização e agricultura. Foram capturados 48 morcegos de diferentes espécies, em onze fragmentos florestais urbanos definidos de acordo com fluxo de pessoas e animais domésticos, para obtenção de amostras biológicas. Essas amostras foram processadas e submetidas aos cultivos microbiológicos, para isolamento e identificação dos gêneros dos fungos. Dos 48 morcegos, 34 (70,83%) foram positivos para pelos menos um gênero de fungo, sendo 18 (37,5%) fêmeas e 16 (33,33%) machos, e os gêneros isolados a partir das amostras biológicas foram Penicillium sp., Scopulariopsis sp., Fusarium sp., Aspergillus sp., Alternaria sp., Cryptococcus sp., Trichosporon sp. e Candida sp., que podem ser causadores de infecções oportunistas. Desse total, a espécie que apresentou maior positividade para pelo menos um gênero de fungo foi Molossus molossus com 21 (43,8%). Nossos resultados demonstram que os morcegos capturados nos fragmentos florestais urbanos na cidade de Sinop - MT, podem atuar como agentes veiculadores de fungos com potencial patogênico, aumentando assim o risco de exposição e aquisição de infecções fúngicas oportunistas por pessoas e animais domésticos.

Higienización y eliminación fúngica en instalaciones industriales para la disminución de riesgos ocupacionales / Sanitization and fungal elimination in industrial facilities to reduce occupational risks

Se ha comprobado que la exposición a bioaerosoles se asocia con varios efectos sobre la salud, como enfermedades pulmonares y alergias. El presente estudio transversal tuvo como objetivo investigar la contaminación por hongos en varias superficies pertenecientes a dos industrias de alimentos. La toma de muestra se realizó en tres semanas, inmediatamente después de realizada la limpieza y desinfección y antes de iniciar la producción. Se recolectaron 400 muestras de superficie (vidrio, acero inoxidable, aluminio, goma y plástico), también se evaluó la eficacia del programa de higienización en el control de la población de hongos. Todas las muestras presentaron recuentos fúngicos <10 ufc/cm2, sin haber diferencias significativas entre los tipos de superficie, sin embargo, las superficies plásticas exhibieron mayor crecimiento logarítmico de los hongos. Se determinó que 49,60% de las especies identificadas correspondieron al género Penicillium. Aunque los niveles de hongos en las industrias estudiadas fueron inferiores a los niveles recomendados por la Organización Mundial de la Salud, algunas medidas de salud ambiental como lavar y desinfectar las superficies después de cada turno de trabajo, y se recomiendan inspecciones periódicas para garantizar la seguridad de los trabajadores y de los productos que allí se manufacturan(AU)

It has been proven that exposure to bioaerosols is associated with several health effects, such as pulmonary diseases and allergies. The present crosssectional study aimed to investigate fungal contamination on various surfaces belonging to two food industries. The sampling was carried out in three weeks, immediately after cleaning and disinfection and before starting production. 400 surface samples were collected (glass, stainless steel, aluminum, rubber and plastic), the effectiveness of the sanitation program in controlling the fungal population was also evaluated. All the samples presented fungal counts <10 cfu / cm2, with no significant differences between the types of surfaces, however, the plastic surfaces exhibited higher logarithmic growth of the fungi. It was determined that 49.60% of the identified species corresponded to the Penicillium genus.Although the levels of fungi in the studied baths were lower than the levels recommended by the World Health Organization, some environmental health measure ssuch as washing and disinfecting surfaces after each working shift and periodic inspections are recommended ensuring the safety of the workers and the products that are manufactured there(AU)

A novel nano-sulphur and essential oil-based room freshener

Introducción: Alternaria spp y Candida spp. son hongos patógenos de ambientes interiores como la casa, la oficina, el aula, etc., causan enfermedades como asma crónica e infecciones sistémicas en individuos inmunodeprimidos a través de la secreción de diversas sustancias tóxicas. Los ambientadores a base de productos químicos disponibles comercialmente que se utilizan para controlar la carga de hongos en el ambiente interior no son beneficiosos para la salud humana. Objetivo: proporcionar una alternativa viable en forma de enfoque basado en nanopartículas para el manejo de hongos transmitidos por el aire. Metodología: aislamiento, identificación microscópica y bioquímica de hongos de interior; Síntesis de nanopartículas de azufre (SNP) mediadas por Azadirachta indica, su detección y caracterización; y evaluación in vitro de SNP contra hongos aislados presentes en el ambiente interior. Resultado: Los hongos aislados se identificaron como especies de Alternaria spp y Candida spp. Los SNP mostraron máximos de absorbancia a 291 nm. El análisis NTA mostró un tamaño medio de 188,4 nm y un potencial zeta de -4,94 mV, lo que representa una síntesis de SNP estables. El patrón XRD confirmó la naturaleza cristalina cúbica centrada en la cara de los SNP. El espectro FTIR representó la presencia de compuestos polihidroxilo, nitrilo, ceto, aromáticos y carboxílicos que estabilizaron los SNP. Los ensayos antifúngicos demostraron la actividad significativa de los SNP formulados y del ambientador infundido con aceite de eucalipto. Conclusión: Los SNP mediados por A. indica se pueden aplicar en la formulación y fabricación de un ambientador ecológico para el manejo de hongos patógenos de interior como Alternaria spp y Candida spp.

Introduction: Alternaria spp. and Candida spp. are the main fungal pathogen of indoor environment like house, office, classroom, etc. These may cause various diseases and infections like systemic infections, or chronic asthma in immunocompromised individuals through secretion of various toxic substances. Chemical-based commercially available room fresheners used to control the fungal load of indoor environment are not beneficial to human health. Objective: was to provide viable alternative in the form of nanoparticle-based approach for the management of air-borne fungi. Methodology: The present study primarily focuses on the isolation, microscopic and biochemical identification of indoor fungi; Azadirachta indica-mediated sulphur nanoparticles (SNPs) synthesis, their detection and characterization; and in vitro assessment of SNPs against isolated fungi present in indoor environment. Result: The isolated fungi were identified as Alternaria spp and Candida spp. The SNPs showed absorbance maxima at 291 nm. NTA analysis showed average size of 188.4 nm, and zeta potential of -4.94 mV which represented synthesis of stable SNPs. XRD pattern confirmed the face centered cubic, crystalline nature of SNPs. FTIR spectrum depicted the presence of polyhydroxyl, nitrile, keto, aromatic and carboxylic compounds which stabilized the SNPs. The antifungal assays demonstrated the significant activity of the formulated SNPs and eucalyptus oil infused air freshener. Conclusion: It can be concluded that A. indica-mediated SNPs can be applied in the formulation and manufacture of an ecofriendly air freshener for the management of indoor fungal pathogens like Alternaria spp. and Candida spp.

Afectación intestinal por un hongo dematiáceo / Intestinal disease by a dematiaceous fungi

Resumen Introducción: Los hongos dematiáceos se caracterizan por la presencia de abundante melanina en su pared celular. Presentan una distribución mundial, siendo más comunes en climas tropicales y subtropicales. Producen infecciones cutáneas y subcutáneas, además de enfermedades alérgicas, neumonías, abscesos cerebrales o infecciones diseminadas. Caso Clínico: Presentamos el caso de un paciente con adenocarcinoma de recto intervenido quirúrgicamente con hallazgo incidental de divertículo de Meckel y en el cual en el estudio anatomopatológico reveló la presencia de un hongo dematiáceo

Introduction: Dematiaceous fungi are characterized by the presence of brown melanine or melanine like pigments in their cell wall. They are generally distributed worldwide, being more common in tropical and subtropical climates. The clinical syndromes are often cutaneous and subcutaneous infections, but can be also responsible of allergic diseases, pneumonias, cerebral abscesses or disseminated infections. Clinical Case: We present the case of a patient with a diagnosis of rectal adenocarcinoma intervening surgically and with an incidental finding of Meckel's Diverticulum. The anatomopathological study revealed the presence of a dematiaceous fungi.

Tolerance of dark septate endophytic fungi (DSE) to 1 agrochemicals in vitro / Tolerancia de hongos endofíticos septados oscuros a agroquímicos in vitro

Abstract Dark septate endophytes (DSE) are a heterogeneous group of fungi, mostly belonging to the Phylum Ascomycota, that are involved in a mutualistic symbiosis with plant roots. The aim of this study is to evaluate the behavior of two strains of DSE isolated from wheat roots of two cropping areas in the province of Buenos Aires, Argentina, against some agrochemicals. Of all the isolates obtained, two strains were identified as Alternaria alternata and Cochliobolus sp. These DSE were found to be tolerant to glyphosate, carbendazim and cypermethrin when evaluated at the recommended agronomic dose (AD), 2 AD and, in some cases, 10 AD. This work contributes to the study of the biology of this group of fungi and their tolerance in the presence of xenobiotics widely used in agriculture.© 2019 Asociaci´on Argentina de Microbiolog´ıa. Published by Elsevier Espa˜na, S.L.U. This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/bync-nd/4.0/).

Resumen Los endófitos septados oscuros (DSE) son un grupo heterogéneo de hongos que participan de una simbiosis mutualista con raíces de plantas, perteneciendo principalmente al Phylum Ascomycota. El objetivo de este estudio fue aislar DSE de raíces de trigo proveniente de dos áreas de cultivo de la provincia de Buenos Aires y evaluar el comportamiento de dos cepas de DSE aisladas de raíces de trigo frente a algunos agroquímicos en dos áreas de cultivo de la provincia de Buenos Aires. De todos los aislamientos obtenidos se seleccionaron dos cepas que se identificaron como Alternaria alternata y Cochliobolus sp. Se encontró que estos DSE son tolerantes al glifosato, el carbendazim y la cipermetrina, evaluados a las dosis agronómicas recomendadas (AD), a 2x AD y, en algunos casos, a 10x AD. Este trabajo contribuye al conocimiento de la biología de este grupo de hongos y su tolerancia a xenobióticos ampliamente utilizados en la agricultura.

Anaphylaxis to Agaricus bisporus ingestion / Anafilaxia por ingestão de Agaricus bisporus

ABSTRACT A 33-year-old male with house dust mite allergic rhinitis and asthma reported an episode of facial and lip angioedema, dyspnea, cough and dysphagia at the age of 25, minutes after eating a mushroom ( Agaricus bisporus ) pizza. He denied any drug intake, hymenoptera stings or other possible triggers, and no identifiable cofactors were present. Since then he avoided all types of mushrooms, however an accidental contact occurred with mushroom sauce that resulted in angioedema of the lip within minutes. The allergy workup included measurements of total IgE and specific IgE to mushroom, and skin prick test to aeroallergens sources, possible food allergen sources and mushroom extract, a prick to prick test with raw and cooked A. bisporus , in addition to a SDS-PAGE and immunoblotting assay. The study revealed a specific IgE to mushroom of 0.76kUA/L positive skin prick test to mushroom extract, and prick to prick test positive to white and brown A. bisporus (raw and cooked). The immunoblotting identified two IgE binding proteins with 10kDa and 27kDa. We report a case of A. bisporus anaphylaxis probably due to primary mushroom sensitization. We detected two IgE-reactive proteins with 10kDa and 27kDa as possible culprit allergens.

RESUMO Paciente do sexo masculino, 33 anos, com asma e rinite alérgica desencadeadas por ácaros, relatou episódio de angioedema facial e labial, dispneia, tosse e disfagia aos 25 anos, minutos após a ingestão de uma pizza de cogumelo ( Agaricus bisporus ). O paciente negou consumo de medicamentos, picadas de himenópteros, ou quaisquer outros possíveis desencadeadores ou cofatores que pudessem estar presentes. Desde então, evita todos os tipos de cogumelos, até a ocorrência de um contato acidental com molho de cogumelo, que resultou em angioedema labial minutos após. O estudo imunoalergológico incluiu doseamento de IgE total e específica para cogumelos, testes cutâneos para aeroalérgenos, possíveis alérgenos alimentares e extrato de cogumelos, teste prick to prick com A. bisporus cru e cozido e teste de SDS-PAGE immunoblotting . O estudo revelou IgE específica para cogumelos de 0,76kUA/L, teste cutâneo positivo para extrato de cogumelos e teste prick to prick positivo para A. bisporus branco e castanho (cru e cozido). O immunoblotting identificou duas proteínas de ligação de IgE, de 10kDa e 27kDa. Relatamos, assim, um caso de anafilaxia por ingestão de A. bisporus , provavelmente explicado por sensibilização primária a cogumelos. Detectamos duas proteínas IgE-reativas de 10kDa e 27kDa como os possíveis alérgenos responsáveis.

Efeito combinado de carvacrol e tiabendazol contra Colletotrichum gloesporioides, Fusarium solani e Alternaria alternata / Combined effect of carvacrol and thiabendazole against Colletotrichum gloesporioides, Fusarium solani and Alternaria alternata

O objetivo deste trabalho foi determinar a atividade antimicrobiana do carvacrol e sua combinação com tiabendazol no controle de fungos patogênicos deteriorantes de frutas (Colletotrichum gloesporioides, Fusarium solani e Alternaria alternata). O carvacrol apresentou uma concentração inibitória mínima (CIM) de 282 a 563 μg mL-1 para os fungos testados. Quando avaliado em conjunto com o tiabendazol apresentou efeito aditivo contra C. gloesporioides e F. solani (FICI 0,5 e 1,0, respectivamente) e sinérgico contra a A.alternata (FICI 0,1). Houve redução da CIM do carvacrol de 50 a 88%. Este estudo mostra o potencial do uso.

Biocontrol potential of saline- or alkaline-tolerant Trichoderma asperellum mutants against three pathogenic fungi under saline or alkaline stress conditions

ABSTRACT Salinity and alkalinity are major abiotic stresses that limit growth and development of poplar. We investigated biocontrol potential of saline- and alkaline-tolerant mutants of Trichoderma asperellum to mediate the effects of salinity or alkalinity stresses on Populus davidiana × P. alba var. pyramidalis (PdPap poplar) seedlings. A T-DNA insertion mutant library of T. asperellum was constructed using an Agrobacterium tumefaciens mediated transformation system; this process yielded sixty five positive transformants (T1-T65). The salinity tolerant mutant, T59, grew in Potato Dextrose Agar (PDA) containing up to 10% (1709.40 mM) NaCl. Under NaCl-rich conditions, T59 was most effective in inhibiting Alternaria alternata (52.00%). The alkalinity tolerant mutants, T3 and T5, grew in PDA containing up to 0.4% (47.62 mM) NaHCO3. The ability of the T3 and T5 mutants to inhibit Fusarium oxysporum declined as NaHCO3 concentrations increased. NaHCO3 tolerance of the PdPap seedlings improved following treatment with the spores of the WT, T3, and T5 strains. The salinity tolerant mutant (T59) and two alkalinity tolerant mutants (T3 and T5) generated in this study can be applied to decrease the incidence of pathogenic fungi infection under saline or alkaline stress.

Effect of fungal incidence on physiological quality of tobacco seeds used in Brazil / Efeito da incidência de fungos na qualidade fisiológica de sementes de tabaco utilizadas no Brasil

The objectives of the present project were to evaluate the sanity and germination of tobacco seed samples cultivated in Brazil and to identify potential pathogenic fungi to the culture. Thirty-four representatives samples of tobacco seeds were evaluated through germination and sanity test by the blotter-test. On the germination test, results were expressed in percentage of seedlings considered normal, abnormal and non-germinated seeds. The percentage of seeds germination varied between 54.5 and 90%. According to sanity test, it was verified the incidence of Alternaria alternata, Cladosporium spp., Fusarium spp., and Phoma spp. The incidence of A. alternata varied from 3 to 67%, and Fusarium spp. varied from 0 to 19%. There was significative positive correlation between the incidence of A. alternata and the percentage of seed germination. Species of Alternaria and Fusarium can be potential pathogens to tobacco culture and an alarm for the necessity of seed treatment of the seeds that are transporting these fungi.(AU)

Os objetivos do presente trabalho foram avaliar a sanidade e a germinação de amostras de sementes de tabaco utilizadas no Brasil e identificar fungos potencialmente fitopatogênicos à cultura. Foram avaliadas 34 amostras representativas de lotes de sementes de tabaco, por meio dos testes de germinação, do método do rolo de germinação, e sua sanidade pelo método do papel de filtro. Os resultados do teste de germinação foram expressos em porcentual de plântulas consideradas normais, anormais e sementes não germinadas. A porcentagem de germinação das sementes variou entre 54,5 e 90%. Mediante o teste de sanidade, foi verificada a incidência dos fungos Alternaria alternata, Cladosporium spp., Fusarium spp. e Phoma spp. A incidência de A. alternata variou de 3 a 67%, e Fusarium spp., de 0 a 19%. Houve correlação significativa positiva entre a incidência de A. alternata e a porcentagem de germinação das sementes. Espécies dos gêneros Alternaria e Fusarium podem ser potenciais patógenos para a cultura do tabaco, alertando para a necessidade do tratamento das sementes portadoras desses fungos.(AU)

Pathogenicity and transmission of fungi detected on Passiflora alata seeds / Patogenicidade e transmissão de fungos detectados em sementes de Passiflora alata

Passion fruit is usually propagated by seeds because of the ease and lower cost in seedling production. However, the seed is the most efficient agent for the spread of pathogens. The damages from seed-borne diseases occur mainly during the germination stages or at the formation of seedlings in nurseries. Considering the need for knowledge on the pathology of sweet passion fruit seeds, the objective was to evaluate the transmission and pathogenicity of the fungi Alternaria sp., Botrytis fabae, Cladosporium cladosporioides, Fusarium spp. and Lasiodiplodia theobromae, known as potentially pathogenic to this crop, and isolated from sweet passion fruit seeds. Therefore, tests on seed health, germination and seedling emergence in a sterilized commercial substrate were conducted using seeds from this species, inoculated with those fungal isolates. Leaves, stems and fruit from this plant were also inoculated with the same fungi. Alternaria sp., Fusarium spp. and L. theobromae were identified in seedlings obtained from inoculated seeds, confirming the transmission of these fungi by seeds. L. theobromae was also considered the most harmful fungus to passion fruit crop, as it causes seed rot and other disease symptoms on the leaves, stem and fruit. These findings inferred that healthy seeds of sweet passion fruit are essential for producing seedlings and to prevent the spread of the diseases caused by these fungi to exempt areas.(AU)

O maracujazeiro geralmente é propagado por meio de sementes em virtude da facilidade e do menor custo na produção de mudas. No entanto, a semente é o agente mais eficiente de disseminação de patógenos, sendo que os danos decorrentes das doenças transmitidas por elas ocorrem principalmente durante os estágios de germinação ou na formação de mudas nos viveiros. Considerando a necessidade de informações acerca da patologia de sementes de maracujá-doce nesse contexto, objetivou-se obter informações sobre a transmissão e a patogenicidade dos fungos Alternaria sp., Botrytis fabae, Cladosporium cladosporioides, Fusarium spp. e Lasiodiplodia theobromae, isolados de sementes de maracujá-doce e potencialmente patogênicos à cultura. Para tanto, testes de sanidade, germinação e emergência de plântulas em substrato comercial esterilizado foram conduzidos com sementes dessa espécie, inoculadas com esses isolados. Folhas, colo e frutos dessa planta também foram inoculados com os mesmos fungos. Alternaria sp., Fusarium spp. e L. theobromae foram identificados em plântulas obtidas de sementes inoculadas, confirmando a transmissão por sementes. L. theobromae foi considerado o mais agressivo à cultura do maracujá, por ter causado podridão nas sementes, além de maiores lesões nas folhas, no colo da planta e nos frutos. Dessa forma, infere-se que a obtenção de sementes de maracujá-doce sadias é imprescindível para a produção de mudas, evitando-se assim a disseminação desses patógenos em áreas isentas.(AU)

Seeds of weeds as an alternative host of phytopathogens / Sementes de plantas daninhas como hospedeiras alternativas de fitopatógenos

Weeds compete with agricultural crops for water, light, nutrients and space, besides having an extensive seed bank. However, another aspect to be considered relates to few studies pointing out weeds as hosts of phytopathogenic fungi. Many fungi, the main cause of diseases in plants, are known to use seeds as an efficient means of survival and dispersal. The objective of this work was to evaluate the health of weed seeds and the pathogenicity of fungi associated to plants of agricultural importance. The seeds were collected manually in Cerrado areas located in the municipality of Gurupi, Tocatins, Brazil. The blotter test method was used to evaluate seed health. The incidence of fungi was evaluated through an individual analysis of seeds using a stereoscopic and an optical microscope. The pathogenicity of fungi from weed seeds was evaluated by inoculation in plants of agronomic interest and, when pathogenic, we inoculated them in the host plant of the fungus. Weed seeds have been identified in fungi of the genus Alternaria, Aspergillus, Bipolaris, Cladosporium, Curvularia, Fusarium, Nigrospora, Papularia, Rhizopus and Pythium. The seeds of Acanthospermum australe, Bidens pilosa, Cenchrus echinatus, Digitaria horizontalis, Echinochloa crus-pavonis, Eleusine indica, Ipomoea sp., Pennisetum setosum, Sida rhombifolia, Spermacoce latifolia, Tridax procumbens and Vernonia polyanthes carry and disseminate fungi that, once inoculated, cause infection in plants of agricultural importance, such as Oryza sativa, Phaseolus vulgaris, Vigna unguiculata, Zea mays and Glycine max.(AU)

As plantas daninhas competem com culturas agrícolas por água, luz, nutrientes e espaço, além de possuírem um extenso banco de sementes. Entretanto, outra vertente a ser considerada é quanto aos poucos estudos relacionando plantas daninhas como hospedeiras de fungos fitopatogênicos. É sabido que muitos fungos, principais causadores de doenças em plantas, utilizam as sementes como meio eficiente de sobrevivência e de dispersão. Dessa forma, o trabalho objetivou avaliar a sanidade de sementes de plantas daninhas e a patogenicidade dos fungos associados às plantas de importância agrícola. As sementes foram coletadas manualmente em áreas de cerrado localizadas no município de Gurupi, Tocantins, utilizando o método blotter test para avaliação da sanidade. A incidência dos fungos foi avaliada com auxílio de microscópio estereoscópico e ótico. A patogenicidade dos fungos oriundos das sementes de plantas daninhas foi avaliada por meio da inoculação em plantas de interesse agronômico e, quando patogênico, a inoculação foi na própria planta daninha hospedeira do fungo. Foram identificados os fungos dos gêneros Alternaria, Aspergillus, Bipolaris, Cladosporium, Curvularia, Fusarium, Nigrospora, Papularia, Rhizopus e Pythium. As sementes de Acanthospermum australe, Bidens pilosa, Cenchrus echinatus, Digitaria horizontalis, Echinochloa crus-pavonis, Eleusine indica, Ipomoea sp., Pennisetum setosum, Sida rhombifolia, Spermacoce latifolia, Tridax procumbens e Vernonia polyanthes transportam e disseminam fungos que, uma vez inoculados, causam infecção em plantas de importância agrícola, como Oryza sativa, Phaseolus vulgaris, Vigna unguiculata, Zea mays e Glycine max.(AU)

Isolation, identification, and biocontrol of antagonistic bacterium against Botrytis cinerea after tomato harvest

ABSTRACT Tomato is one of the most important vegetables in the world. Decay after harvest is a major issue in the development of tomato industry. Currently, the most effective method for controlling decay after harvest is storage of tomato at low temperature combined with usage of chemical bactericide; however, long-term usage of chemical bactericide not only causes pathogen resistance but also is harmful for human health and environment. Biocontrol method for the management of disease after tomato harvest has great practical significance. In this study, antagonistic bacterium B-6-1 strain was isolated from the surface of tomato and identified as Enterobacter cowanii based on morphological characteristics and physiological and biochemical features combined with sequence analysis of 16SrDNA and ropB gene and construction of dendrogram. Effects of different concentrations of antagonistic bacterium E. cowanii suspension on antifungal activity after tomato harvest were analyzed by mycelium growth rate method. Results revealed that antifungal activity was also enhanced with increasing concentrations of antagonistic bacterium; inhibitory rates of 1 × 105 colony-forming units (cfu)/mL antagonistic bacterial solution on Fusarium verticillioides, Alternaria tenuissima, and Botrytis cinerea were 46.31%, 67.48%, and 75.67%, respectively. By using in vivo inoculation method, it was further confirmed that antagonistic bacterium could effectively inhibit the occurrence of B. cinerae after tomato harvest, biocontrol effect of 1 × 109 cfu/mL zymotic fluid reached up to 95.24%, and antagonistic bacterium E. cowanii has biocontrol potential against B. cinerea after harvest of fruits and vegetables.

Chalcones to control Alternaria alternata in murcott tangor fruits / Chalconas no controle de Alternaria alternata em frutos de tangor murcote

Alternaria brown spot (ABS), caused by the fungus Alternaria alternata pathotype tangerine, is one of the main phytosanitary problems for mandarin growers. About 15 applications per year of harmful fungicides are required for controlling ABS disease in citrus orchards. As chalcones seem to be less toxic to humans and environment than the commercial fungicides in use, this study initially aimed at synthesizing 137 chalcones through aldolic condensations between benzaldehydes and acetophenones. The resulting chalcones were screened for activity against A. alternata through a fungal growth assay that was carried out in 96-cell polypropylene plates, using the same concentration to all studied substances. The four active chalcones underwent conidia germination and mycelial growth, which confirmed the antifungal activity of the compounds. These chalcones were then poured onto Murcott tangor fruit that had been inoculated with conidia of the fungus. All four chalcones reduced the ABS progress to values significantly smaller (P0.05) than that observed for the control. Statistical calculations showed that the best results were afforded by two compounds, bearing a 2,4,5-trimethoxyphenyl group at position 3 of prop-2-enal and a 3-nitro- or 3-hydroxyphenyl group at position 1 of the aldehyde. Such compounds reduced the incidence of the disease in Murcott tangor fruit to values that did not differ statistically from those obtained with a commercial fungicide.

A mancha marron de Alternaria (MMA), causada pelo fungo Alternaria alternata patótipo tangerina, é um dos maiores problemas fitossanitários dos produtores de tangerina. Aproximadamente 15 aplicações de fungicidas de elevada periculosidade, por ano, são necessárias para o controle de MMA em várias plantações de citros. Como as chalconas parecem menos tóxicas para os seres humanos e para o ambiente que os fungicidas comerciais atualmente em uso, este estudo teve como objetivo inicial sintetizar 137 chalconas através da condensação aldólica de benzaldeídos com acetofenonas. As atividades das chalconas resultantes contra A. alternata foram avaliadas através do emprego de teste de crescimento fúngico em placas de polipropileno com 96 cavidades, empregando a mesma concentração para todas as substâncias estudadas. As quatro chalconas mais ativas foram submetidas a testes de germinação de conídios e de crescimento micelial, que confirmaram as atividades antifúngicas dos compostos selecionados. Estes foram então, aplicados em frutos de tangor Murcote que tinham sido inoculados com conídios do fungo. Todas as quatro chalconas reduziram o progresso de MMA a valores significativamente inferiores (P0.05) ao observado para o controle. Cálculos estatísticos mostraram que os melhores resultados foram obtidos para dois compostos, que tinham um grupo 2,4,5-trimetoxifenil na posição 3 do prop-2-enal e um grupo 3-nitro- ou 3-hidroxifenil na posição 1 do aldeído. Tais compostos reduziram a incidência da doença em frutos de tangor Murcote a valores que não diferiam estatisticamente do obtido com um fungicida comercial.

Alternariol 9-methyl ether from the endophytic fungus Alternaria sp. Samif01 and its bioactivities

Abstract One bioactive compound, identified as alternariol 9-methyl ether, was isolated from the crude extract of the endophytic fungus Alternaria sp. Samif01 residing in the roots of Salvia miltiorrhiza Bunge. Alternariol 9-methyl ether was active against bacteria with minimum inhibitory concentration values ranging from 25 to 75 µg/mL and median inhibitory concentration (IC50) values ranging from 16.00 to 38.27 µg/mL. The IC50 value of alternariol 9-methyl ether against spore germination of Magnaporthe oryzae was 87.18 µg/mL. Alternariol 9-methyl ether also showed antinematodal activity against Bursaphelenchus xylophilus and Caenorhabditis elegans with IC50 values of 98.17 µg/mL and 74.62 µg/mL, respectively. This work is the first report on alternariol 9-methyl ether and its biological activities from the endophytic fungus Alternaria sp. Samif01 derived from S. miltiorrhiza Bunge. The results indicate the potential of Alternaria sp. Samif01 as a source of alternariol 9-methyl ether and also support that alternariol 9-methyl ether is a natural compound with high potential bioactivity against microorganisms.

Controle da requeima e pinta preta da batata por fungicidas e seu reflexo sobre a produtividade e a qualidade de tubérculos / Control of potato late blight and early blight by fungicides and its effects on yield and quality of tubers

A requeima (Phytophthora infestans) e a pinta preta (Alternaria solani) estão entre as doenças mais frequentes e destrutivas da cultura da batata. Com o objetivo de avaliar a ação de diferentes grupos fungicidas no controle dessas doenças, foram realizados dois experimentos em cultivo comercial de batata (cultivares Agata e Monalisa), localizados em Pilar do Sul, São Paulo, nas safras 2008 e 2009. O delineamento experimental utilizado foi o de blocos ao acaso, com 4 repetições, sendo cada parcela de 20 m2. As aplicações foram realizadas com um pulverizador costal, munido de barra de aplicação e pressão constante de 3 Bar. O volume de aplicação variou de 300 a 500 L.ha-1 em função do desenvolvimento da cultura. As variáveis avaliadas para as duas doenças foram: severidade em folhas, área abaixo da curva de progresso da doença (AACPD), produtividade total e comercial, e classificação de tubérculos. Os menores níveis de doença e aumentos significativos na produtividade comercial e qualidade de tubérculos foram observados nas parcelas tratadas com fluopicolide + propamocarbe, dimetomorfe + ametoctradina, mandipropamida + clorotalonil, mandipropamida, fenamidona + propamocarbe, bentiavalicarbe + fluazinam, seguidos de dimetomorfe + clorotalonil, mefenoxam + clorotalonil e famoxadona + cimoxanil + mancozebe para requeima, e azoxistrobina + difenoconazol, picoxistrobina, piraclostrobina + metconazol, trifloxistrobina + tebuconazol, azoxistrobina, boscalida + piraclostrobina, iprodiona + pirimetanil e ciprodinil para pinta preta.(AU)

Late blight (Phytophthora infestans) and early blight (Alternaria solani) are among the most current and destructive diseases of potato. With the aim of evaluating the efficiency of different fungicides in the control of both diseases, two experiments were carried out in commercial potato crops (cultivars Agata and Monalisa) in Pilar do Sul, São Paulo State, Brazil, during 2008 and 2009 cropping season. Randomized block designs were used, with 4 replications and each parcel of 20 m2. Applications were carried out using coastal sprayer with spray-bar under constant pressure of 3 Bar. The application volume varied from 300 to 500 L per hectare according to the crop development. The evaluated variables for both diseases were: severity on leaves, area under disease progress curve (AUDPC), total and commercial yield, and classification of tubers. For late blight, the lower severity levels and significant increases in commercial yield and quality tubers were observed on the rows treated with fluopicolide + propamocarb, dimetomorph + ametoctradin, mandipropamid + chlorothalonil, mandipropamid, fenamidone + propamocarb, benthiavalicarb + fluazinam, followed by dimetomorph + chlorothalonil, mefenoxam + chlorothalonil e famoxadone + cymoxanil + mancozeb and, for early blight, azoxystrobin + difenoconazole, picoxystrobin, pyraclostrobin +metconazole, trifloxystrobin + tebuconazole, azoxystrobin, boscalid + pyraclostrobin, iprodione + pyrimethanil and cyprodinil.(AU)

Analysis of a suppressive subtractive hybridization library of Alternaria alternata resistant to 2-propenyl isothiocyanate

Background Isothiocyanates (ITCs) are natural products obtained from plants of the Brassicas family. They represent an environmentally friendly alternative for the control of phytopathogenic fungi. However, as it has been observed with synthetic fungicides, the possibility of inducing ITC-resistant strains is a major concern. It is, therefore, essential to understanding the molecular mechanisms of fungal resistance to ITCs. We analyzed a subtractive library containing 180 clones of an Alternaria alternata strain resistant to 2-propenyl ITC (2-pITC). After their sequencing, 141 expressed sequence tags (ESTs) were identified using the BlastX algorithm. The sequence assembly was carried out using CAP3 software; the functional annotation and metabolic pathways identification were performed using the Blast2GO program. Results The bioinformatics analysis revealed 124 reads with similarities to proteins involved in transcriptional control, defense and stress pathways, cell wall integrity maintenance, detoxification, organization and cytoskeleton destabilization; exocytosis, transport, DNA damage control, ribosome maintenance, and RNA processing. In addition, transcripts corresponding to enzymes as oxidoreductases, transferases, hydrolases, lyases, and ligases, were detected. Degradation pathways for styrene, aminobenzoate, and toluene were induced, as well as the biosynthesis of phenylpropanoid and several types of N-glycan. Conclusions The fungal response showed that natural compounds could induce tolerance/resistance mechanisms in organisms in the same manner as synthetic chemical products. The response of A. alternata to the toxicity of 2-pITC is a sophisticated phenomenon including the induction of signaling cascades targeting a broad set of cellular processes. Whole-transcriptome approaches are needed to elucidate completely the fungal response to 2-pITC.

Efecto antifúngico de extractos fenólicos y de carotenoides de chiltepín (Capsicum annum var. glabriusculum) en Alternaria alternata y Fusarium oxysporum / Antifungal effect of phenolic and carotenoids extracts from chiltepin (Capsicum annum var. glabriusculum) on Alternaria alternata and Fusarium oxysporum

En el presente estudio se evaluó el efecto de extractos fenólicos y de carotenoides procedentes de frutos de chiltepín sobre el crecimiento micelial y la germinación de conidios de Alternaria alternata y Fusarium oxysporum, 2 importantes hongos causantes de podredumbres en frutas y hortalizas. Los extractos fenólicos presentaron una inhibición en el crecimiento micelial de A. alternata del 38,46 %, y redujeron significativamente la germinación de conidios al quinto día después del tratamiento al 92 % en relación al control. No se observaron cambios significativos en el crecimiento micelial de F. oxysporum, pero sí se redujo significativamente al 85 % en relación al control, el número de conidios germinados a los 5 días de tratamiento. Los extractos de carotenoides mostraron una inhibición del 38,5 % en el crecimiento micelial y del 85,3 % en la germinación de conidios de A. alternata, 5 días después del tratamiento. Frente a F. oxysporum, dichos extractos presentaron menor inhibición del crecimiento micelial (20,3 %), mientras que hubo una mayor inhibición en la germinación de conidios (96 %). Los extractos fenólicos y de carotenoides de chiltepín pueden ser una alternativa promisoria de importancia agrícola como fungicidas naturales.

The effect of phenolic and carotenoid extracts from chiltepin fruits on mycelial growth and the inhibition of conidial germination of Alternaria alternata and Fusarium oxysporum were investigated in the present work. Phenolic extracts inhibited mycelial growth of A. alternata by 38.46%, and significantly reduced conidial germination on the fifth day after treatment to 92% in relation to control. No significant changes were observed in the inhibition of mycelial growth in Fusarium oxysporum; however, the number of germinated conidia was reduced, showing 85% inhibition five days after treatment in relation to control. Moreover, carotenoid extracts showed 38.5% inhibition of mycelial growth and 85.3% inhibition of conidial germination of A. alternata, five days after treatment. Carotenoid extracts showed less inhibition of mycelial growth (20.3%) in F. oxysporum, with respect to A. alternata; while there was greater inhibition of conidial germination (96%) on the fifth day after treatment. Phenolic and carotenoid extracts from chiltepin may be a promising alternative as a natural fungicide against fungi of agricultural importance.

Análisis de tres extractos naturales para el control de enfermedades (Alternaria alternata, Fusarium solani) bajo diferentes dosis de aplicación en el cultivo de haba (Vicia faba) / Analysis of three natural extracts for the control of diseases (Alternaria alternata, Fusarium solani) under different application doses in the bean crop (Vicia faba)

Actualmente la producción del cultivo de Haba (Vicia faba) en el altiplano Boliviano y en especial en el departamento de La Paz, en cada ciclo de producción sufre de diversos factores que impiden su normal desarrollo ecológico y rendimiento como son las plagas, enfermedades como la Mancha de Chocolate, manchas concéntricas, roya y pudrición radicular, además de los factores climáticos extremos. El objetivo de la presente investigación es conocer la eficiencia que podrían causar de manera favorable los extractos naturales de khoa (Satureja boliviana), saponina de la quinua (Chenopodium quinoa willd) e Itapallo (Urtica urens) en el control de las Manchas concéntricas (Alternaria alternata) y la pudrición radicular (Fusarium solani), mediante diferentes dosis de aplicación con la característica especial de que en este caso los extractos naturales son elaborados desde laboratorio, puesto que de manera artesanal ya fueron comprobados su eficiencia en otras enfermedades en diversos cultivos. Este trabajo fue realizado en la Comunidad de Tagachi que pertenece al Municipio de Puerto Acosta, Provincia Camacho del departamento de La Paz, ubicado a 197 kilómetros de la cede de Gobierno. Para este trabajo se aplicó el diseño experimental bloques al azar con arreglo factorial de dos factores, en las parcelas se evaluaron la Incidencia, Severidad, Eficiencia, Longitud de vainas, rendimiento en vainas, grano seco y su análisis económico. Antes de la evaluación en parcela experimental ya fueron evaluados los comportamiento de los patógenos frente a diversos extractos naturales en laboratorio de la facultad de ciencias puras, en donde concluyen que los extractos de Khoa, Itapallo y Saponina de la quinua mostraron mayor eficiencia frente a las enfermedades de Alternaria alternata, Botrytis fabae, Fusarium solani, Phytóphtora infestans entre otras a una dosis de aplicación similar al del systane. Para la evaluación en campo se determinó las siguientes dosis de aplicación en campo: Se tomó en cuenta las recomendaciones de Porco (2009) donde señala que también se debe tomar en cuenta la altura de las plantas para el cálculo de la dosis y el agua es así que se determina que se utilice un total de 180 g de extractos naturales de Khoa, Saponina de quinua e Itapallo respectivamente, 150% representa a 75.12 g con una frecuencia de aplicación de 3,13 g por unidad experimental, 60 g representa el 100% con una frecuencia de 2,5 g por unidad experimental y 44.88 g representa a 50% con una frecuencia de 1,87 g por unidad experimental en 8 aplicaciones durante el ciclo del cultivo de Haba en tres etapas (Prefloración, floración y Llenado de vainas).. Los extractos naturales de Khoa e Itapallo aplicados al 150% y 100%, causaron mayor eficiencia desde el punto de vista estadístico en el control de la (Alternaria alternata y Fusarium solani), es decir que la aplicación de extractos naturales en dosis elevadas lograron las mejores eficiencias en cuanto al control de la mancha concéntrica y la pudrición radicular, puesto que los resultados obtenidos lograron eficiencias superiores al resto de los tratamientos. Los resultados obtenidos nos muestran que los extractos naturales incidieron en los costos de producción, lo que significa que estos resultados afectan a los beneficios económicos que se lograron en la presente investigación. En conclusión según los resultados obtenidos en la parcela experimental indicamos que los extractos naturales de khoa e Itapallo aplicados a una dosis de 150% tubo más efectividad en el control de las manchas concéntricas y de relativa eficiencia en la enfermedad de pudrición radicular, posteriormente los mismos extractos naturales en dosis bajas también reportaron eficiencias en el control de estas enfermedades pero de menor eficiencia, por consiguiente se recomienda realizar estudios de factibilidad económica en la elaboración y aplicación de estos extractos naturales con el fin de reducir los costos de producción. (AU)

Feohifomicosis causada por Colletotrichum gloeosporioides y Alternaria infectoria en un paciente trasplantado renal / Phaeohyphomycosis caused by Colletotrichum gloeosporioides and Alternaría infectoria in renal transplant recipient

Several species of black fungi have been reported as agents of subcutaneous phaeohyphomycosis. Although most of these fungi are considered opportunistic pathogens, they play an important role in phaeohyphomycosis, a disease considered an emergent mycosis among solid organ recipients. We report a case of phaeohyphomycosis caused by Alternaria infectoria of the left hand and the 4th finger of the right hand of a 68-year-old male who underwent a renal transplant 35 months before. The lesion was treated with surgical excision. One year later, the patient presented a new lesion on the 5th finger of the right hand, but this time caused by Colletotrichum gloeosporioides that was also removed surgically. Both lesions did not relapse after being removed. Antifungal susceptibility testing was performed against five antifungal drugs (amphotericin B, itraconazole, flucytosine, fluconazole and voriconazole). Alternaria infectoria was resistant to all five drugs and C. gloeosporioides was sensitive only to amphotericin B and voriconazole. We emphasize the need of histopathologic and microbiologic studies of new lesions of phaeohyphomycosis, since in this case the same patient was infected twice by two different fungi.

Diversas especies de hongos negros han sido descritos como agentes de feohifomicosis subcutáneas. A pesar que la gran mayoría de estos se consideran como oportunistas, juegan un papel importante en la feohifomicosis, enfermedad considerada una micosis emergente en pacientes trasplantados de órganos sólidos. Reportamos un caso de feohifomicosis subcutánea causada por Alternaria infectoria en el dorso de la mano izquierda y 4° falange de la mano derecha de un paciente de sexo masculino de 68 años de edad, que había sido sometido a un trasplante de riñón 35 meses antes. Se realizó escisión quirúrgica de la lesión. Después de un año, el paciente presentó una nueva lesión, esta vez en la 5° falange de la mano derecha causada por Colletotrichum gloeosporioides, lesión que también fue removida quirúrgicamente. El paciente no presentó nuevas lesiones después de las cirugías. Las pruebas de susceptibilidad antifúngica fueron realizados utilizando cinco antifúngicos (anfotericina B, itraconazol, 5-fluorcitosina, fluconazol y voriconazol). Alternaria infectoria presentó resistencia a los cinco antifíngicos y C. gloeosporioides fue susceptible solamente a anfotericina B y voriconazol. Enfatizamos la necesidad de realizar estudios histopatológicas y microbiológicos de nuevas lesiones de feohifomicosis, ya que el mismo paciente fue infectado por dos hongos diferentes.

Toxigenic profile and AFLP variability of Alternaria alternata and Alternaria infectoria occurring on wheat

The objectives of this study were to evaluate the ability to produce alternariol (AOH), alternariol monomethyl ether (AME) and tenuazonic acid (TA) by A. alternata and A. infectoria strains recovered from wheat kernels obtained from one of the main production area in Argentina; to confirm using AFLPs molecular markers the identify of the isolates up to species level, and to evaluate the intra and inter-specific genetic diversity of these two Alternaria species. Among all the Alternaria strains tested (254), 84% of them were able to produce mycotoxins. The most frequent profile of toxin production found was the co-production of AOH and AME in both species tested. TA was only produced by strains of A. alternata. Amplified fragment polymorphism (AFLPs) analysis was applied to a set of 89 isolates of Alternaria spp (40 were A. infectoria and 49 were A. alternata) in order to confirm the morphological identification. The results showed that AFLPs are powerful diagnostic tool for differentiating between A. alternata and A. infectoria. Indeed, in the current study the outgroup strains, A. tenuissima was consistently classified. Characteristic polymorphic bands separated these two species regardless of the primer combination used. Related to intraspecific variability, A. alternata and A. infectoria isolates evaluated seemed to form and homogeneous group with a high degree of similarity among the isolates within each species. However, there was more scoreable polymorphism within A. alternata than within A. infectoria isolates. There was a concordance between morphological identification and separation up to species level using molecular markers. Clear polymorphism both within and between species showed that AFLP can be used to asses genetic variation in A. alternata and A. infectoria. The most important finding of the present study was the report on AOH and AME production by A. infectoria strains isolated from wheat kernels in Argentina on a semisynthetic media for the first time. Also, specific bands for A. alternata and A. infectoria have been identified; these may be useful for the design of specific PCR primers in order to differentiate these species and to detect them in cereals.