Background: Uganda has a high neonatal mortality rate (27 per 1,000 live births), with birth asphyxia as one of the major contributor. Helping babies breathe (HBB) is an evidence-based program that aims to reduce neonatal mortality in resource-limited settings. Successful resuscitation depends on nurses’ and midwives’ knowledge and skills in neonatal resuscitation, and access to functional neonatal resuscitation equipment. This study aimed to evaluate knowledge and skills retention in neonatal resuscitation after HBB training among nurses and midwives, and the state/availability of neonatal resuscitation equipment. Methods: This study used a cross sectional design. Participants were 75 nurses and midwives from two hospitals in Central Uganda. Data were collected using questionnaires and observation checklists. Ethics approval was obtained from the Uganda Christian University and the research and ethics committees of the two hospital hospitals. Results: Nurses and midwives showed a high level of knowledge (92%). However, neonatal resuscitation skills among 44 observed participants were poor, as 68.2% failed to check equipment and select the correct mask and 45.5% did not make a firm seal when applying the mask. In addition, about 72% of participants did not ventilate at a rate of 40 breaths per minute, and 18.2% failed to assess chest movement. Observation of 44 resuscitations to evaluate the state/availability of neonatal resuscitation equipment showed that 27.3% did not have a suction device, 59.1% did not have a heat source/pre-warmed towels to warm the babies, 50% did not have appropriate self-inflating bags and masks for term and preterm babies, 72.7% had no clock/watch to count heart rate and determine the length of time ventilation was required, and 36.4% did not document that resuscitation was performed. Conclusions: To address Uganda’s neonatal mortality rate, it is necessary to implement regulatory policies for neonatal resuscitation, building nurses and midwives skills for active interventions during neonatal resuscitation as well as regular refresher courses to enhance skills. Key words: helping babies breathe (HBB), neonatal resuscitation, knowledge, skills and equipment