Contents lists available at ScienceDirect International Journal of Food Microbiology journal homepage: www.elsevier.com/locate/ijfoodmicro Fungal community, Fusarium head blight complex and secondary metabolites associated with malting barley grains harvested in Umbria, central Italy Giovanni Beccari a , Maria Teresa Senatore a,1 , Francesco Tini a , Michael Sulyok b , Lorenzo Covarelli a, ⁎ a Department of Agricultural, Food and Environmental Sciences, University of Perugia, Borgo XX Giugno 74, 06121 Perugia, Italy b Centre for Analytical Chemistry, Department of Agrobiotechnology (IFA-Tulln), University of Natural Resources and Life Sciences (BOKU), Vienna, Konrad Lorenz Strasse 20, A-3430 Tulln, Austria ARTICLE INFO Keywords: Fusarium Barley Mycotoxins LC-MS/MS Enniatins Beer ABSTRACT In recent years, due to the negative impact of toxigenic mycobiota and of the accumulation of their secondary metabolites in malting barley grains, monitoring the evolution of fungal communities in a certain cultivation area as well as detecting the different mycotoxins present in the raw material prior to malting and brewing processes have become increasingly important. In this study, a survey was carried out on malting barley samples collected after their harvest in the Umbria region (central Italy). Samples were analyzed to determine the composition of the fungal community, to identify the isolated Fusarium species, to quantify fungal secondary metabolites in the grains and to characterize the in vitro mycotoxigenic profile of a subset of the isolated Fusarium strains. The fungal community of barley grains was mainly composed of microorganisms belonging to the genus Alternaria (77%), followed by those belonging to the genus Fusarium (27%). The Fusarium head blight (FHB) complex was represented by nine species with the predominance of Fusarium poae (37%), followed by Fusarium avenaceum (23%), Fusarium graminearum (22%) and Fusarium tricinctum (7%). Secondary metabolites bio- synthesized by Alternaria and Fusarium species were present in the analyzed grains. Among those biosynthesized by Fusarium species, nivalenol and enniatins were the most prevalent ones. Type A trichothecenes (T-2 and HT-2 toxins) as well as beauvericin were also present with a high incidence. Conversely, the number of samples contaminated with deoxynivalenol was low. Conjugated forms, such as deoxynivalenol-3-glucoside and HT-2- glucoside, were detected for the first time in malting barley grains cultivated in the surveyed area. In addition, strains of F. avenaceum and F. tricinctum showed the ability to biosynthesize in vitro high concentrations of enniatins. The analysis of fungal secondary metabolites, both in the grains and in vitro, revealed also the presence of other compounds, for which further investigations will be required. The combination of microbiological analyses, of molecular biology assays and of multi-mycotoxin screening shed light on the complexity of the fungal community and its secondary metabolites released in malting barley. 1. Introduction Barley (Hordeum vulgare) is one of the most important cereal crops worldwide. With a production of 988,285 tons in 2016 it is also one of the most cultivated cereals in Italy (FAO, 2018). A significant amount (10–15%) of the Italian barley production is used for malt obtainment, mostly destined to the national beer industry. In Italy, beer obtained from malting barley was estimated in 1 296,800 tons in 2014 (FAO, 2018) and this beverage is becoming increasingly important within the national food-processing economy. In fact, over the last three decades https://doi.org/10.1016/j.ijfoodmicro.2018.03.005 Received 17 November 2017; Received in revised form 15 February 2018; Accepted 10 March 2018 ⁎ Corresponding author. 1 Present address: Department of Agricultural Sciences, Alma Mater Studiorum University of Bologna, Viale G. Fanin 44, 40127, Bologna, Italy. E-mail address: lorenzo.covarelli@unipg.it (L. Covarelli). Abbreviations: EU, European Union; FHB, Fusarium head blight; FIESC, Fusarium incarnatum-equiseti species complex; DON, deoxynivalenol; 3AcDON, 3-acetyldeoxynivalenol; 15AcDON, 15-acetyldeoxynivalenol; NIV, nivalenol; MON, moniliformin; ENs, enniatins; BEA, beauvericin; DON-3G, deoxynivalenol-3-glucoside; T-2G, T-2-glucoside; HT-2G, HT-2- glucoside; PDA, potato dextrose agar; SE, standard error; EDTA, ethylenediamine-tetraacetic acid disodium salt dehydrate; TAE, trizma base-glacial acid acetic-ethylenediamine tetra- acetic acid disodium salt dehydrate; tef1α, translation elongation factor 1α; LC-MS/MS, liquid chromatography-tandem mass spectrometry; ESI, electrospray ionization; MRM, multiple reaction monitoring; ENB, enniatin B; ENB1, enniatin B1; ENA1, enniatin A1; ENA, enniatin A; ZEN, zearalenone; TeA, tenuazonic acid; Ten, tentoxin; AOH, alternariol; AME, alternariol methyl ether; Macro, macrosporin; EAs, ergot alkaloids; ENB2, enniatin B2; ENB3, enniatin B3; HIV-1, human immunodeficiency virus type 1 International Journal of Food Microbiology 273 (2018) 33–42 Available online 12 March 2018 0168-1605/ © 2018 Elsevier B.V. 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