The Effect of Ustilago maydis and Delayed Harvesting on A- and B-Type Trichothecene Concentrations in Maize Grain.

The aim of this study was to investigate whether, in the context of a higher incidence of Ustilago maydis and Fusarium spp. at optimal and delayed harvest times, a higher incidence of mycotoxin contamination in maize grains could be expected. The field experiment was carried out at the Lithuanian Research Centre for Agriculture and Forestry experimental fields over three consecutive years (2020-2022). Two maize hybrids (Duxxbury and Lapriora) with different FAO numbers were used. The experimental design in the field was a randomized complete block design. Harvesting took place at three different times: first at physiological maturity, and then 10 (±2) and 20 (±2) days after the first harvest. Each hybrid had four repetitions at different harvest times. The U. maydis infection was only detected in 2021 and after the first harvest cobs were further divided into four different groups with four repetitions: healthy cobs, cobs visually infected with Fusarium spp., cobs visually infected with common smut, and cobs visually infected with both pathogens. No U. maydis-damaged maize cobs were found in 2020 and 2022. The levels of Fusarium microscopic fungi in maize grains were also from 4 to 16 times higher in 2021 than in 2020 and 2022. Harvest delays in 2020 led to a significant deoxynivalenol concentration increase in the Duxxbury hybrid and an HT-2 concentration increase in the Lapriora hybrid. In 2021, deoxynivalenol, 3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, and HT-2 concentrations significantly rose in both hybrids, but the T-2 concentration significantly increased only in the Lapriora hybrid. Deoxynivalenol concentrations were, respectively, 110 and 14.6 times higher than in cobs only infected with Fusarium spp. or U. maydis. Concentrations of 15-acetyl-deoxynivalenol were, respectively, 60, 67, and 43 times higher than in asymptomatic cobs and cobs only infected with Fusarium spp. or U. maydis. Cobs contaminated with both pathogens also had higher concentrations of 3-acetyl-deoxynivalenol. T-2 and HT-2 were detected in maize grains harvested from cobs infected only with Fusarium spp. The presence of U. maydis and Fusarium fungi in maize cobs, along with harvest delays, led to significant increases in mycotoxin concentrations, highlighting the importance of timely harvesting and pathogen management to mitigate mycotoxin contamination in maize grains.

T-2 and HT-2 toxins in harvested oat grains and their prevalence in whole grain flour during storage.

Mycotoxin contamination of food is a constant global concern. There has been a scientific debate in Europe on the validation of accredited detection methods for type A trichothecenes T-2 and HT-2 and the restriction on dangerous concentrations. The issue is of great importance as this type of mycotoxin is frequently found in spring cereals grown in Lithuania. The aim of this study was to optimise and validate a method for the determination of T-2/HT-2 toxin concentrations in oats harvested in 2015-2018 and to observe the changes in the concentrations of both toxins in oat flour during 3- and 6-week storage at different temperatures and increased relative air humidity. All of the oat grain samples (100%) collected in 2015-2018 tested positive for contamination with type A trichothecenes. The highest mean co-contamination by T-2 + HT-2 (260.4 ± 140.9 µg/kg) and the highest concentration (594.6 µg/kg) were determined in 2018 when warm and wet weather conditions prevailed during oat flowering. The effect of long-term storage (6 weeks) on T-2 and HT-2 toxin production manifested itself only when the samples had been stored under cooler conditions (8 °C). The most important factors which impacted the variation of the concentrations of type A trichothecenes in flour were ambient temperature and storage time.

The effect of technological processes on contamination with B-class trichothecenes and quality of spring wheat products from grain harvested at different times.

The study aimed to investigate the effect of technological processes on deoxynivalenol (DON), 3-acetyl-deoxynivalenol (3-ADON) and 15-acetyl-deoxynivalenol (15-ADON) concentrations and quality of spring wheat products from grain harvested at different times. In this study, 408 samples were analysed for DON, 3-ADON and 15-ADON contamination by the HPLC method with UV detection. Delays in harvesting due to cool and rainy weather conditions resulted in increased DON, 3-ADON and 15-ADON levels. The highest DON concentrations were determined in bran. Higher DON concentrations in the bran indicate the protective function of the grain hull. On the other hand, the highest levels of minerals have been found in bran and whole grain flours, highlighting the importance of consuming these milling fractions as a mineral source with sustained health benefits. Our results showed that DON is stable at 170 °C, and high DON levels in whole-meal flour and white flour could not be converted or decomposed during baking. The levels of 3-ADON and 15-ADON in whole-meal flour bread and white flour bread were significantly reduced but not completely removed. The levels of DON and its derivatives 3-ADON and 15-ADON were significantly reduced in starch and gluten produced from contaminated whole meal flour; however, the washing process did not completely eliminate these toxic compounds. The concentrations of mycotoxins in starch and gluten remained relatively high. Negative correlation was found in highly contaminated samples between DON and bread baking properties. Also, inverse relationship was found between high mycotoxin concentrations and mineral element content in white flour.

Modelling the Effects of Weather Conditions on Cereal Grain Contamination with Deoxynivalenol in the Baltic Sea Region.

Fusarium head blight (FHB) is one of the most serious diseases of small-grain cereals worldwide, resulting in yield reduction and an accumulation of the mycotoxin deoxynivalenol (DON) in grain. Weather conditions are known to have a significant effect on the ability of fusaria to infect cereals and produce toxins. In the past 10 years, severe outbreaks of FHB, and grain DON contamination exceeding the EU health safety limits, have occurred in countries in the Baltic Sea region. In this study, extensive data from field trials in Sweden, Poland and Lithuania were analysed to identify the most crucial weather variables for the ability of Fusarium to produce DON. Models were developed for the prediction of DON contamination levels in harvested grain exceeding 200 µg kg-1 for oats, spring barley and spring wheat in Sweden and winter wheat in Poland, and 1250 µg kg-1 for spring wheat in Lithuania. These models were able to predict high DON levels with an accuracy of 70-81%. Relative humidity (RH) and precipitation (PREC) were identified as the weather factors with the greatest influence on DON accumulation in grain, with high RH and PREC around flowering and later in grain development and ripening correlated with high DON levels. High temperatures during grain development and senescence reduced the risk of DON accumulation. The performance of the models, based only on weather variables, was relatively accurate. In future studies, it might be of interest to determine whether inclusion of variables such as pre-crop, agronomic factors and crop resistance to FHB could further improve the performance of the models.

The effect of antifungal extracts on the contamination of grain with microfungi.

The study aimed to analyze the effects of extracts made from buckwheat grain, hulls, and bee products (propolis, bread, and pollen) and extraction solvents on the growth of microfungi on a medium and on buckwheat, wheat, oat, and maize grain. Research findings suggest that bioactive compounds contained in buckwheat grain reduced the amount of Fusarium spp. in the grain kept in the antifungal extract for 90 min at 25°C temperature. Buckwheat hull extract was more effective in inhibiting mycelial growth of mycotoxin-producing Fusarium culmorum and Fusarium graminearum compared with buckwheat grain extract (13%-50% and 14%-36%, respectively). The antifungal activity of extracts of bee products did not depend on the content of phenolic compounds in them; however, it depended on the grain species treated. After treatment of oat, wheat, and maize grain with bee product extracts, the lowest concentration of microfungi was identified on oat grain. More significant analysis results were obtained for the samples where ethanol solvent had been used for the preparation of extracts.

The influence of harvesting time and meteorological conditions on the occurrence of Fusarium species and mycotoxin contamination of spring cereals.

BACKGROUND: The aim was to determine the influence of harvesting time and meteorological conditions on the occurrence of Fusarium spp. and mycotoxins in the grain of spring cereals. A field experiment was performed in 2016-2018 with spring oat (Avena sativa L.) and spring triticale (× Triticosecale Wittm.) plots. Grain samples of oat and triticale were analysed for Fusarium infection and co-contamination with mycotoxins deoxynivalenol (DON), zearalenone (ZEA) and T-2 toxin (T-2). RESULTS: Results from the three-year study showed that the occurrence of Fusarium spp. fungi and mycotoxins produced by them in spring oat and triticale grain was most influenced by the meteorological conditions at harvesting time and crop species. CONCLUSIONS: It was found that in all experimental years, F. poae, F. tricinctum and F. sporotrichioides predominated in oat grains and F. graminearum, F. sporotrichioides and F. avenaceum predominated in spring triticale; as a result, oat grains were more contaminated with T-2 and triticale grains with DON and ZEA. Due to the rainy harvesting period in 2017, the contamination level of Fusarium fungi of grain of both crop species was 100%, and the concentrations of DON and ZEA in the samples of spring triticale were several times higher than those set forth in the EU regulation. Co-occurrence of all three mycotoxins analysed (DON, ZEA and T-2) was identified in these samples. © 2020 Society of Chemical Industry.

The prevalence of deoxynivalenol and its derivatives in the spring wheat grain from different agricultural production systems in Lithuania.

Deoxynivalenol (DON) together with two acetylated derivatives, 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON) occurs in cereal grains and their products. Co-occurrence of DON and acetylated derivatives in cereal grain is detected worldwide. Until now, DON and its derivatives have been considered equally toxic by health authorities. In this study, we analysed 103 samples of spring wheat grain, originating from the fields of different production systems in Lithuania, for the co-occurrence of type-B trichothecenes (DON, 3-ADON, 15-ADON). The samples were classified according to the production system-organic, sustainable and intensive. Mycotoxin levels in the spring wheat grain samples were determined by the HPLC method with UV detection. The type-B trichothecenes were found to be present at higher concentrations in the grain from the intensive production system. Eighty-one percent of the spring wheat grain samples from the intensive production system were co-contaminated with a combination of DON+3-ADON+15-ADON, 1% with DON+3-ADON. Additionally, DON+15-ADON and DON were found in 5% and 10% of the tested samples, respectively. Two percent of the samples were free from mycotoxins. In the grain samples from the sustainable production system, DON and a combination of DON+3-ADON showed a higher incidence - 47% and 23%, respectively. The samples with a combination of DON+3-ADON+15-ADON accounted for 18%. Completely different results were obtained from the analyses of organic grain samples. A large number of the organic spring wheat grain samples were contaminated with DON+3-ADON (55%) or DON (36%). The combination of DON+3-ADON+15-ADON was not present, while DON+15-ADON was present in 9% of the samples tested. The production systems did not lead to significant differences in mycotoxin levels, although a trend toward higher incidence and higher contamination was observed for the samples from the intensive and sustainable production systems.

Occurrence of mycotoxins in spelt and common wheat grain and their products.

Organic farming does not allow the use of conventional mineral fertilizers and crop protection products. As a result, in our experiments we chose to grow different species of cereals and to see how cereal species affect mycotoxin accumulation. This study describes the occurrence of deoxynivalenol (DON), zearalenone (ZEA) and T-2/HT-2 toxin in a survey of spelt and common wheat and their bran as well as flour. The analysis was conducted using an enzyme-linked immunosorbent assay (ELISA) method. The concentrations of DON, ZEA and T-2/HT-2 in Triticum spelta and T. aestivum were influenced by species, cereal type and year interaction. The highest concentrations of these mycotoxins were found in spelt grain with glumes, in spelt glumes and in spring wheat. These results show significantly higher concentrations of Fusarium toxins in glumes than in dehulled grain, which indicates the possible protective effect of spelt wheat glumes. The lowest DON, ZEA and T-2/HT-2 concentrations were determined in spelt grain without glumes. The research shows that it is potentially risky to produce bran from grain in which mycotoxin concentrations are below limits by European Union Regulation No. 1881/2006, since the concentration of mycotoxins in bran can be several times higher than that in grain. As a result, although bran is a dietary product characterised by good digestive properties, it can become a harmful product that can cause unpredictable health damage.

Distribution of trichothecene and zearalenone producing Fusarium species in grain of different cereal species and cultivars grown under organic farming conditions in Lithuania.

Fusarium infection level, DNA quantity of the Fusarium poae, F. sporotrichioides, F. langsethiae, F. culmorum, F. graminearum and F. equiseti as well as deoxynivalenol (DON), zearalenone (ZEN ) and T-2 toxin (T-2) content were investigated in grain from cultivars of different cereal species grown on organic farming sites during 2005-2006. The Fusarium infection level was examined by agar plating of single grains, Fusarium spp. DNA content was determined by real-time PCR and the mycotoxins were analyzed by ELISA. Almost all cereal grain samples grown under organic conditions were infected by Fusarium spp. The grains of winter cereals were less infected with Fusarium compared with those of spring cereals. The presence of F. culmorum, F. equiseti, F. sporotrichioides, F. poae, F. langsethiae in cereal grain depended on the environmental conditions during the experimental years. Higher Fusarium species diversity was found in 2005 when the conditions were more favourable for Fusarium infection in cereal grain, whereas F. poae and F. langsethiae were prevalent in cereal grain in 2006. F. langsethiae, identified in Lithuania for the first time, was more frequent in spring cereals than in winter cereals. Almost all grain samples were found to be contaminated with DON, ZEN, T-2 at low concentrations; however, it is known that the action of toxins at low concentrations is slow, the adverse effects are evidenced only after some time and in different forms, which poses a serious risk to human and animal health.

Fusarium mycotoxins in Lithuanian cereals from the 2004-2005 harvests.

Fusarium mycotoxins deoxynivalenol (DON), T-2 toxin, and zearalenone (ZEN) contamination in 5 kinds of cereal grain harvested in 2004 and 2005 in different regions of Lithuania was examined for their occurrence frequency and level. In all cereal species DON was the most frequently detected mycotoxin with an incidence rate of 98.0-100% and range in positive samples from traces to 691 microg kg(-1) in 2004 and 62.5-94.0%, range from traces to 1,121 microg kg(-1) in 2005, respectively. All the tested oat samples collected in 2004-2005 were found to be contaminated with the T-2 toxin. In one sample from the year 2004 the level of T-2 toxin (121.5 microg kg(-1)) exceeded the allowable level. In 2004, ZEN contamination was more frequent in spring wheat, barley and oats grain, whereas in 2005 this toxin was identified at higher levels only in barley grain (68.0%). In one barley grain sample from 2004, ZEN content (193.4 microg kg(-1)) exceeded the allowable level. Variation in the relative air-humidity exerted some effect on the incidence of Fusarium spp. fungi and mycotoxin content in wheat grain. The weather conditions at harvesting contributed to an increase in the contents of Fusarium fungi and DON and ZEN mycotoxins produced by them in winter wheat grain. This risk factor increases the threat to human and animal health.