Background: Breastfeeding is a critical component of infant nutrition and development. The timing of initiation and the practice of exclusive breastfeeding play a pivotal role in ensuring the health and well-being of infants. In rural areas, where healthcare resources may be limited, understanding the knowledge, attitude, and practices of lactating mothers regarding breastfeeding is essential for designing effective interventions. Materials and Methods: This cross-sectional study was conducted in a tertiary care center in the rural area of Indore. A sample of 350 lactating mothers with infants younger than 6 months was selected using a systematic random sampling technique. Data was collected through structured interviews and questionnaires, focusing on knowledge about the timing of initiation and the practice of exclusive breastfeeding. Attitude and practices were assessed using a Likert scale. Results: The study revealed that 65% of mothers had adequate knowledge regarding the recommended timing of breastfeeding initiation within the first hour after birth. However, only 40% were practicing exclusive breastfeeding, with a significant gap between knowledge and practice. The attitude towards breastfeeding was positive among 85% of mothers. Factors such as maternal education, socioeconomic status, and access to healthcare services were found to influence knowledge and practices. Conclusion: The findings underscore the need for targeted interventions to bridge the gap between knowledge and practice among lactating mothers in rural Indore. Promoting early initiation of breastfeeding and exclusive breastfeeding should be a priority, and strategies should consider factors such as maternal education and socioeconomic status. Improving access to healthcare services and providing education and support to mothers can significantly enhance infant nutrition and health outcomes in this underserved population.