The objectives of the present study were to investigate the change in the number of viable pathogens during preservation of milk obtained from cows with subclinical mastitis and the association between the decreasing ratio of viable bacteria during preservation and the somatic cell count (SCC) and the values of lingual antimicrobial peptide (LAP), lactoferrin (LF) and lactoperoxidase (LPO). After preservation of milk at room temperature for 0, 0.5, 1, 2, 3, 4 and 5 hr, the bacterial colonies in the milk were counted to determine the number of colony forming units (CFUs). Fresh skim milk was used to determine the values of LAP, LPO and LF. Bacteria were not detected in 19.4% of milk samples, and this percentage increased up to 30% after 5 hr of preservation. The number of Staphylococcus aureus and Streptococcus uberis in milk did not change significantly during the 5-hr incubation, whereas significant decreases were observed in the number of coliforms, coagulase-negative staphylococci, yeasts and Corynebacterium bovis. High SCC significantly decreased CFUs of S. aureus and yeast after preservation of milk for 4 to 5 hr. High LF concentration in milk was associated with decrease in CFU of S. aureus during 4-hr preservation. These results suggest that the viable counts of some pathogens in milk decreased during preservation at room temperature after collection, which may be attributed to the leukocytes and antimicrobial components present in milk.