Fumonisin B1, zearalenone and deoxynivalenol production by Fusarium moniliforme,F proliferatum and F graminearum in mixed cultures on irradiated maize kernels

The impact on fungal growth and mycotoxin formation of interactions between fumonisin‐producing isolates of Fusarium moniliforme and F proliferatum and a zearalenone (ZEA)‐ and deoxynivalenol (DON)‐producing isolate of F graminearum inoculated together on irradiated maize at 15 and 25 °C and at 0.98, 0.95 and 0.93 a_w was studied. The presence of F graminearum decreased the fungal populations (CFU g⁻¹ grain) of F moniliforme and F proliferatum under almost all conditions tested. In the presence of F moniliforme, CFUs of F graminearum increased significantly at 25 °C, especially at 0.93 and 0.95 a_w, while the presence of F proliferatum caused them to increase at 15 °C. The presence of F graminearum always inhibited FB₁ production, except at 25 °C and 0.98 a_w where it increased. However, the observed differences were not statistically significant. There was no effect of fungal interaction on ZEA production by F graminearum; however, when paired with F moniliforme and F proliferatum, DON production by F graminearum was significantly stimulated, especially at 0.98 a_w. © 2000 Society of Chemical Industry     

Investigations on Fusarium spp. and their mycotoxins causing Fusarium ear rot of maize in Kosovo

After wheat, maize (Zea mays L.) is the second most important cereal crop in Kosovo and a major component of animal feed. The purpose of this study was to analyse the incidence and identity of the Fusarium species isolated from naturally infected maize kernels in Kosovo in 2009 and 2010, as well as the mycotoxin contamination. The disease incidence of Fusarium ear rot (from 0.7% to 40% diseased ears) on maize in Kosovo is high. The most frequently Fusarium spp. identified on maize kernels were Fusarium subglutinans, F. verticillioides/F. proliferatum and F. graminearum. Maize kernel samples were analysed by LC-MS/MS and found to be contaminated with deoxynivalenol (DON), DON-3-glucoside, 3-acetyl-DON, 15-acetyl-DON, zearalenone, zearalenone-14-sulphate, moniliformin, fumonisin B₁ and fumonisin B₂. This is the first report on the incidence and identification of Fusarium species isolated from naturally infected maize as well as the mycotoxin contamination in Kosovo.     

Influence of water activity,temperature and incubation time on the simultaneous production of deoxynivalenol and zearalenone in corn (Zea mays) by Fusarium graminearum

The production of deoxynivalenol and zearalenone by Fusarium graminearum was studied under different culture conditions (water activity, temperature and incubation time). The maximum levels of both toxins were obtained at the 35th day of incubation, the zearalenone level being much higher than the deoxynivalenol. The culture conditions that gave higher yields of deoxynivalenol were at 22 and at 28 °C (6.0 and 5.5 mg/kg ), after 35 days. At an incubation temperature of 28 °C 16 days, followed by 12 °C, for the same time, the production was low (1.1 mg/kg). The highest level of zearalenone was obtained at 28 °C for 16 days, followed by incubation at 12 °C (36.7mg/kg) at the 35th day. When the temperature was constant at 28 °C, the zearalenone production was lower (3.0 mg/kg) than when incubated at 22 °C (12.3 mg/kg), at the 35th day. Fusarium graminearum did not produce deoxynivalenol and zearalenone at 37 °C.     

Contamination of barley seeds with Fusarium species and their toxins in Spain: an integrated approach

Fusarium is a globally distributed fungal genus that includes different species pathogenic to cereals among others crops. Some of these Fusarium species can also produce toxic compounds towards animals and humans. In this work, the presence of the most important Fusarium toxins was determined in barley seeds from Spain, sampled according to European Union requirements. The results obtained were compared with the presence of mycotoxigenic species considered responsible for their synthesis by using species-specific polymerase chain reaction protocols. Fumonisins B₁ and B₂, zearalenone, trichothecenes type A (T-2 and HT-2) and trichothecenes type B (deoxynivalenol and nivalenol) were analysed by using high-performance liquid chromatography. Deoxynivalenol and zearalenone were detected in 72% and 38% of the barley samples, respectively, at levels below European Union limits in all cases. However, the co-occurrence of both toxins in 34% of the samples suggested that synergistic activity of these two mycotoxins should be evaluated. Nivalenol and HT-2/T-2 were detected at low levels in 17% and 10% of the samples, respectively. Fumonisins occurred in 34% of the samples at levels up to 300?µg/kg. This suggested that they might represent a risk in Spanish barley, and to our knowledge, this is the first report on the presence of fumonisins in barley in this country. The species-specific polymerase chain reaction assays to detect mycotoxin-producing Fusarium species showed a very consistent correlation between F. verticillioides detection and fumonisin contamination as well as F. graminearum presence and zearalenone, deoxynivalenol and nivalenol contamination in barley samples. The approach used in this study provided information of mycotoxin contamination of barley together with the identification of the fungal species responsible for their production. Detection of the species with the current polymerase chain reaction assay strategy may be considered predictive of the potential mycotoxin risk in this matrix.     

Relationship between environmental factors,dry matter loss and mycotoxin levels in stored wheat and maize infected with Fusarium species

This study examined the relationship between storage environmental factors (water activity (a _w) (0.89–0.97) and temperature (15°C–30°C)), colonisation of wheat and maize by Fusarium graminearum and F. verticillioides respectively and the dry matter losses (DMLs) caused and quantified by contamination with deoxynivalenol (DON), zearalenone (ZEA) and fumonisins (FUMs) during storage. Fungal growth was assessed by the amount of CO₂ produced under different interacting conditions of a _w and temperature. DMLs were quantified using the cumulative CO₂ data, and these were shown to increase as temperature and a _w increased. The amount of DON, ZEA (wheat for human consumption) and FUMs (feed maize) produced was significantly affected by the storage conditions. The three toxins however showed different patterns of production. Optimum for DON was at the wettest conditions (0.97a _w) and the highest temperature assessed (30°C), whereas for ZEA this shifted to 25°C. FUMs were produced in higher amounts in maize at 30°C and 0.97a _w; however, at intermediate a _w levels (0.955a _w), the highest production occurred at 25°C followed by 20°C. Polynomial models were developed for the effect of the storage factors on DMLs and toxin production. DMLs under different environmental conditions were significantly correlated with DON and FUMs. DON contamination was above the EU limits in at least 80% of the wheat samples with DMLs >1%, whereas at least 70% of the same samples contained ZEA above the respective EU legislative limits. Similarly, at least 75% of the maize samples with DMLs?≥?0.9% exceeded the EU limits for the sum of FUMs in feed. These results show that it may be possible to use temporal CO₂ production during storage of grains as an indicator of the level of contamination of the grain with mycotoxins.     

Ecophysiology of Fusarium temperatum isolated from maize in Argentina

The effect of water activity (a_w = 0.95, 0.98 and 0.995), temperature (15, 25 and 30°C), incubation time (7, 14, 21 and 28 days), and their interactions on growth and moniliformin (MON), beauvericin (BEA), fusaproliferin (FUS) and fumonisin B₁ (FB₁) production by two strains of Fusarium temperatum isolated from Argentinean maize were determined in vitro on sterile layers of maize grains. The results showed that there was a wide range of conditions for growth and mycotoxins production by F. temperatum. Both strains were found to grow faster with increasing a_w and at 30°C. In relation to mycotoxin production, the two strains produced more FUS than the other mycotoxins regardless of a_w or temperature evaluated (maximum = 50 000 μg g^?1). For FUS, MON and BEA, the maximum levels were observed at 0.98 a_w and 30°C (50 000, 5000 and 2000 μg g^?1 respectively). The lowest levels for these three mycotoxins were detected at 15°C and 0.95 a_w (1700 and 100 μg g^?1 for FUS and MON respectively), and at 0.98 a_w (400 μg g^?1 for BEA). The maximum levels of FB₁ were produced at 15°C and 0.98 a_w (1000 μg g^?1). At all a_w and temperatures combinations evaluated there was an increase in toxin concentrations with time incubation. The maximum levels were detected at 21 days. Statistical analyses of a_w, temperature, incubation time, and the two- and three-way interactions between them showed significant effects on mycotoxins production by F. temperatum. For its versatility on growth and mycotoxin production, F. temperatum represents a toxicological risk for maize in the field and also during grain storage.     

Impact of environment and interactions of Fusarium verticillioides and Fusarium proliferatum with Aspergillus parasiticus on fumonisin B1 and aflatoxins on maize grain

Fusarium verticillioides and F proliferatum isolates were inoculated in mixed cultures with Aspergillus parasiticus on irradiated maize grain at two different inoculum concentrations (2 × 10⁵ and 2 × 10² conidia g^?1 dry maize). The treatments were 0.93–0.98 water activity (a_w) and 15 and 25 °C for 28 days. A complex relationship was found between a_w, temperature, inoculum concentration and the interactions which took place between fumonisin and aflatoxin producers. In general, A parasiticus reduced F verticillioides and F proliferatum populations (by 6–36%) but did not affect fumonisin B₁ production by these species. In contrast, while the Fusarium species were not able to decrease A parasiticus populations, they significantly reduced aflatoxin B₁ accumulation (by 30–93%). © 2001 Society of Chemical Industry     

Occurrence of Fusarium Head Blight species and Fusarium mycotoxins in winter wheat in the Netherlands in 2009

Most recent information on the occurrence of Fusarium Head Blight species and related mycotoxins in wheat grown in the Netherlands dates from 2001. This aim of this study was to investigate the incidence and levels of Fusarium Head Blight species and Fusarium mycotoxins, as well as their possible relationships, in winter wheat cultivated in the Netherlands in 2009. Samples were collected from individual fields of 88 commercial wheat growers. Samples were collected at harvest from 86 fields, and 2 weeks before the expected harvest date from 21 fields. In all, 128 samples, the levels of each of seven Fusarium Head Blight species and of 12 related mycotoxins were quantified. The results showed that F. graminearum was the most frequently observed species at harvest, followed by F. avenaceum and M. nivale. In the pre-harvest samples, only F. graminearum and M. nivale were relevant. The highest incidence and concentrations of mycotoxins were found for deoxynivalenol, followed by zearalenone and beauvericin, both pre-harvest and at harvest. Other toxins frequently found – for the first time in the Netherlands – included T-2 toxin, HT-2 toxin, and moniliformin. The levels of deoxynivalenol were positively related to F. graminearum levels, as well as to zearalenone levels. Other relationships could not be established. The current approach taken in collecting wheat samples and quantifying the presence of Fusarium Head Blight species and related mycotoxins is an efficient method to obtain insight into the occurrence of these species and toxins in wheat grown under natural environmental conditions. It is recommended that this survey be repeated for several years to establish inter-annual variability in both species composition and mycotoxin occurrence.     

Effect of essential oils of cinnamon,clove, lemon grass,oregano and palmarosa on growth of and fumonisin B1 production by Fusarium verticillioides in maize

The ability of cinnamon, clove, lemon grass, oregano and palmarosa essential oils to prevent growth of and fumonisin B₁ (FB₁) production by Fusarium verticillioides at different water activity (0.95 and 0.995 a_w) and temperature (20 and 30 °C) levels in irradiated maize grain was evaluated. All the essential oils inhibited growth of F verticillioides isolates under all conditions tested, but FB₁ production was only inhibited at 30 °C and 0.995 a_w. Moreover, stimulation of toxin production was found under certain environmental conditions. None of the essential oils showed a significantly greater ability to inhibit FB₁ production when compared with the others. At 1000 mg essential oil kg^?1 maize the essential oils showed a greater inhibitory effect on growth of F verticillioides than at 500 mg kg^?1, but there was no difference in FB₁ production between the two levels of essential oil. Copyright © 2004 Society of Chemical Industry     

Reaction of winter wheat (Triticum aestivum L.) cultivars to infection with Fusarium spp.: mycotoxin contamination in grain and chaff

This study compares the susceptibility of winter wheat (Triticum aestivum L.) cultivars to Fusarium head blight (FHB) and accumulation of mycotoxins in kernels and chaff under different climatic conditions in two locations–Cerekwica near Poznan (Central West Poland) and Sitaniec, near Zamosc, Lublin region (South East Poland). Very high variations were found in the concentrations of mycotoxins (zearalenone, ZEA; nivalenol, NIV; deoxynivalenol, DON; moniliformin, MON) in examined fractions: Fusarium-damaged kernels (FDK) and healthy looking kernels (HLK) and in chaff for individual cultivars in both locations. In most cases, significantly higher concentrations of investigated toxins were recorded in wheat from the area of Lublin than from Poznan (p < 0.05). The highest Fusarium infection rates and mycotoxin biosynthesis levels were observed in the Lublin location, with the percentage of the FDK fraction ranging 8.1–81.6. In this region, ZEA concentration (µg g^?1) after inoculation with F. culmorum and F. graminearum ranged from 0.02–0.48 and 0.32–1.04, respectively. In the Poznan area, the toxin concentrations were considerably lower, ranging 0.01–0.10 and 0.03–0.13 µg g^?1 for both isolates, respectively. The concentration of DON was significantly higher than ZEA or NIV levels. The levels of MON accumulation (µg g^?1) in the FDK fraction were between 0.14 and 1.73 (Poznan area) and ND (not detected) to 2.51 (Lublin area). F. avenaceum infection rate ranged 7–35% in samples where the toxin was detected.     

Trichothecenes and zearalenone production by Fusarium equiseti and Fusarium semitectum species isolated from Argentinean soybean

Fusarium equiseti and Fusarium semitectum represent the most abundant species in the Fusarium complex isolated from flowers, soybean pods and seeds in Argentina. The aim of the present study was to assess the production of major type A and type B trichothecenes (diacetoxyscirpenol, neosolaniol, T-2 toxin and HT-2 toxin, nivalenol, deoxynivalenol) and zearalenone by 40?F. equiseti and 22 F. semitectum isolates on rice culture. Mycotoxins were determined by HPLC with fluorescence detection after derivatisation with 1-anthronylnitrile for type A trichothecenes (i.e. diacetoxyscirpenol, neosolaniol, T-2 toxin and HT-2 toxin), by HPLC with UV detection for type B trichothecenes (i.e. nivalenol and deoxynivalenol), and by TLC for zearalenone. A total of 22 of 40?F. equiseti isolates produced diacetoxyscirpenol, nivalenol and ZEA alone or in combination, whereas only two of 20?F. semitectum isolates were nivalenol and ZEA producers. Both Fusarium species did not produce any deoxynivalenol, neosolaniol, T-2 toxin and HT-2 toxin. The variable retention in toxigenicity displayed by both fungal species suggests that these species have a saprophytic lifestyle in the soybean agroecosystem in Argentina.     

THE FUSARIUM MYCOTOXIN DEOXYNIVALENOL AND YEAST GROWTH AND FERMENTATION

At a concentration of 50 μg ml^?1 medium, the trichothecene mycotoxin deoxynivalenol (DON) caused reductions of up to 15% in cell number, dry mass and total protein during growth of Saccharomyces cerevisiae for 24 h at 25°C, but had little effect on viability of cultures. During fermentation of glucose-amended Wickerham medium at 16°C, the presence of 20 μg DON ml^?1 had no effect on the rate of attenuation of glucose or production of ethanol, but lower concentrations of the related mycotoxin diacetoxscirpenol (DAS) strongly affected growth and also caused an initial lag during fermentation.     

Occurrence of free and conjugated Fusarium mycotoxins in cereal-based food

A collection of 84 cereal-based food products in 25 composites, including beer, was screened for the presence of deoxynivalenol, zearalenone, and their respective metabolites deoxynivalenol-3-glucopyranoside, 3-acetyl-deoxynivalenol, zearalenol-4-glucopyranoside, α-zearalenol, β-zearalenol, α-zearalenol-4-glucopyranoside, β-zearalenol-4-glucopyranoside, and zearalenone-4-sulfate. The most abundant analyte was zearalenone-4-sulfate, which was found in 13 composites, albeit in low concentrations. Furthermore, deoxynivalenol was detected in eight, zearalenone in seven, and deoxynivalenol-3-glucopyranoside in two composites. None of the remaining six analytes was found in any matrices, which suggests that, if at all present, the concentrations of these latter metabolites are very low and, hence, do not impose any danger to consumers. The highest mycotoxin content was found in bran flakes with 254 ng g^?1 deoxynivalenol, 6 ng g^?1 zearalenone-4-sulfate, and 44 ng g^?1 zearalenone.     

Effect of fungicides on the development of Fusarium head blight,yield and deoxynivalenol accumulation in wheat inoculated under field conditions with Fusarium graminearum and Fusarium culmorum

The effect of different fungicide treatments on Fusarium head blight (FHB) development, grain yields and deoxynivalenol (DON) accumulation in wheat was evaluated after artificial inoculation under field conditions with a mixture of Fusarium graminearum and F culmorum. The trials were carried out using commercially available products on five different cultivars of soft and durum wheat (Serio, Genio, Bracco, Duilio and Orobel) in two separate experimental fields located in the North of Italy. Treatments with Tiptor^® S (cyproconazole plus prochloraz) and a mixture of Horizon^® 250 EW (tebuconazole) plus Amistar^® 250 SC (azoxystrobin) significantly reduced the FHB disease severity (by between 25 and 77%) and DON content (by between 32 and 89%) in the grain as compared with the inoculated control. Yields (tonne ha^?1) and thousand grain weight (TGW) were higher in plots subjected to fungicide treatments. Tetraconazole (Eminent^® 40 EW) showed a markedly reduced effectiveness compared with the other treatments. Regression analysis showed a strong correlation of disease severity with DON levels (positive correlation) and with yields or TGW (negative correlation) for individual cultivars and locations. Fusarium graminearum, F culmorum and F poae were the species most commonly isolated from all trials, including inoculated and non‐inoculated control plots. Copyright © 2004 Society of Chemical Industry     

Effect of cycling temperatures on the production of deoxynivalenol and zearalenone by Fusarium graminearum NRRL 5883

The effects of three regimens of cycling incubation temperatures and incubation at constant 25 degrees C on the growth of Fusarium graminearum NRRL 5883 and production of deoxynivalenol (DON) and zearalenone (ZEN) on rice were compared. The effects of low-temperature stress were also studied by incubating rice cultures at a constant 15 degrees C for 4 weeks following incubation at constant 25 degrees C for 2 weeks. Both incubation temperature and time significantly (P < or = 0.05) affected growth of F. graminearum NRRL 5883 and production of DON and ZEN. The highest amount of free ergosterol (640 microg/g culture material) that was used as a measure of fungal growth was found in cultures incubated at temperatures cycling between 15 and 30 degrees C during a 6-week period. The highest amounts of DON (1,679 microg/g culture material) and ZEN (603 microg/g culture material) were produced in cultures incubated at a constant 25 degrees C for 2 weeks prior to incubation at a constant 15 degrees C for an additional 4 weeks. Under cycling incubation temperatures, maximum amounts of DON (850 microg/g culture material) and ZEN (98 microg/g culture material) were produced in cultures incubated at temperatures cycling between 15 and 30 degrees C for 6 weeks. Overall, there was no correlation between mold growth and production of either DON or ZEN. However, DON production and ZEN production were correlated.     

Mycoflora and deoxynivalenol in whole wheat grains (Triticum aestivum L.) from Southern Brazil

The fungal species Fusarium graminearum is related to deoxynivalenol (DON) formation. The aim of this study was to evaluate mycoflora and DON occurrence in 53 whole wheat grain samples collected in Southern Brazil during the 2012 crop. Wheat grains showed adequate values of water activity ranging from 0.48 to 0.72, within the required limits of moisture content, ranging from 9.1% to 13.9%. In addition, low counts of fungal colonies, ranging from 10 to 8.2 × 10², were found. For Fusarium genera, there was predominance of Fusarium verticillioides (34%) and F graminearum (30.2%). For Aspergillus species, 37.7% of Aspergillus flavus was determined. Regarding the Penicillium species, Penicillium digitatum (49%) was the most found species. DON was detected in 47.2% (25 out of 53) of the samples analysed, with levels ranging from 243.7 to 2281.3 µg kg^?1 (mean: 641.9 µg kg^?1).     

Production of trichothecenes and zearalenone by Fusarium species isolated from wheat

Fusaria isolates from wheat were tested for ability to produce trichothecenes and zearalenone. Four isolates of F. culmorum out of 13 produced vomitoxin (DON) and 3 Ac-DON, one produced diacetoxysirpenol and 12 zearalenone. Particularly high yield of zearalenone was observed in cultures of sever pathogenic isolates. Higher temperature (20 °C) during first week of incubation favoured yield of zearalenone. About 50% of zearalenone was produced by surface mycelium.     

Fusarium mycotoxin content of UK organic and conventional wheat

Each year (2001–2005), 300 samples of wheat from fields of known agronomy were analysed for ten trichothecenes by gas chromatography-mass spectrometry (GC/MS) including deoxynivalenol (DON), nivalenol, 3-acetyl-DON, 15-acetyl-DON, fusarenone X, T2 toxin, HT2 toxin, diacetoxyscirpenol, neosolaniol and T-2 triol and zearalenone by high-performance liquid chromatography (HPLC). Of the eleven mycotoxins analysed from 1624 harvest samples of wheat, only eight were detected, and of these only five–deoxynivalenol, 15-acetyl-DON, nivalenol, HT-2 and zearalenone–were detected above 100 µg kg^?1. DON was the most frequently detected Fusarium mycotoxin, present above the limit of quantification (10 µg kg^?1) in 86% of samples, and was usually present at the highest concentration. The percentage of samples that would have exceeded the recently introduced legal limits varied between 0.4% and 11.3% over the five-year period. There was a good correlation between DON and zearalenone concentrations, although the relative concentration of DON and zearalenone fluctuated between years. Year and region had a significant effect on all mycotoxins analysed. There was no significant difference in the DON concentration of organic and conventional samples. There was also no significant difference in the concentration of zearalenone between organic and conventional samples, however organic samples did have a significantly lower concentration of HT2 and T2. Overall, the risk of UK wheat exceeding the newly introduced legal limits for Fusarium mycotoxins in cereals intended for human consumption is low, but the percentage of samples above these limits will fluctuate between years.     

Solid substrate bioassay to evaluate impact of Trichoderma on trichothecene mycotoxin production by Fusarium species

BACKGROUND: Fusarium head blight of wheat is a destructive disease in various wheat‐growing regions and leads to significant yield losses for farmers and to contamination of cereal grains with mycotoxins, mainly deoxynivalenol and its derivatives. Toxigenic Fusarium species sporulate on cereal crop residues and produce significant inoculum for epidemics. Reduction of mycotoxin production and pathogen sporulation may be influenced by saprophytic fungal antagonists. RESULTS: Trichoderma isolates were examined in dual culture bioassays on rice with two isolates of Fusarium graminearum Schwabe and two isolates of Fusarium culmorum (W.G. Smith) Saccardo, belonging to three different chemotypes. Production of five trichothecene mycotoxins, deoxynivalenol (DON), 3‐acetyl‐deoxynivalenol (3AcDON), 15‐acetyl‐deoxynivalenol (15AcDON), nivalenol (NIV) and fusarenone X (FUS), was reduced by over 95%. Two Trichoderma isolates partially reduced the amounts of four metabolites when inoculated on autoclaved cultures of the same four Fusarium isolates. However, in the case of the 15AcDON chemotype of F. culmorum culture the content of DON increased and that of 15AcDON decreased. Isolates of Trichoderma varied in their ability to inhibit production of the five trichothecene mycotoxins by Fusarium. Susceptibility of the four Fusarium isolates to antagonistic activity of the same Trichoderma isolate differed significantly. CONCLUSION: Selected non‐toxigenic Trichoderma isolates proved to be useful biocontrol agents against toxigenic Fusarium pathogens of wheat, significantly reducing production of the trichothecene mycotoxins DON, NIV and their acetylated derivatives. Copyright © 2007 Society of Chemical Industry