Background: Toilet training is an important social skill that teaches youngsters how to control the timing of their bladder and bowel movements. It is one of the first steps towards independence that children take. Mother plays a vital role in the training process. Aim of the study: to evaluate effect of a health education program on mothers' performance regarding their toddler children's toilet training. Subjects and Methods: A quasi-experimental study method was used prior to and three months after the intervention. the study was done at four private nurseries in Sohag City such as (al-Mohannadi Group, Toyor Al Ganna, Nasser, and El-Ber). Sample: A purposive sample of 200 mothers with their toddlers was chosen from the prior setting. The study's tools: Three tools were used in this investigation. Tool (1): A structured interviewing questionnaire about personal characteristics and mother's knowledge about toilet training. Tool (II): Mothers' reported practices assessment sheet. Tool (III): Mothers' attitude regarding toilet training scale. Results: 1.5% of the mothers have a good knowledge degree, 35.0% of them had a satisfactory level of practice and 13.0% of the mothers had a positive attitude toward their toddler's toilet training before executing the program. However, After the program was implemented, the mothers' knowledge, practice, and attitude raised significantly. Knowledge, practice, and attitude were all highly significant (p<0.001). Conclusion: the toilet training health education program dramatically enhanced the knowledge, practice, and attitudes of mothers. Recommendations: Publication of the health education program for mothers addressing the toilet training of their toddlers in in different settings.