The increasing contamination of cereals by micromycetes and mycotoxins during malting still poses an unresolved food safety problem. This study characterises the potential of the novel, rapidly developing food production technology of Pulsed Electric Field (PEF) to reduce the viability of Fusarium fungi and the production of mycotoxins during malting. Barley, artificially inoculated with four Fusarium species, was treated by PEF with two different intensities and then malted using a standard Pilsner-type technology. Concentrations of fungi were quantified by RT-PCR, expression of fungal growth-related genes was assessed using mRNA sequencing, and mycotoxin levels were analysed by U-HPLC-HRMS/MS. Despite the different trends for micromycetes and mycotoxins after application of variously intense PEF conditions, significant reductions were generally observed. The greatest decrease was for F. sporotrichioides and F. poae, where up to six fold lower levels were achieved for malts produced from the PEF-treated barley when compared to the control. For F. culmorum and F. graminearum, up to a two-fold reduction in the PEF-generated malts was observed. These reductions mostly correlated with a decrease in relevant mycotoxins, specifically type A trichothecenes.

Crop Research Institute Prague Drnovska 507 73 161 06 Prague Czech Republic

Department of Biotechnology University of Chemistry and Technology Prague Technicka 5 166 28 Prague Czech Republic

Department of Food Analysis and Nutrition University of Chemistry and Technology Prague Technicka 3 166 28 Prague Czech Republic

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Alshannaq A., Yu J.-H. Occurrence, Toxicity, and Analysis of Major Mycotoxins in Food. Int. J. Environ. Res. Public Health. 2017;14:632. doi: 10.3390/ijerph14060632. PubMed DOI PMC

Foroud N.A., Baines D., Gagkaeva T.Y., Thakor N., Badea A., Steiner B., Bürstmayr M., Bürstmayr H. Trichothecenes in Cereal Grains—An Update. Toxins. 2019;11:634. doi: 10.3390/toxins11110634. PubMed DOI PMC

Battilani P., Palumbo R., Giorni P., Dall’Asta C., Dellafiora L., Gkrillas A., Toscano P., Crisci A., Brera C., de Santis B., et al. Mycotoxin mixtures in food and feed: Holistic, innovative, flexible risk assessment modelling approach: MYCHIF. EFSA Support. Publ. 2020;17:1757E. doi: 10.2903/sp.efsa.2020.EN-1757. DOI

Munkvold G.P. Fusarium Species and Their Associated Mycotoxins. Methods Mol. Biol. 2017;1542:51–106. doi: 10.1007/978-1-4939-6707-0_4. PubMed DOI

Hu L., Gastl M., Linkmeyer A., Hess M., Rychlik M. Fate of enniatins and beauvericin during the malting and brewing process determined by stable isotope dilution assays. LWT Food Sci. Technol. 2014;56:469–477. doi: 10.1016/j.lwt.2013.11.004. DOI

Habler K., Hofer K., Geißinger C., Schüler J., Hückelhoven R., Hess M., Rychlik M. Fate of Fusarium Toxins during the Malting Process. J. Agric. Food Chem. 2016;64:1377–1384. doi: 10.1021/acs.jafc.5b05998. PubMed DOI

Prusova N., Dzuman Z., Jelinek L., Karabin M., Hajslova J., Rychlik M., Stranska M. Free and conjugated Alternaria and Fusarium mycotoxins during Pilsner malt production and double-mash brewing. Food Chem. 2022;369:130926. doi: 10.1016/j.foodchem.2021.130926. PubMed DOI

Vegi A., Schwarz P., Wolf-Hall C.E. Quantification of Tri5 gene, expression, and deoxynivalenol production during the malting of barley. Int. J. Food Microbiol. 2011;150:150–156. doi: 10.1016/j.ijfoodmicro.2011.07.032. PubMed DOI

Habler K., Geissinger C., Hofer K., Schüler J., Moghari S., Hess M., Rychlik M. Fate of Fusarium toxins during brewing. J. Agric. Food Chem. 2017;65:190–198. doi: 10.1021/acs.jafc.6b04182. PubMed DOI

Homa K., Barney W.P., Davis W.P., Guerrero D., Berger M.J., Lopez J.L., Wyenandt C.A., Simon J.E. Cold Plasma Treatment Strategies for the Control of Fusarium oxysporum f. sp. basilici in Sweet Basil. HortScience. 2021;56:42–51. doi: 10.21273/HORTSCI15338-20. DOI

Filho F.O., Silva E.O., Lopes M.M.A., Ribeiro P.R.V., Oster A.H., Guedes J.A.C., Zampieri D.S., Bordallo P.D.N., Zocolo G.J. Effect of pulsed light on postharvest disease control-related metabolomic variation in melon (Cucumis melo) artificially inoculated with Fusarium pallidoroseum. PLoS ONE. 2020;15:e0220097. doi: 10.1371/journal.pone.0220097. PubMed DOI PMC

Zuluaga-Calderón B., González H.H.L., Alzamora S.M., Coronel M.B. Multi-step ozone treatments of malting barley: Effect on the incidence of Fusarium graminearum and grain germination parameters. Innov. Food Sci. Emerg. Technol. 2023;83:103219. doi: 10.1016/j.ifset.2022.103219. DOI

Evrendilek G.A., Karatas B., Uzuner S., Tanasov I. Design and effectiveness of pulsed electric fields towards seed disinfection. J. Sci. Food Agric. 2019;99:3475–3480. doi: 10.1002/jsfa.9566. PubMed DOI

Zhong C., Guan X., Fan Z., Song W., Chen R., Wang Y., He S. Pulsed electric field disinfection treatment of Fusarium oxysporum in nutrient solution. Water Supply. 2019;19:2116–2122. doi: 10.2166/ws.2019.090. DOI

Chen R., Zhang D., Zhu N., Liu C., Novac B., Zhong C. Continuous Disinfection of Fusarium oxysporum in Nutrient Solution by Pulsed Electric Field; Proceedings of the 2020 IEEE International Conference on High Voltage Engineering and Application (ICHVE); Beijing, China. 6–10 September 2020; pp. 1–4. DOI

Palicova J., Chrpova J., Tobolkova A., Ovesna J., Stranska M. Effect of pulsed electric field on viability of Fusarium micromycetes. Cereal Res. Commun. :2024. doi: 10.1007/s42976-024-00561-z. DOI

Stranska M., Prusova N., Behner A., Dzuman Z., Lazarek M., Tobolkova A., Chrpova J., Hajslova J. Influence of pulsed electric field treatment on the fate of Fusarium and Alternaria mycotoxins present in malting barley. Food Control. 2023;145:109440. doi: 10.1016/j.foodcont.2022.109440. DOI

Karabin M., Jelinek L., Prusova N., Ovesna J., Stranska M. Pulsed electric field treatment applied to barley before malting reduces Fusarium pathogens without compromising the quality of the final malt. LWT-Food Sci. Technol. 2024;206:116575. doi: 10.1016/j.lwt.2024.116575. DOI

Karlsson I., Mellqvist E., Persson P. Temporal and spatial dynamics of Fusarium spp. and mycotoxins in Swedish cereals during 16 years. Mycotoxin Res. 2023;39:3–18. doi: 10.1007/s12550-022-00469-9. PubMed DOI PMC

Miedaner T., Reinbrecht C., Lauber U., Schollenberger M., Geiger H.H. Effects of genotype and genotype-environment interaction on deoxynivalenol accumulation and resistance to Fusarium head blight in rye, triticale and wheat. Plant Breed. 2001;120:97–105. doi: 10.1046/j.1439-0523.2001.00580.x. DOI

Xu X.M., Monger W., Ritieni A., Nicholson P. Effect of temperature and duration of wetness during initial infection periods on disease development, fungal biomass and mycotoxin concentrations on wheat inoculated with single, or combinations of, Fusarium species. Plant Pathol. 2007;56:943–956. doi: 10.1111/j.1365-3059.2007.01650.x. DOI

Gautam P., Dill-Macky R. Impact of moisture, host genetics and Fusarium graminearum isolates on Fusarium head blight development and trichothecene accumulation in spring wheat. Mycotoxin Res. 2012;28:45–58. doi: 10.1007/s12550-011-0115-6. PubMed DOI

Zhao W., Yang R., Zhang H.Q. Recent advances in the action of pulsed electric fields on enzymes and food component proteins. Trends Food Sci. Technol. 2012;27:83–96. doi: 10.1016/j.tifs.2012.05.007. DOI

Tibola C.S., Fernandes J.M.C., Guarienti E.M., Nicolau M. Distribution of Fusarium mycotoxins in wheat milling process. Food Control. 2015;53:91–95. doi: 10.1016/j.foodcont.2015.01.012. DOI

Edwards S.G., Dickin E.T., MacDonald S., Buttler D., Hazel C.M., Patel S., Scudamore K.A. Distribution of Fusarium mycotoxins in UK wheat mill fractions. Food Addit. Contam. A. 2011;28:1694–1704. doi: 10.1080/19440049.2011.605770. PubMed DOI

Chelkowski J. Mycotoxins in Agriculture and Food Safety. 1st ed. CRC Press; Boca Raton, FL, USA: 1998. Distribution of Fusarium Species and Their Mycotoxins in Cereal Grains. DOI

Lewandowski S.M., Bushnell W.R., Evans C.K. Distribution of mycelial colonies and lesions in field-grown barley inoculated with Fusarium graminearum. Phytopathology. 2007;96:567–581. doi: 10.1094/PHYTO-96-0567. PubMed DOI

Jin Z., Solanki S., Ameen G., Gross T., Poudel R.S., Borowicz P., Brueggeman R.S., Schwarz P. Expansion of Internal Hyphal Growth in Fusarium Head Blight-Infected Grains Contributes to the Elevated Mycotoxin Production During the Malting Process. Molec. Plant-Microbe Interact. 2021;34:793–802. doi: 10.1094/MPMI-01-21-0024-R. PubMed DOI

Munkvold G.P., Proctor R.H., Moretti A. Mycotoxin Production in Fusarium According to Contemporary Species Concepts. Annu. Rev. Phytopathol. 2021;59:373–402. doi: 10.1146/annurev-phyto-020620-102825. PubMed DOI

Xue A.G., Armstrong K.C., Voldeng H.D., Fedak G., Babcock C. Comparative aggressiveness of isolates of Fusarium spp. causing head blight on wheat in Canada. Can. J. Plant Pathol. 2004;26:81–88. doi: 10.1080/07060660409507117. DOI

Wagacha J.M., Oerke E.C., Dehne H.W., Steiner U. Interactions of Fusarium species during prepenetration development. Fungal Biol. 2012;116:836–847. doi: 10.1016/j.funbio.2012.05.001. PubMed DOI

Velluti A., Marín S., Bettucci L., Ramos A.J., Sanchis V. The effect of fungal competition on colonization of maize grain by Fusarium moniliforme, F. proliferatum and F. graminearum and on fumonisin B1 and zearalenone formation. Int. J. Food Microbiol. 2000;59:59–66. doi: 10.1016/S0168-1605(00)00289-0. PubMed DOI

Etcheverry M. Aflatoxin B1, zearalenone and deoxynivalenol production by Aspergillus parasiticus and Fusarium graminearum in interactive cultures on irradiated corn kernels. Mycopathologia. 1998;142:37–42. doi: 10.1023/A:1006972016955. PubMed DOI

Martins M.P., Silva L.G., Rossi A., Sanches P.R., Souza L.D.R., Martinez-Rossi N.M. Global analysis of cell wall genes revealed putative virulence factors in the dermatophyte Trichophyton rubrum. Front. Microbiol. 2019;10:2168. doi: 10.3389/fmicb.2019.02168. PubMed DOI PMC

Djordjevic J.T. Role of phospholipases in fungal fitness, pathogenicity, and drug development—Lessons from Cryptococcus neoformans. Front. Microbiol. 2010;1:125. doi: 10.3389/fmicb.2010.00125. PubMed DOI PMC

Westrick N.M., Park S.C., Keller N.P., Smith D.L., Kabbage M. A broadly conserved fungal alcohol oxidase (AOX) facilitates fungal invasion of plants. Molec. Plant Pathol. 2023;24:28–43. doi: 10.1111/mpp.13274. PubMed DOI PMC

Gai X., Li S., Jiang N., Sun Q., Xuan Y.H., Xia Z. Comparative transcriptome analysis reveals that ATP synthases regulate Fusarium oxysporum virulence by modulating sugar transporter gene expressions in tobacco. Front. Plant Sci. 2022;13:978951. doi: 10.3389/fpls.2022.978951. PubMed DOI PMC

Sella L., Gazzetti K., Faoro F., Odorizzi S., D’Ovidio R., Schäfer W., Favaron F. A Fusarium graminearum xylanase expressed during wheat infection is a necrotizing factor but is not essential for virulence. Plant Physiol. Biochem. 2013;64:1–10. doi: 10.1016/j.plaphy.2012.12.008. PubMed DOI

Chen W., Lee M.K., Jefcoate C., Kim S.C., Chen F., Yu J.H. Fungal cytochrome P450 monooxygenases: Their distribution, structure, functions, family expansion, and evolutionary origin. Genome Biol. Evol. 2014;6:1620–1634. doi: 10.1093/gbe/evu132. PubMed DOI PMC

Cui Y., Peng Y., Zhang Q., Xia S., Ruan B., Xu Q., Yu X., Zhou T., Liu H., Zeng D., et al. Disruption of EARLY LESION LEAF 1, encoding a cytochrome P450 monooxygenase, induces ROS accumulation and cell death in rice. Plant J. 2021;105:942–956. doi: 10.1111/tpj.15079. PubMed DOI

Laugé R., Joosten M.H.A.J., Van den Ackerveken G.F.J.M., Van den Broek H.W.J., De Wit P.J.G.M. The in planta-produced extracellular proteins ECP1 and ECP2 of Cladosporium fulvum are virulence factors. Mol. Plant-Microbe Interact. 1997;10:725–734. doi: 10.1094/MPMI.1997.10.6.725. DOI

Gozzi M., Blandino M., Bruni R., Capo L., Righetti L., Dall’Asta C. Mycotoxin occurrence in kernels and straws of wheat, barley, and tritordeum. Mycotoxin Res. 2024;40:203–210. doi: 10.1007/s12550-024-00521-w. PubMed DOI PMC

Peiris K., Pumphrey M., Dong Y., Dowell F. Fusarium Head Blight Symptoms and Mycotoxin Levels in Single Kernels of Infected Wheat Spikes. Cereal Chem. 2011;88:291–295. doi: 10.1094/CCHEM-08-10-0112. DOI

Janik E., Niemcewicz M., Podogrocki M., Ceremuga M., Stela M., Bijak M. T-2 Toxin-The Most Toxic Trichothecene Mycotoxin: Metabolism, Toxicity, and Decontamination Strategies. Molecules. 2021;26:6868. doi: 10.3390/molecules26226868. PubMed DOI PMC

Ohshima T., Tanino T., Hashimoto Y. Inactivation of a plant pathogenic fungus by pulsed electric field treatment. Int. J. Plasma Environ. Sci. Technol. 2020;14:e02007. doi: 10.34343/ijpest.2020.14.e02007. DOI

Chrpova J., Šíp V., Matejova E., Sýkorova S. Resistance of Winter Wheat Varieties Registered in the Czech Republic to Mycotoxin Accumulation in Grain Following Inoculation with Fusarium culmorum. Czech J. Genet. Plant Breed. 2007;43:44–52. doi: 10.17221/1910-CJGPB. DOI

Dzuman Z., Zachariasova M., Lacina O., Veprikova Z., Slavikova P., Hajslova J. A rugged high-throughput analytical approach for the determination and quantification of multiple mycotoxins in complex feed matrices. Talanta. 2014;121:263–272. doi: 10.1016/j.talanta.2013.12.064. PubMed DOI

Ovesna J., Vacke J., Kucera L., Chrpova J., Novakova I., Jahoor A., Šíp V. Genetic analysis of resistance in barley to barley yellow dwarf virus. Plant Breed. 2000;119:481–486. doi: 10.1046/j.1439-0523.2000.00522.x. DOI

Leišová L., Kučera L., Chrpová J., Sýkorová S., Šíp V., Ovesná J. Quantification of Fusarium culmorum in Wheat and Barley Tissues Using Real-Time PCR in Comparison with DON Content. J. Phytopathol. 2006;154:603–611. doi: 10.1111/j.1439-0434.2006.01154.x. DOI

Yli-Mattila T., Paavanen-Huhtala S., Jestoi M., Parikka P., Hietaniemi V., Gagkaeva T., Sarlin T., Haikara A., Laaksonen S., Rizzo A. Real-time PCR detection and quantification of Fusarium poae, F. graminearum, F. sporotrichioides and F. langsethiae in cereal grains in Finland and Russia. Arch. Phytopathol. Plant Protect. 2008;41:243–260. doi: 10.1080/03235400600680659. DOI

Köhl J., Lombaers C., Moretti A., Bandyopadhyay R., Somma S., Kastelein P. Analysis of microbial taxonomical groups present in maize stalks suppressive to colonization by toxigenic Fusarium spp.: A strategy for the identification of potential antagonists. Biol. Control. 2015;83:20–28. doi: 10.1016/j.biocontrol.2014.12.007. DOI

Bluhm B.H., Cousin M.A., Woloshuk C.P. Multiplex real-time PCR detection of fumonisin-producing and trichothecene-producing groups of Fusarium species. J. Food Prot. 2004;67:536–543. doi: 10.4315/0362-028X-67.3.536. PubMed DOI

European Brewing Convention Analytica EBC. Chapter: 4.12.2—Diastatic Power of Malt by Segmented Flow Analysis. [(accessed on 28 November 2023)]. Available online: https://brewup.eu/ebc-analytica/malt/diastatic-power-of-malt-by-segmented-flow-analysis/4.12.2.

Megazyme Ltd. α-Amylase Assay Kit (Ceralpha Method) [(accessed on 28 November 2023)]. Available online: https://www.megazyme.com/documents/Assay_Protocol/K-CERA_DATA.pdf.

Megazyme Ltd. β-Glucanase Assay Kit (Malt and Microbial) [(accessed on 28 November 2023)]. Available online: https://d1kkimny8vk5e2.cloudfront.net/documents/Assay_Protocol/K-MBGL_DATA.pdf.

Pulsed Electric Field Treatment Modulates Gene Expression and Stress Responses in Fusarium-Infected Malting Barley

What Happens Inside the Germinating Grain After Microbial Decontamination by Pulsed Electric Field? Data-Driven Multi-Omics Helps Find the Answer

Pulsed Electric Field Induces Significant Changes in the Metabolome of Fusarium Species and Decreases Their Viability and Toxigenicity