Assessment of Human Mycotoxin Exposure in Hungary by Urinary Biomarker Determination and the Uncertainties of the Exposure Calculation: A Case Study

Urinary biomarkers of mycotoxin exposure were evaluated in the case of healthy people (n = 41) and coeliac patients (n = 19) by using a multi-biomarker LC-MS/MS immunoaffinity based method capable to analyse biomarkers of nine mycotoxins, i.e., fumonisin B1 (FB1), fumonisin B2 (FB2), deoxynivalenol (DON), zearalenone (ZEN), ochratoxin A (OTA), Aflatoxin B1 (AFB1), T-2 toxin, HT-2 toxin and Nivalenol (NIV). Urinary biomarker concentrations were used to calculate the probable daily intake (PDI) of fumonisin B1, deoxynivalenol, zearalenone and ochratoxin A and compared with their tolerable daily intake (TDI). The human urinary excretion rate values reported in the literature and the 24 h excretion rate measured in piglets were used to estimate and compare the PDI values of the four mycotoxins. The highest mean biomarker concentrations were found for DON (2.30 ng/mL for healthy people and 2.68 ng/mL for coeliac patients). Mean OTA concentration was significantly higher (p < 0.001) in healthy people compared to coeliac patients. PDI calculated with piglets excretion data exceeded the TDI values by a much smaller percentage than when they were calculated from human data, especially for FB1. The uncertainties arising from the different calculations can be well perceived on the basis of these data.

Fumonisin B1 Accumulates in Chicken Tissues over Time and This Accumulation Was Reduced by Feeding Algo-Clay

The toxicokinetics of the food and feed contaminant Fumonisin B (FB) are characterized by low oral absorption and rapid plasma elimination. For these reasons, FB is not considered to accumulate in animals. However, recent studies in chicken and turkey showed that, in these species, the hepatic half-elimination time of fumonisin B1 (FB1) was several days, suggesting that FB1 may accumulate in the body. For the present study, 21-day-old chickens received a non-toxic dose of around 20 mg FB1 + FB2/kg of feed to investigate whether FB can accumulate in the body over time. Measurements taken after four and nine days of exposure revealed increased concentrations of sphinganine (Sa) and sphingosine (So) over time in the liver, but no sign of toxicity and no effect on performances were observed at this level of FB in feed. Measurements of FB in tissues showed that FB1 accumulated in chicken livers from four to nine days, with concentrations of 20.3 and 32.1 ng FB1/g observed, respectively, at these two exposure periods. Fumonisin B2 (FB2) also accumulated in the liver, from 0.79 ng/g at four days to 1.38 ng/g at nine days. Although the concentrations of FB found in the muscles was very low, an accumulation of FB1 over time was observed in this tissue, with concentrations of 0.036 and 0.072 ng FB1/g being measured after four and nine days of exposure, respectively. Feeding algo-clay to the chickens reduced the accumulation of FB1 in the liver and muscle by , approximately 40 and 50% on day nine, respectively. By contrast, only a weak non-significant effect was observed on day four. The decrease in the concentration of FB observed in tissues of chickens fed FB plus algo-clay on day nine was accompanied by a decrease in Sa and So contents in the liver compared to the levels of Sa and So measured in chickens fed FB alone. FB1 in the liver and Sa or So contents were correlated in liver tissue, confirming that both FB1 and Sa are suitable biomarkers of FB exposure in chickens. Further studies are necessary to determine whether FB can accumulate at higher levels in chicken tissues with an increase in the time of exposure and in the age of the animals.

Dried fruits marketed in Russian: toxigenic mold contamination

Introduction. Dried fruits are a valuable source of dietary fibre, many vitamins and minerals in the population’s diet. However, the high content of readily available carbohydrates makes this type of product vulnerable to mould contamination. The greatest danger among which are toxigenic species. But there is practically no scientific information about the contamination presented on the Russian market dried fruits with moulds producing mycotoxins. That does not allow judging about this aspect of food safety. Materials and methods. Contamination with moulds and bacteria of 57 samples of dried fruits of 7 species popular in Russia was studied using cultural methods of analysis. Monospore isolates of moulds were isolated from dried fruits; in vitro mycotoxins production studied; by UHPLC-MS / MS analyzed mycotoxins in the multidetection mode. Results and discussion. In general, the microbial contamination of dried fruits was low: 87.7% of the samples met the established microbiological standards, in most cases, moulds caused it. At the same time, the highest frequency and levels of contamination were found in dates. Aspergillus sp. dominated in the micoflora of all types of dried fruits. Among the isolated 33 strains of moulds, 45.5% turned out to be toxigenic and, in vitro, were capable of biosynthesis of significant amounts of several types of mycotoxins, including emergent mycotoxins. Fumonisin- and ochratoxin-producing activities have been found in Aspergillus strains of the Nigri section. In model experiments, the accumulation of mycotoxins in individual strains exceeded the level normalized in grain products, including (in μg/kg): for aflatoxins B1 - more than 32000 and B2 - 3230; fumonisin B2 - more than 3100; ochratoxin A up to 4.3; for emergent accumulation reached: sterigmatocystin up to 6218220 and citreoviridine - 153. Conclusion. Moulds are the main type of microflora that contaminates dried fruits. The ability of mould isolates from dried fruits to form mycotoxins has been established, among which highly toxigenic strains have been identified. This indicates the presence of a potential risk of contamination of this type of food with unregulated mycotoxins and a possible increase in their content in the diets of consumers. The results obtained substantiate the need for extensive monitoring of mycotoxin producers in dried fruits. This is important for predicting the risk of toxin formation and identifying the relationship of specific mycotoxins with certain types of dried fruits. The presence of toxigenic activity of moulds isolated from dried fruits has been shown in Russia for the first time.

Potential Health Risk Associated with Mycotoxins in Oat Grains Consumed in Spain

Spain is a relevant producer of oats (Avena sativa), but to date there has been no study on the occurrence/co-occurrence of mycotoxins in oats marketed in Spain. The present study is addressed to overcome this lack of knowledge. One hundred oat kernel samples were acquired across different Spanish geographic regions during the years 2015–2019 and analyzed for mycotoxin content using an ultra-high performance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC–ESI–MS/MS) method and matrix-matched calibration. The focus was on the regulated mycotoxins although other relevant mycotoxins were considered. The percentage of incidence (levels ≥ limit of detection), mean and range (ng/g) of mycotoxins were as follows: zearalenone (66%, mean 39.1, range 28.1–153), HT-2 toxin (47%, mean 37.1, range 4.98–439), deoxynivalenol, (34%, mean 81.4, range 19.1–736), fumonisin B1 (29%, mean 157.5, range 63.2–217.4), and T-2 toxin, (24%, mean 49.9, range 12.3–321). Fumonisin B2, 3-acetyldeoxynivalenol, aflatoxins B1, B2, and G2, and ochratoxin A were also detected at low levels, but aflatoxin G1 was not. The maximum limits established by the European Commission for unprocessed oats were not exceeded, except for zearalenone (in one sample), and the sum of aflatoxins (in two samples). Mycotoxin co-occurrence at quantifiable levels in the same sample (two to five combinations) was found in 31% of samples. The most common mixtures were those of HT-2 + T-2 toxins alone or together with deoxynivalenol and/or zearalenone.

Bio-Preservative Potential of Microorganisms Isolated from Red Grape against Food Contaminant Fungi

Fungal spoilage is one of the main reasons of economic losses in the food industry, especially in the wine sector. Consequently, the search for safer and new preservation techniques has gained importance in recent years. The objective of this study was to investigate the antifungal and anti-mycotoxigenic activity from 28 microorganisms (MO) isolated from red grape. The antifungal activity of a cell free supernatant of fermented medium by the isolated MO (CFS) was tested with the agar diffusion method and the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) assay. Additionally, different antifungal compounds from the CFS were identified and quantified (organic acids, phenolic compounds, and volatile organic compounds). Finally, the most active CFS were tested as red grape bio-preservative agents. Results evidenced that CFS fermented by the strain UTA 6 had the highest antifungal activity, above all isolates, and produced a wide pool of antifungal compounds. The use of UTA 6 CFS as bio-preservative agent showed a reduction of 0.4 and 0.6 log10 spores per gram of fruit in grapes contaminated by A. flavus and B. cinerea, respectively. Moreover, UTA 6 CFS treatment reduced the occurrence of aflatoxin B1 and fumonisin (B2, B3, and B4) production in grapes contaminated by 28–100%.

Simultaneous determination of 13 mycotoxins in feedstuffs using QuEChERS extraction

Mycotoxins are secondary metabolites produced by various fungi and are known to have a significant negative impact on human and animal health. When feedstuffs are contaminated with mycotoxins, their toxicities may be caused a variety of diseases. In this study, the residual mycotoxins in feedstuffs were analyzed using LC–MS/MS incorporated with QuEChERS extraction. Analytical method validation was performed for LOD, LOQ, linearity, and recoveries with consideration of matrix effects prior to the residual analysis. They were all reached to the accepted range of validation level. Using 39 feedstuff samples (5 g) for mycotoxin analysis, nine samples were contaminated by four major mycotoxins such as fumonisin B1 (FB1), deoxynivalenol, fumonisin B2, and zearalenone. Among them, FB1 was detected at the highest concentration as 18.0943 mg/kg. The total sum of fumonisins in 39 samples did not exceed the maximum residual level (MRL) criterion set by Korean Food and Drug Administration. Altogether, intensive management of mycotoxins in Korean feedstuffs should be implemented with proper and routine monitoring, even their residual concentrations are not exceeded over the MRL levels because of high frequent detection found in this study.

Occurrence of Fusarium spp. in Maize Grain Harvested in Portugal and Accumulation of Related Mycotoxins during Storage

Maize is an important worldwide commodity susceptible to fungal contamination in the field, at harvest, and during storage. This work aimed to determine the occurrence of Fusarium spp. in maize grains produced in the Tagus Valley region of Portugal and the levels of related mycotoxins in the 2018 harvest and during their storage for six months in barrels, mimicking silos conditions. Continuous monitoring of temperature, CO2, and relative humidity levels were done, as well as the concentration of mycotoxins were evaluated and correlated with the presence of Fusarium spp. F. verticillioides was identified as the predominant Fusarium species. Zearalenone, deoxynivalenol and toxin T2 were not found at harvest and after storage. Maize grains showed some variability in the levels of fumonisins (Fum B1 and Fum B2). At the harvest, fumonisin B1 ranged from 1297 to 2037 µg/kg, and fumonisin B2 ranged from 411 to 618 µg/kg. Fumonisins showed a tendency to increase (20 to 40%) during six months of storage. Although a correlation between the levels of fumonisins and the monitoring parameters was not established, CO2 levels may be used to predict fungal activity during storage. The composition of the fungal population during storage may predict the incidence of mycotoxins.

Mycotoxin Contamination of Selected Organic Enrichment Materials Used in Pig Farming

Abnormal behavior, such as tail biting, is a fundamental problem in pig husbandry worldwide, and the application of enrichment materials, particularly organic materials, is one of the most promising preventive and curative measures. However, the potential health risks posed by these materials, such as being an additional source of mycotoxins, have not been sufficiently studied to date. Therefore, 21 different organic enrichment materials were tested for mycotoxin contamination with a liquid chromatography tandem mass spectrometry multi-mycotoxin method. Concerning the legally regulated mycotoxins in the EU, aflatoxin B1 and ochratoxin A were not detected in any of the tested materials. Fumonisin B2 was detected in straw meal made of wheat, rye, and triticale, but the level (0.014 mg/kg) was very low. The level of deoxynivalenol in maize pellets (5.01 mg/kg) and maize silage (2.12 mg/kg) exceeded the guidance value for pig feed. Zearalenone was present at high levels in maize pellets (1.21 mg/kg), hay (0.30 mg/kg), and maize silage (0.25 mg/kg). Maize products showed high levels of mycotoxins presenting a health risk for pigs and cannot be recommended as enrichment material.

Post-Flood Impacts on Occurrence and Distribution of Mycotoxin-Producing Aspergilli from the Sections Circumdati, Flavi, and Nigri in Indoor Environment

Mycotoxin-producing Aspergilli (Circumdati, Flavi, and Nigri), usually associated with contaminated food, may also cause respiratory disorders and are insufficiently studied in water-damaged indoor environments. Airborne (N = 71) and dust borne (N = 76) Aspergilli collected at post-flood and control locations in Croatia resulted in eleven different species based on their calmodulin marker: A. ochraceus, A. ostianus, A. pallidofulvus, A. sclerotiorum, and A. westerdijkiae (Circumdati); A. flavus (Flavi); and A. tubingensis, A. welwitschiae, A. niger, A. piperis, and A. uvarum (Nigri). Most of the airborne (73%) and dust borne (54%) isolates were found at post-flood locations, and the highest concentrations measured in indoor air (5720 colony-forming units (CFU)/m3) and dust (2.5 × 105 CFU/g) were up to twenty times higher than in the control locations. A. flavus dominated among airborne isolates (25%) at the unrepaired locations, while 56% of the dust borne Aspergilli were identified as A. tubingensis and A. welwitschiae. The ability of identified isolates to produce mycotoxins aflatoxin B1 (AFB1), fumonisin B2 (FB2), and ochratoxin A were assessed by LC-MS analysis. All ochratoxin A (OTA)-producing Circumdati belonged to A. westerdijkiae (13.7 ± 15.81 µg/mL); in the section, FlaviA. flavus produced AFB1 (2.51 ± 5.31 µg/mL), while A. welwitschiae and A. niger (section Nigri) produced FB2 (6.76 ± 13.51 µg/mL and 11.24 ± 18.30 µg/mL, respectively). Water damage dominantly supported the occurrence of aflatoxigenic A. flavus in indoor environments. Yet unresolved, the causal relationship of exposure to indoor Aspergilli and adverse health effects may support the significance of this research.

The Effect of Agrotechnical Factors on Fusarium Mycotoxins Level in Maize

The occurrence of diseases in the cultivation of maize (Zea mays L.) leads to the loss of grain yield and the simultaneous deterioration of its quality. Fungi of the genus Fusarium spp. pose the greatest threat to maize cultivation. These fungi occur at all stages of the plant’s development, causing Fusarium seedling blight, root rot, foot rot, and ear rot, including grains. Therefore, the aim of the conducted field studies was to determine the influence of selected agriculture factors on mycotoxin contents in maize grain and ear core. Moreover, it should be noted that the presence of mycotoxins in food products in terms of legal regulations is quite a fundamental barrier both in domestic and international trade. The field experiment was carried out at the Department of Agronomy of Poznań University of Life Sciences, on the fields of the Experimental and Educational Unit in Swadzim, Poland, in 2013–2014, in four field replicates. The study involved the following factors: soil sowing preparation method, types of variates, and fertilization method, all of which shape mycotoxin accumulation. The results indicated that the main effects of years were significant for all observed traits for both the core and grain. The concentrations of deoxynivalenol, nivalenol, and zearalenone were higher in 2014 than in 2013 for both core and grain. The concentrations of fumonisin B1 and fumonisin B2 were higher in the first year of study for core and grain. The direct sowing of maize significantly affects plants’ health status, expressed by an increase in mycotoxin accumulation. Traditional hybrid SY Cooky characterized higher values of observed traits than “stay-green” hybrid Drim (except fumonisin B2 for cob). The positive effect of the row method of NP fertilizer application is based on a decrease in mycotoxin content.