The aim of this communication is to report, for the first time, the occurrence of Amblyomma triste in Santa Fe province, Argentina, and to add a new isolation place for Amblyomma tigrinum. Both species of ticks are vectors of Rickettsia parkeri, a spotted fever group rickettsia. Ticks were recovered from tourists in August 2014 and December 2015 at the Federico Wildermuth Foundation (31° 59'S, 61° 24'O), San Martin Department, Santa Fe province. Five adult ticks were morphologically identified as A. tigrinum (3 females and 1 male) and A. triste (1 female). This is the first finding including both Amblyomma maculatum group species, A. triste and A. tigrinum, together in the same locality in Argentina. This finding suggests that this site might have favorable features for the development of both species of R. parkeri vector. Further studies including sampling of a larger number of ticks and detection of R. parkeri DNA are needed to better document the epidemiology of this rickettsia in Santa Fe.