Botulinum toxin (BTX) is a potent neurotoxin produced by Clostridium botulinum and has wide usage in different areas. The current study aimed to analyze the effects of C. botulinum toxin on the central nerve system in chick embryos. Forty fertile Hubbard Broil eggs, all at Stage 8 of development, were divided into four equal groups: Group 1 embryos (n=10), the control group, were explanted and grown for 18 h in a nutrient medium (thin albumin). Group 2 embryos (n=10) were grown in medium containing 5 U BTX, Group 3 embryos (n=10) in a medium containing 10 U BTX and Group 4 embryos (n=10) in medium containing 20 U BTX. After the incubation period, 80% of Group 1 and 2 embryos and 90% of Group 3 and 4 embryos had intact neural tubes (P>0.05). The results of this study suggest that BTX had no additional effect on neural tube development in early chick embryos.