Ecoagriculture is an approach to landscape management that simultaneously advances agricultural production, conservation of biodiversity and ecosystem services, and sustainable rural livelihoods. In this dissertation, I address gaps in the science and application of ecoagriculture by reviewing key antecedents of this approach, enumerating a framework for ecoagriculture practice, and evaluating ecoagriculture outcomes—particularly biodiversity conservation—in pasture-dominated landscapes in northern Latin America. Ecoagriculture builds upon multiple disciplines and traditions of practice including landscape and ecosystem approaches, adaptive management, and integrated natural resource management. These antecedents inform the Landscape Measures approach, a structure for guiding multi-stakeholder planning, decision-making, and monitoring of ecoagriculture objectives at a landscape scale. Pilot applications of this approach in Honduras and Kenya suggest its value for promoting landscape multifunctionality by documenting landscape dynamics, fostering constructive dialogue among diverse stakeholders, and improving empirical bases for collective decision-making. Previous research has indicated that a substantial proportion of native species may persist in Neotropical agricultural mosaics. However, the consistency of these findings—and thus their applicability to unstudied locales—remains unknown. I conducted a coordinated multi-site study to evaluate the consistency of relationships between bird and butterfly assemblage characteristics and land use practices in four landscapes spanning four countries in northern Latin America: Colombia, Costa Rica, Honduras, and Nicaragua. Bird and butterfly species richness and abundance were significantly related to land use across all four landscapes, and were generally positively correlated with degree of tree cover. In the Honduras landscape, I also investigated the relative influence of land use, vegetation, and landscape composition and structure on bird and butterfly assemblages. These assemblages were more strongly related to land use and plot-scale habitat features than to gradients of landscape composition and structure. Across all landscapes, habitat heterogeneity fostered high levels of beta diversity and high overall species richness, but only a modest presence of forest-dependent and high conservation value species. The findings underscore the importance of considering multiple conservation values when evaluating conservation outcomes in ecoagriculture landscapes. These values include protecting species of intermediate conservation concern and species that support rural livelihoods—not only globally threatened biodiversity.