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Background Despite being a cost-effective means of improving the childhood health indices, exclusive breastfeeding (EBF) remains low in the low middle-income countries. Hence, we evaluated the determinants of EBF among mothers of infants less than six months in Southwestern Nigeria. Methods This was a cross-sectional descriptive study that involved 271 mothers of infants aged less than six months attending the immunization clinic of the Bowen University Teaching Hospital, Ogbomoso, Nigeria. Pretested semi-structured questionnaires were used to get relevant information from the mothers who were recruited using convenience sampling method. Descriptive statistics was carried out while chi square test and binary logistic regression were used for inferential statistics. Results The mean age (±SD) of the respondents was 30.4 ± 5.0 years. The EBF rate in this study was 46.1% (125/271); 40.6% of mothers breastfed their infants within an hour of birth, with most (91.1%) breastfeeding their babies on demand. Factors associated with EBF included mothers' age > 30 years (OR 2.080, 95% CI 1.274-3.395). After controlling for potential confounders, family size > 4, (adjusted OR 2.053, 95% CI 1.120-3.762) and having vaginal delivery (adjusted OR 2.769, 95% CI 1.585-4.829) were the significant determinants of EBF practices among the study participants. Conclusion EBF practice was average in the studied population. Family size >4 and vaginal delivery were the determinants of EBF. There is a need to sustain the promotion of appropriate breastfeeding practices., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2021, Olasinde et al.)