The effect of combinations of temperature (2 degrees, 3 degrees, 4 degrees, 5 degrees, 8 degrees and 10 degrees C), pH (5.0-7.2) and NaCl (0.1-5.0% w/w) on growth from spores of non-proteolytic Clostridium botulinum types B, E and F was determined using a strictly anaerobic medium. Inoculated media were observed weekly for turbidity, and tests were made for the presence of toxin in conditions that approached the limits of growth. Growth and toxin production were detected at 3 degrees C in 5 weeks, at 4 degrees C in 3/4 weeks and at 5 degrees C in 2/3 weeks. The resulting data define growth/no growth boundaries with respect to low temperature, pH, NaCl and incubation time. This is important in assessment of the risk of growth and toxin production by non-proteolytic Cl. botulinum in minimally processed chilled foods.