Fusarium head blight is one of the most noxious cereal diseases. Worldwide, F. graminearum (FG) and the mycotoxin deoxynivalenol (DON) is the most dominant species/mycotoxin in barley and wheat. Barley is often produced as on farm feed and thus routine mycotoxin analyses similar to those of cereals for human consumption are not performed. Hence, an early prediction of mycotoxin levels is important for farmers to minimise the risk of contaminated feed but also of contaminated cereals entering the cereal supply chain. Therefore, climate chamber experiments with artificial FG infection of barley investigating the influence of different temperatures (10 °C, 15 °C, 20 °C) and durations (4 h, 8 h, 12 h) at 99% relative humidity were conducted to accumulate data to develop a forecasting system. An up to three times higher DON contamination in the 15 °C treatments for the feed barley variety Ascona was detected compared with the 10 °C and 20 °C treatments. For the malting barley variety Concerto, the prolonged humidity durations had a stronger effect under all tested temperatures and resulted in up to two times higher DON contaminations. In addition, field experiments where spore deposition during anthesis as well as disease incidence, fungal amount and mycotoxins were observed, showed that the overwintered straw treatment resulted, depending on the year, in a three times higher FG incidence and DON content compared with the control and freshly inoculated straw treatment.