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Decimal reduction time (D-value) was calculated for six non-O157 Shiga toxin–producing Escherichia coli(STEC) in a laboratory medium and ground beef. For the laboratory medium, an overnight culture of each strain of STEC was divided into 10-mL sample bags and heated in a water bath for a specific time on the basis of the temperatures. Survival curves were generated by plotting the surviving bacterial population against time, and a linear-log primary model was used to estimate the D-values from survival curves. The z-values (the temperature raised to reduce the D-value by one-tenth) were calculated by plotting the log D-values against temperature. Similarly, for ground beef, six fat contents, 5, 10, 15, 20, 25, and 30% of ground beef were formulated for this study. Inoculated meat was divided into 5-g pouches and submerged in a water bath set at specific temperatures (55, 60, 65, 68, and 71.1°C). The average D-value for these strains in a laboratory medium was 17.96 min at 55°C, which reduced significantly (P< 0.05) to 1.58 min at 60°C, and then further reduced (P< 0.05) to 0.46 min at 65°C. In ground beef, a negative correlation (P< 0.05) between fat content of ground beef and D-values was observed at 55°C. However, at temperatures greater than 60°C, there was no impact (P> 0.05) of fat content of ground beef on the thermal resistance of non-O157 STECs. Irrespective of the fat content of ground beef, the D-values ranged from 15.93 to 11.69, 1.15 to 1.12, and 0.14 to 0.09 min and 0.05 at 55, 60, 65, and 68°C, respectively. The data generated from this study can be helpful for the meat industry to develop predictive models for thermal inactivation of non-O157 STECs in ground beef with varying fat content.