Background: Despite its strong growth in many parts of the world, mobile health access is still limited in low-and middle-income countries. Among the many factors restricting implementation are the lack of information security, insufficient evidence base, low sensitization, and user acceptance. Limited evidence has been obtained on current practices, perceptions and user acceptability in such settings. The aim of this study was therefore to evaluate the knowledge, attitude and perceptions on mobile health use among health workers and veterinary officers in Uganda. Material and Methods: A cross section study was carried out, targeting health practitioners in both hospitals and veterinary laboratories/clinics. A structured questionnaire was used to collect data from the Central, Eastern, Northern and Western representative regions. Interviews with selected health workers were also conducted as well as a focused group discussion. Results: Out of the one hundred and twenty health practitioners that were targeted, a total of eighty health workers and seven veterinary practitioners participated in the study of which 46% were male and 54% female. Majority of the health workers had encountered mHealth but had never used it, whereas the fifteen practitioners who had used it before the survey did not use it for disease diagnosis in hospitals but used it for ordering medicine online, for patient consultations with the doctors, result interpretation, tracking women menstrual cycles, TB assessment and phone in gloves resuscitating the baby. Discussion and Conclusion: Participants expressed significant interest in mobile health as it addresses key challenges including challenges with management of patient data, and long patient queues, which would ultimately improve service delivery. However, there is some scepticism about access as many rural facilities lack access to smartphones and stable internet.