Fossils of megaherbivores from eight late Pleistocene 14 C-and OSL-dated doline infillings of Ajoie (NW Switzerland) were discovered along the Transjurane highway in the Swiss Jura. Carbon and oxygen analyses of enamel were performed on forty-six teeth of large mammals (Equus germanicus, Mammuthus primigenius, Coelodonta antiquitatis, and Bison priscus), coming from one doline in Boncourt (~80 ka, marine oxygen isotope stage MIS5a) and seven in Courtedoux (51-27 ka, late MIS3), in order to reconstruct the paleoclimatic and paleoenvironmental conditions of the region. Similar enamel δ 13 C values for both periods, ranging from −14.5 to −9.2‰, indicate that the megaherbivores lived in a C 3 plant-dominated environment. Enamel δ 18 O PO4 values range from 10.9 to 16.3‰ with a mean of 13.5 ± 1.0‰ (n = 46). Mean air temperatures (MATs) were inferred using species-specific δ 18 O PO4 -δ 18 O H2O -calibrations for modern mammals and a present-day precipitation δ 18 O H2O -MAT relation for Switzerland. Similar average MATs of 6.6 ± 3.6°C for the deposits dated to~80 ka and 6.5 ± 3.3°C for those dated to the interval 51-27 ka were estimated. This suggests that these mammals in the Ajoie area lived in mild periods of the late Pleistocene with MATs only about 2.5°C lower than modern-day temperatures.