Lack of hygiene and puerperal mastitis are common causes of bacterial diseases in nursing neonates. The aim of this study was to isolate microorganisms from milk samples of healthy female Jindo dogs with suckling puppies and to investigate antimicrobial susceptibility against the isolated bacteria. Milk samples were collected from 120 udders of 12 lactating Jindo dogs that were 2~4 years old without any clinical diseases including mastitis. Bacteria were isolated from 64 milk samples (53.3%), either singly (76.6%) or in combination (23.4%). Staphylococcus (S.) spp. was the most common microorganisms (74.7%) isolated from canine milk, followed by Haemophillus spp. (10.9%), Streptococcus spp. (9.6%), Gardnerella spp. (2.4%) and Moraxella spp. (2.4%). The most frequently isolated organism was S. warneri (31.3%). Antimicrobial susceptibility of these bacteria was tested with 17 antimicrobial agents by Kirbyand Bauer standardized disc diffusion method. Results indicated that bacteria isolated from healthy canine milk were mostly susceptible to amoxicillin-clavulanic acid, cephalothin and ceftiofur, but were resistant to erythromycin, neomycin and tetracycline.