BACKGROUND AND PURPOSE Studying the effects of toxic and infective compounds on the respiratory system requires a reliable method for delivering inoculum into the distal region of the lung. Although transoral intratracheal inoculation methods have been well documented for adult rats, to the authors' knowledge, a reliable method has not been validated for neonatal rats. The purpose of the study reported here was to develop a simple method for transoral inoculation in rat neonates. METHODS Seven-day-old Fischer 344 rats were anesthetized with halothane, and a spinal needle was inserted in the tracheal lumen, by use of illumination and a modified otoscope. Meconium was injected into the lungs as a marker, and the neonates were kept under close observation. After euthanasia at 24 h, lungs were removed and fixed in formalin, and the microscopic distribution of the inoculum was assessed in the left, right cranial, middle, median, and caudal lung lobes. RESULTS Microscopic examination of lungs indicated that intratracheal inoculation was achieved in 100% of neonatal lungs and the inoculum was consistently distributed in the alveoli of all pulmonary lobes. Important complications or mortality were not observed in the neonates. CONCLUSIONS Intratracheal inoculation of neonatal rats is possible by use of a modified otoscope for transoral illumination. This technique is simple and reproducible and ensures, without complications, widespread distribution of inoculum in the lungs of neonatal rats.