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To determine the effect of interacting conditions of water activity (aw, 0·99–0·85), temperature (15, 25°C) and time (40 days) on growth and production of the mycotoxins deoxynivalenol (DON) and nivalenol (NIV) by Fusarium culmorum on a wheat-based agar medium.Fusarium culmorum grew optimally at 0·995aw and minimally at 0·90 at both 15 and 25°C. No growth was observed at < 0·90aw. Overall, temperature, aw and their interaction had a statistically significant effect on the growth rate of F. culmorum. Production of both DON and NIV were over a much narrower range (0·995–0·95aw) than that for growth. The highest concentrations of DON and NIV levels were produced at 0·995aw and 0·981aw at 25°C, respectively, after 40 days of incubation. Statistically, aw, temperature and incubation time, and aw × temperature and temperature × incubation time had a statistically significant effect on DON/NIV production.This is the first detailed report on the two-dimensional environmental profiles for DON/NIV production by F. culmorum in the UK.As part of a hazard analysis critical control point (HACCP) approach, this type of information is critical in monitoring critical control points for prevention of DON/NIV entering the wheat production chain.