Brucellosis is one of the most common zoonotic diseases caused by Brucella. The disease is located genital organs in the females of mammals such as cattle, sheep, goats, dogs and pigs and cause to infertility, mastitis, preterm birth, placenta retention and metritis; in men it causes orchitis (gonorrhea inflammation). The bacteria can also spread from animals to humans and cause disease in humans. The aim of this study was to determine the prevalence of Brucellosis in milk collected from villages of Tokat province and districts. For this purpose, 161 cows, 58 sheep and 33 goat milk samples were investigated by Brucellosis agent with Milk Ring Test (MRT) and direct sowing method. At the end of the study, 24 of 161 cow's milk, 20 of 58 sheep's milk and 5 of 33 goat's milk were found to be positive by MRT. Of the 252 raw milk samples, 49 (n = 19.41%) were positive for MRT. Brucella melitensis biotype 3 were isolated from the 2 milk samples after the culturing from 49 samples with positive MRT. It was determined that 2 milk samples isolated from Brucea belong to 2 different sheep from the same herd. Thus, Brucella was isolated in 0.79% of all studied samples, 4.08% of MRT positive samples and 3.45% of sheep milk samples. As a result of our first study with milk collected from the villages and towns of Tokat province and districts, 19.41% of the samples were positive for MRT and 0.79% of the samples were isolated by direct planting method. Thus, in this study, it was determined that there is a certain amount of Bruceila agent in Tokat province and its districts and it shows that dairy products may pose a risk for Brucellosis.