Rhipicephalus sanguineus is a species complex of ticks that vector disease worldwide. Feeding primarily on dogs, members of the complex also feed incidentally on humans, potentially transmitting disease agents such as Rickettsia rickettsii, R. conorii, and Ehrlichia species. There are two genetic Rh. sanguineus lineages in North America, designated as the temperate and tropical lineages, which had occurred in discrete locations, although there is now range overlap in parts of California and Arizona. Rh. sanguineus in Europe are reportedly more aggressive toward humans during hot weather, increasing the risk of pathogen transmission to humans. The aim of this study was to assess the impact of hot weather on choice between humans and dog hosts among tropical and temperate lineage Rh. sanguineus individuals. Ticks in a two-choice olfactometer migrated toward a dog or human in trials at room (23.5°C) or high temperature (38°C). At 38°C, 2.5 times more tropical lineage adults chose humans compared with room temperature, whereas temperate lineage adults demonstrated a 66% reduction in preference for dogs and a slight increase in preference for humans. Fewer nymphs chose either host at 38°C than at room temperature in both lineages. These results demonstrate that risk of disease transmission to humans may be increased during periods of hot weather, where either lineage is present, and that hot weather events associated with climatic change may result in more frequent rickettsial disease outbreaks.