Background: Neural tube defects are prevalent malformations of the central nervous system in infants, often fatal, and considered global medical-social problems. Due to National Laws fortifying wheat flour in the bread industry, 100% of the population of this province has consumed wheat flour enriched with folic acid since June 2007. Objectives: This study aimed to determine the prevalence of neural tube defects following the fortification of flour with folic acid in the north of Iran. Methods: This descriptive-analytical study examined all neural tube defect cases in Golestan province, northern Iran, from March 2016 to March 2018. Data on the parent's age, ethnicity, mother's education, residency, and types of neural tube defects were recorded for each subject. Results: The prevalence rate of NTDs was 1.04 per 1000 births. The highest frequency was observed in the Turkmen ethnic group, with a prevalence rate of 1.251 per 1000 births. Geographically, most cases were in the West of Golestan province, with a prevalence rate of 1.876 per 1000 births. Additionally, the highest prevalence of NTD in Kalaleh City was reported as 1.67 per 1000 births. Among NTD cases, the highest frequency was related to mothers aged less than 35 years (82.72%). Conclusions: The prevalence of neural tube defects during 2016 - 2018 has increased compared to the previous study conducted in this region from 2006 - 2009. Future studies should investigate the reasons behind this increase.