Our database comprises records of pairwise ecological interactions between species in Do?ana National Park from a systematic literature review spanning from 1900 to 2023. This survey includes a diverse range of interaction types, such as predation, competition, pollination, mycorrhizae, among others, and covers taxa from all kingdoms (Plant, Animal, Fungi, Bacteria, Chromista, Archaea, Protozoa, Viruses). Each record represents an encounter between two species at a specific location and time and includes information on the interaction features, biotic partners and environmental context. The database consolidates data from multiple sources, including 'sampling event' and 'occurrence' data, enabling comprehensive coverage of all interaction data extracted from the literature. These data are valuable for scientific research focused on ecological interactions within natural ecosystems, including big-data approaches for generating species distribution models and macro-ecological analyses. The diverse nature of our database allows for an extensive exploration of the complexity and diversity of ecological interactions, with implications for a wide range of ecological studies.