Loxoscelism is a health problem caused by the bite of spiders of the genus Loxosceles. In Chile all cases are attributable to Loxosceles laeta. It has been suggested that the spitting spider Scytodes globula may be a predator of L. laeta and control its population, which is only possible if they share the microhabitat. This study compared the thermal preferences and tolerances of the two species. Later, spiders acclimated to 15°C and 25°C were exposed to decreasing and increasing temperatures to determine the lower and upper critical temperatures. The preferred temperatures were lower during the morning, but there were no differences between the species. The thermal niche breadths were similar for the species, with a large overlap. Both species showed tolerance to extreme temperatures, but L. laeta showed greater tolerance to low temperatures. Both species showed acclimation of the lower critical temperatures to changes in acclimation temperatures. The similarity of preferred and tolerated temperatures was partly an expected fact, since the species share the same macrohabitat; these spider species are very common in domestic environments of central Chile. However, the results imply that their microhabitat choices are also very similar, indicating a high probability of meeting and predation, which could have important consequences in loxoscelism epidemiology. •Scytodes globula may be a predator of Loxosceles laeta, a species which causes great proportion of loxoscelism in America.•Preferred temperatures of both species were lower during the morning, without differences between species.•Thermal niche breadths were similar for both species, with a large overlap.•Both species showed a high tolerance to extreme temperatures as adaptation to xeric environments.•Microhabitat choices were similar, determining high probability of meeting and predation, which could have important consequences in loxoscelism epidemiology.