The aim of this study was to compare Kangal shepherd dogs with and without ankyloglossia in terms of various physiological and behavioral changes. A total of 24 dogs were used, 11 of which have had ankyloglossia and 13 of which did not have ankyloglossia. Heart rates, respiratory rates, and body temperatures of the dogs were measured at three different environmental temperatures (18, 23, and 28 degrees C), resting state, and different exercise durations (5, 10, and 15 min). In addition, resting state and some behavioral parameters were recorded with a video camera. Mann-Whitney U, Kruskal-Wallis, and Wilcoxon tests were used to analyze the data. While no statistical difference was detected in the two groups in terms of all parameters during the resting state at all temperatures, the difference between the resting state and exercising was statistically significant (P < 0.05, P < 0.01). Except for 15 min of exercise for the respiration rates, statistical differences were detected in both groups in parameters at both different temperatures and different exercise durations at each temperature. While some of the stress behavior parameters were not detected at any of the temperatures in normal dogs, the number of the dogs showing other stress behavioral parameters increased in both groups as the temperature increased, but the number was higher in the dogs with ankyloglossia. This study is the first study on physiological and stress behavioral parameters in Kangal shepherd dogs with ankyloglossia.