Milk pH is increased in lactating dairy cattle with subclinical mastitis (SCM) and intramammary infection (IMI). We hypothesized that milk pH testing provides an accurate, low-cost, and practical on-farm method for diagnosing SCM and IMI. The main objective was to evaluate the clinical utility of measuring milk pH using 3 tests of increasing pH resolution: Multistix 10 SG Reagent Strips for Urinalysis (Multistix strips, Bayer HealthCare Inc., Elkhart, IN), pH Hydrion paper (Microessential Laboratory, Brooklyn, NY), and Piccolo plus pH meter (Hanna Instruments, Woonsocket, RI), for diagnosing SCM and IMI in dairy cattle. Quarter foremilk samples were collected from 115 dairy cows at dry off and 92 fresh cows within 4 to 7 d postcalving. Quarter somatic cell count (SCC) was measured using a DeLaval cell counter (DeLaval, Tumba, Sweden), with SCM defined as SCC >200,000 cells/mL and IMI defined as SCC >100,000 cells/mL and the presence of microorganisms at ≥10 cfu/mL of milk. Milk pH was measured at 37°C using the 3 test methods. The Hydrion pH paper performed poorly in diagnosing SCM and IMI. Receiver operating curve analysis provided optimal pH cutpoints for diagnosing SCM for the pH meter (dry off, ≥6.67; freshening, ≥6.52) and Multistix strips (dry off and freshening, ≥7.0). Test performance of the pH meter and Multistix strips was poor to fair based on area under the receiver operating curve, sensitivity, specificity, positive likelihood ratio, and kappa coefficient. The pH meter and Multistix strips performed poorly in diagnosing IMI at dry off and freshening. We concluded that milk pH does not provide a clinically useful method for diagnosing SCM or IMI in dairy cattle.

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Subclinical mastitis (SCM) and intramammary infection (IMI) increase esterase activity in the glandular secretions of dairy cattle. Our objective was to evaluate the clinical performance of 3 commercially available esterase tests for diagnosing SCM and IMI. Foremilk samples were collected from 380 quarters (96 cows) at dry-off and from 329 quarters (83 cows) within 4 to 7 d after calving. Quarter somatic cell count (SCC) was measured using the reference method (DeLaval cell counter; De Laval International AB, Tumba, Sweden) with SCM defined as SCC >200,000 cells/mL. Bacterial culture of foremilk samples was used to diagnose IMI based on the growth of ≥100 cfu/mL. The SCC was estimated using 3 PortaSCC tests (PortaCheck, Moorestown, NJ) from the measured esterase activity and the California Mastitis Test (CMT). Clinical performance was evaluated using logistic regression to determine the area under the receiver operating characteristic curve (AUC) and identify test sensitivity (Se) and specificity (Sp) at the optimal cut-point for diagnosing SCM and IMI. Test agreement was also evaluated using the kappa coefficient (κ) and weighted κ. The PortaSCC color test was the best-performing PortaSCC test for diagnosing SCM at dry-off (AUC = 0.90, Se = 0.91, Sp = 0.81, κ = 0.71) and at freshening (AUC = 0.86, Se = 0.74, Sp = 0.95, κ = 0.72), at an optimal cut-point of ≥250,000 cells/mL but required 45 min to produce a result. For comparison, the CMT required 2 min to produce a result and a CMT score of trace or higher was superior to the PortaSCC color test for diagnosing SCM at dry-off (AUC = 0.95, Se = 0.95, Sp = 0.86, κ = 0.81) and freshening (AUC = 0.88, Se = 0.79, Sp = 0.95, κ = 0.76). The PortaSCC quick test was the best-performing PortaSCC test for diagnosing IMI at dry-off (AUC = 0.81, Se = 0.81, Sp = 0.78 κ = 0.40) and required 5 min to produce a result, whereas the PortaSCC color test was the best performing PortaSCC test for diagnosing IMI at freshening (AUC = 0.80, Se = 0.75, Sp = 0.79 κ = 0.38). For comparison, the CMT was inferior to the PortaSCC quick test for diagnosing IMI at dry-off (AUC = 0.73, Se = 0.76, Sp = 0.60, κ = 0.20) but was equivalent to the PortaSCC color test at freshening (AUC = 0.79, Se = 0.58, Sp = 0.93, κ = 0.50). The PortaSCC color and quick tests and CMT were considered good tests for diagnosing SCM and IMI because clinically useful tests typically have an AUC >0.80 and κ >0.6. Based on the test sensitivity, cost, and analysis time, there does not appear to be a persuasive reason to select the PortaSCC tests over the traditional CMT for diagnosing SCM and IMI.