Fever in newborn remains one of the major reasons of pediatric consultations. Utilization of the healthcare facility is sometimes late leading to the increased risk of death. The aim of our study was to evaluate knowledge and practices of nursing mothers attending the immunization ward and to identify influencing factors. This was a cross-sectional descriptive and analytical study conducted in February 2014 in Biyem-Assi health district, center region, Cameroon. The interview was done using a standard paper-based questionnaire. Data entry and analysis was performed using EPI Info software version 3.5.3. A total of 275 mothers was interviewed. The median age was 26, IQR [23; 30) years. About 82.9% (n=228) had attended at least secondary school and 1.5% had not been to school. About 54.6% (n=149/273) of children present for immunization were females; median age was 03 months, IQR [2; 9]. Half (137) of our respondents had good knowledge of fever in newborn. Concerning management of fever in newborn, 62.9% (n=173) of mothers reported self-medication compared with 22.9% (n=102) who, in case of fever, went to a healthcare facility. Level of education and occupation appeared to be the key factors that influenced knowledge and practices on fever in newborns. The level of knowledge and practices of mothers in Biyem-Assi health district concerning fever in the newborn seems to be quite good. However, given the factors influencing them, there is need to further educate mothers on screening and appropriate management of fever.