Background: The neonatal period is the most vulnerable time for child morbidity and mortality. Asphyxia due to poor resuscitation techniques contributed significantly to this vulnerability. Therefore, this study is aimed to assess the extent of knowledge and practice towards neonatal resuscitation among nurses and midwives in public hospitals of south Wollo northeast Ethiopia. Method: Institutional based cross-sectional study design was employed among 143 study participants selected by a simple random sampling method from each hospital. Self-administered questionnaire and interview-based guide were used to collect data. Data were coded and entered into EPI data software version 3.1 and was exported into SPSS version 20 for analysis. Logistic regression with backward LR method was performed to see the possible associations of factors with the outcome variables. Finally, p-values of less than 0.05 in multivariate analysis were declared having a significant association with the outcome variable. Result: One hundred and forty-three participants were included with a response rate of 100%. Only 32.9% and 24.5% of the participants had good knowledge and practice toward neonatal resuscitation respectively. After adjustment was made for covariates; lack of training (AOR: 3.44, 95% CI:1.54-7.68), absence of guideline (AOR: 3.8, 95% CI: 1.68-8.58) and lack of supportive supervision (AOR: 2.49, 95% CI:1.1-5.6) were significantly associated with poor knowledge score, whereas absence of guidelines (AOR: 2.83, 95% CI:1.12-7.15) and lack of supportive supervision (AOR: 5.28, 95% CI: 1.86-14.96) were significantly associated with the poor practice of the participants. Specialization with neonatology (AOR: 0.19, 95% CI: 0.047-0.8) was associated with good practice level of neonatal resuscitation. Conclusion: Knowledge and practice of nurses and midwives toward neonatal resuscitation were suboptimal. The training was found to be a single predictor for knowledge while supportive supervision and availability of resuscitation guidelines were predictors for both knowledge and practice toward neonatal resuscitation. Therefore, regular training and supportive supervision need to be strengthened to fill the identified gaps. Other observational and qualitative studies are recommended to explore factors that contributed to poor knowledge and practice toward neonatal resuscitation.