Background: Basic food of infant is mother’s milk. Breast feeding should be initiated within the first half an hour of birth. The first milk is the most suitable feed for the newborn. Colostrum is very important for growth, development and protection from infections in infants. The importance of colostrum is still not known to many mothers and caregivers. This study aimed to compare the knowledge and practice regarding colostrum feeding among mothers in urban and rural area.Methods: A descriptive approach and comparative survey design was adopted for the study. Data was collected from 150 mothers (75 each from urban and rural area) having infants of birth to 6 months age. Data was collected using a performa with items on socio-demographic varibles, a structured questionnaire to assess knowledge and a practice questionnaire regarding colostrum feeding.Results: 46.7% urban and 48% rural had moderate level of knowledge and 22.7% mothers from rural had poor knowledge. The urban mothers were more aware about colostrum feeding and its importance than rural mothers. Majority of the mothers (92% urban, 89.3% rural) practiced colostrum feeding. Comparison of knowledge and practice showed statistically significant difference between mothers of urban and rural area.Conclusions: Knowledge regarding colostrum feeding was inadequate and improper breast feeding practices were found in both urban and rural area. Urban area mothers had more knowledge and better feeding practices compared to mothers of rural area. There is an ongoing need for individual and community awareness programmes regarding importance of colostrum feeding.