Heat treatment is the most applied process to ensure the quality and safety of milk and milk products. Several research groups have published data on heat inactivation of Mycobacterium avium subspecies paratuberculosis (MAP) in milk under various pasteurization conditions. In order to determine the effectiveness of pasteurization and homogenization to inactivate MAP in raw milk a series of experiments were performed. Based on the knowledge of critical factors influencing the heat inactivation experiments were designed. To mimic industrial pasteurization a pilot plant scale turbulent flow was used at temperatures between 60 - 90 °C and holding times of 6, 10 and 15 sec and the heat resistance of MAP in milk was determined.MAP infected feces of cows with clinical symptoms of Johne's disease was used to contaminate the raw milk with high concentrations of MAP (initial levels in raw milk from 10² to 3.5 * 10⁴ cells per ml). The different levels of MAP in milk were submitted to an array of time-temperature combinations. MAP levels were monitored by real time PCR and survival was determined by growth in MGIT media.Regression analysis of al data point allowed modeling of the heath inactivation of MAP. Model simulations showed that at the standard high-temperature and short time pasteurization conditions applied in the Dutch dairy industry a >6 log reduction of MAP is achieved. Additional homogenization did not contribute significantly to the inactivation.