Inhibitory effect of the essential oil of Curcuma longa L. and curcumin on aflatoxin production by Aspergillus flavus Link.

Aflatoxins are highly toxic, mutagenic, teratogenic and carcinogenic mycotoxins. Consumption of aflatoxin-contaminated food and commodities poses serious hazards to the health of humans and animals. Turmeric, Curcuma longa L., is a native plant of Southeast Asia and has antimicrobial, antioxidant and antifungal properties. This paper reports the antiaflatoxigenic activities of the essential oil of C. longa and curcumin. The medium tests were prepared with the oil of C. longa, and the curcumin standard at concentrations varied from 0.01% to 5.0%. All doses of the essential oil of the plant and the curcumin standard interfered with mycotoxin production. Both the essential oil and curcumin significantly inhibited the production of aflatoxins; the 0.5% level had a greater than 96% inhibitory effect. The levels of aflatoxin B(1) (AFB(1)) production were 1.0 and 42.7 µg/mL, respectively, for the samples treated with the essential oil of C. longa L. and curcumin at a concentration of 0.5%.

The neem [Azadirachta indica a: juss (meliaceae)] oil reduction in the in vitro production of zearalenone by Fusarium graminearum

Zearalenone, a mycotoxin produced by fungi of the genus Fusarium, including F. graminearum, triggers reproduction disorders in certain animals and hyperestrogen syndromes in humans. Current research investigates three concentrations of neem oil extract (0.1, 0.25 and 0.5 percent) in reducing the production of zearalenone. Neem oil extract decreased zearalenone amount in the three concentrations but highest inhibition (59.05 percent) occurred at 0.1 percent.

The neem [Azadirachta indica a. juss (meliaceae)] oil reduction in the in vitro production of zearalenone by Fusarium graminearum.

Zearalenone, a mycotoxin produced by fungi of the genus Fusarium, including F. graminearum, triggers reproduction disorders in certain animals and hyperestrogen syndromes in humans. Current research investigates three concentrations of neem oil extract (0.1, 0.25 and 0.5%) in reducing the production of zearalenone. Neem oil extract decreased zearalenone amount in the three concentrations but highest inhibition (59.05%) occurred at 0.1%.

Effect of neem leaf extract and neem oil on Penicillium growth, sporulation, morphology and ochratoxin A production.

In vitro trials were conducted to evaluate the effect of Azadirachtaindica (neem) extracts on mycelial growth, sporulation, morphology and ochratoxin A production by P. verrucosum and P. brevicompactum. The effect of neem oil extract from seeds and leaf was evaluated at 0.125; 0.25 and 0.5% and 6.25 and 12.5 mg/mL, respectively, in Yeast Extract Sucrose (YES) medium. Ochratoxin A production was evaluated by a thin-layer chromatography technique. Oil extracts exhibited significant (p ≤ 0.05) reduction of growth and sporulation of the fungi. No inhibition of ochratoxin A production was observed. Given its accessibility and low cost, neem oil could be implemented as part of a sustainable integrated pest management strategy for plant disease, as it has been shown to be fungitoxic by inhibition of growth and sporulation.

Inhibition of citrinin production in Penicillium citrinum cultures by neem [Azadirachta indica A. Juss (Meliaceae)].

The efficacy of different concentrations of aqueous neem leaf extract (3.12 to 50 mg/mL) on growth and citrinin production in three isolates of Penicillium citrinum was investigated under laboratory conditions. Mycotoxin production by the isolates was suppressed, depending on the concentration of the plant extract added to culture media at the time of spore inoculation. Citrinin production in fungal mycelia grown for 21 days in culture media containing 3.12 mg/mL of the aqueous extract of neem leaf was inhibited by approximately 80% in three isolates of P. citrinum. High-performance liquid chromatography was performed to confirm the spectrophotometric results. Vegetative growth was assessed, but neem extract failed to inhibit it. Neem leaf extract showed inhibition of toxin production without retardation in fungal mycelia growth.

Efeito dos extratos aquoso e oleoso de Nim [Azadirachta indica A. Juss (Meliaceae)] na produção de patulina em maçãs contaminadas por Penicillium expansum / Effect of aqueous and oily extracts of Neem [Azadirachta indica A. Juss (Meliaceae)] on patulin production in apples contaminated with Penicillium expansum

A Região Sul do Brasil é grande produtora de maçã, sendo 80 por cento destinada ao consumo in natura. As micotoxinas são metabólitos secundários de fungos presentes na cadeia alimentar como contaminantes, causando diversos efeitos toxicológicos e imunológicos. Considerando que a patulina seja uma micotoxina produzida por Penicillium expansum, principal contaminante da maçã propôs-se investigar a ação de extratos aquoso e oleoso de Nim (Azadirachta indica) em maçã artificialmente contaminada. Foram testados dois tipos de extratos: o extrato aquoso obtido de maceração de folhas de Nim nas concentrações de 5, 10, 20 e 30 por cento, e o extrato oleoso comercial (DalNeem®), obtido de sementes de Nim, nas concentrações de 0,125, 0,25, 0,5, 1, 2 e 5 por cento. Os extratos aquosos não alteram a produção da patulina em maçãs contaminadas artificialmente por P. expansum, mas o extrato oleoso obtido das sementes da planta diminuiu acentuadamente a produção de patulina, inclusive em concentrações inferiores a 0,5 por cento.

The South region of Brazil is a major apple producer, where 80 percent is destined to the "in natura" consumption. The mycotoxins are fungal secondary metabolites present as contaminants in the food chain, causing several toxicological and immunological effect. Patulin is a mycotoxin mainly produced by Penicillium expansum and well known as the main contaminant in apples. Due to this contamination, the aim of this work was to access the effect of Neem extract (Azadirachta indica) on artificially contaminated apples. Two types of Neem extracts were tested: the aqueous extracts of Neem leafs at 5, 10, 20 and 30 percent concentrations, and commercial seed oil of Neem (DalNeem®) at 0.125, 0.25, 0.5, 1, 2 and 5 percent in water. Although the addition of Neem aqueous extracts in artificially contaminated apples with P. expansum was unable to affect the patulin production, the Neem seed oil extract at concentrations as lower as 0.5 percent, caused a pronounced diminution over patulin production.

Production and characterization of galactose oxidase produced by four isolates of Fusarium graminearum

A screening aimed to find new galactose oxidase producer isolates and to evaluate the production among Fusarium graminearum strains was conducted. Thirty-five isolates out of 39 analysed produced the enzyme at several levels. The data indicated a wide distribution of galactose oxidase within F. graminearum and also revealed new producer isolates. The enzyme produced by different isolates showed similar thermal activity and stability and were active on same substrates. However, the optimum pH ranged from 7.0 to 7.5. Thus, all evaluated isolates were suitable for the production of galactose oxidase.

Uma análise visando encontrar novos isolados produtores de galactose oxidase (GO) e avaliar a distribuição da produção entre cepas de Fusarium graminearum foi conduzida. Trinta e cinco isolados de 39 testados produziram a enzima em diferentes níveis. Os dados indicaram uma ampla distribuição da produção da galactose oxidase por F. graminearum e revelaram novos isolados produtores. A enzima produzida por diferentes isolados apresentou atividade e estabilidade térmicas similares e foi ativa sobre os mesmos substratos. No entanto, o pH ótimo variou de 7,0 a 7,5. Assim, todos os isolados são adequados para a produção de galactose oxidase.

Production of mycotoxins by Fusarium graminearum isolated from small cereals (wheat, triticale and barley) affected with scab disease in Southern Brazil

Fungos do gênero Fusarium são bem conhecidos como patógenos para plantas e como produtores de micotoxinas. O objetivo deste trabalho foi avaliar qualitativamente a produção "in vitro" de desoxinivalenol e de zearalenona, em 24 diferentes isolados de Fusarium graminearum coletados a partir de cereais associados à doença Giberela na Região Sul do Brasil. Os isolados foram cultivados em arroz, durante 14 dias, a 28ºC. Os cultivos foram extraídos com metanol:água (40:60, v/v) e analisados por cromatografia em camada delgada. Outros tricotecenos (diacetoxiscirpenol, fusarenona-X, neosolaniol e nivalenol) e zearalenol, freqüentemente produzidos por Fusarium, também foram avaliados. Nas condições utilizadas, foi possível determinar o perfil de produção dessas micotoxinas, sendo que 67 por cento dos isolados produziram zearalenona e 33 por cento dos isolados produziram desoxinivalenol. Também foram detectadas as presenças de zearalenol, diacetoxiscirpenol e fusarenona. Finalmente, em nenhum dos isolados estudados foram encontrados nivalenol e neosolaniol.

Effect of aqueous extract from Neem (Azadirachta indica A. Juss) on hydrophobicity, biofilm formation and adhesion in composite resin by Candida albicans.

OBJECTIVE: Azadirachta indica, a Meliaceae family tree, has been used in India for many years in the treatment of several diseases in medicine and dentistry. Current research analyses the effects of the leaf aqueous extract from Azadirachta indica (Neem) on the adhesion, cell surface hydrophobicity and biofilm formation, which may affect the colonisation by Candida albicans. METHODS: Azadirachta indica extract was tested in vitro on strains of Candida albicans 12A and 156B. Changes in hydrophobicity were reported in assays of yeast adhesion to hydrocarbons, in biofilm formation with glucose and in the adhesion of the microorganisms on light cured composite resin. Assays involved enumeration of candidal colony-forming units together with scintillation counting of radiolabelled Candida and compared to a solution of chlorhexidine digluconate 0.125% widely used in dentistry. RESULTS: Yeast growth in Neem extract was not inhibited in concentrations ranging from 0.1mg/ml. A statistically significant increase (p<0.05) in cell surface hydrophobicity was evident for the two strain tested and there was also an associated increase in biofilm formation after contact with Neem extract in concentration 0.01 g/ml. Decrease in adhesion capacity of cells to composite resin was also recorded. CONCLUSION: An anti-adhesive mechanism of action by Azadirachta indica is proposed based on the results observed.

Inhibition of patulin production by Penicillium expansum cultured with neem (Azadirachta indica) leaf extracts.

Aqueous extract of the leaves of neem [Azadirachta indica A. JUSS (Meliaceae)] was tested in vitro for antifungal activity against Penicillium expansum. Patulin production was inhibited during cultivation, when concentrations higher than 50 mg/ml of neem leaf extract was added to the culture medium. Analyses of mycotoxin production were performed by TLC and HPLC. Fungal growth and colony characteristics, in the presence of the extract, were investigated and compared with extract-free cultures.

Identification of deoxynivalenol, 3-acetyldeoxynivalenol, and zearalenone in galactose oxidase-producing isolates of Fusarium graminearum.

Fusarium graminearum strains are well known for their role as plant pathogens and for their production of mycotoxins, and less known for their secretion of galactose oxidase, a well-studied and useful enzyme. Three galactose oxidase-producing isolates of F. graminearum were grown on rice to identify the production of zearalenone and trichothecenes through the use of thin layer chromatography and gas chromatography coupled to mass fragmentation. Detection and identification of deoxynivalenol, 3-acetyldeoxynivalenol, and zearalenone were accomplished.

Biomass and exopolysaccharide production in submerged cultures of Pleurotus ostreatoroseus Sing. and Pleurotus ostreatus "florida" (Jack.: Fr.) Kummer.

The species of the genus Pleurotus produce large amounts of biomass and exopolysaccharide (EPS) in submerged cultures, which may be used for biotechnological purposes. In the present work two Brazilian autochthonous strains of edible Pleurotus (P. ostratoroseus Sing. and P. ostreatus "florida") were used. The fungi grown in liquid Potato Dextrose medium (PD) were used as inocula to cultivate those microorganisms in the POL culture medium. After a 9-day incubation, the optimal growth time for biomass production, P. ostreatus "florida" presented higher biomass production (22.8 g d.w./l) than P. ostreatoroseus (16.8 g d.w./l). After a 7-day incubation, the optimal growth time for EPS production, P. ostreatoroseus produced higher amounts of crude EPS (5.8 g d.w./l) than P. ostreatus "florida" (1.4 g d.w./l). Relative carbohydrate composition for P. ostreatoroseus and P. ostreatus "florida" EPS were: glucose (95.5-87.7), galactose (traces - 4.9), mannose (traces - 3.1), xylose (1.3-2.5), and arabinose (3.2-1.8). Lower ammonium sulfate concentration in the POL culture medium increased the exopolysaccharides production by P. ostreatoroseus.

The presence of partially 3-O-methylated mannogalactan from the fruit bodies of edible basidiomycetes Pleurotus ostreatus 'florida' Berk. and Pleurotus ostreatoroseus Sing.

The partially 3-O-methylated mannogalactans were isolated from the fruiting bodies of edible basidiomycetes Pleurotus ostreatus 'florida' Berk. and Pleurotus ostreatoroseus Sing. They were obtained via successive aqueous extraction, freeze thawing, and precipitation with Fehling solution and then investigated using (13)C- and (1)H-nuclear magnetic resonance spectroscopy (including COSY, TOCSY and HMQC techniques), methylation analysis and Smith degradation. The main chain consisted of (1-->6)-linked alpha-D-galactopyranosyl residues containing 3-O-Me-alpha-D-galactopyranoses, a part of these units being substituted in the position O-2 with beta-D-mannopyranose residues. The heteropolysaccharides found were similar with differences only in the levels of the 3-O-Me-alpha-D-galactopyranoses residues. The presence of partially 3-O-methylated mannogalactan appears to be typical of Pleurotus spp.

A partially 3-O-methylated (1-->4)-linked alpha-D-galactan and alpha-D-mannan from Pleurotus ostreatoroseus Sing.

The two main water-soluble extracellular polysaccharides produced by the basidiomycete fungus Pleurotus ostreatoroseus Sing were isolated and purified. They were characterized using 13C, 1H, and 1H, 13C HMQC NMR spectroscopy, methylation analysis, and Smith degradation. One was a mannan having a main chain of (1-->6)-linked alpha-D-mannopyranosyl residues, almost all of which were branched at O-2 with side chains of different lengths, containing 2-O- and 3-O-linked mannopyranosyl units. The other was a partially 3-O-methylated (1-->4)-linked alpha-D-galactopyranan, a structure that has not been previously described.

Alternative method of inoculum and spawn production for the cultivation of the edible brazilian mushroom Pleurotus ostreatoroseus SING.

Efficiency of solid and liquid inocula and their use for spawn production were compared so that improved cultivation conditions for the edible mushroom Pleurotus ostreatoroseus could be tested. Solid and liquid inocula were prepared respectively with Potato Dextrose Agar (PDA) and Liquid Potato Dextrose (LPD). Wheat grains and cotton residues were used as substrates for spawn preparation. Inoculum types did not affect the development of P. ostreatoroseus, and LPD spawns were cheaper, more homogenous, less contaminated. Decomposition activity of mushroom growth, as a percentage of organic matter loss (OML), was higher in the wheat grain spawn and was not influenced by the inoculum type. Advantages in the use of cotton residue for spawn production were longer storage time, lower contamination and reduced costs. The cotton residue substrate may be also used for the production of mushroom fruiting bodies.

Production of mycotoxins by galactose oxidase producing "Fusarium" using different culture media

The original isolate of the galactose oxidase producing fungus "Dactylium dendroides, and other five galactose oxidase producing "Fusarium" isolates were cultivated in different media and conditions, in order to evaluate the production of 11 mycotoxins, wich are characteristic of the genus "Fusarium": moniliform, fisaric acid, deoxyvalenol, fusarenone-X, nivalenol, 3-acetyldeoxynivalenol, neosolaniol, zearakenol, zeralenone, acetyl T-2, and iso T-2. The toxicity of the culture extractes to "Artemia salina" larvae was tested

Incidence of aflatoxins and Aspergillus flavus in peanuts consumed in Maringá city, Brazil

It is know that the presence of Aspergillus flavus in stored grains indicates the possibility of aflatoxin contamination. In the present work the occurrence of aflatoxins and aflatoxigenic fungi in peanut samples collected in Maringága city was analysed. Of the 72 samples studied, 48.6 per cent were found positive. Aflatoxin B1, B2, G1, and G2 had maximal levels about 679, 192, 680 and 320 ppb. respectively by fungi of A.flavus group and evaluation in 83.3 per cent of them showed that all of the isolated strains had the capacity to produce aflatoxin B1