The high level of pathogenic microbial contamination in milk can reduce milk quality. Milk as a source of animal protein is produced by dairy cattle with complete nutritional content. The complete nutritional content causes milk to be an ideal medium for bacterial growth. The bacteria that are found in milk are generally the result of cross-contamination. The contamination can occur due to the wrong handling of milk, both from the milking process to the processing process. Method to minimize the number of microbes due to contamination in milk have been carried out, one of which is heat treatment, but in practice, the wrong application of heat treatment can cause protein damage in milk. Another method that can be done is to use ozone technology. Ozone is one of the sterilization methods using ozone (O 3 ) to oxidize bacterial cell walls. Sterilization can injure or kill bacteria that found in milk. This research carried out through several stages, one of which is done by adding 2%, 4%, 6% and 8% fat and protein, each sample injected with Escherichia coli, Staphylococcus aureus and Listeria monocytogenes bacteria. This study aims to test the resistance of pathogenic bacteria that are commonly found in milk. The results showed an increase in the resistance of bacteria to ozone.