The U.S. beef industry must provide documentation to the U.S. Department of Agriculture, Food Safety and Inspection Service (USDA-FSIS) that the antimicrobial interventions implemented or any subsequent changes in the process are effective under the actual conditions that apply in its operation. The main objective of this study was to determine whether surface pH after application of diluted lactic acid solution on surfaces of fresh meat can be used as a control measure indicator for the reduction of E. coli O157:H7 and Salmonella. Samples (240 each) of lean and adipose beef tissues were inoculated with cocktail mixtures of Escherichia coli O157:H7 and Salmonella. Application parameters were varied such that lean and adipose tissues were spray treated with either 2 or 4.5% lactic acid solution at either 38 or 60°C for 1 to 10 s. Lean and adipose tissues were collected before and after spray treatments for enumeration of the pathogens. Based on the conditions of this study, there was no difference between spray treatments at 38 or 60°C, but 4.5% lactic acid solution reduced pathogens more effectively (P ≤ 0.05) than did 2% lactic acid solution. Spray treatment with lactic acid solution for 1 to 10 s reduced surface pH values of lean tissues (3 to 3.8) and adipose tissues (2.75 to 3.65). At surface pH values of 3.0 and 2.75, lactic acid reduced E. coli O157:H7 on surfaces of lean and adipose tissues by approximately 1.60 and 1.54 log CFU/cm2, respectively. At surface pH values of 3.8 and 3.65, lactic acid reduced E. coli O157:H7 on lean and adipose tissues by approximately 0.3 and 0.42 log CFU/cm2, respectively. The surface pH values after lactic acid treatment and the reductions of both pathogens showed a strong linear relationship; this indicates that a surface pH of 3.1 would provide at least 1-log reduction of E. coli O157:H7 and Salmonella, regardless of lactic acid application parameters. Therefore, surface pH after spray treatment with lactic acid could be used to validate pathogen reduction.