Among the 251 described species of ticks from the genus Ixodes, only eight were previously reported in Brazil, Ixodes amarali, Ixodes aragaoi, Ixodes auritulus, Ixodes fuscipes, Ixodes loricatus, Ixodes luciae, Ixodes paranaensis and Ixodes schulzei. Of those species, I. loricatus is considered commonly found, whereas I. auritulus and I. aragaoi were registered just one time in Rio Grande do Sul (RS) state, Southern Brazil. This paper aims to update the Ixodes species occurring in RS through the investigation of the ixodofauna of small rodents. Data from 314 wild rodents were analyzed from nine municipalities of Pampa biome and five from the Atlantic Rainforest in RS. Rodents belonging to the following species were infested by Ixodes spp. ticks: Akodon azarae, Akodon montensis, Akodon paranaensis, Akodon reigi, Calomys laucha, Delomys dorsalis, Deltamys kempi, Holochilus brasiliensis, Oligoryzomys flavescens, Oligoryzomys nigripes, Oxymycterus nasutus, Scapteromys tumidus (Cricetidae) and Cavia aperea (Caviidae). The tick identification was performed based on morphological dichotomous keys, their updates and by molecular techniques. Considering the ticks from the rodents and those collected directly from the environment, 34 specimens of Ixodes spp. were collected. Our results improved to five the list of Ixodes species that occur in RS: I. loricatus, I. auritulus, I. aragaoi, I. fuscipes and I. longiscutatus. Moreover, the finding of I. longiscutatus increases to nine the number of Ixodes species in Brazil. The occurrence of two species of the Ixodes ricinus complex (I. aragaoi and I. fuscipes) highlights the potential impact of Ixodes spp. ticks on public health.