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Abstract Background Stepfamilies remain poorly understood in Japanese society, and the support needs of stepfamily mothers are unclear. This study aimed to identify characteristics of stepfamilies and maternal mental health as compared with non-stepfamilies in Japan to utilize as a primary resource for providing effective support through community-based health care for stepfamilies. Methods From December 2011 to July 2012, we conducted this questionnaire survey with mothers at 3- and 4-month checkups for infants. The response rate was 75.1%. The sample for analysis included responses of 2246 mothers, excluding single mothers. Results Respondents comprised 47 (2.1%) stepfamilies and 2199 (97.9%) non-stepfamilies. There were significantly higher rates of parents with not more than a high school education and ≥3 children among stepfamilies compared with non-stepfamilies. Stepfamily mothers had significantly higher rates of feeling a lack of economic resources, absence of participation in childbirth education classes, smoking during pregnancy, and unplanned pregnancy. Furthermore, they also had significantly higher rates of depression and a lack of confidence in the parent role. Maternal depression was associated with factors such as maternal age, self-perceived health, stress level, confidence in breastfeeding, confidence in the parent role, and number of children. Conclusions These findings suggest that stepfamilies exhibit many characteristics related to social disadvantage and problems with community-based health care in Japan. Healthcare providers should be aware of stepfamily mothers’ support needs and should put in place a support system for stepfamilies. Moreover, compared with non-stepfamily mothers, stepfamily mothers have a significantly higher prevalence of depression. However, stepfamily composition does not necessarily increase the risk of maternal depression. Therefore, healthcare providers should put in place a system for obtaining more thorough information about stepfamilies and conduct an early assessment to identify their support needs.