Aims: This study investigated the in vitro effects of water activity (a(w); 0.85-0.987) and temperature (10-40degreesC) on growth and ochratoxin A (OTA) production by two strains of Aspergillus carbonarius isolated from wine grapes from three different European countries and Israel on a synthetic grape juice medium representative of mid-veraison (total of eight strains). Methods and Results: The synthetic grape juice medium was modified with glycerol or glucose and experiments carried out for up to 56 days for growth and 25 days for OTA production. The lag phase prior to growth, growth rates and ochratoxin production were quantified. Statistical comparisons were made of all factors and multiple regression analysis used to obtain surface response curves of a(w) x temperature for the eight strains and optimum growth and OTA production by A. carbonarius. The lag phase increased from <1 day at 25-35degreesC and 0.98 a(w) to >20 days at marginal temperatures and water availabilities. Generally, most A. carbonarius strains grew optimally at 30-35degreesC, regardless of solute used to modify a(w), with no growth at <15degreesC. The optimum a(w) for growth varied from 0.93 to 0.987 depending on the strain, with the widest a(w) tolerance at 25-30degreesC. There was no direct relationship among growth, environmental factors and country of origin of individual strains. Optimum conditions for OTA production varied with strain. Some strains produced optimal OTA at 15-20degreesC and 0.95-98 a(w). The maximum OTA produced after 10 days was about 0.6-0.7 mug g(-1), with a mean production over all eight strains of 0.2 mug g(-1) at optimum environmental conditions. Conclusions: This work demonstrates that optimum conditions for OTA production are very different from those for growth. While growth rates differed significantly between strains, integration of the OTA production data suggests possible benefits for use of the information on a regional basis. Significance and Impact of the Study: Very little detailed information has previously been available on the ecology of A. carbonarius. This knowledge is critical in the development and prediction of the risk models of contamination of grapes and grape products by this species under fluctuating and interacting environmental parameters.