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Interest for breeding mouflons in Europe is growing by the day, so it is important to extend our knowledge regarding their physiology. To establish reference intervals of biochemical values for mouflon (Ovis orientalis musimon), serum samples from 35 males and 26 females were analyzed using the automatic analyzer Olympus AU 640 for 22 serum biochemistry parameters. All mouflons aged from 1 to 4 years were apparently healthy and in good body condition and kept at an average altitude of 87 m (45A degrees 4’N, 18A degrees 6’E). Obtained results were tested according to age and sex categories by the repeated measurement model with Tukey’s post hoc test. Data showed a significantly higher (P < 0.01) concentration of total serum bilirubin and chloride in adult pregnant female mouflons compared to adult male mouflons. A significantly higher (P < 0.01) value of sodium and a lower (P < 0.01) value of serum phosphate in adult pregnant female mouflons compared to adult male mouflons have been determined. These data provide the baseline information about serum biochemical parameters which can be used to manage the health in mouflons.