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This study aimed to evaluate the relative contribution of psychosocial variables to donation behavior in a sample of primary healthcare users in a Brazilian municipality. Donor recruitment and retention continue to pose significant challenges to blood collection services worldwide and is especially important among low and middle income countries (LMICs), challenged with higher rates of stigma and low public awareness about donation. A cross-sectional study with randomized stratified sampling of 1055 primary healthcare users was conducted across 12 healthcare facilities in Ribeirão Preto, state of São Paulo. Participants completed measures assessing psychosocial variables, including religiosity, knowledge regarding blood donation process, overall quality of life, blood donation of peers, sociodemographic variables, and the fear of blood, injections, and fainting. The associations between psychosocial variables and donation behaviour was examined using structural equation modelling. Blood donation was more frequent in males and among individuals with higher socioeconomic and educational levels. The structural model indicated associations between blood donation behaviour and fear, knowledge, age, sex, socioeconomic status, and educational level. Fear of blood, injections and vasovagal reactions, and a lack of knowledge of the donation process were revealed as important barriers to the decision to donate blood. In addition, there is evidence that considering the population of primary healthcare users, the women, the youngers, and those with lower socioeconomic status and lower educational level are less likely to donate blood., (Copyright © 2019 Elsevier Ltd. All rights reserved.)