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Background: The success of national immunization programmes depends largely on effective logistics management of the vaccine cold chain system. This study assessed cold chain equipment functionality, healthcare workers’ knowledge and practice of the logistics management of vaccine cold chain system in Ile-Ife, Nigeria.Methods: A descriptive study was conducted in immunization clinics of 35 health facilities in Ife East and Central Local Government Areas (LGA) in Ile-Ife. There were 100 immunization service providers in the (LGAs) and they were all recruited into the study. Information was obtained with the aid of an interviewer-administered, structured questionnaire. Cold chain equipment functionality was assessed using a checklist. Data were analyzed using SPSS version 20.0.Results: Eleven (31.4%) of the facilities had functional refrigerators for storing vaccines, 16 (45.7%) had cold boxes while 13 (37.1%) had thermometers for vaccine temperature monitoring. Fifty-four (54.0%) of the healthcare workers were aware of the “shake test” and 19 (19.0%) could correctly interpret colour changes on a vaccine vial monitor. Consumption record was considered by 69 (69.0%) of healthcare workers when making vaccine requisitions while the required lead time was considered by 24 (24.0%) of them. Only 29 (29.0%) of healthcare workers kept records of vaccines stock-on-hand.Conclusion: Adequate training and supportive supervision is essential to improve healthcare workers’ knowledge and cold chain practices. Relevant cold chain equipment should be provided to boost storage capacity across health facilities.Keywords: Vaccine cold chain; Logistics management; Nigeria