<p>Chiral fungicides are commonly applied as racemic mixtures, although their enantiomers may differ in environmental behavior and biological uptake. This study investigated the enantioselective bioaccumulation of three widely used triazole fungicides—in four aquatic species representing different ecological traits. Two standardized test organisms (<i>Danio rerio</i> and <i>Daphnia magna</i>) were evaluated together with two native South American taxa (<i>Cnesterodon decemmaculatus</i> and <i>Hyalella curvispina</i>) to compare stereoselective patterns between model and regionally relevant species. Enantiomer-specific bioconcentration factors (BCFs), enantiomeric fractions (EFs), and differences between organism and water-phase stereochemical composition were jointly evaluated. Overall bioconcentration was low to moderate across all species; however, enantiomer-resolved analyses revealed species- and compound-dependent differences that were not evident from racemic assessments alone. Fish generally exhibited higher total BCF values, whereas invertebrates showed greater variability in stereoselective enrichment patterns despite lower overall accumulation. Native taxa also displayed distinct enantioselective behaviors not always mirrored by standardized species. These findings demonstrate that stereochemical discrimination may occur even under conditions of relatively limited bioaccumulation and support the inclusion of enantiomer-resolved approaches and regionally relevant organisms in comparative ecotoxicological assessments of chiral fungicides.</p> Graphical abstract <p></p>