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ABSTRACT: The aims of this study were to compare the viability and biomass production of B. amyloliquefaciens and M. oleovorans in different growth media, and the efficiency of a freeze-drying method as a possible formulation process. B. amyloliquefaciens and M. oleovorans were grown in 100 ml of four different media. Media water activity was modified at 0.99, 0.98, 0.97 and 0.96. Nutrient yeast dextrose broth (NYDB) and molasses soy powder (MSB) media were selected and survival levels of cells were determined before and after the freeze-drying process. B. amyloliquefaciens showed the highest survival after freeze-drying when grown in NYDB medium at 0.99 a(w), whereas, at 0.98, 0.97 and 0.96 a(w), the highest survival was obtained in MSB medium. M. oleovorans showed the highest survival in MSB medium at 0.99 a(w). MSB medium was select for biomass production due to high growth and survival after freeze-drying.
Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment 01/2011; 29(2):287-92.
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ABSTRACT: To assess the impact of Bacillus amyloliquefaciens and Microbacterium oleovorans on bacterial and fungal groups associated to the roots of field-grown maize.
Identification and count of bacterial and fungal culturable populations associated to the roots of maize seedlings, changes in culturable community structure according to the richness and diversity indexes concept and shifts in microbial activity through analysis of cellulolytic, ammonification and nitrification potentials were determined, in relation to kernel treatment with biological control agents. Following the treatment of maize kernels with B. amyloliquefaciens at 10(7) CFU ml(-1), an increase in bacterial diversity was observed at the rhizoplane of resultant seedlings. Bacterial richness was significantly increased at the root inner tissues of seedlings treated with Mic. oleovorans. Fusarium, Aspergillus, Penicillium and Trichoderma were the main fungal genera isolated and there population sizes were unequally affected by the addition of biocontrol agents.
Numbers and types of isolated bacteria and fungi changed in response to the addition of biocontrol agents, while microbial activity remained unchanged with respect to control.
This study provides an insight of the effects of proven biocontrol agents on micro-organisms naturally associated to the target crop.
Letters in Applied Microbiology 05/2009; 48(4):493-9. · 1.62 Impact Factor
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ABSTRACT: Natural phytochemicals may be an alternative to synthetic chemicals for controlling fungal growth and mycotoxin production in stored maize. A key to progress in this field is to select the best natural maize phytochemicals to be applied in a storage maize ecosystem. This research was undertaken to evaluate the effects of the natural phytochemicals trans-cinnamic acid (CA) and ferulic acid (FA) alone at concentrations of 20 to 30 mM and in 5 combinations on Aspergillus flavus Link and A. parasiticus Speare populations and aflatoxin B(1) production. Studies on Aspergillus population and aflatoxin B(1) production were carried out in maize grain in relation to a water activity a(w) of 0.99, 0.97, 0.95, and 0.93. CA and FA at concentrations of 25 to 30 mM, respectively, and CA-FA mixture T9 (25 + 30 mM) were the treatments most effective at inhibiting A. flavus and A. parasiticus population at all a(w) assayed after 11 d of incubation. At all a(w) values, the mixture CA-FA T9 (25 + 30 mM) completely inhibited (100%) aflatoxin B(1) production by both strains at a(w)= 0.99, 0.97, 0.95, and 0.93. Decreased aflatoxin B(1) levels in comparison with the control were observed with mixtures CA-FA T6 (10 + 25 mM), T7 (20 + 20 mM), and T8 (20 + 30 mM) of both strains in the majority of a(w) assayed. The data show that CA and FA could be considered as effective fungitoxicants for A. flavus and A. parasiticus in maize in the a(w) range 0.99 to 0.93. The information obtained shows promise for controlling aflatoxigenic fungi in stored maize.
Journal of Food Science 07/2007; 72(5):M180-5. · 1.66 Impact Factor
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ABSTRACT: To examine sclerotium characteristics of two Aspergillus flavus and two A. parasiticus strains at different growth media and water stress.
The effects of growth media and water activity (0.999, 0.971, 0.955 and 0.937) on characteristics of sclerotia production (number, size and volume) of four isolates of Aspergillus section Flavi were examined. There was total inhibition under the driest conditions (0.955 and 0.937). When an osmotic potential of 0.971 was generated in Czapek agar (CD) and maize meal extract agar with sucrose and sodium nitrate (MMEA S/N), an increase in sclerotial size and volume was observed. The amount of sclerotia produced by cultures at 0.999 a(w) value was higher on CD.
The data show that the sclerotia characteristics of A. flavus and A. parasiticus have been influenced by water availability and growth media composition.
The information obtained shows that if we know the nutritional and water stress requirements for sclerotia production, it could be possible to develop effective prevention strategies to inhibit the survival of these fungi in grain.
Letters in Applied Microbiology 03/2007; 44(2):149-54. · 1.62 Impact Factor
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ABSTRACT: Antagonist activity of Kluyveromyces spp. isolates on Aspergillus section Flavi was studied.
The screening of isolates were made through studies of growth at different water activities and temperatures, index of dominance (I(D)), ecological similarity, antifungal activity and impact on aflatoxin B1 accumulation. High optical density was obtained at 25 and 30 degrees C and 48 h of incubation. Cell growth decreases with decrease in water activity. The predominant interaction was mutual intermingling at a(w) = 0.982 and 0.955, while at a(w) = 0.999 and 0.937 mutual inhibition for contact was exhibited. All isolates were catabolically identical to Aspergillus section Flavi and compete by nutritional source. At high water activities yeasts showed inhibitory activity on Aspergillus strains, inhibition percentages varied between 75 and 100%. The isolates Y9, Y14, Y16, Y22, Y25 and Y33 showed antifungal activity and inhibitory activity on aflatoxin B1 accumulation at all water activities assayed from all Aspergillus section Flavi strains.
The data show that the isolates selected in a wide range of environmental conditions could exert their roll like biological control agents for Aspergillus section Flavi in storage maize ecosystem.
Isolates of Kluyveromyces spp. may have practical value in the postharvest control of storage maize.
Letters in Applied Microbiology 02/2004; 38(4):257-64. · 1.62 Impact Factor
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ABSTRACT: The effect of osmotic and matric potential stress on growth and sugar alcohols (polyols: glycerol, erythritol, arabitol and mannitol) and sugars (trehalose and glucose) accumulation in toxigenic and nontoxigenic colonies of Aspergillus flavus and A. parasiticus was evaluated.
Growth of Aspergillus section Flavi with significant reductions at 20 and 30 degrees C was more sensitive to changes in matric potential, between 60 and 100% in the range of -7 to -14 MPa. No significant differences were found between toxigenic and nontoxigenic strains for both species. Total polyol accumulation in unamended maize meal agar medium (-0.75 MPa water potential) was higher at 30 than 20 degrees C. The major change in concentrations of endogenous sugars and total polyols was in matrically amended medium (with PEG 8000) at -7 and -10 MPa. Accumulation of glucose, arabitol, mannitol and erythritol content of A. flavus and A. parasiticus mycelial colonies was greater in normal unstressed maize meal agar medium (-0.75 Mpa) at 20 degrees C. This was modified by solute and matric stress.
The data showed relative sensitivity to osmotic and matric potential, and temperature, and the impact on growth rates, polyol and sugar accumulation in mycelia of A. flavus and A. parasiticus.
The matric potential effects on growth may be of particular importance for growth and survival in environments with low-matric potential stress. The tolerance of spoilage fungi such as Aspergillus section Flavi to such modifications could increase the potential for spoilage and mycotoxin production in such substrates. This knowledge is important for understanding the relative ecological fitness of these aflatoxigenic species and in the development of prevention strategies for their control.
Journal of Applied Microbiology 02/2004; 96(5):965-72. · 2.34 Impact Factor
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ABSTRACT: The effect of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), trihydroxybutyrophenone (THB) and propyl paraben (PP) (at concentrations of 1, 10 and 20 mmol l(-1)) on germination, growth and aflatoxin B1 production by Aspergillus section Flavi was evaluated.
Studies on the percentage of spore germination, elongation rate, growth rate and aflatoxin B1 production were carried out in vitro in relation to water activity (aw) at 0.982, 0.937, 0.809 and 0.747 values. At 0.809 and 0.747aw values none of the isolates was able to germinate. Overall, PP and BHA were the antioxidants most effective at inhibiting germination of both species. In the presence of the lowest concentration of BHA and PP (1 mmol l(-1)) the conidial germination percentage ranged from 2 to 19% after 15 h of incubation at the highest water activity tested. BHA and PP at 10-20 mmol l(-1) completely inhibited conidial germination. The antioxidants more efficient in controlling Aspergillus elongation rate were PP, BHT and BHA. All strains were much more sensitive to all antioxidants tested on the percentage of spore germination and growth rate at 0.937aw. The antioxidants PP and BHA completely inhibited aflatoxin B1 production by all strains when added at 1 mmol l(-1). Decreased aflatoxin B1 levels in comparison with the control, were observed with BHT at 1, 10 and 20 mmol(-1) with the strain T20 at 0.982aw. In contrast, stimulation was observed with the antioxidant THB at 10 and 20 mmol l(-1) at 0.937aw with the strains T20 and T23. The effect of BHA and PP at 1 mmol l(-1) on lag phase and growth rate was maintained in the pH range between 6 and 8. At all pH values the inhibitory effect of BHA was higher than PP. No aflatoxin B1 was detected at all pH values.
The data show that BHA and PP could be considered as effective fungitoxicants for A. flavus and A. parasiticus.
The information obtained show promise for controlling growth and aflatoxin B1 in stored maize. Futher studies should be carried out to examine the potential for antioxidants, such as BHA and PP to effectively control both growth and aflatoxin production.
Journal of Applied Microbiology 02/2003; 95(2):279-87. · 2.34 Impact Factor
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ABSTRACT: Populations of Aspergillus section Flavi were studied from a commercial field of maize in Río Cuarto, Córdoba, Argentina.
The Aspergillus species were isolated from soil, debris and insects during three periods: pre-planting, growing maize and post-harvest. The colony count from non-rhizospheric soil in the pre-planting period was higher than in growing maize and the post-harvest period. Debris samples analysed during all periods showed similar infection percentages for Aspergillus section Flavi. The samples of insects collected during the maize-growing period showed a lower percentage of Aspergillus isolates than the samples from soil and debris. Aflatoxigenic strains were present in lower levels in each component of the agroecosystem studied. All the strains that produced sclerotia were L strains.
In this field agroecosystem, the only strains with a high probability for transfer to the storage agroecosystem were L strains with low toxigenic potential.
Maize pre-harvest contamination with aflatoxigenic inoculum was not significant.
Letters in Applied Microbiology 02/2002; 34(5):343-8. · 1.62 Impact Factor
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ABSTRACT: Production of aflatoxin B1 and fumonisin B1 in pure and mixed cultures of Aspergillus flavus and Fusarium proliferatum were determined on irradiated maize seeds inoculated with different spore concentrations at 0.97 water activity (a(w)) and a temperature of 25 degrees C. The highest levels of aflatoxin B1 were produced by A. flavus at the lowest levels of inoculum (10(3) spore ml(-1)). There was no spore concentration influence on fumonisin B1 production after 10, 20 and 35 days of incubation. When A. flavus was co-inoculated with F. proliferatum, aflatoxin B1 production was inhibited. The higher the inocula levels of Fusarium produced, the higher the inhibition and this inhibition increased during the incubation period. Total inhibition was reached at 35 days of incubation. There was no interaction influence on fumonisin B1 production at all inoculum levels assayed. These results suggest that under optimal environmental conditions of substrate, water activity and temperature, the interaction between A. flavus and F proliferatum could produce inhibition of aflatoxin B1 and stimulation of fumonisin B1.
Natural Toxins 02/1999; 7(6):331-6.
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ABSTRACT: A study has been carried out in Argentina on samples of corn genotypes from a breeding station as well as in commercially available corn meal. All samples were analyzed for fungal infection and aflatoxin B1. Mycological analysis of corn genotypes showed the presence of three principal genera of filamentous fungi Fusarium (100%), Penicillium (67%) and Aspergillus (60%). In the genus Fusarium three species were identified, F. moniliforme (42%), F. nygamai (56%) and F. proliferatum (1.8%). Eight species of Penicillium were identified, the predominant species isolated were P. minioluteum, P. funiculosum and P. variabile. In the genus ranked third in isolation frequency, two species were identified, A. flavus and A. parasiticus, the percentage of infection was 78% and 21%, respectively. Only one corn genotype was contaminated with aflatoxin B1 at a level of 5 ppb. The corn meal samples showed great differences in fungal contamination, the values ranging from 1 x 10(1) to 7 x 10(5) cfu g-1. Fusarium (68%), Aspergillus (35%) and Penicillium (21%) were the most frequent genera isolated. Among the genus, Aspergillus, A. parasiticus (38%) was the most frequent species isolated. All the samples of corn meal were negative to aflatoxin B1. These results indicate a low degree of human exposure to aflatoxins in Argentina through the ingestion of maize or corn meal.
Mycopathologia 02/1999; 147(1):37-41. · 1.65 Impact Factor
