National guidelines for cystic fibrosis recommend cleaning and disinfecting nebulizers after each use. We tested two groups of five reusable breath-enhanced nebulizers after 0, 5, 10, 15, 20, 30, 60, 90, 120, 150, and 180 sterilization (baby bottle sterilizer) or cleaning cycles. The nebulizers were operated for 7 min (6 L/min) after loading albuterol (2.5 mg/3 mL), and they were evaluated with and without breathing simulation after cleaning/sterilization (0-180 and 0-60 cycles, respectively). Over the course of 180 cleaning/sterilization cycles, the mean (SD) solution output was 1.33 mL (0.12 mL)/1.29 mL (0.08 mL); the nebulizer mass remaining in the nebulizer was 61.5% (5.2%)/63% (4%); sputtering time was 4.7 min (0.8 min)/4.8 s (0.6 min); inspiratory filter was 19% (3%)/18.5% (2.4%); expiratory filter was 6.7% (1.1%)/6.7% (0.8%); and difference in drug output calculated using the solution output and nebulizer mass was 6.8% (4%)/5.2% (2.9%). Thermal disinfection with a baby-bottle sterilizer did not alter the performance of a reusable breath-enhanced nebulizer. The nebulizer test performed without breathing simulation underestimated its performance. The calculation of the drug output based on the solution output resulted in its overestimation.