Despite early diagnosis and appropriate antibiotic therapy, ventilator-associated pneumonia (VAP) remains the leading cause of death from hospital-acquired infection in ventilator-dependent patients. Strategies to prevent bacterial colonization of the trachea and lungs are the key to decrease mortality, hospital length of stay, and cost. It is well established that the VAP can result from entry of infected oropharyngeal/gastric secretions into the lower airways. Aspiration may occur during 1) intubation, 2) mechanical ventilation through leakage around the tracheal tube cuff, 3) suctioning of the tracheal tube when bacteria can detach from the biofilm within the tube, or 4) areosolization of bacterial biofilm during mechanical ventilation through the tracheal tube or the ventilator circuit biofilm. From experimental studies in sheep, we drew 3 relevant conclusions: 1) The tracheal tube and neck should be oriented horizontal/below horizontal to prevent aspiration of colonized secretions and subsequent bacterial colonization of the lower respiratory tract. 2) Continuous aspiration of subglottic secretions (CASS) can lower bacterial colonization of the respiratory tract, but at the price of severe tracheal mucosal damage at the level of the suction port. 3) Coating the interior of the tracheal tube with bactericidal agents can prevent bacterial colonization of the tube surface and of the entire respiratory circuit, during 24 hours of mechanical ventilation.