During the period from November 1988 to May 1989, causative bacteria in a total of 172 clinical mastitis cases observed in 66 farms in 5 districts with 6 areas in Japan were examined and frequencies of their occurrences were determined. Susceptibilities (in MICs) of the isolates to 6 beta-lactam antibacterial agents were also determined. As a result, coagulase-negative staphylococci (CNS) was identified in 94 out of 172 cases (54.7%) and were the most prevalent. Corynebacterium spp., Gram-negative bacteria and Staphylococcus aureus were found in 52 (30.2%), 49 (28.5%) and 43 (25.0%) cases, respectively. Four species of Streptococcus family (S. agalactiae, S. dysgalactiae, S. uberis and S. bovis) were identified in a total of 58 cases (33.7%). Susceptibility testing of CNS to cefoperazone (CPZ), cefazolin (CEZ), benzylpenicillin (PCG), ampicillin (ABPC), methicillin (DMPPC) and cloxacillin (MCIPC) showed that all MIC80's (inhibiting bacterial growth of 80% of all isolates) were within a range from 0.10 to 3.13 micrograms/ml and that there was no marked differences in antibacterial effects among the antibiotics used. The highest antibacterial effect on S. aureus was exhibited by MCIPC, which inhibited the growth of all isolates at 0.39 microgram/ml. The MICs of DMPPC against all isolates of S. aureus were 3.13 micrograms/ml or less and no methicillin-resistant S. aureus (MRSA) was detected. There was no difference in antibacterial activities against Streptococcus family between penicillin antibiotics (DMPPC, MCIPC, ABPC and PCG) and cephem antibiotics (CPZ and CEZ), both of which showed excellent antibacterial activities. Cephem antibiotics exhibited higher activities against Gram-negative bacteria than penicillin antibiotics. Especially CPZ, the third generation cephem, showed excellent antibacterial activity against Escherichia coli, Klebsiella spp., Enterobacter spp., as well as against other Enterobacteriaceae.