To provide additional insights on pathogen survival, we evaluated the relative efficacy of acidification (pH 2.7), thermophilic treatment (55 °C), and low temperature pasteurization (68 °C) on the inactivation of E. coli O157:H7, Salmonella, and Listeria monocytogenes in ground beef. A series of experiment was conducted under biosafety level 3 environments for assessing the impacts of heat and low pH on pathogen survival. Results showed that 5-log reductions of E. coli O157:H7 could take more than 2640 min at 55 °C, 134 min at 68 °C and 120 min under pH 2.7. Compared to E. coli O157:H7, the 5-log reduction of Salmonella was obtained in 4836, 126, 86 min at 55 °C, 68 °C, and pH 2.7, respectively. The 5-log reduction of Listeria was achieved in 4704, 200, and 115 min under 55 °C, 68 °C, and pH 2.7, respectively. The results of this study will provide additional insights for developing improved methods for controlling pathogens in ground beef.