The Serranias Costeiras of the Ribeira de Iguape River basin comprise landscapes with steep slopes, where many streams are contained in protected areas. We evaluated the importance of these protected areas for stream fish diversity. We sampled the ichthyofauna during the dry season of 2010, 2018 and 2019 in 36 stream stretches. We used beta diversity measures and estimated species richness and dark diversity in streams from two types of protected areas (full protection, FP and sustainable use,

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... U) and outside (Out). The altitude-width and velocity gradient of the streams explained the species turnover. The PA type promoted the richness difference, with FP streams having less species richness than SU and outside. Streams from FP presented lower species richness and dark diversity. These results indicate that the few species in FPs are well protected. The FP streams contain a relevant proportion of the regional species pool and, therefore, are essential for conserving fish stream diversity in the study region. On the other hand, streams from SU or Out have higher species richness, but their fish fauna is more vulnerable. Due to longitudinal stream connectivity, we highlight the importance of rethinking the limits of protected areas.