We report the finding of Amblyomma aureolatum (Acari: Ixodidae) parasitizing margay (Leopardus wiedii) in the locality of Gravataí (29° 47' 12.9" S and 50° 53' 44.1" W; 241 m alt.), State of Rio Grande do Sul, Southern Brazil. This is the first report of this species of Ixodidae on margay in Southern Brazil. This finding indicates an increased number of hosts for this tick species.