Aims: The aim of this study was to investigate changes in Salmonella and total viable count (TVC) survival on beef carcass surfaces stored for 72 h under different combinations of relative humidity (i.e. RH 75% or 96%) and temperature (5°C or 10°C).Methods and Results: The influence of low water activity (aw) and temperature on the survival and growth of Salmonella enterica serovar Typhimurium DT104 and the aerobic mesophilic flora on meat pieces from different sites on beef carcasses was investigated, under controlled conditions (75% or 96% RH; 5 or 10°C) in an environmental cabinet. Salmonella counts declined during storage at low aw (75% RH) conditions at 5°C or 10°C. Salmonella counts increased during storage at high aw (96% RH) at 10°C only. At 5°C, TVCs increased during storage at high aw, but not at low aw. TVCs increased on all samples from carcasses stored at high or low aw at 10°C, except those samples taken from areas of surface fat.Conclusions: This suggests that substrate composition dictates growth rates under low aw conditions. The results are discussed in terms of the possible protective effects of substrate osmolyte accumulation in bacterial survival and/or growth.Significance and Impact of the Study: The data obtained in this study provides useful insights on the influence of aw and temperature on pathogen survival on meat surfaces at chill temperature.