Effects of vacuum and pulsation ratio on udder health were studied for 36 first parity animals in a 60-day trial. Treatments consisted of vacuum at 33.3, 41.6, and 50 kPa at pulsation ratios of 50:50, 60:40, and 70:30 at 60 pulsations per min. All teats were exposed to a culture broth of Staphylococcus aureus after machine removal. Numbers of intramammary infections and somatic cells were determined. Analysis for number of intramammary infections indicated no significant differences among treatments for number of infected available quarters. Trend for intramammary infections was that as pulsation ratio widened, the number of infected available quarters increased, especially at the ratio 70:30. As vacuum increased, number of infected available quarters increased. Least squares means of Wisconsin Mastitis Test scores were 6.29, 5.57, and 6.68 for 33.3 kPa; 12.18, 3.82, and 7.86 for 41.6 kPa; and 9.11, 6.40, and 15.02 for 50 kPa at pulsation ratios 50:50, 60:40, and 70:30. There were significant differences among treatments for vacuum and pulsation ratio. Wisconsin Mastitis Test data indicated an interaction between vacuum and pulsation ratio. Optimum predicted vacuum and pulsation ratio based on Wisconsin Mastitis Test data were 27.2 kPa and 62:38.