Studies were conducted to examine the effect of water activity (a(w), 0.97, 0.95, 0.93), pH (4.5, 6.0) and preservative type and concentration (potassium sorbate (KS), calcium propionate (CaP), 300 and 3000 ppm) on colonisation and ochratoxin A (OTA) production by three strains of Penicillium verrucosum on bread analogues. At 25 degrees C, colony extension from a point source was similar for all three strains at the different environmental treatments examined, with optimum growth at 0.97 and 0.96 a(w) and pH 6. Temporal OTA production over 36 days varied significantly with strain, pH and a(w) level. Maximum OTA was produced at approximately 28-36 days for two of the strains with an optimum at 0.93 a(w) at pH 6.0. One strain produced 75-80 microg g(-1) analogue under these conditions. However, at pH 4.5, optimum OTA was produced at 0.95 a(w) after a similar time period on bread analogues, with maximum amounts of about 25-35 microg g(-1) bread. KS and CaP incorporation inhibited growth completely at pH 4.5 and 3000 ppm at all three a(w) levels. However, at suboptimal concentration, little growth inhibition occurred. At pH 6, even with 3000 ppm, growth of all three strains occurred. Where effective inhibition of growth was achieved (3000 ppm/pH 4.5, all a(w) levels), no OTA was detected. However, when 300 ppm was used, up to 7 microg g(-1) was detected in bread analogues at pH 4.5. At pH 6, OTA was produced at all treatment conditions by all three strains, although preservative-treated bread had less present. Statistical analysis showed that many of the one-, two- and three-way interactions had a significant effect on growth and OTA production. These results suggest that there is a significant risk from OTA contamination if lower concentrations of existing preservatives are used in wheat-based bakery products contaminated by spoilage mycotoxigenic moulds such as P. verrucosum.