Animal health professionals act as the first line of defence in zoonoses transmission. By handling animals on daily basis, they are at risk of contracting zoonotic diseases. However, lack of data on animal health professional risk perception on zoonotic disease and preventive practices may hamper decision making and policies to fight against transmissible disease at the human-animal and environmental interface. Therefore, a questionnaire-based survey of 272 animal health professional was done between July and December 2022 to assess their knowledge, risk perception on zoonosis and the implementation of preventive practices in four regions of Cameroon. Data obtained was recorded into binary scale and the sum of sufficient responses recorded for each participant was divided by the total number of items within the category. A one-way ANOVA was performed to identify factors associated with sufficient knowledge on zoonosis. The respondents mean knowledge scores of 0.79 ± 0.15 towards zoonosis was obtained. Gender and educational level significantly influence the knowledge scores on zoonosis. More than half (60.5%) of veterinarians perceived wild animals as the highest zoonotic diseases transmitter species, while for 50.26% of para-veterinarians, the highest risk was related to pets. All respondents considered there was low risk of contracting a zoonotic infection during assistance in parturition and when handling apparently healthy animals. Less than one third of the veterinarians reported using adequate personal protective equipment for surgery (17.3%), necropsy (4.7%) and delivery (11.1%), and not even one para-veterinarian reported using adequate personal protective equipment for the same cases. Factors such as regions, service sector, risk perception when handling apparently healthy animals and when getting in contact with animal saliva and animal faeces were significantly associated with preventive practices against zoonotic infection risks. Reducing the risk of occupationally acquired zoonotic infections requires continuous education and sensitization of all stakeholders