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Abstract Aim Freshwater species face significant challenges from direct and indirect anthropogenic impacts, leading to a global decline in freshwater biodiversity. Protected areas are a key tool for conservation, but their effectiveness in covering freshwater biodiversity remains uncertain. This study assesses the protection coverage of freshwater macroinvertebrates, vertebrates, and macrophytes in Cuba against the 17% and 30% conservation targets set by the Convention on Biological Diversity. Location Caribbean biodiversity hotspot, including freshwater ecosystems across the Cuban archipelago. Methods We analyzed the distribution of 182 freshwater macroinvertebrates, 26 vertebrates, and 19 macrophyte species using an ensemble of four species distribution modeling techniques: Maxent, Boosted Regression Trees (BRT), Random Forest (RF), and Spatial Stream Network (SSN). We evaluated species overlap with Cuba’s current protected areas and conducted spatial conservation prioritization exercises that (i) included (lock-in) and (ii) excluded (free-choice) existing protected areas. Results Our analysis revealed that 41% (90 species) and 71% (161 species) failed to meet the 17% and 30% conservation targets, respectively. Many of the insufficiently protected species are globally threatened or endemic to the Cuban archipelago, heightening their extinction risk. Conservation planning that includes current protected areas requires significantly larger areas to meet the 30% representation target due to redundancy in existing protections. Conversely, excluding current protected areas achieves conservation goals more efficiently with fewer resources. Both approaches highlight the need to improve connectivity, particularly in upstream regions that are often neglected under the current protected area configuration. Main conclusions Freshwater biodiversity in Cuba is poorly represented within existing protected areas. Meeting the 30% conservation target would require protecting an additional 30–70% of area, with a focus on headwaters and underrepresented taxa. Expanding Cuba’s National System of Protected Areas (SNAP) to prioritize fresh-water species, particularly endemics, is essential. A spatial conservation planning approach that integrates both lock-in and free-choice strategies can optimize resource use while enhancing connectivity across key rivers and tributaries.