Introduction: The expansion of conventional agriculture has caused the loss of natural habitats for the fauna, but the coffee agroecosystems of Oaxaca can provide an alternative habitat for mammals, which is associated with the cultural value that human communities assign to them. Objective: To assess the diversity and cultural value of medium and large mammals associated with the coffee plantation, and what they represent for the inhabitants of San Gabriel Mixtepec, Oaxaca, Mexico. Method: A mixed method was used. For the diversity analysis, the mammal species were identified through camera traps. To determine the Cultural Importance Index (IIC), semi-structured interviews and participatory workshops were carried out with the residents, who also helped to identify the different uses of the medium and large mammals. Results: A total of 26 species grouped into seven orders, 14 families, and 23 genera were recorded. Alpha diversity was greater in the dry season than in the wet season. Some species were detected in only one of the two seasons. Six hundred and thirty-nine records of cultural uses of mammals were obtained; use as food was the most mentioned. Twelve types of uses were identified. The species with the greatest cultural importance were Odocoileus virginianus (IIC = 0.92), with 11 uses, followed by Dasypus novemcinctus (IIC = 0.61), with 10 uses, Conepatus leuconotus (IIC = 0.60), with eight uses and Didelphis virginiana (IIC = 0.52), with 11 uses. Conclusion: The results show the importance that human residents assign to mammals associated with coffee agroecosystems, both for their use for subsistence and to complement the family economy. Furthermore, the residents themselves emphasize the importance of their traditional knowledge and the need to adopt sustainable management practices for the mastofauna, because they recognize the importance it has in the community and the benefits it can have on the environment.