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This study was conducted to assess the levels of motivation of 14 motivating factors on academic librarians in four Nigerian Federal Universities based on gender and years of service. The population comprised of 66 librarians from the universities of Abeokuta, Calabar, Ibadan, and Uyo. An 18-item structured questionnaire, with section A soliciting for respondents’ biodata and section B soliciting for respondents’ opinion regarding how much they were motivated by each of the 14 motivating factors, was adopted for data collection. Data were subjected to Analysis of Variance and Duncan’s Multiple Range Test. With regards to gender, results indicated that male librarians were similarly motivated by each of the 14 factors while the female librarians were significantly motivated by Professional development/Growth and Promotion. For years of service, the librarians who have worked for 1 – 10 years and 21 years and above were similarly motivated by all the factors while Training, Professional development/Growth, Conducive environment, Job security, Promotion, and Career advancement significantly motivated those who have worked for 11 – 20 years. It is concluded that the female librarians were better focused on the factors that motivate them than the males; their primary motivator was promotion. Motivation of librarians also varied with years of service.