Benthic macroinvertebrate in four rivers, three in the Pelotas River basin (Divisa, Marco and Silveira rivers, in the headwaters of the Uruguai River) and one in the Taquari-Antas system (Antas River), a tributary in the Guaiba basin, in the state of Rio Grande do Sul, were identified. Two samples were collected in summer, autumn and spring, with one replicate in each river. The total of 28,961 specimens included members of Platyhelminthes, Annelida, Acarina, Insecta, Crustacea and Mollusca. The Silvelra and Marco rivers showed significant differences in the indices of Shannon-Weaver (HI, Simpson's Reciprocal (1/D), Margalef (DMg) and Equitability (E). The Silveira River showed the highest means of diversity and the EPT index (Ephemeroptera, Plecoptera, Trichoptera). Comparison among the diversity indices, considered individually, were insufficient to show differences in community structure, for the purpose of ecological characterization of the rivers. The EPT values characterized the Divisa River as having the highest abundance (73%), followed by the Marco (71%), Antas (48%) and Silveira (36%). These results suggest that the Silveira River is subject to moderate environmental stress, from human impact, although it showed the highest diversity of the major macrobenthic groups.