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HODULÍKOVÁ LUCIA, SKLÁDANKA JIŘÍ, MLEJNKOVÁ VERONIKA, KNOT PAVEL, KLUSOŇOVÁ IVA, HORKÝ PAVEL, KONEČNÁ KLÁRA, KNOTOVÁ DANIELA, NEDĚLNÍK JAN, SLÁMA PETR. 2016. Effect of Soil Contamination of Fodder and Wilting on the Occurrence of Fungi and Mycotoxins in Alfalfa Silages. Acta Universitatis Agriculturae et Silviculturae Mendelianae Brunensis, 64(5): 1529–1536. The aim of the work is to assess the effect of soil contamination of fodder and wilting on the occurrence of fungi in the biomass of alfalfa and subsequently consider the mycotoxin content in alfalfa silage. The alfalfa biomass of Jarka variety, harvested from two experimental plots in different climatic conditions, was evaluated. The total number of microorganisms and fungi were determined in silage biomass and silage. The content of deoxynivalenol and zearalenone in silages was treated with bacterial enzyme inoculant and chemical preservative, which is an acid. The total number of microorganisms was not influenced by the soil contamination of fodder. The occurrence of fungi was increased from 126,807 to 321,992 CFU·g-1. The total number of microorganisms and fungi was raised by wilting. The occurrence of fungi was increased from 113,909 to 334,890 CFU·g-1. Deoxynivalenol (increased from 101.9 to 131.5 ppb) was especially determined in alfalfa silage. The content of zearalenone reached up to 0.5442 ppb. The alfalfa silages, produced from wilted silage, contained lower levels (P < 0.05) of deoxynivalenol and zearalenone than alfalfa silages produced from no-wilted biomass. The content of mycotoxins was affected neither biological nor chemical preservative.