Abstract: Aim In this study, we examined the effects of non-native leaf litter on the functioning of an Atlantic Forest stream ecosystem. Methods Were tested two predictions: (i) Leaf litter from the native trees with high nutritional quality will have higher decomposer’s activity and faster litter decomposition; (ii) Given the presence of anti-grazing defenses, we also hypothesized that non-native leaf litter would be colonized by fewer invertebrates and that native species would be more species-rich. For this, in a forest stream (Florianópolis, SC, Brazil) we conduct the experiment to understand the decomposition and biological colonization of leaf litter among two non-native (Eucalyptus sp. and Pinus radiata D. Don) and two native trees (Ficus eximia Schott and Alchornea triplinervia (Spreng) Mull. Arg). Results Our predictions were partially corroborated. The percentage of dry mass remaining was lower for the native leaf litter. The invertebrate abundance and richness, and functional feeding groups vary between native and non-native leaf litter. Invertebrate abundance was higher in non-native Eucalyptus detritus, largely due to the high larval abundance of Chironomidae (Diptera). Conclusions Our results indicate that the presence of non-native riparian species can modify leaf decomposition and aquatic invertebrate communities in subtropical streams, with potential consequences for ecosystem functioning.