Heat- and drought-tolerant sorghum is an attractive alternative to maize, yet its silage mycotoxin trade-offs remain unclear. We compared maize (Zea mays L.), sorghum (Sorghum bicolor (L.) Moench; cultivars Goliath and Sucrosorgo 506) and 1:1 maize-sorghum silages after 60 d ensiling (microsilo scale), with or without a commercial multi-strain lactic acid bacteria (LAB) inoculant. All silages fermented well (pH 4.06 - 4.21; lactic acid 18.1 - 27.9 g/kg dry matter). Maize silage showed the highest Fusarium toxins-deoxynivalenol (DON) 499 - 516 and nivalenol (NIV) 173 - 216 μg/kg dry matter. In sorghum-only silages, DON was below quantification limits, and NIV was not detected. This benefit came with a new hazard: in silages of maize and sorghum mixture (0.51 - 3.72 μg/kg dry matter) and was absent from maize-only silage. Maize and sorghum mixing reduced DON (80.4 - 180 μg/kg dry matter) while keeping OTA low (0.54 - 1.82 μg/kg dry matter). LAB inoculation sharply suppressed yeasts and ethanol in maize silage (4.7 × 107 → 1.4 × 105 CFU/g; 8.26 → 5.03 g/kg dry matter) but showed substrate-dependent effects in sorghum-based silages and did not reliably reduce mycotoxins. Sorghum substitution by maize therefore shifts, rather than removes, hygienic risk - from Fusarium toxins toward OTA-associated storage mould contamination-underscoring the need for stringent anaerobiosis and OTA monitoring when diversifying forages under warm growing conditions.

Enterococci are among the most frequently isolated environmental bacteria that cause mastitis in cows. This study aimed to determine the prevalence of virulence genes, as well as phenotypic and genotypic antibiotic resistance, among eighty enterococcal isolates from cases of bovine mastitis in Polish herds. The presence of virulence and antibiotic resistance genes was determined by PCR. E. faecalis isolates were found to carry more virulence genes than E. faecium isolates, including the efaAfs (100%), ace (98.1%), gelE (86.5%), asa1 (63.5%), esp (57.7%) and cylA (17.3%) genes. The efaAfm gene was the only virulence gene detected in E. faecium isolates. This study revealed that E. faecalis showed a higher virulence gene burden. The ermB gene was present in 90.9% of the Enterococcus spp. that were phenotypically resistant to erythromycin. Almost all tetracycline-resistant Enterococcus isolates carried the tet(M) gene (94.3%), either alone or in combination with the tet(L) and tet(O) genes. Three isolates harboured vanC genes and were susceptible to vancomycin (MIC = 4 μg/mL). The results confirm the high level of antimicrobial resistance of enterococci isolated from cows with mastitis and indicate the genes that may be responsible for this resistance.

[This corrects the article DOI: 10.3389/fpubh.2025.1536836.].

Mycotoxins are secondary metabolites produced by molds, primarily by species of Penicillium, Fusarium, and Aspergillus genera. They are widespread contaminants of foods, feeds and various feed materials. This study aimed to evaluate the levels of mycotoxin contamination in raw materials and animal feed products in Poland between 2021 and 2024. A total of 2,411 samples were collected from farmers and producers across the country, including 1,102 feed samples for swine, poultry and cattle; 519 maize samples; 569 small grain samples; 183 total mixed ration (TMR) samples; and 38 corn silages samples. Samples were examined for several mycotoxins commonly found in Europe, including aflatoxins, ochratoxin A, zearalenone, deoxynivalenol, nivalenol, T-2 toxin, H-2 toxin and fumonisins, Trichothecenes, zearalenone, and fumonisins were analyzed using the LC-MS/MS technique, while ochratoxin A and aflatoxins were analyzed using the HPLC-FLD method. Deoxynivalenol and HT-2 toxin were the most prevalent in feed samples (100 % and 99.9 % occurrence, respectively). In maize, deoxynivalenol and zearalenone were also dominant (99.5 % and 99.7 % occurrence, respectively). The highest concentrations of deoxynivalenol and zearalenone were 11,033 μg/kg and 4,487 μg/kg, respectively, observed in maize. Two maize samples exceeded the maximum recommended concentration level for zearalenone as set by the European Union. These findings confirm that mycotoxins remain widespread contaminants in feed and feed materials, underscoring the need for continuous monitoring and preventive measures.

Poultry farms are hostpost for fungi and mycotoxins proliferation. Still, seasonal dynamics on of these hazards on farm and their impacts on animals/workers health, and lastly on food safety remain unexplored. This study provides the first comprehensive assessment of seasonal (summer/winter) fungal/mycotoxins exposure in poultry farms environment (Electrostatic dust cloths (EDC), feed, bed) and broiler chicken tissues (breast, liver) to identify potential exposure risks for broiler chickens/workers health and food safety. Viable mycotoxigenic fungi and mycotoxins profile (LC-MS/MS) was characterized and cytotoxicity assessment was performed using A549 (human alveolar epithelial) and SK (swine kidney) cells. Zootechnical parameters were also measured to evaluate broiler chickens' growth performance. Summer conditions favour the proliferation of mycotoxigenic fungi belonging to Penicillium and Aspergillus genera, where co-contamination of hazardous mycotoxins (ZEN/ TRCs/FBs) was prevalent. Considering cytotoxicity assessment, EDC samples induce high toxicity in 47 % of A549 cells during summer, whereas on bed samples, high toxicity was obtained during winter on both cell lines (100 % SK cells; 83 % A549 cells). Our results evidence diverse exposure pathways (inhalation, dermal contact) and seasonal health risks. Regarding mycotoxins in biological samples, ZEN was detected in 53.3 % of liver samples, highlighting chronic exposure risks. Thus, future research should focus on toxicokinetics/toxicodynamics of co-occurring mycotoxins in animal production environments and their subsequent human exposure through the food chain. Given that climate change may exacerbate seasonal fungal/mycotoxin contamination, understanding these interactions is crucial for improving risk assessment frameworks and to implement protection measures on the farm level and along the food chain.

IntroductionWaste management represents an occupational setting where fungi are significant contaminants. This study aimed to assess the exposure of waste workers to mycotoxins through a human biomonitoring study.MethodsA total of 33 workers and 19 controls provided spot urine samples to determine 10 mycotoxins’ urinary biomarkers using liquid chromatography coupled with mass spectrometry. Risk characterization was performed using hazard quotient and margin of exposure assessments.ResultsThe results indicated that workers were exposed to six out of the 10 mycotoxins tested, with the following detection rates: deoxynivalenol (91%, 30/33), ochratoxin A (33%, 11/33), zearalenone (17%, 5/33), α-zearalenol (12%, 4/33), β-zearalenol (12%, 4/33), and HT-2 toxin (3%, 1/33). Within controls and outwith controls, were exposed to 5/10 and 2/10 mycotoxins, respectively. All participants exhibited hazard quotients for deoxynivalenol and zearalenone below one, indicating that the exposure is unlikely to pose a health risk. However, when considering the margin of exposure determined for ochratoxin A, 18% of the total participants presented results below 200 for non-neoplastic effects, and 100% of the total participants presented values below 10,000 for neoplastic effects, suggesting potential health concerns that require further assessment.DiscussionThis study highlights the need for future research on occupational exposure to mycotoxins in waste management settings.

The aim of the study was to evaluate the potential of using five selected species of entomopathogenic fungi (Beauveria bassiana, B. brongniartii, Conidiobolus coronatus, Isaria fumosorosea, and Metarhizium robertsii) in the bioregulation of the dispersive stages of the parasitic nematode-Ascaris suum. Experimental cultures of each of the selected entomopathogenic fungi, as well as a control culture without fungi, were incubated with A. suum eggs at 26 °C for 28 days. Development of the A. suum eggs was observed using a light microscope on the 7th, 14th, 21st, and 28th days of incubation. The API-ZYM® test was used to determine, semiquantitatively, the activity of 19 hydrolytic enzymes from the entomopathogenic fungi. The cytotoxicity of the fungi was determined using tetrazole salt MTT. It was found that none of the five tested strains of entomopathogenic fungi showed an ovicidal effect, and none of them colonized the A. suum egg shells. However, ovistatic activity was observed mainly until the 14th day of incubation by I. fumosorosea, M. robertsii, and B. bassiana. In the MTT test, M. robertsii showed moderate cytotoxicity, while the other species showed low cytotoxicity. Among the strains tested, I. fumosorosea showed the highest spectrum of hydrolase production (13 out of 19 enzymes gave a positive reaction from 3 to 5; 20-40 nM or more). The absence of morphological changes in the A. suum egg shells suggests that the antagonistic effect of the studied entomopathogenic fungi may be due to their cytotoxicity, associated with the production of secondary metabolites-toxins (M. robertsii) and enzymatic activity (I. fumosorosea).

Streptococcus agalactiae can produce a wide variety of virulence factors, including toxins and proteins which facilitate adhesion to and colonization and invasion of the host cells. There are few reports on the characteristics of field isolates from bovine mastitis in Poland. Thus, the aim of this study was to determine the occurrence of types of hemolysis on blood agar, virulence factor genes, and cytotoxicity of S. agalactiae isolates derived from cows with mastitis across Poland. The study included 68 isolates. Virulence genes were tested using standard PCR, and cytotoxicity was determined using methylthiazol tetrazolium (MTT) and lactate dehydrogenase (LDH) tests. Among the tested isolates, 89.7% were β-hemolytic, 8.8% γ-hemolytic, and 1.5% alpha-hemolytic. The only genes detected in all isolates were the cfb, cspA, hylB, and sip genes. Cytotoxicity assessment based on the LDH test revealed that isolates were cytotoxic only to Vero cells. However, according to the results obtained from the MTT test, more than half of the isolates exhibited low cytotoxicity to both SK and Vero cells, whereas the other isolates showed moderate or no cytotoxicity to both cell lines. Our research confirms the prevalence of various virulence genes in S. agalactiae isolated from Polish dairy herds, which have previously been found in isolates recovered from human and animal infections. For the first time, the presence of bac- and scpB-positive isolates of S. agalactiae was determined in Polish dairy cattle, and the cytotoxicity of bovine isolates was assessed. IMPORTANCE We believe that this manuscript is one of the few reports on the characteristics of field S. agalactiae isolates derived from cases of bovine mastitis in cows in Poland in terms of the occurrence of virulence genes and cytotoxicity. For the first time, the presence of bac- and scpB-positive isolates of S. agalactiae was determined in Polish dairy cattle, and the cytotoxicity of bovine isolates was assessed.

Mycotoxins are secondary metabolites of molds and mainly produced by species of the genera Aspergillus, Penicillium and Fusarium. They can be synthesized on the field, during harvest as well as during storage. They are fairly stable compounds and difficult to remove. Among several hundreds of mycotoxins, according to the WHO, ochratoxin A, aflatoxins, zearalenone, deoxynivalenol, patulin, fumonisins as well as T-2 and HT-2 toxins deserve special attention. Cytotoxicity is one of the most important adverse properties of mycotoxins and is generally assessed via the MTT assay, the neutral red assay, the LDH assay, the CCK-8 assay and the ATP test in different cell lines. The apoptotic cell ratio is mainly assessed via flow cytometry. Aside from the assessment of the toxicity of individual mycotoxins, it is important to determine the cytotoxicity of mycotoxin combinations. Such combinations often exhibit stronger cytotoxicity than individual mycotoxins. The cytotoxicity of different mycotoxins often depends on the cell line used in the experiment and is frequently time- and dose-dependent. A major drawback of assessing mycotoxin cytotoxicity in cell lines is the lack of interaction typical for complex organisms (for example, immune responses).

Abstract In the past, pigs were commonly fed with acorns, and this was of remarkable economic importance. Currently this habit is continued in some areas, especially for production of prime-quality Iberian ham. Mature acorns, after shedding and during storage in unsuitable conditions, can be quickly infected with spores of many moulds, which cause mummification, blackening, dehydration, and nutrient loss. This study aimed to evaluate the quality of acorns of pedunculate oak (Quercus robur L.). The samples were collected in 2017 in southern Wielkopolska (central part of Poland), as feed material. In mouldy acorns a very high number of fungi was found (2.6 × 106 cfu/g), and 97% of them represented pathogenic Penicillium spp. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis showed in mouldy acorns high concentrations of mycophenolic acid (14580 μg/kg) and patulin (50 μg/kg). The dominant mould species, Penicillium expansum, showed a high cytotoxicity of swine kidney cells using assay based on the conversion of the tetrazolium salt, 3-(4,5, dimethylthiazol-2-yl)-2-5 diphenyltetrazolium (MTT). This raises the question if the pathogenic metabolites of moulds present in acorns can be dangerous for livestock, especially pigs, and people, as acorns are beginning to be seen as an interesting and functional part of their diet.

Mycotoxins are secondary metabolites produced by several fungi, mainly from Penicillium, Fusarium, and Aspregillus genera. They can contaminate many raw materials both in the field and during storage under very different conditions. This study aimed to evaluate mycotoxin contamination levels of raw materials and products for animals in Poland in the years 2015-2020. Producers, farmers, and veterinary doctors delivered a total of 3980 samples (642 maize samples, 2027 feed samples, 990 small grain samples, 142 maize silage samples and 179 TMR samples). Samples were analyzed for the occurrence of several mycotoxins, including aflatoxins, fumonisins, ochratoxin A, deoxynivalenol, nivalenol, T-2 toxin, H-2 toxin, and zearalenone. Trichothecenes, zearalenone, and fumonisins were determined using the LC-MS/MS technique. Ochratoxin A and aflatoxins were determined using the HPLC-FLD method. Deoxynivalenol and zearalenone were the most common contamination in maize samples (97.3% and 98.4%, respectively) and feed samples (99.7% and 100% samples, respectively). They were also present in all maize silage and TMR samples. The highest concentration levels of deoxynivalenol and zearalenone were 16,889 μg/kg in the wheat sample and 1420 μg/kg in the maize sample. Additionally, in 51 samples, mycotoxins level (mainly deoxynivalenol and zearalenone levels) was above recommended levels set by the European Union. The present study showed that both feeding and raw materials are contaminated by mycotoxins, often by more than one.

Milk and dairy products are consumed by people around the world every day. Unfortunately, these products can be contaminated by various types of fungi that can produce various pathogenic mycotoxins. Consumption of contaminated food products can lead to the accumulation of hazardous substances in human and animal bodies causing health problems. For this reason, people should limit the consumption of products contaminated by fungi. However, sometimes these products are intentionally contaminated to improve the quality of consumer products. The aim of this study is to collect data on contamination of few popular dairy products and milk by fungi.

Project Waste Workers’ Exposure to Bioburden through Filtering Respiratory Protective Devices” (IPL/2018/WasteFRPD_ESTeSL).

As a valuable food base and safe shelter, large agricultural fields create favourable living conditions for wild boars for most of the year. The occurrence of mould in these fields, causing hormonal disturbances, may lead to a lower age of sexual maturity and a prolonged breeding season, and therefore, the population growth in farmlands can be markedly higher than that in extensive woodlands. This study was initiated because of reproductive cycle disturbances in wild boar populations, which were presumably linked with habitat and food types, especially rotten maize. To determine if the major sources of zearalenone (ZEN) in female wild boars are maize cobs infected with mycotoxins, we compared the concentrations of ZEN and its metabolites in organs, tissues and body fluids of wild boars hunted in extensive maize fields and in extensive woodlands where no large maize fields were located within a distance of several km. Samples of blood, bile, liver, kidneys, muscles, urine, stomach and colon contents as well as ovaries from young female wild boars (40-60 kg) were collected in 2011-2014 and prepared for liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The results showed that ZEN was present in most of the samples, from both farmlands and woodlands, but its concentration within individual types of the analysed body fluids or tissues differed depending mainly on habitat type. In nearly all the analysed cases, higher concentrations of ZEN and its derivatives were detected in the samples collected from wild boars in farmlands, suggesting that wild boars living and feeding in extensive maize fields take in markedly higher amounts of mycoestrogens.

The aim of this study was to determine the degree of mold contamination and mycotoxin levels in commercially available green coffee products and dietary supplements with green coffee extract. The study included 34 samples from green coffee products: raw beans (n = 16), ground coffee (n = 15) and instant coffee (n = 3), as well as 22 samples from dietary supplements in form of capsules (n = 19), tablets (n = 2) and sachets (n = 1). Total mold count was determined with spread-plate method. Anamorphic mold were identified based on their microscopic morphology and the type of sporulation. Concentrations of mycotoxins, ochratoxin A and citrinin, were quantified by means of HPLC-fluorescence detection. Molds, typically Aspergillus spp. and Penicillium spp., were found in 94% of green coffee beans, 100% of ground and instant coffee samples, and 55% of dietary supplement samples. None of the samples contained detectable levels of citrinin. Ochratoxin A (0.4 ng/g) was detected in only one sample of raw green coffee beans, but in up to 40% and 67% of ground and instant coffee samples, respectively. Mean concentrations of ochratoxin A in ground and instant coffee samples were 3.28 ng/g and 4.09 ng/g, respectively, and maximum concentrations amounted to 6.65 ng/g and 7.44 ng/g, respectively. Ochratoxin A (mean concentration 9.60 ng/g, maximum level 31.4 ng/g) was also detected in up to 58% of the supplement capsules, but in none of tablets and sachets.

One of the solutions for decreasing the workers' exposure to bioburden is the use of Filtering Respiratory Protective Devices (FRPD). As such it is important to determine whether these devices are fulfilling their protective role. This is the basis of the current study, aimed at characterizing bioburden retained by 120 FRPD (both in interior layers and in exhalation valves) through culture based-methods and molecular tools and also via analysis of antifungal resistance and mycotoxins profile. Our results show that Gram - Bacteria are present at a higher prevalence than total bacteria in both matrixes. Regarding fungal identification, Chrysonilia sitophila presented the highest prevalence on interior layers (55.1% on malt extract agar (MEA) supplemented with chloramphenicol (0.05%); 59.6% on dichloran-glycerol agar (DG18)), whereas on exhalation valves Aspergillus sp. presented the highest prevalence on MEA (6.8%) and C sitophila on DG18 (36.3%). Among Aspergillus genera, section Fumigati was the one with the highest prevalence in both matrices. Aspergillus sp. was the most prevalent on exhalation valves (75.0% ITRA) in the screening of azole resistance. Fumigati section was the most abundant Aspergillus sp. detected on the interior layers (33.33%, 40 samples out of 120) and on the exhalation valves (1.66%, 2 samples out of 120). The interior layers and exhalation valves from workers with more waste contact showed an increased exposure to bioburden. This study showed that FRPD can have high levels of bioburden, toxigenic fungal strains and Aspergillus sections with reduced susceptibility to the tested azoles and can be used as a passive sampling method since it mimics the results obtained by active methods in previous studies. The gathered information will be useful to prioritize multiple interventions on workers' education or even on FRPD replacement frequency. (C) 2019 Elsevier Ltd. All rights reserved.

The aim of this study was to compare the chemical composition of European soy seeds. A mycological and toxigenic screening was carried out on 18 varieties of soy seeds harvested in Poland. Moreover, the level of soybean meal (SBM) substitution by raw soybean seeds was analysed in terms of its effect on young pigs' performance (body weight gain, feed intake, feed utilisation) along with apparent total tract digestibility (ATTD) of dry matter and crude protein in the diets. In a 28-day trial, 48 male pigs were tested using a marker method with TiO2. In their diets, SBM was replaced by soy seeds in the amounts of 0%, 5%, 10%, 15%, 20% and 25%. In the last 3 days of the experiment, samples of excreta from each animal separately were collected three times per day. The chemical composition of soybean seeds differed in terms of their contents of crude protein, ether extract, neutral detergent fibre and raffinose family oligosaccharides, as well as the trypsin inhibitor activity. Seeds were also contaminated with fungi, yeast and mycotoxins, mainly zearalenone and deoxynivalenol. The ATTD of crude protein ranged from 70.6% to 77.6% and that of dry matter from 93.5% to 94.6%, with no differences between the groups being found (p > 0.05). Pigs' performance parameters were reduced strongly with increasing amounts of raw seeds in the diets (p < 0.05). The results indicate that only a 5% addition of raw soy seeds in pigs' diet is recommended.

In Portugal, mechanical protection gloves (MPG) are of mandatory use and during their use sweat is released and, consequently, the humidity of the material increases leading to conditions favorable to the growth of microorganisms. However, no studies have been conducted in MPG to assess the bioburden. This study intended to determine the bioburden present in MPG and their biological effects, and to discuss the possibility to use MPG as a passive method to assess occupational exposure to microbial contamination. Fungal burden was characterized through molecular tools for fungal toxigenic species, and antifungal resistance and mycotoxins profiles were determined. Cell viability was determined in swine kidney (SK) monolayer and hepatocellular carcinoma (Hep G2) cell lines. All MPG samples presented Gram-negative bacteria. The fungal contamination ranged from 0 CFU. m(-2) in both MEA and DG18, to 5.09 x 10(6) and 2.75 x 10(6) and the most commonly fungi found was Aspergillus spp. (50.46%). Azole resistant Aspergillus sections were found in azole supplemented media. Aspergillus sections (Circumdati, Flavi, Fumigati and Versicolores) were detected by molecular tools in 66 out of 67 samples. The most reported mycotoxin was mycophenolic acid (89.6%). HepG2 cells appear to be more sensitive to MPG contamination, with high cytotoxicity (IC50 < 0.05 mm2/ml) observed for 18 out of 57 gloves. MPG can be used in passive sampling to assess occupational exposure to bioburden in waste sorting industries and contribute for risk characterization. Some contaminants of MPG had cytotoxic potential and affected the biology of hepatic cells more than renal cells.

The bioburden in a Hospital building originates not only from patients, visitors and staff, but is also disseminated by several indoor hospital characteristics and outdoor environmental sources. This study intends to assess the exposure to bioburden in one central Hospital with a multi-approach protocol using active and passive sampling methods. The microbial contamination was also characterized through molecular tools for toxigenic species, antifungal resistance and mycotoxins and endotoxins profile. Two cytotoxicity assays (MTT and resazurin) were conducted with two cell lines (Calu-3 and THP-1), and in vitro pro-inflammatory potential was assessed in THP-1 cell line. Out of the 15 sampling locations 33.3% did not comply with Portuguese legislation regarding bacterial contamination, whereas concerning fungal contamination 60% presented I/O > 1. Toxigenic fungal species were observed in 27% of the sampled rooms (4 out of 15) and qPCR analysis successfully amplified DNA from the Aspergillus sections Flavi and Fumigati, although mycotoxins were not detected. Growth of distinct fungal species was observed on Sabouraud dextrose agar with triazole drugs, such as Aspergillus section Versicolores on 1 mg/L VORI. The highest concentrations of endotoxins were found in settled dust samples and ranged from 5.72 to 23.0 EU.mg(-1). While a considerable cytotoxic effect (cell viability < 30%) was observed in one HVAC filter sample with Calu-3 cell line, it was not observed with THP-1 cell line. In air samples a medium cytotoxic effect (61-68% cell viability) was observed in 3 out of 15 samples. The cytokine responses produced a more potent average cell response (46.8 +/- 12.3 rho g/mL IL-1 beta; 90.8 +/- 58.5 rho g/mL TNF-alpha) on passive samples than air samples (25.5 +/- 5.2 rho g/mL IL-1 beta and of 19.4 +/- 5.2 rho g/mL TNF-alpha). A multi-approach regarding parameters to assess, sampling and analysis methods should be followed to characterize the biorburden in the Hospital indoor environment. This study supports the importance of considering exposure to complex mixtures in indoor environments.

Rye (Secale cereale L.) is one of the most important cereals and is used in both the food and feed industries. It is produced mainly in a belt extending from Russia through Poland to Germany. Despite the great economic importance of this cereal, there is little research on rye contamination with mycotoxins. In this study, the occurrence of Fusarium mycotoxins (deoxynivalenol, nivalenol, 3-acetyl-deoxynivalenol, monoacetoxyscirpenol, diacetoxyscirpenol, T-2 toxin, HT-2 toxin, and zearalenone), as well as ochratoxin A, in 60 winter rye samples of four varieties (KWS Binntto, KWS Serafino, Dańkowskie Granat and Farm Saved Seed) cultivated in three consecutive growing seasons in five different regions of Poland was determined using liquid chromatography with tandem mass spectrometry and fluorescence detection. Deoxynivalenol, T-2 toxin, HT-2 toxin, and zearalenone had the highest occurrence in samples (90%, 63%, 57%, and 45% positive results, respectively). The mean concentrations of these analytes were 28.8 µg/kg (maximum 354.1 µg/kg), 0.98 µg/kg (maximum 6.63 µg/kg), 2.98 µg/kg (maximum 29.8 µg/kg), and 0.69 µg/kg (maximum 10.2 µg/kg), respectively. The mean concentrations for individual mycotoxins were highest in the 2016/2017 growing season. In the 2016/2017 growing season, at least two mycotoxins were detected in 95% of the samples, while in the 2018/2019 growing season, 70% of samples contained one or no mycotoxins. The frequencies of mycotoxin occurrence in different rye varieties were similar. Although a high frequency of mycotoxin occurrence was noted (especially deoxynivalenol), their concentrations were low, and none of the analyzed rye samples exceeded the maximum acceptable mycotoxin level set by the European Commission.