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1.
The aim of this study was to determine the effects of water activity (aw) (0.99-0.90), temperature (15, 25 and 30 °C) and their interactions on growth and alternariol (AOH) and alternariol monomethyl ether (AME) production by Alternaria alternata on irradiated soya beans. Maximum growth rates were obtained at 0.980 aw and 25 °C. Minimum aw level for growth was dependent on temperature. Both strains were able to grow at the lowest aw assayed (0.90). Maximum amount of AOH was produced at 0.98 aw but at different temperatures, 15 and 25 °C, for the strains RC 21 and RC 39 respectively. Maximum AME production was obtained at 0.98 aw and 30 °C for both strains. The concentration range of both toxins varied considerably depending on aw and temperature interactions. The two metabolites were produced over the temperature range 15 to 30 °C and aw range 0.99 to 0.96. The limiting aw for detectable mycotoxin production is slightly greater than that for growth. Two-dimensional profiles of aw × temperature were developed from these data to identify areas where conditions indicate a significant risk from AOH and AME accumulation on soya bean. Knowledge of AOH and AME production under marginal or sub-optimal temperature and aw conditions for growth can be important since improper storage conditions accompanied by elevated temperature and moisture content in the grain can favour further mycotoxin production and lead to reduction in grain quality. This could present a hazard if the grain is used for human consumption or animal feedstuff.  相似文献   

2.
Alternaria alternata is a toxigenic fungus, predominantly responsible for Blackmould of ripe tomato fruits, a disease frequently causing substantial losses of tomatoes, especially those used for canning. The objective of this study was to determine the effect of water activity (aw, 0.904, 0.922, 0.954, 0.982) and temperature (6, 15, 21 and 35 °C) on germination and radial growth rate on a synthetic tomato medium of a cocktail inoculum of five strains of A. alternata isolated from tomato fruits affected by Blackmould. The shortest germination time (1.5 days) was observed at 0.982 aw, both at 21 °C and 35 °C. The germination time increased with a reduction on aw. The fastest growth rate was registered at 0.982 aw and 21 °C (8.31 mm/day). Growth rates were higher when aw increased. No growth or germination was observed at the lowest aw level evaluated (0.904) after 100 days of incubation at 6 °C and 15 °C. A temperature of 6 °C caused a significant reduction in growth rates, even at the optimum aw level. The knowledge on the ecophysiology of the fungus in this substrate is necessary to elaborate future strategies to prevent its development and evaluate the consumer health risk.  相似文献   

3.
Fusarium langsethiae has been isolated from infected cereals in central and northern Europe where it has been identified in the last decade as the main species involved in the occurrence of high levels of T-2 and HT-2 toxins, mainly in oats. The efficacy of three fungicides (prochloraz, tebuconazole, fenpropimorph) for controlling growth of two strains of F. langsethiae isolated from oats was examined at 0.96 and 0.98 aw at 15, 20 and 25 °C on oat-based media. The concentrations necessary for 50 and 90% growth inhibition (ED50 and ED90 values) were determined. The effect on the trichothecene type A mycotoxins T-2 and HT-2 was also determined. Without fungicides both strains grew faster at 0.98 than at 0.96 aw and the influence of temperature on growth rates was 25 > 20 > 15 °C. Prochloraz and tebuconazole were more effective than fenpropimorph against F. langsethiae. Strain, temperature and type of fungicide significantly influenced the ED50 and ED90 values for growth. The concentration ranges under different environmental conditions were: prochloraz (0.03-0.1 and 0.3-1.5), tebuconazole (0.06-0.9 and 1.3-8.2), and fenpropimorph (22-59 and 125-215 mg l−1). Production of T-2 and HT-2 toxins was influenced by temperature, aw, type of fungicide and dose. Levels of T-2 were usually higher than those of HT-2 under the same conditions. The biosynthesis of T-2 toxin increased after 10 day incubation, but was reduced with decreasing temperature and increasing fungicide dose. At 0.98 aw T-2 levels increased in cultures containing fenpropimorph while at 0.96 aw the toxin concentrations increased in response to the other two fungicides. Low doses of prochloraz or tebuconazole enhanced toxin production when compared with untreated cultures for strain 2004-59 at 0.96 aw and 20-25 °C. HT-2 was hardly detectable in the treatments with prochloraz or tebuconazole at 0.98 aw. This is the first study on the effect of these anti-fungal compounds on control of growth of F. langsethiae and on production of T-2 and HT-2 toxins in oat-based media.  相似文献   

4.
The aim of this study was to determine the effects of water activity (aw) (0.92-0.98), temperature (5-45 °C) and incubation time (5-60 days) on growth and ochratoxin A (OTA) production by Aspergillus niger and Aspergillus carbonarius on maize kernels using a simple method. Colony diameters of both strains at 0.92 aw were significantly lower than those at 0.96 and 0.98 aw levels. The optimum growth temperature range for A. niger was 25-40 °C and for A. carbonarius 20-35 °C. A. niger produced OTA from 15 to 40 °C, and the highest OTA level was recorded at 15 °C. The concentration of OTA produced at 0.92 aw was significantly lower than those at 0.96 and 0.98 aw. A. carbonarius produced OTA from 15 to 35 °C and the maximum concentration was achieved at 15 °C, although not differing statistically from the concentration detected at 20 °C. At 0.98 aw the OTA concentration was significantly higher than at 0.96 and 0.92 aw. Our results show that maize supports both growth and OTA production by A. niger and A. carbonarius. The studied strains were able to produce OTA in maize kernels from the fifth day of incubation over a wide range of temperatures and water availabilities. Although the limit of quantification of our method was higher than that required for the analysis of OTA in food commodities, it has proved to be a useful and rapid way to detect OTA production by fungi inoculated onto natural substrates, in a similar way as for pure culture. Both species could be a source of OTA in this cereal in temperate and tropical zones of the world.  相似文献   

5.
The aim of this study was to determine the influence of different carbon and nitrogen sources on the production of the mycotoxins alternariol (AOH), alternariol monomethyl ether (AME) and tenuazonic acid (TA) by Alternaria alternata at 28 °C using a semi-synthetic medium (modified Czapek-Dox broth) supplemented with nitrogen and carbon sources. Additionally the effect of shaken and static cultivation on mycotoxin production was tested. Initial experiments showed a clear dependency between nitrogen depletion and mycotoxin production. To assess whether nitrogen limitation in general or the type of nitrogen source triggers the production, various nitrogen sources including several ammonium/nitrate salts and amino acids were tested. In static culture the production of AOH/AME can be enhanced greatly with phenylalanine whereas some nitrogen sources seem to inhibit the AOH/AME production completely. TA was not significantly affected by the choice of nitrogen source. In shaken culture the overall production of all mycotoxins was lower compared to static cultivation. Furthermore tests with a wide variety of carbon sources including monosaccharides, disaccharides, complex saccharides such as starch as well as glycerol and acetate were performed. In shaken culture AOH was produced when glucose, fructose, sucrose, acetate or mixtures of glucose/sucrose and glucose/acetate were used as carbon sources. AME production was not detected. The use of sodium acetate resulted in the highest AOH production. In static culture AOH production was also stimulated by acetate and the amount is comparable to shaken conditions. Under static conditions production of AOH was lower except when cultivated with acetate. In static cultivation 9 of 14 tested carbon sources induced mycotoxin production compared to 4 in shaken culture. This is the first study which analyses the influence of carbon and nitrogen sources in a semi-synthetic medium and assesses the effects of culture conditions on mycotoxin production by A. alternata.  相似文献   

6.
Zearalenone (ZON) distribution was measured in main fractions of malted barley, dependent on incubation time (17, 26, 34 days), water activity (0.95 and 0.98) and temperature (20 °C and 30 °C). Malted samples were sterilised and inoculated with Fusarium graminearum. ZON levels were higher (p < 0.01) in bran and germ than flour under almost all growing conditions. Incubation at 30 °C resulted in generally lower ZON levels in germ and bran, regardless of aw and incubation time. After 34 days, ZON levels in flour from samples that were incubated at the higher temperature rose significantly. At 20 °C ZON concentration showed a bell-shaped concentration profile with increasing incubation time in bran and germ, whilst ZON levels in flour increased at aw 0.98 and dropped at the lower aw. The results indicate the importance of storage conditions for ZON levels in commercially relevant grain fractions of malted barley and help predict existing mycotoxin levels or manipulate storage conditions to reduce ZON content.  相似文献   

7.
The aim of this study was to assess the impact of suboptimal environmental conditions on the intraspecific variability of A. carbonarius growth and OTA production using thirty isolates of A. carbonarius.Three aw/temperature conditions were tested, one optimal (0.98aw/25 °C) and two suboptimal: 0.90aw/25 °C and 0.98aw/37 °C as suboptimal water activity and temperature, respectively, which might take place through over ripening and dehydration of grapes. For each condition, 12 Petri dishes were inoculated, and colony growth and OTA production were measured over time.ANOVA revealed significant differences among μ and λ within the 30 assayed isolates. Coefficients of variation (CV%) revealed a wider dispersion of growth rates at 0.90aw/25 °C compared to 0.98aw/25 °C, and a more than 4-fold higher CV at 0.98aw/37 °C compared to 0.98aw/25 °C. However, dispersion of lag phases was similar at 0.98aw/25 °C and 0.90aw/25 °C and wider at 0.98aw/37 °C.There were significant differences (p < 0.05) among OTA levels (ng/mm2) for the different conditions, values being lower under marginal conditions, and particularly at 0.98aw/37 °C. Coefficients of variation (CV%) revealed a wider dispersion of OTA production at 0.90aw/25 °C compared to 0.98aw/25 °C, while CV at 0.98aw/37 °C was similar to that at 0.98aw/25 °C.In order to address the strain variability in growth initiation and prove the well-established notion of reducing OTA in foods by preventing fungal growth, a greater number of strains should be included when developing models for conditions that are suboptimal both for aw for OTA production and temperature levels for growth.  相似文献   

8.
The objective of the present study was to evaluate the effects of different gamma radiation doses on the growth of Alternaria alternata and on the production of toxins alternariol (AOH), and alternariol monomethyl ether (AME) in sunflower seed samples. After irradiation with 2, 5 and 7 kGy, the spore mass was resuspended in sterile distilled water and the suspension was inoculated into sunflower seeds. The number of colony-forming units per gram (CFU/g) was determined after culture on Dichloran Rose Bengal Chloramphenicol and Dichloran Chloramphenicol Malt Extract Agar. The presence of AOH and AME was investigated by liquid chromatography coupled to mass spectrometry. The radiation doses used resulted in a reduction of the number of A. alternata CFU/g and of AOH and AME levels when compared to the nonirradiated control group. Maximum reduction of the fungus (98.5%) and toxins (99.9%) was observed at a dose of 7 and 5 kGy, respectively. Under the present conditions, gamma radiation was found to be an alternative for the control of A. alternata and, consequently, of AOH and AME production in sunflower seeds.  相似文献   

9.
Although Fusarium species remain a main source of mycotoxin contamination of wheat, in recent years, due to the evident climatic changes, other mycotoxigenic fungi have been recognised as important wheat contaminants. Alternaria species, especially A. alternata, have been found as contaminants of wheat as well as wheat-based products. Under favourable conditions A. alternata very often produce alternariol (AOH), alternariol monomethyl ether (AME), tenuazonic acid (TeA) and others Alternaria toxins. The aim of the present study was to examine the presence of three Alternaria toxins (AOH, AME and TeA) in wheat samples harvested during three years (2011–13). To this end, 92 samples were collected during wheat harvesting from different growing regions of the Autonomous Province of Vojvodina, which represents the most important wheat-growing area in Serbia. The presence of Alternaria toxins was analysed by HPLC with electrospray ionisation triple quadrupole mass spectrometry (LC-ESI-MS/MS). Among all the analysed wheat samples, 63 (68.5%) were contaminated with TeA, 11 (12.0%) with AOH and 6 (6.5%) with AME. Furthermore, the maximum and mean toxin concentrations were 2676 and 92.4 µg kg?1, 48.9 and 18.6 µg kg?1, and 70.2 and 39.0 µg kg?1 for TeA, AOH and AME, respectively. Co-occurrence of three Alternaria toxins in wheat samples was detected in six samples; a combination of two toxins was found in two samples; and 64 samples contained one toxin. The results showed that among 92 analysed wheat samples, only 20 (21.7%) samples were without Alternaria toxins. The presence of Alternaria toxins was also investigated in terms of weather conditions recorded during the period of investigation, as well as with the sampling region. This study represents the first preliminary report of the natural occurrence of Alternaria toxins in wheat (Triticum aestivum) from Serbia.  相似文献   

10.
Lipoxidation in almond-derived products was investigated using the chemiluminescence (CL) and thiobarbituric acid-reactive substances (TBARS) methods to detect the first and later reaction products, respectively. The effects of light during storage at 5 °C, 22 °C and 40 °C were studied, as well as the effects of combined heat/water activity treatments in the 60–120 °C and 0.38–0.72 range. During storage, light was found to enhance the CL and TBARS values, and specific responses were observed in almond paste and the final Calisson product. During the heating of almond paste, as the initial water activity (aw) increased, the CL rate constants increased during heating to 60 °C and 80 °C, but interestingly, these values decreased during further heating to 120 °C, whereas the maximum TBARS rate constants occurred at aw 0.57 at all the heating temperatures tested. The activation energies, based on the CL and TBARS values, decreased specifically when the aw increased from 0.38 to 0.72, giving overall values ranging from110 kJ mol−1 to 60 kJ mol−1. Likewise, in the same water activity range, the temperature-dependent rate constant enhancing factor (Q10) decreased from 3.3 to 1.6.  相似文献   

11.
The aim of this work was to evaluate the application of a convective multi-flash drying process (CMFD) to producing dehydrated and crisp fruits. To accomplish this process, samples of banana (Musa sapientum L.) or mango (Mangifera indica L.) were heated to 60 °C by hot air, and a vacuum pulse was applied, which resulted in dehydration by a combination of convective drying and flash evaporation. Banana processed by CMFD had a moisture content of 0.293 g/g (dry basis) and aw = 0.272 after 3 h of processing. Mango had a moisture content of 0.09 g/g and aw = 0.359 after 4 h of processing. Puncture tests on fruits dehydrated by CMFD and on commercial freeze-dried fruits showed strain-force curves with many peaks (jagged curves). For CMFD much smaller global shrinkage was observed. These results indicate that the CMFD process can be applied for producing crispy fruits and is an alternative to the freeze-drying process.  相似文献   

12.
The aim of this study was to evaluate the efficacy of ferulic acid (1, 10, 20 and 25 mM) at different water activity (aw) values (0.99, 0.98, 0.96 and 0.93) at 25 °C on growth and fumonisin production by Fusarium verticillioides and Fusarium proliferatum on maize based media. For both Fusarium species, the lag phase significantly decreased (p ≤ 0.001), and the growth rates increased (p ≤ 0.001) at the lowest ferulic acid concentration used (1 mM), regardless of the aw. However, high doses of ferulic acid (10 to 25 mM) significantly reduced (p ≤ 0.001) the growth rate of both Fusarium species, regardless of the aw. In general, growth rate inhibition increased as ferulic acid doses increased and as media aw decreased. Fumonisin production profiles of both Fusarium species showed that low ferulic acid concentrations (1–10 mM) significantly increased (p ≤ 0.001) toxin production, regardless of the aw. High doses of ferulic acid (20–25 mM) reduced fumonisin production, in comparison with the controls, by both Fusarium species but they were not statistically significant in most cases. The results show that the use of ferulic acid as a post-harvest strategy to reduce mycotoxin accumulation on maize needs to be discussed.  相似文献   

13.
S. Kavitha 《LWT》2007,40(7):1280-1286
The influence of water activity (aw) (0.7,0.8 and 0.9) and temperature (80° and 120 °C) on the degradation of meat flavor precursor inosine monophosphate (IMP) was studied in a meat fiber model system. Breast and leg muscle from Indian domesticated layer chicken (Gallus gallus) were washed repeatedly with 0.1 mol/l phosphate buffer of pH 6 to obtain pigment free and with minimum content of natural IMP in muscle fiber. The freeze-dried breast and leg meat fiber had a protein content of 86.5±0.48% and 85.6±0.50%, respectively. The IMP contents (mg/100 g) of leg muscle fiber (7.3±0.60) was higher (P?0.05) than in the breast meat fiber (5.1±1.20). The degradation of IMP was temperature-dependent (P?0.05) in both types of meat fiber systems. In the samples of aw 0.8, the IMP degradation in breast meat fiber system was lower (P?0.05) than in aw at 0.7 and 0.9 samples, when treated at 80 °C, whereas, there was no significance difference (P>0.05) in the degradation of IMP at aw 0.7 and 0.9 when heated at 120 °C. The degradation of IMP in leg meat fiber model system at higher aw (0.9) was more (P?0.05) as compared to lower aw (0.7 and 0.8) at 80 °C, while the samples treated at 120 °C, the degradation of IMP at aw 0.8 and 0.9 was more (P?0.05) than at aw 0.7.  相似文献   

14.
In the present investigation, fresh and dried tomato samples from markets and packinghouses located in Apulia region (southern Italy) were analysed for Alternaria toxins. All samples proved to be contaminated by tenuazonic acid (TeA); in particular, dried tomatoes were contaminated in the range 425–81,592 µg/kg, while fresh tomatoes were in the range 11–4560 µg/kg. The second most abundant toxin was alternariol monomethyl ether (AME), followed by tentoxin (TEN) and alternariol (AOH). Overall dried tomatoes were more contaminated than fresh ones, although this seemed not directly related to the presence of sodium chloride, utilized in the drying process. Five representative Alternaria isolates within those collected from samples proved to be one Alternaria arborescens (A215) and four Alternaria alternata. Within the latter species, one strain belonged to morphotype tenuissima (A216), and three to alternata (A214, A217 and A218). They were confirmed to produce TeA, AOH, and AME in vitro. This study demonstrates the possible risk for consumers’ health related to the consumption of contaminated fresh and dried tomatoes, and thus the need for suitable control strategies.  相似文献   

15.
16.
    
Zusammenfassung Es wird eine Phasenumkehr-HPLC-Methode beschrieben, die es gestattet, dieAlternaria-Toxine Alternariol (AOH), Alternariolmonomethylether (AME), Altenuen (ALT) und Altertoxin I (ATX-I) in Obst- und Gemüseerzeugnissen mit guter Empfindlichkeit zu bestimmen. Die Ergebnisse werden dünnschichtchromatographisch abgesichert. Sie wurde an ca. 50 Handelsprodukten Bowie an ca. 20 verschimmelten Früchten erprobt. Keines der Handelsprodukte war mit den o. g. Mykotoxinen kontaminiert, während zwei der verschimmelten Proben (Äpfel) 160 ppb AOH, bzw. 250 ppb AME enthielten.
Determination of alternaria toxins in fruit and vegetable products
Summary A reverse phase HPLC method for the determination of theAlternaria mycotoxins alternariol (AOH), alternariol monomethyl ether (AME), altenuene (ALT) and altertoxin I (ATX-I) in fruit and vegetable products is described. The procedure has good sensitivity. The results are corroborated by TLC. About 50 commercial products and about 20 mouldy fruits were analysed. None of the commercial products was contaminated with the toxins whereas two of the mouldy samples (apples) contained 160 ppb AOH respectively 250 ppb AME.
  相似文献   

17.
Little is known about the mould, Xeromyces bisporus, unique in its strong xerophilicity and ability to grow at water activity (aw) 0.62, lower than for any other known organism. The linear growth rates of one fast and one slow-growing strain of X. bisporus were assessed at 20, 25, 30 and 37 °C on solid agar media containing a mixture of glucose and fructose to reduce aw to 0.94, 0.88, 0.84, 0.80, 0.76 and 0.66. Growth rates of xerophilic species closely related to X. bisporus, viz. Chrysosporium inops, C. xerophilum and Monascus eremophilus, were also assessed. Optimal conditions for growth of both X. bisporus strains were approx. 0.84 aw and 30 °C, despite FRR 2347 growing two- to five-fold faster than CBS 185.75. X. bisporus FRR 2347 even grew well at 0.66 aw (0.48 mm/day). C. inops and C. xerophilum were more tolerant of high aw than X. bisporus, and could be differentiated from each other based on: the faster growth of C. xerophilum; its preference for temperatures ≥ 30 °C and aw ≥ 0.94 (c.f. ≤ 25 °C and ~ 0.88 aw for C. inops); and its ability to grow at 0.66 aw, which is the lowest aw reported to date for this species. M. eremophilus grew slowly (max. 0.4 mm/day) even in its optimal conditions of ~ 0.88 aw and 25 °C. To investigate the competitive characteristics of X. bisporus at low aw, both X. bisporus strains were grown in dual-culture with xerotolerant species Aspergillus flavus and Penicillium roqueforti, and xerophilic species A. penicillioides, C. inops, C. xerophilum and Eurotium chevalieri, on glucose-fructose agar plates at 0.94, 0.84, 0.80 and 0.76 aw and at 25 °C. Growth rates and types of interactions were assessed. Excretion of inhibitory substances acting over a long-range was not observed by any species; inhibitors acting over a short-range that temporarily slowed competitors' growth or produced a protective zone around the colony were occasionally observed for A. penicillioides, C. inops and C. xerophilum. Instead, rapid growth relative to the competitor was the most common means of dominance. The xerotolerant species, A. flavus and P. roqueforti were dominant over X. bisporus at 0.94 aw. E. chevalieri was often dominant due to its rapid growth over the entire aw range. At aw < 0.80, X. bisporus was competitive because it grew faster than the other species examined. This supports the concept that its ideal environmental niche is sugary foods with low aw.  相似文献   

18.
C.M. Lee  A.V.A. Resurreccion 《LWT》2006,39(8):872-882
Consumer acceptance and intensity ratings of roasted peanuts stored at temperatures of 23, 30, 35, and 40 °C, and water activities (aw) of 0.33, 0.44, 0.54, 0.67 and 0.75 were determined over time. Consumer acceptance ratings, including overall, appearance, color, and texture were affected by storage water activity and time, but not by storage temperature. Consumer intensity ratings of crunchiness were affected by storage water activity and time, but not storage temperature. Aroma acceptance, flavor acceptance, and roasted peanutty and stale/oxidized/rancid intensity ratings of roasted peanuts were dependent on storage temperature, water activity and time.Shelf-life of roasted peanuts was predicted by all consumer attributes (R2>0.60) and was best predicted by aroma acceptance (R2=0.75). Using contour plots with ratings >5.0 for all acceptance attributes, the shelf-life of roasted peanuts stored at 23 °C and between 0.33 and 0.75 aw was limited by overall acceptance and decreased by approximately 50% with each 0.1 aw increase. At accelerated temperatures of 30, 35 and 40 °C, shelf-life of roasted peanuts was predominantly limited by flavor acceptance (>5.0), and to a lesser extent by overall acceptance (>5.0). Shelf-life of roasted peanuts stored at accelerated temperatures decreased by 50% or more with each 0.1 aw increase.  相似文献   

19.
Apple pomace (mixed Red Delicious and Golden Delicious varieties) obtained from the industrial production of puree was both vacuum-dried at 40 °C and air-dried at 60 °C, ground and stored at water activity (aw) in the range 0.11–0.75, for 9 months, at 30 °C. The aims were to investigate the effect of drying and long-term storage on phytochemical contents. Air-drying at 60 °C was better than vacuum-drying at 40 °C in terms of anthocyanin and flavanol retention; no adverse effect of drying was observed for flavonols, dihydrochalcones and hydroxycinnamic acids. The maximum stability occurred for all apple phytochemicals at the lowest aw. At aw 0.75 degradation of all phytochemicals occurred with the following stability ranking: phloridzin > chlorogenic acid > quercetin 3-O-galactoside > epicatechin > procyanidin B2 and cyanidin 3-O-galactoside. A promising feature of pomace is its exclusive high content of phloridzin, which showed relatively high stability during long-term storage.  相似文献   

20.
This study has examined the effect of water activity (aw, 0.995-0.90) and temperature (10-37 °C) on the lag phases prior to growth, growth rates and used models to develop two dimensional profiles for optimum and marginal conditions for two strains of Fusarium langsethiae from four northern European countries (UK, Norway, Sweden, and Finland) on an oat-based medium. Results showed that the optimum aw for growth was at 0.98-0.995 and 25 °C. The limit for growth of the strains was at 0.92-0.93 aw with minima of 10 °C. No growth occurred at 37 °C. The lag phases prior to growth were lowest under optimum conditions and extended to > 10 days at marginal conditions. Statistical analyses of intra and inter-strain differences in terms of both lag phases prior to growth and growth rates were not statistically significant. However, aw and temperature were statistically significant factors. Two dimensional profiles for strains from each country of origin were built to identify optimum and marginal conditions for F. langsethiae for the first time. These environmental profiles will be beneficial for improving the ecological knowledge of this species which is able to produce trichothecene mycotoxins in a range of temperate cereals.  相似文献   

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