The effects of antibiotic treatment on the results of protected specimen brushing (PSB) in ventilator-associated pneumonia were prospectively assessed by performing this procedure before antibiotic treatment, and 12, 24, 48 and 72 h after initiation of antibiotic treatment, in 35 ventilated patients who developed pneumonia during mechanical ventilation. The number of micro-organisms isolated, their concentration (colony-forming units (cfu) mL(-1)), and the number of cases with a positive PSB (> or =10(3) cfu x mL(-1)) were evaluated. Within 12 h of the initiation of effective antibiotic treatment a rapid, significant decrease in the numbers of organisms isolated, their individual concentrations and the percentage of positive PSB results were observed. Certain bacterial species (Streptococcus pneumoniae, Haemophilus influenzee) appeared to be more vulnerable to antibiotics than others (Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumanni). This data confirms that prior antibiotic treatment, even after only a few hours of activity, significantly decreases the sensitivity of protected brush specimen; this effect appears to be particularly marked among the species involved in early ventilator associated pneumonia.