Latin America’s tremendous socio-cultural and biological diversity has evolved along tightly intertwined, far-reaching river networks. Decisions taken by any one country, may have strong impacts on the regional and even global biodiversity conservation agenda, such as the Convention on Biological Diversity. Here we propose and illustrate four perspectives that complement the actions suggested by Azevedo-Santos et al. (2021) to contribute to the effective conservation of freshwater biodiversity in Latin America, particularly in the context of the post-2020 Global Biodiversity Framework, currently under negotiation among the parties. Our suggestions put forward cross-border perspectives, urging governments to engage in actions, objectives, monitoring elements and post-2020 indicators that consider the reality of and threats to transnational ecosystems such as many river basins of Latin America.