Using ethnobotanical techniques, the medicinal flora used by the inhabitants of San Rafael Coxcatlán, Puebla was determined. During the field work, two types of interviews were applied (free listing and semi-structured) to 60 informants, who supplied consistent information concerning the use of 46 species of medicinal plants. Further analysis showed 13 categories of different medicinal use. An informant consensus factor was calculated and 16 species were selected due to their utilization in the treatment of diseases of possible bacterial origin. Of these 16 plants, sequential extractions were made with hexane, ethyl acetate and methanol. The obtained extracts were used to assess their antibacterial activity against 14 bacterial strains; 75% of the plants presented antibacterial activity. The medicinal species Jatropha neopauciflora Pax (Euphorbiaceae) and Juliania adstringens (Schldl.) Schldl. (Julianiaceae) were those that showed the biggest activity. Moreover, these species also had the highest informant consensus factor values.