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Objective To assess if positive end‐expiratory pressure (PEEP) titration improves gas exchange and respiratory mechanics, without hemodynamic impairment in horses during anesthesia. Design Prospective, randomized study. Animals Thirteen isoflurane‐anesthetized healthy horses. Methods After 60 minutes of anesthesia with spontaneous breathing, mechanical ventilation was initiated with an inspiratory‐expiratory ratio of 1: 2, PEEP of 5 cmH 2 O, tidal volume of 10–20 mL kg− 1 and respiratory rate adjusted to maintain normocapnia. Constant PEEP of 5 cmH 2 O was continued (control group; n= 6) or titrated (PEEP group; n= 7) by increasing and decreasing PEEP from 5 to 20 cmH 2 O at 15‐minute intervals. The horses were instrumented with an arterial catheter to measure blood pressure and allow collection of blood for pH and blood gas analysis and a Swan‐Ganz catheter for measurement of cardiac …