Abstract The village of Zapotitlán de las Salinas is situated in the Valley of Tehuacán-Cuicatlán, Puebla, Mexico. Plant species used by the local inhabitants to treat gastrointestinal diseases were identified using ethnobotanical, ethnographic and taxonomic methods. Out of 119 interviews, 44 plant species were registered, of which the following are the most frequently used (listed in descending order): Lippia graveolens H.B. et K. (Verbenaceae), Lantana achyranthifolia Desf. (Verbenaceae), Turnera diffusa (Willd.) ex Schult. (Turneraceae), Lippia oaxacana Rob. et Greenm. (Verbenaceae), Gymnolaena oaxacana (Greenm.) Rydb. (Asteraceae), Cordia curassavica (Jacq.) Roem. et Schult. (Boraginaceae), Lantana camara L. (Verbenaceae) and Acalypha hederacea Torrey (Euphorbiaceae). From these plants, hexane, chloroform and ethanol extracts were prepared in order to assess their antibacterial activity against 14 bacterial strains causing the most common gastrointestinal diseases in Mexican population. All hexane extracts showed antibacterial activity against Gram-positive and Gram-negative bacteria. There is a correlation between the frequency of mention (of plant use) and the antibacterial activity. In conclusion, the knowledge of plants most frequently used for gastrointestinal infections in Zapotitlán de las Salinas is supported by scientific rationale.