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This field trial was conducted to evaluate two antibiotics at a close-up period in curing the existing IMI and to prevent new clinical and subclinical Intramammary infection (IMI). Two hundred and twelve Holstein cows were assigned to one of three treatment groups: TYLO, MARB and CONT. Cows in TYLO group received 10 mg/kg Tylosin for three days at the close-up period (21 days before calving), cows in MARB group received single SC injection of 8 mg/kg SC marbofloxacin at the close-up period and cows in CONT group remained untreated. Milk samples were collected for somatic cell count, microbial culture and Total oxidant/antioxidant capacity before drying-off, and 3 and 7days post calving. Antimicrobial susceptibility test and RAPT-PCR were performed on S.aureus isolates. No significant differences were detected in total cure rate within the groups, but S. aureus cure rates in TYLO and MARB were significantly higher than in CONT (74 and 73.5 % Vs 58.1 %). No significant differences in total new IMI were observed. Furthermore, the rate of new S. aureus IMI was higher in both treatment groups than in CONT group. Clinical mastitis rate in TYLO (3.8 %) and MARB (5.8 %) was significantly lower than CONT (11.3 %). Paired S. aureus isolates from dry-off and post-calving have been clustered into 9 different RAPD types (A-I). 8 paired strains collected at dry-off were identical to those at post-calving,and 35 strains had more than 60 % dissimilarity. Administration of Tylosin or Marbofloxacin is not useful in all cases;however, they have the potential to reduce the incidence of post-calving clinical mastitis and improve S.aureus cure rate if used selectively.