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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, Bryła, Marcin, Stępień, Łukasz, Modrzewska, Marta, Waśkiewicz, 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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Variation in Tocochromanols Level and Mycotoxins Content in Sweet Maize Cultivars after Inoculation with Fusarium verticillioides and F. proliferatum

2022, Waśkiewicz, Agnieszka, Muzolf-Panek, Małgorzata, Stępień, Łukasz, Czembor, Elżbieta, Uwineza, Pascaline Aimee, Górnaś, Paweł, Bryła, Marcin

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

2024, Gwiazdowska, Daniela, Waśkiewicz, Agnieszka, Juś, Krzysztof, Marchwińska, Katarzyna, Frąk, Szymon, Popowski, Dominik, Pawlak-Lemańska, Katarzyna, Uwineza, Pascaline Aimee, Gwiazdowski, Romuald, Padewska, Daria, Roszko, Marek, Bryła, 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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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, Juś, Krzysztof, Marchwińska, Katarzyna, Gwiazdowski, Romuald, Waśkiewicz, 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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Lamium album Flower Extracts: A Novel Approach for Controlling Fusarium Growth and Mycotoxin Biosynthesis

2023, Uwineza, Pascaline Aimee, Urbaniak, Monika, Stępień, Łukasz, Gramza-Michałowska, Anna, Waśkiewicz, Agnieszka

Lamium album is a medicinal flowering plant that is rich in bioactive compounds with various biological properties. Fusarium species, known for causing significant crop losses and mycotoxin contamination, pose threats to food safety and human health. While synthetic fungicides are commonly employed for fungal management, their environmental impact prompts the ongoing development of alternative methods. This study aimed to evaluate the efficacy of L. album flower extracts in inhibiting the in vitro growth and biosynthesis of mycotoxins by Fusarium culmorum and F. proliferatum strains. The extracts were obtained by supercritical fluid extraction using CO2 (SC-CO2). The effects of various concentrations (2.5, 5, 7.5, and 10%) were assessed on a potato dextrose agar (PDA) medium using the “poisoning” technique. L. album flower extracts reduced mycelium growth by 0 to 30.59% for F. culmorum and 27.71 to 42.97% for F. proliferatum. Ergosterol content was reduced by up to 88.87% for F. culmorum and 93.17% for F. proliferatum. Similarly, the amounts of synthesized mycotoxins produced by both strains were also lower compared to control cultures. These findings are a preliminary phase for further in vivo tests planned to determine the fungistatic effect of L. album flower extracts on cereal substrates as seedlings incubated in controlled environments and under field conditions. Their phytotoxicity and biological stability, as well as the possibility of formulating a bio-preparation to protect cereals against Fusarium infections, will be evaluated.

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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, Marchwińska, Katarzyna, Juś, Krzysztof, Uwineza, Pascaline Aimee, Gwiazdowski, Romuald, Waśkiewicz, 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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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, Stępień, Łukasz, Gramza-Michałowska, Anna, Waśkiewicz, 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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Recent Research on Fusarium Mycotoxins in Maize—A Review

2022, Bryła, Marcin, Pierzgalski, Adam, Zapaśnik, Agnieszka, Uwineza, Pascaline Aimee, Ksieniewicz-Woźniak, Edyta, Modrzewska, Marta, Waśkiewicz, Agnieszka