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Background: Poor resuscitation contributes significantly to neonatal deaths globally. Helping Babies Breathe (HBB) is a new evidence-based neonatal resuscitation curriculum for low-resource settings. Objective: We sought to characterize knowledge changes after national-level HBB training in Ethiopia, factors correlated with successful training, resuscitation skills and trainees’ perceptions. Methods: Trainees completed multiple-choice questionnaires (MCQ) before and after a 2-day course. After training, bag-mask ventilation (BMV) skills were assessed and feedback questionnaires completed. Results: Resuscitation knowledge improved from 8.7/10 (SD 1.4) to 9.4/10 (SD 1.1; p = 0.003). Correct MCQ responses relating to essential aspects of resuscitation increased 68–79%. Pre-training knowledge differences between physicians and non-physicians disappeared. MCQ scores increased as trainer:trainee ratio decreased (p = 0.004). Mean post-HBB BMV scores [5.7/7 (SD 1.6)] were not impacted by trainer:trainee ratio. Conclusions: Ethiopian HBB training improved neonatal resuscitation knowledge and was well received. Lower trainer:trainee ratio was associated with increased MCQ scores. HBB eliminated baseline knowledge differences between Ethiopian healthworker cadres. [ABSTRACT FROM PUBLISHER]