Amblyomma rotundatum Koch is a parthenogenetic tick usually associated with reptiles and amphibians. However, relatively few studies on occurrences of ticks in wild reptile populations in Brazil have been produced. The aim of this study was to analyze the presence of ticks associated with reptile species in the Grussaí restinga, in the municipality of São João da Barra, state of Rio de Janeiro, Brazil. Between December 2010 and January 2011, 131 individuals belonging to nine species of reptiles of the order Squamata were sampled: the lizards Tropidurus torquatus (n = 51), Hemidactylus mabouia (n = 25), Mabuya agilis (n = 30), Mabuya macrorhyncha (n = 6), Cnemidophorus littoralis (n = 5) and Ameiva ameiva (n = 10); and the snakes Philodryas olfersii (n = 2), Oxyrhopus rhombifer (n = 1) and Micrurus corallinus (n = 1). The only tick species found to be associated with any of the reptiles sampled was A. rotundatum. One adult female was detected on one individual of the lizard A. ameiva, one nymph on one individual of the lizard T. torquatus and four nymphs on one individual of the snake P. olfersii. This study is the first record of parasitism of A. rotundatum involving the reptiles T. torquatus and P. olfersii as hosts. Our results suggest that in the Grussaí restinga habitat, A. rotundatum may use different species of reptiles to complete its life cycle.