Anaesthetic agents have been implicated in the development of postoperative pneumonia, but the direct effect of volatile anaesthetics on bacterial growth has given contradictory results. The effects of isoflurane on the growth of Staphylococcus aureus and Escherichia coli were investigated under conditions similar to those of clinical practice, using standardized microbiological methods. An open anaesthetic circuit was adjusted to the normal ventilatory settings of an adult patient. A spray, installed on the inspiratory side of the circuit, ensured the delivery of isoflurane at 1.5 minimal alveolar concentration. The bacterial strains studied were both wild-type and reference strains. Bacterial inoculums were prepared to obtain a bacterial exponential growth of 103 colony-forming units per mL in 10mL of nutritive broth. Each strain was studied with and without exposure to isoflurane and measured by the usual criteria of bacterial growth, and by bacterial regrowth after exposure to isoflurane. Under experimental conditions close to clinical practice, exposure to isoflurane did not alter bacterial growth of S aureus and E. coli, or their bacterial regrowth when isoflurane exposure is over.