Objectives: The objectives of this research were to test knowledge, attitudes and practice of risks, ways of transmission, behaviour and prevention of blood-borne infections among voluntary blood donors of Clinical Hospital Centre Osijek and to test whether there is any difference regarding age, gender and degree of education. Study design: cross-sectional study Participants and Methods: The study involved voluntary blood donors of Clinical Hospital Centre Osijek. Socio-demographic data and data on knowledge, attitudes and practices of voluntary blood donors about the blood-borne infection were collected via newly formed questionnaire. Results: 130 voluntary blood donors participated in the study. More than 80% of the correct answers were given to 5 out of 7 questions that are related to knowledge of the participants about the risks and the ways of transmitting blood-borne infections. There are no significant differences in knowledge regarding age, gender and education. The attitudes of voluntary blood donors on the prevention of blood-borne infections were positively developed, except for statement about vaccination against hepatitis B, where 35 (27%) participants stated that they did not agree or disagree, and 9 (7%) participants said that they did not agree or they completely disagree with that statement. Women are more likely to agree with statements about attitudes related to blood-borne infections. There are no significant differences in the behaviour of the participants regarding gender, age and education. Conclusion: Knowledge, attitudes and practice of voluntary blood donors are satisfying. Women have more positive attitudes about the prevention of blood-borne infections then men. About 1/4 of the respondents did not have a positive attitude towards vaccination against HBV.