Background; Neural tube defects are a group of severe human congenital malformations caused by the defective closure of neural tube, which ultimately will form a baby’s brain and spine, between 21 to 28 days of conception. It is the second most common type of congenital anomaly and the leading cause of infant mortality and disability in the world. Complex interactions between genetic and environmental factors are responsible for this. This study aims to identify the risk factors associated with neural tube defects, in Tibebe Ghion Specialized Hospital, Bahir Dar, Ethiopia. Methods: an unmatched case control study was conducted among 256 study participants (43 cases and 213 controls), in TGSH, from March 2022 to September 2022. The cases were selected with physicians’ diagnosis of confirmed Neural tube defects, and the controls are selected using systematic sampling method, among the neonates admitted to Tibebe Ghion Specialized Hospital, within the time period. The data was collected using pretested structured questionnaire. The data analysis was done using IBM SPSS 25. Binary logistic regression was done to determine the association between independent variables and the outcome variable. Those variables with p value ≤ 0.25 in Bivariate logistic regression were entered to multivariate logistic regression analysis to control confounders. Variables with p value <0.05 were considered as statistically significant. Results: After controlling the covariates, rural residency (AOR: 6.35, 95% CI: 2.14, 18.81) and pesticide use (AOR: 3.94, 95% CI: 1.08, 14.40) were the identified risk factors for NTDs. Conclusion and recommendation– Urgent attentions needs to be given for pesticide exposure, to decrease NTDs affected birth. Further studies, which are supported with laboratory measurement of serum pesticide level, are needed. Keywords: Neural tube defects, Pesticides, Neonates, Ethiopia