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In order to monitor change of T lymphocyte subpopulation in milk derived from infected bovine mammary glands. 30 lactating cows with mastitis were compared to 10 normal controls. The results showed that CD4+ T cells and CD8+ T cells were significantly higher in milk derived from infected bovine mammary glands compared to that of normal controls(P<0.0001). IL-6, IL-1 and IFN-γ were found in normal and infected milk samples;IL-1a( 26/30), IL-1β( 28/30),IL-12 (29/30) and TNF-a (30/30) were only found in infected milk samples;Neither IL-2 nor IL-4 was found in any milk samples. In conclussion, we can monitor quality of fresh milk by analysis of CD4+ T cells and CD8+ T cells in milk using flow cytometric.