Objective: This study was conducted to evaluate the effect of water restriction (WR) on physiological and blood parameters in lactating dairy cows reared under Mediterranean climate. Methods: The trial lasted 16 days preceded by two weeks of adaptation to the experimental condition in spring 2014 on 6 dairy cows in mid-lactation. These cows were allowed water ad libitum for 4 days (W100) (hydration period), then split into 2 groups, one group has received 25% and the other 50% of water compared to their mean water consumption during the hydration period; then rehydrated for 4 days. Feed intake and physiological parameters: respiratory rate (RR), heart rate (HR), and rectal temperature (RT) were recorded twice a day. Blood was collected once a day and analyzed for serum concentration of glucose (Glc), triglycerides (TG), cholesterol (Chol), urea (Ur), creatinine (Crea), and total protein (TP) by enzymatic colorimetric method and cortisol (Cort) by radioimmunoassay. Results: Total dry matter intake (TDMI) was affected by WR. A decrease in TDMI was observed in WR groups compared to W100 group (effect, group, period, day, groupxday, periodx day: p<0.001). Also, WR resulted in a significant increase in RR, HR, RT in WR groups than in W100 group (effect, group: p<0.001). In addition, an increase in the serum concentration of Glc, TG, Chol, Ur, Crea, TP, and Cort was noted in WR groups (effect, group, period, day: p<0.001). Conclusion: This study has shown the ability of cows raised in a Mediterranean climate to cope with different levels of WR and thus reach a new equilibrium. As result, elucidates the important role of water as a limiting factor for livestock in environments with low water availability.