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Exclusive breastfeeding is recommended as the best feeding approach for new born infantsdue to its enormous benefits to the mother and child. This study assessed the sociodemographic,physiological and individual factors influencing exclusive breastfeedingamong lactating mothers attending Fort Portal regional referral hospital in Western Uganda.The study population was drawn from lactating mothers attending Fort Portal regionalreferral hospital. A cross-sectional study design involving 201 lactating for 6-12 monthswhich employed both quantitative and qualitative methods in data collection was used forthis study. Simple random sampling technique was used to sample the respondents (lactatingmothers) at each facility. A Well tested and validated questionnaires and focus group guidewere used to collect data elicit information from the lactating mothers. Data from the surveywas statistically analyzed using the Statistical Package for Social Sciences (SPSS) (version22.0). The study showed that initiation of breastfeeding after birth was done within the firsthour of birth by most of the mothers 126 (62.7%), majority of the mothers had fed theirbabies other food substances with breast milk 132 (65.7%). The socio-demographic factorswhich significantly influenced exclusive breastfeeding among the lactating mothers were themarital status (X2 =17.715, p =<0.001), the level of education (X2 = 10.680, p = 0.014) andoccupation (X2 = 14.431, p = 0.002). The physiological factors which influence exclusivebreastfeeding among the lactating mothers were the mode of delivery (X2 = 12.258, p<0.001),the HIV status of the mothers (X2 = 12.913, p <0.001) and the breastfeeding disordersexperienced (X2 = 16.538, p = 0.001). Mothers who had delivered vaginally were twice morelikely to practice exclusive breastfeeding (OR = 2) and HIV negative. The rate of exclusivebreastfeeding among lactating mothers attending Fort Portal regional referral hospital is low;this is mainly influenced by individual, physiological factors and to a smaller extent, sociodemographicfactors. Hence health facility deliveries should be promoted by the districthealth authorities to enhance child education after delivery.