Milk is the preferred nutrient for microbes to grow. Under normal conditions the number of microflora is 103CFU/mL of milk. In cows infected with inflammation (masitis) can be found bacteria types Corybacterium, Staphylococcus sp, Streptococcus sp, and Enterobacteriaceae whose number can reach 105 CFU/mL of milk. The purpose of this study was to determine the total number of bacteria in fresh cow's milk in the Kalijambe area, Sragen. The method in this study uses microbiological tests with three tests, namely breed, MBRT and rezasurin. There are 4 types of fresh cow's milk samples used in this study, namely fresh cow's milk A, fresh cow's milk B, fresh cow's milk C and fresh cow's milk D, where in each type of milk there is one repetition (in duplicate). ). Based on the three tests that have been carried out, it can be concluded that milk samples A1, A2, B1, and B2 are suitable for consumption and milk samples C1, C2, D1 and D2 are not suitable for consumption because the amount of bacterial contamination exceeds the minimum limit of SNI 3141.1-2011 (1x106CFU/ml) . The results of the "t" test showed that the total number of microbes in fresh milk in 4 samples (A, B, C, D) was significantly different (P < 0.05), so it can be concluded that the quality of fresh milk at Kalijambe farm, Sragen is good for milk A and B (according to SNI 2011), while for milk C and D are not good (not in accordance with SNI 2011).