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The present study was conducted to investigate certain stress parameters of Ram-Ewe mixed transport group compared with single sex groups in indigenous Red Karaman and Imroz sheep. Sheep from each breed were divided into three transport groups: a. Ram group (8 rams from each breed), b. Ewe group (8 ewes from each breed) and c. Ram-Ewe mixed group (4 rams and 4 ewes from each breed). Sheep from six different sub-groups were transported for 75 min and then lairaged for 2-h, separately. The influence of transport group on concentrations of plasma Cortisol, CK, LDH, glucose, and neutrophile: lymphocyte ratio were not significant (P>0.05). The differences among Ram, Ewe and Ram-Ewe mixed groups for percentages of "standing", "lying" and "walking" behaviours, and frequencies of "butting other animals" and "urination" during 2-h lairage period were significant. Ram group exhibited higher "lying" behaviour and lower "standing" behaviour than those of Ewe group and Ram-Ewe mixed group (P<0.01). "Time spent for walking" percentage was lower in Ram group than that of Ram-Ewe mixed group (P<0.05). Moreover, "butting other animals" was observed more frequently in Ram group than Ewe group (P<0.01). Sheep from Red Karaman breed had higher plasma Cortisol level measured after transport (P<0.001) and after lairage (P<0.01) compared with those of Imroz breed. The percentage of "standing" behaviour (PO.01) and the frequency of "vocalisation" behaviour (PO.001) observed in Red Karaman sheep were higher than those of Imroz sheep. As a consequence, transportation resulted with stress responses in Red Karaman and Imroz sheep, but Ram-Ewe mixed transportation did not cause additional stress responses compared with single sex groups. Therefore, it was concluded that the mixed transportation of rams and ewes during the non-breeding season did not cause any negative result in terms of animal welfare.