Microorganisms from corn stigma with biocontrol potential of Fusarium verticillioides / Microrganismos do estigma do milho com potencial de biocontrole de Fusarium verticillioides

The mycotoxigenic fungus Fusarium verticillioides is the primary maize pathogen and causes the maize stalk and ear rot diseases with significant economic losses. Furthermore, the excessive use of fungicides to control F. verticillioides constitutes threats to the environment and human health. Thus, sustainable alternatives such as biological control are needed to minimize the hazards associated with the current method. Although much is known about the vulnerability of the maize silks as a gateway for several fungal pathogens invading the developing grains, studies on the chemical properties of silk extracts and their resident microbiota are scarce. This study isolated and characterized bacteria and fungi that colonize the maize stigma to assess new potential biocontrol agents. The samples were collected from maize fields in the Brazilian localities of Sete Lagoas-MG, Sidrolândia-MS, Sertaneja-PR, and Goiânia-GO. One hundred sixty-seven microorganisms were isolated, 46% endophytic and 54% epiphytic. First, the antagonist activity was evaluated by the agar disc diffusion method performed in triplicate, and 83% of the isolates showed antagonist activity against F. verticillioides. Then, the 42 most efficient isolates were identified based on the partial sequencing of the bacterial 16S rRNA gene and fungi ITS region. The bacteria belong to the genera Bacillus (57.1%), Burkholderia (23.8%), Achromobacter (7.1%), Pseudomonas (2.4%), and Serratia (2.4%), while the fungi are Penicillium (2.4%), Candida (2.4), and Aspergillus (2.4%). The results showed that microorganisms from maize stigma might represent new promising agents for F. verticillioides control.

O fungo micotoxigênico Fusarium verticillioides é o principal patógeno do milho e causa doenças do colmo e da podridão da espiga com perdas econômicas significativas. Além disso, o uso excessivo de fungicidas no controle de F. verticillioides constitui uma ameaça ao meio ambiente e à saúde humana. Assim, alternativas sustentáveis, como o controle biológico, são necessárias para minimizar os riscos associados ao método atual. Este estudo isolou e caracterizou bactérias e fungos que colonizam o estigma do milho para avaliar novos agentes de biocontrole em potencial. As amostras foram coletadas em campos de milho nas localidades brasileiras de Sete Lagoas-MG, Sidrolândia-MS, Sertaneja-PR e Goiânia-GO. Cento e sessenta e sete microrganismos foram isolados, 46% endofíticos e 54% epifíticos. O teste de antagonismo empregando a técnica de disco de difusão em meio sólido, mostrou que 83% dos isolados apresentaram atividade antagonista contra F. verticillioides. Em seguida, 42 isolados mais eficientes foram identificados a partir do sequenciamento parcial do gene 16S rRNA bacteriano e da região ITS de fungos. Os isolados bacterianos pertencem ao gênero Bacillus (57,1%), Burkholderia (23,8%), Achromobacter (7,1%), Pseudomonas (2,4%) e Serratia (2,4%), enquanto os fungos são Penicillium (2,4%), Candida (2.4), e Aspergillus (2,4%). Os resultados mostraram que microrganismos do estigma do milho podem representar novos agentes promissores para o controle de F. verticillioides.

Antimicrobial photodynamic therapy with phenothiazinium photosensitizers in non-vertebrate model Galleria mellonella infected with Fusarium keratoplasticum and Fusarium moniliforme.

Fusarium keratoplasticum and Fusarium moniliforme are filamentous fungi common in the environment and cause mycosis in both animals and plants. Human infections include mycetoma, keratitis and onychomycosis, while deeper mycosis occurs in immunocompromised patients. Most of the Fusarium spp. are frequently resistant to treatment with currently used antifungals. The frequent occurrence of antifungal resistance has motivated the study of antimicrobial photodynamic therapy as an alternative treatment for fungal infections. Many studies have investigated the in vitro use of antimicrobial photodynamic therapy to kill fungi, but rarely in animal models of infection. Thus, here we employed the invertebrate wax moth Galleria mellonella to study the in vivo effects of antimicrobial photodynamic therapy with three different phenothiazinium photosensitizers, methylene blue, new methylene blue N and the pentacyclic S137 against infection with microconidia of Fusarium keratoplasticum and Fusarium moniliforme. The effect of antimicrobial photodynamic therapy using these photosensitizers and light-emitting diodes with an emission peak at 635 nm and an integrated irradiance from 570 to 670 nm of 9.8 mW cm-2 was investigated regarding the toxicity, fungal burden, larval survival and cellular immune response. The results from this model indicate that antimicrobial photodynamic therapy with methylene blue, new methylene blue N and S137 is efficient for the treatment of infection with F. keratoplasticum and F. moniliforme. The efficiency can be attributed to the fungal cell damage caused by antimicrobial photodynamic therapy which facilitates the action of the host immune response.

Photodynamic treatment with phenothiazinium photosensitizers kills both ungerminated and germinated microconidia of the pathogenic fungi Fusarium oxysporum, Fusarium moniliforme and Fusarium solani.

The search for alternatives to control microorganisms is necessary both in clinical and agricultural areas. Antimicrobial photodynamic treatment (APDT) is a promising light-based approach that can be used to control both human and plant pathogenic fungi. In the present study, we evaluated the effects of photodynamic treatment with red light and four phenothiazinium photosensitizers (PS): methylene blue (MB), toluidine blue O (TBO), new methylene blue N (NMBN) and the phenothiazinium derivative S137 on ungerminated and germinated microconidia of Fusarium oxysporum, F. moniliforme, and F. solani. APDT with each PS killed efficiently both the quiescent ungerminated microconidia and metabolically active germinated microconidia of the three Fusarium species. Washing away the unbound PS from the microconidia (both ungerminated and germinated) before red light exposure reduced but did not prevent the effect of APDT. Subcelullar localization of PS in ungerminated and germinated microconidia and the effects of photodynamic treatment on cell membranes were also evaluated in the three Fusarium species. APDT with MB, TBO, NMBN or S137 increased the membrane permeability in microconidia and APDT with NMBN or S137 increased the lipids peroxidation in microconidia of the three Fusarium species. These findings expand the understanding of photodynamic inactivation of filamentous fungi with phenothiazinium PS.

Cytotoxic activity and induction of inflammatory mediators of the methanol:chloroform extract of Fusarium moniliforme.

BACKGROUND: Fusarium moniliforme is a phytopathogenic facultative fungus with a cosmopolitan distribution in all types of climates, and has a wide host range, including, among others, bean, rice, wheat and sorghum crops. There is a current lack of knowledge regarding the potential of these fungi, so it is considered to be of great importance to obtain information related to the biological activity of extracts and secondary metabolites. AIMS: An evaluation of the role of methanol:chloroform extract of F. moniliforme in the production of inflammatory cytokines and their cytotoxic activity. METHODS: The production of nitric oxide was analyzed by the Griess method, the production of cytokines using ELISA, and the effects of the extract on cell cycle and induction of apoptosis by flow cytometry. RESULTS: The extract of F. moniliforme was seen to be able to stimulate nitric oxide (NO) production in J774A.1 cells, as well as to produce cytokines such as, IL-1ß, IL-6, and TNF-α. It was also observed that the extract of F. moniliforme produces activity on cell cycle modulation and apoptosis when tested in carcinogenic cell lines. CONCLUSIONS: The results obtained from this study open the possibility of obtaining and identifying metabolites of the extract of F. moniliforme that can be evaluated for possible use in cancer therapy.

A new alternative to produce gibberellic acid by solid state fermentation

Gibberellic acid (GA3) is an important hormone, which controls plant's growth and development. Solid State Fermentation (SSF) allows the use of agro-industrial residues reducing the production costs. The screening of strains (four of Gibberella fujikuoroi and one of Fusarium moniliforme) and substrates (citric pulp, soy bran, sugarcane bagasse, soy husk, cassava bagasse and coffee husk) and inoculum preparation study were conducted in order to evaluate the best conditions to produce GA3 by SSF. Fermentation assays were carried out in erlenmeyers flasks at 29°C, with initial moisture of 75-80 percent. Different medium for inoculum production were tested in relation to cells viability and GA3 production by SSF. F. moniliforme LPB 03 and citric pulp were chosen for GA3 production. The best medium for inoculum production was citric pulp extract supplemented with sucrose. GA3 production by SSF reached 5.9 g /kg of dry CP after 3 days of fermentation.

O ácido giberélico (GA3) é um importante hormônio vegetal. A fermentação no estado sólido (FES) utiliza resíduos agro-industriais reduzindo os custos de produção. Neste trabalho a seleção de cepas (quatro de Gibberella fujikuoroi e uma de Fusarium moniliforme) e substratos (polpa cítrica, casca de soja, bagaço de cana, farelo de soja, bagaço de mandioca e casca de café) e o estudo da preparação do inóculo foram conduzidos para otimizar as condições de produção de GA3 por FES. Os ensaios foram realizados em frascos de erlenmeyer a 29°C, com umidade inicial de 75-80 por cento. Diferentes meios para a produção do inóculo foram testados em relação à viabilidade das células e produção de GA3 por FES. F. moniliforme LPB03 e polpa cítrica foram escolhidos. O melhor meio para a produção de inóculo foi o extrato de polpa cítrica. A produção por FES alcançou 5.8 g de GA3/kg de polpa cítrica após 3 dias de fermentação.

Efeito do genótipo de milho e da aplicação foliar de fungicidas na incidência de grãos ardidos / Genotypes and fungicides effects after maize leaves spraying on ear rot incidence

Este trabalho foi realizado visando avaliar a incidência de grãos ardidos em genótipos de milho sob aplicação foliar de fungicidas. Foi conduzido um experimento em Iraí de Minas - MG, com posterior análise dos grãos em Uberlândia - MG. O delineamento experimental foi o de blocos casualizados, em esquema fatorial 8x5x2, correspondentes a híbridos (AG 6018, AG 8060, AG 7000, Penta, Strike, Speed, Fort e Master), fungicidas (Piraclostrobin + Epoxiconazole, Hidróxido de Cobre, Azoxystrobin + Ciproconazole, Azoxystrobin e controle) e épocas de aplicação (45 e 45-60 D.A.S.), respectivamente, com 4 repetições. A quantificação da incidência de grãos ardidos foi determinada pelo método de separação visual de grãos com sintomas de descoloração, causada pela infecção por fungos na lavoura. O teste padrão ("Blotter Test') permitiu detectar a presença dos patógenos Fusarium moniliforme e Penicillium digitatum. Os resultados foram submetidos à análise de variância e teste de médias pelo programa SISVAR (teste de Scott-Knott, a 5% de probabilidade). As épocas de aplicação não influenciaram na incidência de grãos ardidos ao contrário do uso de fungicidas via foliar que resultou em uma menor incidência de grãos ardidos. O híbrido AG6018 apresentou menores valores para grãos ardidos. Já o AG8060 apresentou menor incidência de Fusarium moniliforme nos grãos. Os híbridos Penta, Strike, AG6018, Master e Speed apresentaram menor incidência de Penicillium digitatum nos grãos. Quanto aoFusarium moniliforme, a aplicação de fungicidas diminuiu a incidência do patógeno. Nas análises para o fungo Penicillium digitatum a aplicação de fungicidas via foliar interagindo com os diferentes genótipos de milho, influenciaram na incidência do patógeno.

This study evaluated the incidence of ear rot in maize genotypes in relation to fungicide spraying.,The trial was carried out in Iraí de Minas, with complement in Uberlândia - MG. The experimental design was randomized blocks, in a 8 x 5 x 2 factorial scheme, corresponding to hybrids (AG 6018, AG 8060, AG 7000, Penta, Strike, Speed, Fort e Master), fungicides (Piraclostrobin + Epoxiconazole, Copper Hydroxide, Azoxystrobin + Ciproconazole,Azoxystrobin and non-sprayed control) and spraying times (45 or 45-60 D.A.S.), respectively. All treatments with four replicates. Rotten kernels were determined by visual separation of the kernels with, which was caused by fungi in the . A Blotter Test was done in the laboratory allowed to identify the pathogens Fusarium moniliforme and Penicillium digitatum, which are able to form mycelia on the plant material. The results were submitted to analyses of variance and means separation with the program SISVAR (Scott-Knott test, at 5% probability). The spraying times did not affect ear rot incidence. Fungicide leaf spraying resulted in lower ear rot incidence. The hybrid AG6018 presented lowest incidence of ear rot. Lowest incidence of Fusarium moniliforme was found on AG8060. The hybrids Penta, Strike, AG6018, Master and Speed presented lowest incidence of Penicillium digitatum on the kernels. Fungicide spraying decreased Fusarium moniliforme incidence. Fungicide leaf spraying from strobilurines plus triazols decreased rotten kernels and the interaction of fungicides with maize hybrids affected the incidence of Penicillium digitatum.

Desfolha, população de plantas e precocidade do milho afetam a incidência e a severidade de podridões de colmo

The relationship between source and sink interferes on maize (Zea mays) stalk rot occurrence. This study was conducted in Lages, SC, Brazil, to evaluate the effect of defoliation, plant population, and hybrids on the intensity (incidence or severity) of maize stalk rots. The experiment was set in a randomized complete block design (three replications) with a split-split-plot arrangement. The hybrids P32R21 (very early), Premium (early) and C333B (late) were tested without defoliation and with 50% defoliation on ear formation stage at plant populations of 25,000, 50,000, 75,000, and 100,000 plants ha-1. The incidence and severity of Colletotrichum graminicola, Diplodia maydis and Fusarium moniliforme stalk rots were evaluated. The hybrid P32R21 was the most affected by stalk rots. Defoliation enhanced rots on P32R21 and Premium. Enhancements in plant population increased stalk rots intensity on intact plants but decreased rot occurrence when hybrids were defoliated.

As relações entre fonte e dreno interferem sobre a ocorrência de podridões de colmo na cultura do milho (Zea mays). O estudo foi conduzido em Lages, SC, para avaliar o efeito da desfolha, população de plantas e híbridos sobre as podridões do colmo do milho. O delineamento experimental foi de blocos ao acaso com parcelas sub-subdivididas e três repetições. Os híbridos P32R21 (superprecoce), Premium (precoce) e C333B (tardio) foram testados sem desfolha e com 50% de desfolha no estágio de espigamento nas populações de 25.000, 50.000, 75.000 e 100.000 plantas ha-1. Avaliaram-se a incidência e a severidade das podridões do colmo causadas por Colletotrichum graminicola, Diplodia maydis e Fusarium moniliforme. O híbrido P32R21 foi o mais afetado pelos fungos causadores de podridões. Nos híbridos P32R21 e Premium, as plantas desfolhadas foram mais severamente infectadas pelos patógenos do que as intactas. O aumento da população de plantas aumentou a intensidade (incidência ou severidade) de podridões de colmo nas plantas intactas e diminuiu a ocorrência nas desfolhadas.

Desfolha, população de plantas e precocidade do milho afetam a incidência e a severidade de podridões de colmo

The relationship between source and sink interferes on maize (Zea mays) stalk rot occurrence. This study was conducted in Lages, SC, Brazil, to evaluate the effect of defoliation, plant population, and hybrids on the intensity (incidence or severity) of maize stalk rots. The experiment was set in a randomized complete block design (three replications) with a split-split-plot arrangement. The hybrids P32R21 (very early), Premium (early) and C333B (late) were tested without defoliation and with 50% defoliation on ear formation stage at plant populations of 25,000, 50,000, 75,000, and 100,000 plants ha-1. The incidence and severity of Colletotrichum graminicola, Diplodia maydis and Fusarium moniliforme stalk rots were evaluated. The hybrid P32R21 was the most affected by stalk rots. Defoliation enhanced rots on P32R21 and Premium. Enhancements in plant population increased stalk rots intensity on intact plants but decreased rot occurrence when hybrids were defoliated.

As relações entre fonte e dreno interferem sobre a ocorrência de podridões de colmo na cultura do milho (Zea mays). O estudo foi conduzido em Lages, SC, para avaliar o efeito da desfolha, população de plantas e híbridos sobre as podridões do colmo do milho. O delineamento experimental foi de blocos ao acaso com parcelas sub-subdivididas e três repetições. Os híbridos P32R21 (superprecoce), Premium (precoce) e C333B (tardio) foram testados sem desfolha e com 50% de desfolha no estágio de espigamento nas populações de 25.000, 50.000, 75.000 e 100.000 plantas ha-1. Avaliaram-se a incidência e a severidade das podridões do colmo causadas por Colletotrichum graminicola, Diplodia maydis e Fusarium moniliforme. O híbrido P32R21 foi o mais afetado pelos fungos causadores de podridões. Nos híbridos P32R21 e Premium, as plantas desfolhadas foram mais severamente infectadas pelos patógenos do que as intactas. O aumento da população de plantas aumentou a intensidade (incidência ou severidade) de podridões de colmo nas plantas intactas e diminuiu a ocorrência nas desfolhadas.

Production of fumonisins by strains of Fusarium moniliforme according to temperature, moisture and growth period

Production of fumonisins B1 (FB1) and B2 (FB2) by two Brasilian strains (LAMIC 2999/96 and 113F) and one American strain (NRRL 13616) of Fusarium moniliforme were evaluated in laboratory cultures subjected to different temperatures (20, 25, and 30ºC), and moisture contents (25, 34, and 42%) on corn substrate. The cultures were grown during 10, 20, 30, 45, and 60 days, totalizing 135 treatments with two repetitions for each one. The fumonisins were extracted with acetonitrile/water. The clean-up with end-capped C18 silica (C18ec) cartridges and fumonisin derivatization with o-phtaldialdeyde were carried out through an automated sample processor system (ASPEC), followed by quantification of the toxins through HPLC. Fumonisin production varied widely, reaching average yields from 0.25 to 5515.45 µg/g of FB1 and from 0.15 to 3032.10 µg/g of FB2. In the present work, the factors strain, temperature, moisture and days of fungal culture were evaluated, and all of them had a bearing on the amounts of fumonisins produced. The highest FB1 average yields were obtained by the strain 113F, under the following conditions: 34% moisture content, 60 culture days, and temperature of 25ºC. The highest FB2 average yield was obtained by the same strain with cultures over 45 days, 42% moisture content, at the temperature of 25ºC. Via regression analysis, the ideal temperature for fumonisins production was, calculated as 24.5 and 24.3ºC (± 2ºC) for FB1 and FB2, respectively.

Produção de fumonisinas B1 (FB1) e B2 (FB2) a partir de duas cepas brasileiras (LAMIC 2999/96 e 113F) e uma cepa americana (NRRL 13616) de Fusarium moniliforme foi avaliada em culturas de laboratório submetidas a diferentes temperaturas (20, 25 e 30ºC) e a diferentes teores de umidade (25, 34 e 42%) em substrato de milho. As culturas foram realizadas em períodos de 10, 20, 30, 45 e 60 dias, totalizando135 tratamentos com duas repetições para cada um. As fumonisinas foram extraídas com acetonitrila/água. A limpeza foi realizada empregando cartuchos de sílica C18 encapada (C18ec) e a derivação com o-ftalodialdeído foram realizadas por um sistema processador automático de amostras (ASPEC), seguidas por quantificação das toxinas por CLAE. A produção de fumonisinas variou muito, atingindo rendimentos médios de 0,25 a 5515,45 µg/g de FB1 e de 0,15 a 3032,10 µg/g de FB2. Neste trabalho, os fatores como cepa, temperatura, umidade e dias de cultura fúngica foram avaliados, e todos estes influenciaram nas quantidades de fumonisinas produzidas. As mais altas produções de FB1 foram obtidas pela cepa 113F nas seguintes condições: teor de umidade de 34%, 60 dias de cultura, temperatura de 25ºC. A maior produção média de FB2 foi obtida pela mesma cepa com culturas durante 45 dias, a um teor de umidade de 42%, à temperatura de 25ºC. A temperatura ideal para produção de fumonisinas foi calculada por meio de análise de regressão, sendo 24,5ºC e 24,3ºC (±2ºC) para FB1 e FB2, respectivamente.

Occurrence of Fusarium moniliforme and fumonisins in kernels of maize hybrids in Venezuela

Fusarium moniliforme (FM) incidence and fumonisins (FUM) content were determined in white kernel samples of maize hybrids, harvested in farms of Guárico and Portuguesa states, Venezuela. FM incidence was estimated in subsamples placed on a malt-salt-agar medium. Values in the range of 0-15 % were considered low, from 15-30% intermediate, and more than 30% high. FUM content was quantified through specific immunoassay (Fumonitest, VICAM Sci. Tech., USA) and values above 1 ppm were considered high. The hybrids were compared using a completely randomized design and results were subjected to analysis of variance and mean comparisons. The correlation between FUM content and FM incidence was also estimated. FM and FUM were detected in all of the 89 samples. Mean incidence of FM among hybrids ranged from 26.0 to 72.0% in Portuguesa state (PS) and from 28.7 to 55.7 in Guárico state (GS). Significant differences among hybrids were determined in farms of El Playón and Turén, PS, and El Sombrero and El Socorro, GS. The incidence of FM was high in all hybrids. FUM contents ranged from 0.80 to 10.00 ppm in PS, and from 1.50 to 13.00 ppm in GS. Significant differences among hybrids were found in El Playón, PS, and in both locations of GS. Only in Ospino, PS, the content of FUM was correlated with FM incidence. This indicates the natural occurence of FM strains with different toxigenic capabilities. The results show that Guárico and Portuguesa states are hazardous regions regarding FM and FUM in maize kernels.

A incidência de Fusarium moniliforme (FM) e o conteúdo de fumonisinas (FUM) foram determinados em amostras de grãos brancos de híbridos de milho, coletados em fazendas dos estados Guárico e Portuguesa, Venezuela. Incidência de FM foi calculada em submostras colocadas em ágar malte-sal. Valores na faixa de 0-15% foram considerados baixos, de 15-30% intermediários, e acima de 30% altos. O conteúdo de FUM foi quantificado através de imunoensaio específico (Fumonitest, VICAM Sci. Tech., USA.) e valores acima de 1 ppm foram considerados altos. Os híbridos foram comparados utilizando um esquema completamente aleatório. A correlação entre conteúdo de FUM e incidência de FM também foi calculada. A presença de FM e de FUM foi detectada em todas as 89 amostras. A incidência média de FM nos híbridos variou de 26,0 a 72,0% no estado Portuguesa (PS) e de 28,7 a 55,7% no estado Guárico (GS). Diferenças significativas entre híbridos foram determinadas em fazendas de El Playón e Turén (PS) e El Sombrero e El Socorro (GS). A incidência de FM foi alta em todos os híbridos. Os teores de FUM variaram de 0,80 a 10,00 ppm em PS e de 1,50 a 13,00 ppm em GS. Diferenças significativas entre híbridos foram achadas em El Playón (PS) e em ambas localidades de GS. Somente em Ospino (PS), o teor de FUM correlacionou-se com a incidência de FM. Isto indica a ocorrência natural de cepas de FM com capacidades toxigenicas diferentes. Os resultados mostraram que os estados Guárico e Portuguesa são regiões de risco em relação à FM e FUM em grãos de milho.

Symptomless infection of banana and maize by endophytic fungi impairs photosynthetic efficiency.

Very little is known about the physiological interactions between plant hosts and symptomless endophytic fungi despite their widespread occurrence. We investigated the impact of two such fungi, Colletotrichum musae and Fusarium moniliforme, upon the photosynthetic capacity of two crop plants, banana and maize, respectively. Endophyte-free plants were obtained first and then infected with the fungi. Measurements of total chlorophyll content revealed very little difference between endophyte-free and infected plants of banana, whereas in maize they showed 50% reductions in the endophyte-infected plants. The maximum photochemical capacity (Fv /Fm ) was measured in order to determine if the plants had any photoinhibitory effect caused by biotic or abiotic factors. After 45 d of growth, endophyte-free banana plants had similar values of Fv /Fm to plants typical of nonstressed conditions, whereas the endophyte-infected plants showed a reduction of approx. 15%. Unlike banana, infected maize plants displayed values of Fv /Fm similar to those of control and endophyte-free plants, indicating that the maximum photochemical capacity was not affected by infection. The light response curves of both species showed that the photosynthethic capacity was severely reduced in endophyte-infected plants, reaching saturation at c. 400 µmol m-2 s-1 whereas the control and endophyte-free plants were saturated at much higher photon flux densities. In banana the effect seemed to be due to an impairment of electron transport in the thylakoid membranes. By contrast, reduction of the photosynthetic capacity in maize was due to a reduction in chlorophyll content, leading to a decrease in the electron transport components and a consequent reduction in carbohydrate synthesis. It is possible that the reduction in the maximum yield of photosynthesis in both crops was caused by toxins produced by the fungi. Nevertheless there were no major macroscopic effects on the plants to indicate disease symptoms.

Mycoflora and aflatoxin/fumonisin production by fungal isolates from freshly harvested corn hybrids

The mycoflora of 3 hybrids of freshly harvested corn grains collected from three regions of the state of São Paulo, Brazil (Assis, Capão Bonito and Ribeirão Preto) was investigated. A total of 66 samples were analyzed focusing on the influence of abiotic factors (moisture content, water activity, temperature and rainfall) on both the prevalence of Aspergillus flavus and Fusarium moniliforme, and the ability of these genera isolates to produce aflatoxins and fumonisins, respectively. In the three surveyed regions, the fungal population comprised mainly Fusarium spp., Penicillium spp., Aspergillus spp. and 2 others filamentous fungal genera, which were isolated from corn kernels showing water activity of 0.30 to 0.99 and moisture content of 5.0% to 20.2%. Among the genera Fusarium and Aspergillus, the most frequent species were F. moniliforme and A. flavus, respectively. Concerning the toxigenic potential of F. moniliforme, all isolated strains (40) produced fumonisins at 20 mug/g to 2168 mug/g (FB1) and/or 10 mug/g to 380 mug/g (FB2). From the 10 A. flavus isolates, 6 strains (60.0%) produced aflatoxins at 615 mug/kg to 30.750 mug/kg (AFB1) and/or 11 mug/kg to 22 mug/kg (AFB2).

A microbiota fúngica de 66 amostras de três híbridos de grãos de milho recém-colhido, provenientes de 3 regiões do Estado de São Paulo - Brasil (Assis, Capão Bonito e Ribeirão Preto), foram analisadas perante a influência dos fatores abióticos (teor de umidade, atividade de água, precipitação pluvial e temperatura média) na frequência de isolamento de fungos, bem como a potencialidade toxigênica das cepas de Aspergillus flavus e Fusarium moniliforme quanto à produção de aflatoxinas e fumonisinas, respectivamente. As análises microbiológicas demonstraram predominância de Fusarium spp., Penicillium spp. e Aspergillus spp. e outros dois gêneros de fungos filamentosos, isolados de grãos com atividade de água entre 0,30 e 0,99 e teor de umidade entre 5,0% e 20,2%. Entre Fusarium spp, F.moniliforme foi a mais frequentemente isolada, enquanto que, em relação ao gênero Aspergillus, predominou A. flavus nas três regiões. Todas as cepas de Fusarium moniliforme isoladas (40), produziram fumonisinas, que variaram de 20 mg/g a 2168 mg/g (FB1) e 10 mg/g a 380 mg/g (FB2). Referente a 10 cepas de Aspergillus flavus isoladas, 6 cepas (60,0%) produziram aflatoxinas, que variaram de 615 mg/kg a 30.750 mg/kg (AFB1) e 11 mg/kg a 22 mg/kg (AFB2).

Tratamento fungicida de sementes de milho contra fungos do solo e o controle de Fusarium associado às sementes

The efficiency of captan (120, 90 and 60g.a.i. 100kg-1), thiram (140), formulated as WP and SC, thiabendazole (20), thiram SC + thiabendazole (75 + 10), and carboxin + thiram (75 + 75) was studied for the treatment of corn seeds of the cultivar BR 106. The following characteristics were evaluated: seed health, seedling emergence in the field, in sterile soil, in the field cold test, and in soil artificially infected with the following pathogens: Fusarium moniliforme var. subglutinans, Phytium aphanidermatum, and Rhizoctonia solani. The efficiency of the fungicides varied with all evaluated characteristics, except for thiabendazole that was inefficient in relation to the main characteristics. Seed germination was not affected by the presence of F. moniliforme var.subglutinans, but in cold and humid soil the fungi P. aphanidermatum, F. moniliforme var. subglutinans, and R. solani, caused reduction in corn seed germination.

As sementes de milho infectadas por fungos constitue importantes fontes de inóculos, cujos patógenos podem causar podridões de sementes, morte de plântulas em pré e pós-emergência e podridões radiculares. Ademais, em condição de solo frio e úmido há maior exposição das sementes ao ataque de fungos do solo. Assim, estudou-se a eficiência dos fungicidas captan (120,0; 90,0 e 60,0 g i.a. 100 kg-1 de sementes), thiram (140,0) formulado em PM e SC, thiabendazole (20,0), thiram + thiabendazole (75,0 + 10,0) e carboxin + thiram (75,0 + 75,0) no tratamento de sementes de milho da cultivar BR 106. Foram avaliadas as características : sanidade de sementes; emergência de plântulas no campo, em solo esterilizado; bem como as emergências nos testes de frio em solo de campo, em solo esterilizado, em solo infectado separadamente com Fusarium moniliforme var. subglutinans, Pythium aphanidermatum e Rhizoctonia solani. A eficiência dos fungicidas variou de acordo com as características avaliadas, com exceção do thiabendazole, que foi ineficiente para quase todas as características. A presença de F. moniliforme var. subglutinans em sementes de milho não afetou a germinação, porém em solos frios e úmidos os fungos P. aphanidermatum, F. moniliforme var. subglutinans e R. solani promeveram redução na germinação.

Tratamento fungicida de sementes de milho contra fungos do solo e o controle de Fusarium associado às sementes

The efficiency of captan (120, 90 and 60g.a.i. 100kg-1), thiram (140), formulated as WP and SC, thiabendazole (20), thiram SC + thiabendazole (75 + 10), and carboxin + thiram (75 + 75) was studied for the treatment of corn seeds of the cultivar BR 106. The following characteristics were evaluated: seed health, seedling emergence in the field, in sterile soil, in the field cold test, and in soil artificially infected with the following pathogens: Fusarium moniliforme var. subglutinans, Phytium aphanidermatum, and Rhizoctonia solani. The efficiency of the fungicides varied with all evaluated characteristics, except for thiabendazole that was inefficient in relation to the main characteristics. Seed germination was not affected by the presence of F. moniliforme var.subglutinans, but in cold and humid soil the fungi P. aphanidermatum, F. moniliforme var. subglutinans, and R. solani, caused reduction in corn seed germination.

As sementes de milho infectadas por fungos constitue importantes fontes de inóculos, cujos patógenos podem causar podridões de sementes, morte de plântulas em pré e pós-emergência e podridões radiculares. Ademais, em condição de solo frio e úmido há maior exposição das sementes ao ataque de fungos do solo. Assim, estudou-se a eficiência dos fungicidas captan (120,0; 90,0 e 60,0 g i.a. 100 kg-1 de sementes), thiram (140,0) formulado em PM e SC, thiabendazole (20,0), thiram + thiabendazole (75,0 + 10,0) e carboxin + thiram (75,0 + 75,0) no tratamento de sementes de milho da cultivar BR 106. Foram avaliadas as características : sanidade de sementes; emergência de plântulas no campo, em solo esterilizado; bem como as emergências nos testes de frio em solo de campo, em solo esterilizado, em solo infectado separadamente com Fusarium moniliforme var. subglutinans, Pythium aphanidermatum e Rhizoctonia solani. A eficiência dos fungicidas variou de acordo com as características avaliadas, com exceção do thiabendazole, que foi ineficiente para quase todas as características. A presença de F. moniliforme var. subglutinans em sementes de milho não afetou a germinação, porém em solos frios e úmidos os fungos P. aphanidermatum, F. moniliforme var. subglutinans e R. solani promeveram redução na germinação.

Equine leukoencephalomalacia: report of five cases / Leucoencefalomalécia eqüina: relato de 5 casos

This paper reports the occurrence of 5 cases of Equine Leukoencephalomalacia associated with the ingestion of mouldy corn during the winter of 1990 in three properties of the State of São Paulo, in the municipal districts of Poá, Pirassununga and Santana do Parnaíba. The animals showed in every case the following clinical signs: blindness, ataxia, dysphagya, tendency to circle and convulsions. The necropsy revealed congestion of brain and areas of cavitation within the hemispheres of subcortical white matter. The histopathological examination show edlesions characterized by liquefactive necrosis, multifocal haemorrhages and cellular infiltration, predominantly of polimorphonuclear (neutrophils and eosinophils) associated with mononuclear cells in the white matter. Microbiological analysis showed a mycoflora composed, predominantly, by Fusarium moniliforme (49.4%), followed by genera Aspergillus (25.8%) and Penicillum (24.8%).

Relatam-se 5 casos de leucoencefalomalécia em equinos associados com a ingestão de milho mofado, ocorridos durante o inverno de 1990, provenientes de 3 propriedades no Estado de São Paulo, nos municípios de Poá, Pirassununga e Santana do Parnaíba. Os animais apresentaram sinais clínicos semelhantes como ataxia, cegueira, disfagia, andar em círculos e convulsões. Ao exame necroscópico, os cérebros encontravam-se congestos, revelando ao corte áreas de cavitação ao nível da substancia branca sub-cortical. O exame histopatológico exibiu lesões ao nível de substancia branca, caracterizadas por necrose de liquefação, hemorragias focais e infiltrados celulares nos espaços de Virchow-Robin, compostos predominantemente porpolimorfonucleares (eosinófilos e neutrófilos), associados a mononucleares. Análises microbiológicas demonstram que Fusarium moniliforme foi o fungo mais freqüente (1.9 x 10 6 UFC/g de alimento).