This study was performed to examine the bacteriological findings in 58 mammary secretions from 15 heifers at 4-5 weeks before parturition, and to evaluate whether a high prevalence of S. aureus infection in lactating cows affects the occurrence of S. aureus infection in prepartum heifers in the dairy herd. A total of 86.7%(13/15) of the heifers and 60.3%(35/58) of quarter milk samples from the heifers were bacteriologically positive. No S. aureus isolate was detected in mammary secretions from the heifers. Coagulase-negative staphylococcus (CNS) species were predominantly detected in 54.3%(19/35) and Streptococci other than Streptococcus agalactiae were isolated from 22.9%(8/35) of the quarters. A high S. aureus prevalence in the herd may not necessarily be a decisive factor for S. aureus infection in heifers.