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This study aimed to determine the lactoferrin concentration from healthy and subclinical mastitic buffaloes. Lactoferrin levels were determined in a total of 475 mammary quarter milk samples, 391 of which were healthy and 84 suffering from subclinical mastitis. The mean milk lactoferrin level in logarithmic form for healthy and subclinical mastitic milk samples was 1.34±0.27 and 1.85±0.47, respectively. A significant difference (p<0.001) was determined between the two groups in terms of the lactoferrin levels. The milk lactoferrin levels in healthy buffaloes were significantly higher in the middle and late periods of lactation than the peak period (p<0.05). The mean milk lactoferrin levels in subclinical mastitic buffaloes infected with S. aureus or S. agalactiae were significantly higher than those from in buffaloes infected with Coagulase-Negative Staphylococci (CNS) or Candida spp. (p<0.05). An increase in milk lactoferrin levels was observed in parallel with the increase in somatic cell scores. In conclusion, milk lactoferrin levels in buffaloes were found to increase with the lactation stage, the presence of bacterial infection and the somatic cell count. This is the first study to determine the relationship between lactoferrin concentrations and bacterial infection in milk obtained from mammary quarters with subclinical mastitis in Anatolian buffaloes.