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Toxico-pathological effects of ochratoxin A and its diastereoisomer under in ovo conditions and in vitro evaluation of the toxicity of these toxins against the embryo Gallus gallus fibroblast cell line

2023, BryƂa, Marcin, Damaziak, Krzysztof, TwaruĆŒek, Magdalena, Waƛkiewicz, Agnieszka, StępieƄ, Ɓukasz, Roszko, Marek, Pierzgalski, Adam, SoszczyƄska, Ewelina, Ɓukasiewicz-Mierzejewska, Monika, Chmiel, Marta, WĂłjcik, Wojciech

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Antioxidant, Antimicrobial and Antibiofilm Properties of Glechoma hederacea Extracts Obtained by Supercritical Fluid Extraction, Using Different Extraction Conditions

2022, Gwiazdowska, Daniela, Uwineza, Pascaline Aimee, Frąk, Szymon, Juƛ, Krzysztof, MarchwiƄska, Katarzyna, Gwiazdowski, Romuald, Waƛkiewicz, Agnieszka

Glechoma hederacea var. longituba is a herbaceous plant from the Lamiaceae family, used in herbal medicine. In this work, we aimed to assess the total phenolic content, antioxidant, antimicrobial and antibiofilm activity of extracts obtained from G. hederacea via supercritical dioxide extraction with methanol as a co-solvent under different extraction conditions. The results showed that the activity of the obtained SC-CO2 extracts is strongly dependent on the extraction temperature. Significantly higher total polyphenol content, as well as antioxidant and antimicrobial activity towards bacteria and yeasts, was observed in the extract obtained at 40 °C, compared to extracts obtained at 50 °C and 60 °C; however, antifungal activity against filamentous fungi was not dependent on the extraction conditions. Antimicrobial activity also depended on the microorganism type. Higher sensitivity was exhibited by Gram-positive bacteria than by Gram-negative bacteria, with S. aureus and P. aeruginosa being the most sensitive species among each group. The most susceptible fungi were Candida albicans and Sclerotinia sclerotiorum. The antibiofilm activity was differentiated and depended on the extraction conditions, the microorganism and the method of biofilm treatment. All tested extracts inhibited biofilm formation, with the extract obtained at 40 °C showing the highest value, whereas only extract obtained at 60 °C efficiently removed mature biofilm.

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Fusarium Species Shifts in Maize Grain as a Response to Climatic Changes in Poland

2024, Czembor, Elzbieta, FrasiƄski, Seweryn, Urbaniak, Monika, Waƛkiewicz, Agnieszka, Czembor, Jerzy H., StępieƄ, Ɓukasz

Maize, along with wheat and rice, is the most important crop for food security. Ear rots caused by Fusarium species are among the most important diseases of maize. The distribution of Fusarium species provides essential epidemiological information for disease management. The effect of weather conditions, climate change and geographic localization on the Fusarium population in Poland was evaluated between 2015 and 2018. Grain samples (n = 233) were collected from hybrids at 16 locations (L1–L16). The differences in altitude between locations ranged from 39 to 243 m above sea level, longitude varied between 15°55â€Č and 23°12â€Č E, and latitude spanned from 50°12â€Č to 54°01â€Č N. Isolates were identified using molecular techniques. The highest Fusarium species frequency was recorded for 2016 (30.70%) and 2017 (28.18%), and the lowest for 2018 (5.36%). F. verticillioides and F. temperatum were the most frequent. Altitude has an effect on F. vericillioides frequency: F. graminearum showed a negative correlation with both latitude and longitude. In Polish conditions, from silking to harvesting, the number of days with higher precipitation and lower temperatures is associated with an increased frequency of F. verticillioides, F. temperatum, F. graminearum and F. avenaceum. This suggests that the Fusarium presence in Poland is significantly influenced not only by climate change but also by extreme weather changes.

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Antimicrobial and Antibiofilm Activity of Origanum vulgare Extracts Obtained by Supercritical Fluid Extraction Under Various Extraction Conditions

2024, Gwiazdowska, Daniela, Waƛkiewicz, Agnieszka, Juƛ, Krzysztof, MarchwiƄska, Katarzyna, Frąk, Szymon, Popowski, Dominik, Pawlak-LemaƄska, Katarzyna, Uwineza, Pascaline Aimee, Gwiazdowski, Romuald, Padewska, Daria, Roszko, Marek, BryƂa, Marcin

Sustainable management of agri-food product safety presents a major challenge requiring extensive action to ensure food safety and consumer health. The pursuit of environmentally friendly solutions that will constitute an alternative to the chemical compounds commonly used in agriculture and the food industries is one of the most important problems. One solution is plant extracts containing various biologically active compounds and exhibiting antimicrobial activity. This study aims to determine the biological activity of extracts obtained from Origanum vulgare L. (leaves) by supercritical CO2 (SC-CO2) extraction using different reaction conditions and compositions. In vitro studies revealed antimicrobial activity against selected bacteria (including Salmonella Enteritidis, Listeria monocytogenes, and Staphylococcus aureus) and fungi (Fusarium spp.), depending mainly on the microorganism species; however, extraction conditions also influenced these properties. The microscopic observations established by optical and fluorescence microscopy showed the changes in the fungal cell’s viability and morphology. There was no observed significant release of intracellular material as stated based on ICP-MS analysis of sodium and potassium concentration. Antibiofilm properties of extract obtained by extraction at 40 °C were also demonstrated against S. aureus, P. aeruginosa, and L. monocytogenes, with stronger properties observed against Gram-positive bacteria. Phytochemical characterization of the extracts was determined using a liquid chromatography system with an orbitrap mass spectrometer (LC/MS), identifying, i.e., phenolic acids: protocatechuic, hydroxybenzoic, caffeic, and rosmarinic; flavonoids: luteolin, naringenin, and kaempferol; and terpenoids: oleanolic and ursolic acids.

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Impact of Dietary Salicylates on Iron, Zinc, and Copper Status in Preeclampsia Model Rats Induced by L-NAME

2025, Cholik, Rafsan Syabani, Skrypnik, Katarzyna, Waƛkiewicz, Agnieszka, KaraĆșniewicz-Ɓada, Marta, Suliburska, Joanna

Abstract Low-dose aspirin prophylaxis is recommended for women at high risk of preeclampsia. It has been suggested that dietary salicylates may have a similar effect. Despite the known anti-inflammatory properties of salicylates, their influence on trace elements in preeclampsia remains unclear. This research investigated the effect of dietary salicylates and aspirin on iron, zinc, and copper status in rats with NG-nitro-L-arginine methyl ester (L-NAME)–induced preeclampsia. The study involved pregnant Sprague Dawley rats divided into six groups: control group (CH), preeclamptic rats (CP), preeclamptic rats with a low dose of dietary salicylate (LSP), preeclamptic rats with a high dose of dietary salicylate, preeclamptic rats with a low dose of aspirin (LAP), and preeclamptic rats with a high dose of aspirin. The content of trace elements in diets, liver, kidney, heart, spleen, pancreas, femur, brain, and hair was measured using flame atomic absorption spectrometry. Salicylate concentrations in diets, serum, and urine were analyzed using HPLC and UHPLC-MS/MS systems. Administration of L-NAME resulted in elevated blood pressure across groups, and only the LAP group had blood pressure levels comparable to the CH group. Preeclampsia significantly decreased serum hepcidin levels, while salicylates abolished this effect. Salicylate administration significantly decreased iron levels in hair and increased maternal zinc concentrations in the brain. Dietary salicylates markedly increased zinc levels in the placenta. In conclusion, L-NAME–induced preeclampsia decreases maternal serum hepcidin. Treatment with salicylates modulates iron and zinc status in preeclamptic rats, with specific effects on hepcidin levels.

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Biological Activity and Chemical Composition of Propolis from Various Regions of Poland

2023, WoĆșniak, Magdalena, Sip, Anna, MrĂłwczyƄska, Lucyna, Broniarczyk, Justyna, Waƛkiewicz, Agnieszka, Ratajczak, Izabela

Propolis is one of the bee products, with multiple biological properties used in numerous applications. The research objective was to determine the chemical composition and biological properties (antibacterial, antifungal, antiviral, antioxidant, and cytoprotective activity) of propolis extracts collected from various regions of Poland. The results indicated that the total content of phenols (116.16–219.41 mg GAE/g EEP) and flavonoids (29.63–106.07 mg QE/g EEP) in propolis extracts depended on their geographic origin. The high content of epicatechin, catechin, pinobanksin, myricetin, and acids: vanillic and syringic in propolis samples was confirmed by chromatographic analysis. Moreover, the presence of caffeic acid phenethyl ester was confirmed in all samples. The origin of propolis also influenced the biological properties of its extracts. The propolis extracts were characterized by moderate DPPH free radical scavenging activity (29.22–35.14%), and relatively low ferrous iron chelating activity (9.33–32.32%). The results indicated also that the propolis extracts showed high activity in the protection of human red blood cells against free radicals generated from 2,2’-azobis(2-methylpropionamidine) dihydrochloride (AAPH). The extracts exhibited diversified activity against the tested pathogenic bacteria and limited activity against fungal strains. The research of selected propolis extracts showed that only 2 of 5 examined samples showed moderate activity against HPV (human papillomaviruses) and the activity depended on its geographical distribution.

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The Concentration-Dependent Effects of Essential Oils on the Growth of Fusarium graminearum and Mycotoxins Biosynthesis in Wheat and Maize Grain

2022, Gwiazdowska, Daniela, MarchwiƄska, Katarzyna, Juƛ, Krzysztof, Uwineza, Pascaline Aimee, Gwiazdowski, Romuald, Waƛkiewicz, Agnieszka, Kierzek, Roman

The presence of Fusarium fungi and their toxic metabolites in agricultural crops contributes to significant quantitative and qualitative losses of crops, causing a direct threat to human and animal health and life. Modern strategies for reducing the level of fungi and mycotoxins in the food chain tend to rely on natural methods, including plant substances. Essential oils (EOs), due to their complex chemical composition, show high biological activity, including fungistatic properties, which means that they exhibit high potential as a biological plant protection factor. The aim of this study was to determine the fungistatic activity of three EOs against F. graminearum, and the reduction of mycotoxin biosynthesis in corn and wheat grain. All tested EOs effectively suppressed the growth of F. graminearum in concentrations of 5% and 10%. Cinnamon and verbena EOs also effectively reduced the ergosterol (ERG) content in both grains at the concentration of 1%, while at the 0.1% EO concentration, the reduction in the ERG amount depended on the EO type as well as on the grain. The degree of zearalenone (ZEA) reduction was consistent with the inhibition of ERG biosynthesis, while the reduction in deoxynivalenol (DON) was not consistent with this parameter.

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Pleurotus spp.—an effective way in degradation mycotoxins? A comprehensive review

2025, Zapaƛnik, Agnieszka, BryƂa, Marcin, SokoƂowska, Barbara, Waƛkiewicz, Agnieszka

Abstract Mycotoxins—secondary metabolites produced by filamentous fungal species—occur as a global problem in agriculture due to the reduction in crop quality and the negative effects on human and animal health. There is a need to develop environment-friendly methods of detoxification. In recent years, a number of biological methods for the removal/degradation of mycotoxins have been described. One of them—particularly interesting due to its high effectiveness—is mycoremediation, which involves the ability of Pleurotus spp. mushrooms to remove toxic contaminants from the environment and food. Pleurotus spp. biosynthesizes ligninolytic enzymes, such as laccase and manganese peroxidase that are the main factors of enzymatic degradation of various pollutants, including mycotoxins. The degradation process of mycotoxins (especially aflatoxins) with the participation of isolated enzymes reaches approximately 30–100%, depending on the culture conditions, substrate, and mediators used. In the food industry, their application may include, among others, the detoxification of animal feed from mycotoxins or fermentation products (e.g., juices and wines). While these applications are promising, they require further research to expand toxicological knowledge and optimize their use. This review presents current research on this new and very promising topic related to the use of edible Pleurotus spp. mushrooms in the process of biological degradation of toxic fungal metabolites.

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Ochratoxin A and 2â€ČR-Ochratoxin A in Selected Foodstuffs and Dietary Risk Assessment

2022, Zapaƛnik, Agnieszka, BryƂa, Marcin, Waƛkiewicz, Agnieszka, Ksieniewicz-WoĆșniak, Edyta, Podolska, GraĆŒyna

The aim of this study was to estimate the contamination of grain coffee, roasted coffee, instant coffee, and cocoa purchased in local markets with ochratoxin A (OTA) and its isomerization product 2â€ČR-ochratoxin A (2â€ČR-OTA), and to assess risk of dietary exposure to the mycotoxins. OTA and 2â€ČR-OTA content was determined using the HPLC chromatography with immunoaffinity columns dedicated to OTA. OTA levels found in all the tested samples were below the maximum limits specified in the European Commission Regulation EC 1881/2006. Average OTA concentrations calculated for positive samples of grain coffee/roasted coffee/instant coffee/cocoa were 0.94/0.79/3.00/0.95 ”g/kg, with the concentration ranges: 0.57–1.97/0.44–2.29/0.40–5.15/0.48–1.97 ”g/kg, respectively. Average 2â€ČR-OTA concentrations calculated for positive samples of roasted coffee/instant coffee were 0.90/1.48 ”g/kg, with concentration ranges: 0.40–1.26/1.00–2.12 ”g/kg, respectively. In turn, diastereomer was not found in any of the tested cocoa samples. Daily intake of both mycotoxins with coffee/cocoa would be below the TDI value even if the consumed coffee/cocoa were contaminated with OTA/2â€ČR-OTA at the highest levels found in this study. Up to now only a few papers on both OTA and 2â€ČR-OTA in roasted food products are available in the literature, and this is the first study in Poland.

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Variation in Tocochromanols Level and Mycotoxins Content in Sweet Maize Cultivars after Inoculation with Fusarium verticillioides and F. proliferatum

2022, Waƛkiewicz, Agnieszka, Muzolf-Panek, MaƂgorzata, StępieƄ, Ɓukasz, Czembor, ElĆŒbieta, Uwineza, Pascaline Aimee, GĂłrnaƛ, PaweƂ, BryƂa, Marcin

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Efficacy of Lamium album as a natural fungicide: impact on seed germination, ergosterol, and mycotoxins in Fusarium culmorum-infected wheat seedlings

2024, Uwineza, Pascaline Aimee, Urbaniak, Monika, StępieƄ, Ɓukasz, Gramza-MichaƂowska, Anna, Waƛkiewicz, Agnieszka

Fusarium culmorum is a major wheat pathogen, and its secondary metabolites (mycotoxins) cause damage to plants, animals, and human health. In the era of sustainable agriculture, eco-friendly methods of prevention and control are constantly needed. The use of plant extracts as biocontrol agents has gained popularity as they are a source of active substances that play a crucial role in fighting against phytopathogens. This study evaluated the impact of Lamium album on wheat seed germination and seedling growth. In a pot experiment, the effect of L. album on wheat seedlings artificially inoculated with F. culmorum was evaluated by measuring seedling growth parameters, and by using chromatographic methods, ergosterol and mycotoxins levels were analyzed. The results showed that the phytotoxic effect of L. album flower extracts on wheat seed germination and seedling growth was concentration dependent. The radicle length was also reduced compared to the control; however, L. album did not significantly affect the dry weight of the radicle. A slight phytotoxic effect on seed germination was observed, but antifungal effects on artificially infected wheat seedlings were also confirmed with the reduction of ergosterol level and mycotoxins accumulation in the roots and leaves after 21 days of inoculation. F. culmorum DNA was identified in the control samples only. Overall, this study is a successful in planta study showing L. album flower extract protection of wheat against the pathogen responsible for Fusarium crown and root rot. Further research is essential to study the effects of L. album extracts on key regulatory genes for mycotoxin biosynthetic pathways.

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Interfungal antagonism between Trichoderma and Fusarium proliferatum — metabolomic and DNA-based analyses

2025, Modrzewska, Marta, Popowski, Dominik, BƂaszczyk, Lidia, StępieƄ, Ɓukasz, Waƛkiewicz, Agnieszka, Urbaniak, Monika, BryƂa, Marcin

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In Vitro Effects of Lemon Balm Extracts in Reducing the Growth and Mycotoxins Biosynthesis of Fusarium culmorum and F. proliferatum

2022, Uwineza, Pascaline Aimee, Urbaniak, Monika, BryƂa, Marcin, StępieƄ, Ɓukasz, Modrzewska, Marta, Waƛkiewicz, Agnieszka

The objectives of this research were to obtain the extracts of lemon balm (Melissa officinalis) using supercritical CO2 (SC-CO2) and methanol as co-solvent and evaluate the antifungal activity of those extracts against two selected strains of Fusarium species (Fusarium culmorum and Fusarium proliferatum). The extraction conditions were set at 40 and 60 °C and 250 bar. The obtained extracts were characterized in terms of antifungal activity on potato dextrose agar media (PDA). The results showed that the extraction parameters had different effects on mycelium growth and mycotoxins biosynthesis reduction. All studied lemon balm extracts (1, 2.5, 5, 7.5, and 10%) inhibited the growth of F. proliferatum and F. culmorum mycelia compared to the control. The lemon balm extracts significantly reduced ergosterol content and synthesized mycotoxins in both tested strains. These findings support the antifungal activity of lemon balm extracts against F. proliferatum and F. culmorum. However, more research on other Fusarium species is needed, as well as in vivo applications, before considering lemon balm extracts as a natural alternative to synthetic fungicides.

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Protein and sugar content of tubers in potato plants treated with biostimulants

2022, GƂosek-Sobieraj, MaƂgorzata, Wierzbowska, Jadwiga, Cwalina-Ambroziak, BoĆŒena, Waƛkiewicz, Agnieszka

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The Effect of the Extraction Method on the Content of BioactiveCompounds and the Biological Activity of Nigella sativa Extracts

2025, Gwiazdowski, Romuald, Juƛ, Krzysztof, Kubiak, Krzysztof, BiegaƄska-Marecik, RĂłĆŒa, Waƛkiewicz, Agnieszka, Gwiazdowska, Daniela

Ensuring food safety and consumer health are crucial elements of sustainable food safety management, requiring the use of substances that inhibit the growth of undesirable microorganisms at various stages of production. The negative impact of many of these on human health and the environment has led to increased interest in alternative solutions, such as plant extracts. The aim of this study was to determine the biological activity of extracts obtained from Nigella sativa seeds using various methods, including Soxhlet and ultrasound-assisted extraction (UAE) using hexane and methanol and supercritical CO2 extraction (SFE) assisted with methanol. The content of polyphenolic compounds, their composition, and biological activity depended on the extraction method and solvent type. All extracts exhibited antimicrobial activity against Gram-positive bacteria (Listeria monocytogenes, Priestia megaterium, and Staphylococcus aureus), Gram-negative bacteria (Salmonella Enteritidis, Pseudomonas aeruginosa, and Escherichia coli), yeasts (Candida albicans and Rhodotorula mucilaginosa), and filamentous fungi (Alternaria brassicicola, Pythium sp., Fusarium culmorum, and F. graminearum). The MIC values were in the range of <0.125 to 2 mg/mL for bacteria and 1 to 8 mg/mL for fungi, depending on the extract. Microscopic observations performed using optical and fluorescence microscopy showed changes in the viability and morphology of the fungal cells. TPC values ranged from 9.877 mg/g in hexane extract obtained by ultrasound-assisted extraction to 39.064 mg/g in extract obtained by Soxhlet method with methanol. No negative effects of the extracts on wheat seed germination were observed. Analysis of the composition of polyphenolic compounds revealed the presence of vanillic acid, hydroxybenzoic acid, syringic acid, protocatechuic acid, and p-coumaric, catechin, and epicatechin acids in all extracts. The extracts obtained with methanol, both by the Soxhlet method and by ultrasound-assisted extraction, also contained gallic acid, myricetin, luteolin, apigenin, and sinapic acid. In all extracts, thymoquinone ranged from 8.344 mg/g to 63.125 mg/g of extract, which was detected with the highest concentration in hexane extracts.

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Zawartoƛć związków fenolowych oraz pierwiastków w orzechach ziemnych

2022, Stanicka, Karolina, WoĆșniak, Magdalena, Krueger, MichaƂ, Waƛkiewicz, Agnieszka, Kędzia, ElĆŒbieta, Ratajczak, Izabela

Introduction. Peanuts also known as arachidic nuts, are a valuable raw material commonly used in the food industry, including in confectionery, for the production of oil or peanut butter. Peanuts are a valuable source of protein and are rich in unsaturated fatty acids. Peanuts also owe their health-promoting properties to the presence of phenolic compounds, which are characterized by wide biological activity, including antibacterial and antifungal activity, as well as catch free radicals, which are the cause of many civilization diseases. Aim. The aim of the study was to determine the content of phenolic compounds as well as macro- and microelements in peanuts. Material and methods. Peanuts were used in the research, in which the concentration of phenolic compounds was determined by high-performance liquid chromatography and the content of macro- and microelements by X-ray fluorescence spectroscopy. Results. The presence of phenolic compounds in peanuts was confirmed, their concentration was very diverse and ranged from 10.85 ng/g for caffeic acid to 5818.67 ng/g for syringic acid. The analysis of the content of macro- and microelements also showed their different content, from a trace amount to 3.55 ÎŒg/g for magnesium. Conclusions. The obtained results of the research on the content of phenolic compounds as well as macro- and microelements in arachidic nuts available on the Polish market indicate that they are a valuable source of ingredients with a positive effect on the human body, and thus their consumption may have an impact on preventing the development of many civilization diseases.

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Fusion of DSC and FTIR data with physicochemical profiling to distinguish berry seed oils by extraction methods

2025, Rajagukguk, Yolanda Victoria, Grygier, Anna, Siger, Aleksander, Waƛkiewicz, Agnieszka, RyszczyƄska, Sylwia, Tomaszewska-Gras, Jolanta

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Dietary salicylates affect calcium and magnesium status in preeclampsia model rats induced by NG-nitro-L-arginine-methyl ester (L-NAME)

2025, Cholik, Rafsan Syabani, Skrypnik, Katarzyna, KaraĆșniewicz-Ɓada, Marta, Waƛkiewicz, Agnieszka, Suliburska, Joanna

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Research Project

Naturalne substancje bioaktywne rodzimych roƛlin zielarskich w biologicznej ochronie zbĂłĆŒ - od kompleksowej analizy ich skƂadu i aktywnoƛci po modelowanie parametrĂłw procesu

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Field Assessment of Lamium album in Reducing Mycotoxin Biosynthesis in Winter Wheat Infected by Fusarium culmorum

2024, Uwineza, Pascaline Aimee, Kwiatkowska, Maria, Gwiazdowski, Romuald, StępieƄ, Ɓukasz, BryƂa, Marcin, Waƛkiewicz, Agnieszka

Fungicides play a crucial role in conventional agriculture for disease control, but their prolonged use raises health and environmental concerns. Fusarium culmorum (F. culmorum), a major wheat pathogen causing Fusarium head blight (FHB) and Fusarium crown rot (FCR), poses significant mycotoxigenic threats. The application of natural plant extracts has been proven to fight against phytopathogenic fungi. This study aimed to a field experiment that was carried out at the Field Experimental Station of the Institute of Plant Protection—National Research Institute in Winna GĂłra, Poland, during the 2022/2023 season to evaluate the potential of Lamium album (L. album) flower extract as a foliar spray against mycotoxigenic fungi in two winter wheat varieties: Arkadia and Julius. The supercritical carbon dioxide extraction method (SC-CO2) was employed to obtain the L. album flower extract. Ergosterol (ERG) and mycotoxin accumulation in the harvested wheat grains were analyzed using chromatography-based methods. The results demonstrated a notable reduction in ERG content in the field plots treated with L. album flower extract, from 26.07 ”g/g (control group) to 8.91 ”g/g (extract-treated group) for Arkadia and from 70.02 ”g/g (control group) to 30.20 ”g/g (extract-treated group) for Julius. The treatment with L. album reduced mycotoxin biosynthesis in both varieties, with deoxynivalenol (DON) and zearalenone (ZEN) production significantly decreased. Additionally, Arkadia exhibited greater resistance to Fusarium infection, and the antifungal effect of L. album was more pronounced than in the Julius variety, which proved to be more sensitive. In conclusion, L. album flower extract exhibited promising antifungal effects in field experiments to fight against F. culmorum in winter wheat varieties, suggesting a potential alternative to synthetic fungicides. However, as complete prevention of mycotoxin contamination was not achieved, further research is warranted to optimize extract concentrations and conduct long-term analyses to consider this plant extract as a sustainable control agent.