Background: Weaning is the introduction of food and fluids than breast milk and the transition to a solid diet along with breast milk. It’s gradually starting around the age of 6 months. Weaning is a best process involving nutrition; immunological, biochemical and psychological adjustments. Purpose: The aim of the study was to explore the level knowledge and practice of lactating mothers’ regarding weaning in Bangladesh. Methods: A descriptive study was conducted at three EPI centers in Noakhali Municipality, Noakhali. A purposive sampling technique was used to recruit the sample that has 6 - 12 months of aged baby. The estimated sample size was 120 lactating mothers. Researcher was use developed questionnaires these are socio-demographic characteristic of mothers, mothers’ knowledge related information regarding weaning, mothers’ practice related information regarding weaning. Data was analyzed by using descriptive statistics, t- test and Pearson co-relation. Significance level 0.05 was considered in this study. Results: The mean age of the mothers was 25.18, (SD ± 4.32). Most of the subjects were Muslim. The results revealed that the lactating mothers reported moderate level of total knowledge, with the mean score was 16.13 ± 4.32 and the low level of total practice, mean score was 2.83 ±.27. There were significant factors related to knowledge level were: non-Muslim mothers’ (p = .05), higher educated (p = .03), get information from relative/others (p = .05), and start weaning (p = .01). Significant factors related to practice were older mothers’ age (p = .002), having baby more than one (p = .05) and having older child age (p = .001), higher educated (p = .008), and period of weaning (p = .01). There was no relationship between total knowledge and total practices of weaning among lactating mothers’ (r = .02, p = .82). Discussion: The findings of this study indicated that the mothers who had higher education, more than two babies, and non-Muslim mothers especially were high knowledge and practice regarding weaning. Therefore, the nurses and other health care provider should consider the following characteristics like low educated mothers, prime mothers, starting of weaning, and Muslim mothers especially. Thus, we can prevent of our infant mortality and morbidity regarding weaning practices in Bangladesh as well as abroad.