RESUMO
Maize (Zea mays L.) is an important crop in Argentina. Aspergillus section Flavi can infect this crop at the pre-harvest stage, and the harvested grains can be contaminated with aflatoxins (AFs). During the production of bioethanol from maize, AF levels can increase up to three times in the final co-products, known as, dry and wet distiller's grain with solubles (DDGS and WDGS), intended for animal feed. Fungal enzymes like laccases can be a useful tool for reducing AF contamination in the co-products obtained from this process. The aim of the present study was to evaluate the ability of laccase enzymes included in enzymatic extracts (EE) produced by different species in the Basidiomycota phylum to reduce AF (AFB1 and AFB2) accumulation under the conditions of in vitro assays. Four laccase activities (5, 10, 15, and 20 U/mL) exerted by nine isolates were evaluated in the absence and presence of vanillic acid (VA), serving as a laccase redox mediator for the degradation of total AFs. The enzymatic stability in maize steep liquor (MSL) was confirmed after a 60 h incubation period. The most effective EE in terms of reducing AF content in the buffer was selected for an additional assay carried out under the same conditions using maize steep liquor obtained after the saccharification stage during the bioethanol production process. The highest degradation percentages were observed at 20 U/mL of laccase enzymatic activity and 1 mM of VA, corresponding to 26% for AFB1 and 26.6% for AFB2. The present study provides valuable data for the development of an efficient tool based on fungal laccases for preventing AF accumulation in the co-products of bioethanol produced from maize used for animal feed.
Assuntos
Aflatoxinas , Basidiomycota , Animais , Zea mays , Descontaminação , Lacase , Ácido VanílicoRESUMO
Maize (Zea mays L.) is an important crop in Argentina. Aspergillus flavus may infect this crop at growing stage and the harvested kernels can be contaminated with aflatoxins (AFs), whose levels may increase during storage. In Argentina, silo bags, a hermetic type of storage system, are widely used. Biocontrol based on competitive exclusion by atoxigenic A. flavus strains is a useful tool for AFs management at pre-harvest stage. The aim of the present study was to evaluate the effect of pre-harvest biocontrol treatments on aflatoxin B1 (AFB1) accumulation in maize stored in silo bags during 3 and 6 months. Three bioformulations based on A. flavus AFCHG2 and ARG5/30 strains were applied during field trials as single and mixed inocula. Harvested kernels were stored in non-hermetic and hermetic silo bags. At initial time (t0), 3 and 6 months (t3 and t6) the following parameters were evaluated: percentage of damaged kernels, moisture content, water activity, Aspergillus section Flavi incidence, relative humidity, O2 and CO2 levels into the silo bags, and AFB1 levels. The biocontrol strains included in the 3 bioformulations were able to infect maize kernels during the field trial and displaced native toxigenic isolates. At t0 control plots showed 10.9 ± 0.4 µg/kg of AFB1 while no AFs were detected in all the treatments. Along the storage assay AFB1 levels varied from not detected (<1 µg/kg) to 20.1 ± 0.8 µg/kg. Hermetic bags were better than non-hermetic bags in preventing AFB1 accumulation. Both single and mixed inocula were effective to control AFB1 accumulation in maize kernels during 3 and 6 months. AFB1 was not detected in kernels from the treatment at field stage with AFCHG2 + ARG5/30 after 6 months of storage into hermetic bags. The application of the biocontrol agents at field stage is an appropriate tool to reduce AFB1 accumulation under storage in hermetic silo bags. This is the first report on biocontrol strategy based on native atoxigenic strains applied at pre-harvest stage to reduce AFB1 accumulation during storage in Argentina.
Assuntos
Aflatoxinas , Aflatoxina B1 , Argentina , Aspergillus flavus , Zea maysRESUMO
Fusarium subglutinans and F. temperatum are two important fungal pathogens of maize whose distinctness as separate species has been difficult to assess. We isolated strains of these species from commercial and native maize varieties in Argentina and sequenced >28,000 loci to estimate genetic variation in the sample. Our objectives were to measure genetic divergence between the species, infer demographic parameters related to their split, and describe the population structure of the sample. When analyzed together, over 30% of each species' polymorphic sites (>2,500 sites) segregate as polymorphisms in the other. Demographic modeling confirmed the species split predated maize domestication, but subsequent between-species gene flow has occurred, with gene flow from F. subglutinans into F. temperatum greater than gene flow in the reverse direction. In F. subglutinans, little evidence exists for substructure or recent selective sweeps, but there is evidence for limited sexual reproduction. In F. temperatum, there is clear evidence for population substructure and signals of abundant recent selective sweeps, with sexual reproduction probably less common than in F. subglutinans. Both genetic variation and the relative number of polymorphisms shared between species increase near the telomeres of all 12 chromosomes, where genes related to plant-pathogen interactions often are located. Our results suggest that species boundaries between closely related Fusarium species can be semipermeable and merit further study. Such semipermeability could facilitate unanticipated genetic exchange between species and enable quicker permanent responses to changes in the agro-ecosystem, e.g., pathogen-resistant host varieties, new chemical and biological control agents, and agronomic practices.
Assuntos
Fusarium , Argentina , Ecossistema , Fusarium/genética , Fluxo Gênico , Doenças das Plantas , Zea maysRESUMO
Mycotoxins are secondary metabolites produced by fungal species that mainly belong to Aspergillus, Fusarium, Penicillium and Alternaria, which can grow in a variety of crops including cereals, oilseeds and fruits. Consequently, their prevalence in foods and by-products not only affects human and animal health but also causes important losses in both domestic and international markets. This review provides data about toxigenic fungal species and mycotoxin occurrence in different crops commonly grown in Argentina. This information will be relevant to establish adequate management strategies to reduce the impact of mycotoxins on human food and animal feed chains and to implement future legislation on the maximum permitted levels of these fungal metabolites.
Assuntos
Fusarium , Micotoxinas , Animais , Argentina , Contaminação de Alimentos/análise , Fungos , HumanosRESUMO
Fusarium head blight (FHB) is a devastating disease that causes extensive yield and quality losses to wheat and other small cereal grains worldwide. Species within the Fusarium graminearum complex are the main pathogens associated with the disease, F. graminearum sensu stricto being the main pathogen in Argentina. Biocontrol can be used as part of an integrated pest management strategy. Phytohormones play a key role in the plant defense system and their production can be induced by antagonistic microorganisms. The aims of this study were to evaluate the effect of the inoculation of Bacillus velezensis RC 218, F. graminearum and their co-inoculation on the production of salicylic acid (SA) and jasmonic acid (JA) in wheat spikes at different periods of time under greenhouse conditions, and to evaluate the effect of B. velezensis RC 218 and Streptomyces albidoflavus RC 87B on FHB disease incidence, severity and deoxynivalenol accumulation on Triticum turgidum L. var. durum under field conditions. Under greenhouse conditions the production of JA was induced after F. graminearum inoculation at 48 and 72 h, but JA levels were reduced in the co-inoculated treatments. No differences in JA or SA levels were observed between the B. velezensis treatment and the water control. In the spikes inoculated with F. graminearum, SA production was induced early (12 h), as it was shown for initial FHB basal resistance, while JA was induced at a later stage (48 h), revealing different defense strategies at different stages of infection by the hemibiotrophic pathogen F. graminearum. Both B. velezensis RC 218 and S. albidoflavus RC 87B effectively reduced FHB incidence (up to 30%), severity (up to 25%) and deoxynivalenol accumulation (up to 51%) on durum wheat under field conditions.
Assuntos
Bacillus , Ciclopentanos/metabolismo , Fusarium/metabolismo , Oxilipinas/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Ácido Salicílico/metabolismo , Streptomyces , Tricotecenos/metabolismo , Triticum/metabolismo , Agentes de Controle Biológico , Grão Comestível/química , Grão Comestível/metabolismo , Grão Comestível/microbiologia , Triticum/química , Triticum/microbiologiaRESUMO
Aspergillus section Nigri are described as the main source of ochratoxin A (OTA) contamination in grapes and wine worldwide. The knowledge of the factors affecting grape contamination by species included in this section and OTA production is essential to be able to reduce their presence, not only to improve wine quality, but also to maintain their safety. Therefore, the aims of this study were to determine the incidence of Aspergillus section Nigri species harvested in different grape-growing regions from Argentina, their ability to produce OTA, to correlate with meteorological conditions and geographical coordinates with their prevalence and to evaluate the OTA natural occurrence in grapes and wines. The morphological identification showed that Aspergillus niger aggregate species were the most prevalent ones, followed by Aspergillus carbonarius and Aspergillus uniseriate. These populations were confirmed through using AFLP markers and sequencing and, Aspergillus tubingensis was separated from A. niger aggregate. Climatic factors, altitude, longitude and latitude have influenced on the distribution of species included in the section. A. carbonarius and A. niger were OTA producers but differed in their OTA producing ability. Temperature was the factor which influenced the most over the highest incidence of A. carbonarius in La Rioja and San Juan regions. The trellis system in vineyards and drip irrigation also influenced the species isolation. The OTA levels detected in grapes and wines were low, but grape variety was more important in susceptibility to fungal infection and OTA levels.
Assuntos
Aspergillus/isolamento & purificação , Biodiversidade , Contaminação de Alimentos/análise , Ocratoxinas/biossíntese , Vitis/microbiologia , Argentina , Aspergillus/classificação , Aspergillus/genética , Aspergillus/metabolismo , Frutas/microbiologia , Vinho/microbiologiaRESUMO
Ochratoxin A (OTA) is a mycotoxin commonly present in cereals, grapes, coffee, spices, and cocoa. Even though the main objective of the food and feed chain processors and distributors is to avoid the extended contamination of plant-derived foods and animal feeds with mycotoxins, until now, complete OTA removal from foods and feedstuffs is not feasible. Prevention through pre-harvest management is the best method for controlling mycotoxin contamination. However, in the case that the contamination occurs after this stage, the hazards associated with OTA must be managed through post-harvest strategies. Due to the increasing number of fungal strains resistant to chemical fungicides and the impact of these pesticides on the environment and human health, maximum levels of chemical residues have been regulated in many products. Alternative methods are necessary to substitute or complement treatments with fungicides to control fungi under field or storage conditions. Yeasts are considered one of the most potent biocontrol agents due to their biology and non-toxic properties. Epiphytic yeasts are the major component of the microbial community on the surface of grape berries and they are evolutionarily adapted to this ecological niche. Nowadays, several yeast species included in different genera are considered as potential biocontrol agents to control both, growth of ochratoxigenic Aspergillus species and OTA accumulation.
Assuntos
Contaminação de Alimentos/prevenção & controle , Ocratoxinas/metabolismo , Vitis/metabolismo , Antioxidantes/farmacologia , Fungicidas Industriais/farmacologia , Controle Biológico de VetoresRESUMO
The efficacy of two strains of Kluyveromyces thermotolerans in preventing the growth and ochratoxin A (OTA) accumulation of ochratoxigenic fungi both "in vitro" and "in situ" was evaluated. The data from this study showed that both yeast strains were able to control Aspergillus carbonarius and A. niger aggregate species growth and ochratoxin A accumulation. The inhibitory effects were dependent on the ochratoxigenic species, yeast strains, a(w) and temperature evaluated and their interactions. Over all conditions assayed, ochratoxin A accumulation was reduced from 3% to 100% and the growth rate from 11% to 82.5%, depending on conditions. These results are promising for future development of a bio-pesticide.
Assuntos
Aspergillus niger/crescimento & desenvolvimento , Agentes de Controle Biológico , Contaminação de Alimentos/prevenção & controle , Kluyveromyces/fisiologia , Ocratoxinas/análise , Vitis/microbiologia , DNA Fúngico/genética , Kluyveromyces/genética , Kluyveromyces/isolamento & purificação , Polimorfismo de Fragmento de Restrição , Análise de Sequência de DNA , TemperaturaRESUMO
In this study, ochratoxin A (OTA) occurrence in Argentinean musts, wines and dried vine fruits was evaluated, alongside with the performance of OchraStar(TM) columns for OTA extraction. In all the three matrices analyzed, the OchraStar(TM) columns showed good performance. The analysis of natural occurrence of OTA in the red must and the red wine samples showed low incidence with low levels of mean OTA contamination (0.12 ng/mL and 0.37 ng/mL, respectively), while 60% of the dried vine fruit samples were contaminated with OTA, in levels ranging from 0.26 to 20.28 ng/g.