Background and Aims: Poultry vaccines are intensively used to prevent and control infectious poultry diseases. The aim of vaccination is to reduce the occurrence of clinical disease. Several factors affect vaccination programmers which emphasize the crucial responsibility of person designing the vaccination program. There are limited studies evaluating the knowledge and attitude of veterinarians toward veterinary vaccination. A veterinarian working in veterinary vaccine supplier and administrator centers have a significant role in assisting farmers and poultry owners for vaccination and disease control decisions. The objective of this study was to evaluate the perception, knowledge, and attitude of a veterinarian regarding poultry vaccination. Semi-structured interviews were conducted in 31 centers in different provinces of Iran. The data were analyzed using Excel and SPSS. The analysis revealed that most of the veterinarians are highly familiar with veterinary vaccines, vaccination programs, side effects and effect of maternal antibody on vaccination. However, most of them believed that poultry owners have low knowledge about vaccination and decided to put the opinion of poultry owners in low priority during decision making. Effectiveness of the vaccine and scientific evaluation of new vaccine was the main criteria for choosing a vaccine for the herd. Most respondents believed that proper vaccination program and matching between field and vaccine are the main factors for successful vaccination. Furthermore, they believed that the most effective route of vaccination for ND, IBD, and IB are eye drop, drinking water and spray respectively. Further evaluation of poultry owners regarding their knowledge of vaccination is recommended.