Background: The transmission of Human Immunodeficiency Viruses significantly affected byrnthe knowledge and practice of the working population. The utilization of recommended healthrnservices for the prevention HIV is essential among a growing working population in floriculturernsectors.rnObjectives: To assess HIV Prevention Practice and determinant factors among daily laborers ofrnfloriculture in Holeta Town. rnMethods: Institutional based cross sectional study was conducted from March 2013 to Aprilrn2013 to assess HIV prevention practice and determinant factors among daily laborers inrnfloriculture of Holeta Town. Two stage sampling technique was deployed; both quantitative andrnqualitative data were used. A total of 649 daily laborers were selected randomly from the list ofrnfloriculture workers based on the proportion to the size of daily laborer. The data was collectedrnusing a pre-tested and structured questionnaire. Qualitative data was obtained using focus grouprndiscussion. Quantitative data was entered by using EPI info version 3.5.3 then exported to SPSSrn16 version for cleaning and analysis. Logistic regression was employed to identify factorsrninfluencing prevention practice. Qualitative data were transcribed and translated into English,rnand then analyzed manually by grouping the ideas into similar thematic sets. Results: Among 649 who were interviewed, 466(71.8%) were females. The majority of thernstudy participants 591(91%) were knowledgeable towards prevention methods. 210(32.4%) dailyrnlaborers used abstinence as a preventive methods, where as 54.2% laborers used faithful as arnpreventive methods, among sexually active daily laborers 156(35.6%) used condom as arnpreventive methods. Educational status, marital status, age, income, residence and alcoholrndrinking were associated with prevention practice. Study participants who attended primaryrneducation abstained more frequently than those who were illiterate (AOR=1.59; 95% CI (0.78,rn3.22) and those who attended secondary and above educational level utilize condom morernfrequently than those who were illiterate (AOR=2.76; 95%CI (1.35, 5.62). Study participantsrnwho lived in urban less abstained than those who lived in rural (AOR= 0.58; 95% CI (0.37, 0.93)rnwhereas those who lived in urban used condom more frequently than those who lived in ruralrn(AOR=2.976; 95% CI (1.68, 5.26). Study participants who were currently married less likelyrnused condom than those who were not married (AOR=0.27; 95% CI (0.15, 0.50).rnConclusion: The majority study groups were knowledgeable on means of HIV prevention butrnonly a few of them use preventive practice. We also conclude that this study has shown thatrnincomes, age, educational status, marital status, residential areas and alcohol drinking werernfactors which influence preventive practice of HIV.