Food safety is a constant concern in the food industry, especially regarding the control of pathogens responsible for foodborne diseases (FBDs). The geopropolis produced by Melipona stingless bees has gained interest due to its natural antimicrobial potential. This study aimed to evaluate the antimicrobial activity of the geopropolis from the stingless bee Melipona quadrifasciata against seven bacterial strains: Staphylococcus aureus ATCC 25923, S. aureus (field strains S4 and S31), Salmonella Typhimurium ATCC 14028 and the field strain L14A, Salmonella Heidelberg (field strain), and Listeria monocytogenes ATCC 19111. The raw geopropolis was selected to remove impurities, frozen, ground, and homogenized. Two grams of the material were weighed and mixed with 25 mL of 80% cereal alcohol, followed by extraction in a water bath at 70 °C for 30 minutes under intermittent agitation. The extract was filtered and incorporated into Mueller–Hinton agar at final concentrations of 10% and 5%. Each Petri dish was perforated with seven wells, into which 10 µL of bacterial suspension standardized at 0.5 on the McFarland scale was inoculated. The plates were incubated at 37 °C for 24 hours. Staphylococcus aureus (ATCC and field strains) was completely inhibited at both concentrations tested. Salmonella Typhimurium (ATCC 14028 and L14A) and Salmonella Heidelberg were inhibited only at 10% concentration. Listeria monocytogenes ATCC 19111 was eliminated at both concentrations. The geopropolis of Melipona quadrifasciata demonstrated relevant antimicrobial activity against bacteria of importance in FBDs, particularly S. aureus and L. monocytogenes. The results suggest the potential use of geopropolis as an auxiliary agent in microbiological control, contributing to food safety and the fight against bacterial resistance.