Background: Nurses are particularly vulnerable to nosocomial tuberculosis (TB) infection because, being in the frontline of healthcare provision, they are frequently exposed to patients with infectious TB disease. Although cost-effective measures are available for TB infection control (TBIC), they are often poorly implemented. Knowledge of TBIC is known to positively influence the practice of the measures. There is, however, paucity of data on the knowledge and practices regarding TBIC among nurses in Nigeria. This study was aimed at determining the levels of TBIC-related knowledge and practices of nurses in Ibadan, and their associated socio-demographic factors. Methods: This cross-sectional study utilized a self-administered questionnaire to collect data from 200 nurses in two secondary health facilities, in May 2014. The mean knowledge and practice scores of the nurses were determined and logistic regression was utilized to explore the association between the scores and socio-demographic characteristics. Results: The respondents had mean knowledge and practice scores of 68.2 and 79.9% respectively. Using cut-off points of 80 and 100% for good knowledge and practice scores respectively, small proportions of the nurses had good scores-knowledge (10.5%) and practice (6%). Knowledge was not significantly associated with the socio-demographic characteristics of the nurses. Work experience was the only factor that was significantly associated with practices, with the more experienced nurses (> 18 years of work experience) having lower odds of obtaining good practice scores (OR 0.25, 95% CI 0.06-0.94). There was also no significant association between knowledge and practice scores (the nurses were yet to be trained on the newly-introduced TBIC package at the time of the study). Conclusions: The study revealed that small proportions of the nurses had good knowledge and practice scores. Its findings will be useful for the designing of interventions to improve TBIC among nurses and other healthcare workers, and to benchmark evaluation of the interventions. It is recommended that nurses should be trained on TBIC to equip them with necessary knowledge and skills. This, together with appropriate policy directives, and adequate monitoring and supervision will contribute to optimal implementation of TB preventive measures.