Corynebacterium pseudotuberculosis is the causative agent of caseous lymphadenitis (CLA) in small ruminants. This study was designed to evaluate the efficacy of 0.5% and 1% formalin killed prototype vaccine of C. pseudotuberculosis in bucks and does. A total of 24 healthy bucks and does were equally divided into four groups of 6 animals. Groups A and B were vaccinated with 0.5% and 1% formalin killed prototype vaccine and groups C and D were kept as positive (+) and negative (-) control groups, inoculated with plain vaccine adjuvant. After booster vaccination at the 6th week, groups A, B and C were experimentally inoculated with 2mL of 106 cfu of live C. pseudotuberculosis subcutaneously. On the 12th week, all animals were euthanized and samples of reproductive organs and pituitary glands of both bucks and does were collected. Bacterial isolation and identification was done on the basis of morphological characteristics and PCR detection. Based on the bacterial culture results, 3 samples each were Positive for C. pseudotuberculosis colonies from bucks in groups A and B, while 4 samples were positive from does in group A and only 1 from does in group B. All bucks and does from groups A and B showed negative PCR amplifications in all samples. This study showed that 1% formalin killed prototype vaccine of C. pseudotuberculosis is effective in preventing the spread of the bacteria to the reproductive organs and pituitary glands of both bucks and does. Thus, it will be a suitable preventive measure against CLA in small ruminants.