Neural Tube Defects (NTDs) are congenital structural abnormalities of the brain (anencephaly) and vertebral column (spina bifida) that represented as one of the most common congenital malformations of neonates worldwide. A fouryear and eight months (January 1 st , 2007 to August 31 st , 2011) retrospective record review study on prevalence, trend, and associated demographic factors of NTD was conducted at Orotta National Referral Maternity Hospital, Asmara, Eritrea. The prevalence, trend and associated demographic factors were assessed by data extraction from hospital delivery register and patient cards. Associations between variables of maternal age, parity, and infant gender and primary outcomes were determined using χ 2 analyses and Poisson regression modeled cumulative incidence and controlled for confounders. Out of 39, 803 total deliveries 185 neonates were found to have NTDs related abnormalities, but 156 neonates identified with complete and welldocumented registers, medical records and charts showing a prevalence of 3.9 per 1000 deliveries. The most commonly identified NTDs were Anencephaly 75(48.1%), Hydrocephalus 29 (18.6%), Spina bifida 27 (17.3%), and Multiple NTDs 25 (16.0%). Marital status, ethnicity, religion, and maternal history of abortion were found to have no enough evidence, but women who delivered neonates with NTDs who had parity less than four 125 (80.1%) and age below 30 years 108 (69.2%)were observed to have higher chances. All the Anencephalic neonates were delivered by vaginal delivery, Majority (66 (88%)) with a weight of less than 2000 grams and were female by sex 55(74.3%); whereas 13(44.8%) Hydrocephalic neonates were delivered by Caesarian section and 26 (92.9%) had a weight greater than 2000 grams. The Appearance Pulse Grimace Activity Respiration(APGAR) score results were founded to be zero for all anencephalic neonates and for 17 (74%) of multiple NTD; while 40 percent of and 46 percent of neonates with spina bifida and hydrocephalus, respectively, had no APGAR at five minutes. Over the years of the study a significant increase in the trend of NTDs were observed, mainly that of anencephaly. The increase was 1.5 percent (p < 0.05). The results of Poisson analysis indicated an exceptional statistically significant increase of anencephaly only. The trend of NTDs in neonates was observed to increase significantly, mainly anencephaly. The study findings noted younger women (less than 30 years) and with parity less than four were at higher risk than those of older age and higher parity. NTDs can be prevented and reduced with folic acid supplementation and fortification of principal foods.