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Zygosaccharomyces rouxii is the main spoilage yeast of grape juice concentrates. Detection and identification of Z. rouxii during the production of grape juice concentrate is critical to prevent spoilage in the final product. In this work, three grape juice concentrate processing plants were assessed by identifying osmophilic yeasts in juices and surfaces during different stages of a complete production line. Subsequently, molecular typing of Z. rouxii isolates was done to determine the strain distribution of this spoilage yeast. Osmotolerant yeast species, other than Z. rouxii, were mainly recovered from processing plant environments. Z. rouxii was only isolated from surface samples with grape juice remains. Z. rouxii was largely isolated from grape juice samples with some degree of concentration. Storage of grape juice pre-concentrate and concentrate allowed an increase in the Z. rouxii population. A widely distributed dominant molecular Z. rouxii pattern was found in samples from all three processing plants, suggesting resident microbes inside the plant. Fil: Rojo, María Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Torres Palazzolo, Carolina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Cuyo; Argentina Fil: Cuello, Raúl Andrés. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: González, Magalí Lucía Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Guevara, F.. Universidad de Santiago de Chile; Chile Fil: Ponsone, Maria Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Mercado, Laura Analia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Cuyo; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Martínez, C.. Universidad de Santiago de Chile; Chile Fil: Combina, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina

Mycotoxins including aflatoxins, deoxynivalenol, fumonisins and ochratoxin A are among the main fungal secondary metabolites detected as natural contaminants in South America in different commodities such as peanuts (aflatoxins), cereals (deoxynivalenol and fumonisins) or grapes (ochratoxin A). Different strategies including crop rotation, tillage practices, fungicide application and planting less susceptible cultivars are used in order to reduce the impact of these mycotoxins in both food and feed chains. The development of fungicide resistance in many fungal pathogens as well as rising of public concern on the risks associated with pesticide use led to the search for alternative environmentally friendly methods. Biological control of plant pathogens and toxigenic fungi offers an alternative that can complement chemical control in the frame of an integrated pest management to reduce the impact of mycotoxins in the food and feed chains. The advances made in Argentina on reducing the impact of toxigenic fungi and mycotoxins in peanut, grapes and cereals using the biocontrol strategy are summarised. Native bacteria, yeasts and filamentous fungi have been selected to evaluate them as potential biocontrol agents. Field trials showed that Bacillus subtilis RC 218 and Brevibacillus sp. RC 263 were effective at reducing deoxynivalenol accumulation in wheat. The application of Clonostachys rosea isolates on wheat stubble reduced Fusarium colonisation on the stubble. Bacillus amyloliquefaciens and Microbacterium oleovorans showed good activity to control both Fusarium verticillioides growth and the accumulation of fumonisins at pre-harvest stage in maize. Control of toxigenic Aspergillus flavus and aflatoxin accumulation in peanuts was achieved using a native atoxigenic Aspergillus flavus strain based on competitive exclusion of the toxigenic strains. Kluyveromyces thermotolerans strains were used as biocontrol agents to reduce the impact of Aspergillus section Nigri and ochratoxin A accumulation in grapes. Fil: Chulze, Sofia Noemi. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología. Cátedra de Micología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina Fil: Palazzini, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología. Cátedra de Micología; Argentina Fil: Torres, Adriana Mabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología. Cátedra de Micología; Argentina Fil: Barros, Germán Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología. Cátedra de Micología; Argentina Fil: Ponsone, Maria Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Instituto Nacional de Tecnología Agropecuaria; Argentina Fil: Geisen, R.. Max Rubner-Institut; Alemania Fil: Schmidt Heydt, M.. Max Rubner-Institut; Alemania Fil: Köhl, J.. Wageningen UR. Plant Research International, Wageningen; Países Bajos

In a previous study, it was demonstrated that Kluyveromyces thermotolerans strains can reduce both growth and ochratoxin A (OTA) accumulation by Aspergillus section Nigri strains. There is no information about the mechanisms related to this reduction. A viable hypothesis can be that the presence of biocontrol agents can affect OTA biosynthesis by influencing the transcriptional activity of the polyketide synthase (pks) gene, one of the key enzymes in the OTA biosynthetic pathway. The aims of this work were to determine the effect of two selected strains of K. thermotolerans as potential biocontrol agents and to evaluate if their presence can affect the otapks gene expression of ochratoxigenic Aspergillus and Penicillium species. Growth, OTA and ochratoxin B (OTB) biosynthesis by the fungal strains at the phenotypic and molecular levels were monitored. The results obtained showed that both K. thermotolerans strains evaluated had a strong influence on growth, OTA and OTB biosynthesis, and expression of the mycotoxin biosynthesis genes. However, no direct correlation between the influence of the biocontrol yeasts on pks gene expression, OTA and OTB production could be found. These results could indicate an inhibitory mechanism by the yeasts, which apparently involve a post-transcriptional mechanism. The data obtained could imply that the production of mycotoxins can be regarded as a kind of adaptation mechanism to environmental stress conditions by these mycotoxigenic species. Fil: Ponsone, Maria Lorena. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Kuhn, Yamila Gisele. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Schmidt Heydt, M.. Max Rubner Institut; Alemania Fil: Geisen, R.. Max Rubner Institut; Alemania Fil: Chulze, Sofia Noemi. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

The aims of this work were to identify the Aspergillus sections Nigri and Flavi, and to evaluate the natural occurrence of ochratoxin A (OTA) and aflatoxins in green and roasted coffee bean samples. The capacity to produce these toxins by Aspergillus species was also studied. Fifty samples of Colombian coffee beans (25 green and 25 roasted) were obtained from a processor plant located in the south of Córdoba province (Argentina). OTA and aflatoxin analysis were performed by high performance liquid chromatography (HPLC). OTA production by strains belonging to Aspergillus niger aggregate were cultivated using YES medium and detected by HPLC. Aflatoxin production was tested in strains belonging to section Flavi on malt extract agar and was detected by thin liquid chromatography (TLC). From green coffee samples, the predominant species isolated belonged to A. niger aggregate, 60 and 55%, in dichloran rose bengal chloramphenicol agar (DRBC) and dichloran 18% glycerol agar (DG18) respectively. While A. flavus strains were isolated in 14% from DRBC and A. parasiticus strains in 12% and 28% in DRBC and DG18, respectively. From roasted coffee samples, A. flavus was the most predominant fungi, isolated in similar percentages from both media (28%); followed by A. niger aggregate isolated in 28 and 14% in DRBC and DG18, respectively. In green and roasted coffee samples mean colony counts ranged from 2×103 to 3.5×104 colony forming units per gram of sample (cfu/g). OTA and aflatoxins were not detected in any sample analyses (1 (AFB1) and 80% of the A. parasiticus strains were AFB1 and aflotoxin G1 producers. The high percentage of A. flavus and A. parasiticus aflatoxin-producing strains suggest a potential risk for contamination in coffee with aflatoxins.

Aspergillus section Nigri are described as the main source of ochratoxin A (OTA) contamination in grapes and wine worldwide. The grape-growing area in Argentina has a wide latitudinal extension with ecological variations that allow the classification of well-demarcated regions. The aims of this study were: to determine the effect of eco-physiological parameters on growth of Aspergillus tubingensis and Aspergillus carbonarius and to evaluate the interaction between these species on OTA production in synthetic grape juice medium under different water activity (aw) and temperature conditions. The results showed that optimal growth conditions for A. tubingensis and A. carbonarius were 0.980 awand 28 °C, however A. tubingensis grew faster than A. carbonarius at all temperatures and awtested. OTA production by A. carbonarius was favoured at 20 °C and 0.950-0.965 aw, during 14 days of incubation. The effect of A. tubingensis on OTA production by A. carbonarius was mainly dependent on temperature. At 35 °C, A. tubingensis reduced the OTA production while this effect was not observed at 20 °C. More OTA could be produced as a defence reaction against fungal competitors to maintain niche colonisation, but in this study no effects by a related species were observed. Fil: Chiotta, María Laura. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Sosa, Débora María. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ponsone, Maria Lorena. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Chulze, Sofia Noemi. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Peanut is an important food commodity in Argentina. Last year Cordoba Province accounted for approximately 96% of the total Argentinian production. Few surveys of peanuts for the natural occurrence of ochratoxins and ochratoxin-producing fungi have been reported. The objectives of this study were to investigate the occurrence of Aspergillus section Nigri and ochratoxin A (OTA) in storage peanuts during a three-month-period. The capacity to produce OTA by Aspergillus section Nigri was also studied. A total of 100 samples were collected from May to July 2004. The frequency of contaminating fungi were determined by surface-disinfection the seeds and plating onto several agar types. Detection of OTA in seed samples was performed using an HPLC method. Strains belonging to Aspergillus section Nigri or Flavi were detected in all seeds samples. From the section Nigri, the species belonging to A. niger aggregate were isolated in 100% of the samples. The main ochratoxigenic specie, A. carbonarius, was present at low levels throughout the study period. OTA was found in 50% of the peanut samples, with mean levels ranging from 5.6 to 130 ng g(-1). The mean value of OTA obtained after the first month of storage (30 ng g(-1)) was significantly higher from those obtained after the second (6.5 ng g(-1)) and third (13 ng g(-1)) month (p