Recent studies indicate that milk from healthy mothers may harbor potential probiotics. Nonetheless, the distribution of bacterial profiles in human milk samples in Taiwan is not fully understood. Therefore, with the aim to address this question, in this study, milk samples were collected from 33 healthy mothers (D1 to D33) visiting our hospital during a 6-month period. The milk microbiota was analyzed by a molecular approach (Illumina MiSeq sequencing). The results indicate that the milk samples have a unique profile and patterns of bacterial abundance levels. Moreover, in colostrum and transitional-milk samples, we detected 154 and 127 bacterial species, respectively, and these sets shared 42.6% of the bacterial species. The most common bacterial species among all milk samples were Staphylococcus epidermidis, Streptococcus lactarius, and Staphylococcus hominis, suggesting that the skin contamination route plays an important role in the composition of the milk microbiota. Nevertheless, four Lactobacillus species, Lactobacillus helveticus, Lactobacillus iners, Lactobacillus zeae, and Lactobacillus gasseri, were present in only 7 samples (21% prevalence), and bifidobacterial species were quite rare taxa among the present samples. The Staphylococcus aureus was detected in a total of 15 samples (45% prevalence), suggesting that this species may be commonly present in milk samples. In conclusion, each milk sample revealed a unique profile and patterns of bacterial abundance levels, and our data do not support the idea that lactobacilli and bifidobacteria are common and abundant in modern milk samples. Because none of the donors of the milk samples showed mastitis or any discomfort during the sampling process or at follow-up inspection, the microbiota of these milk samples is not likely to negatively affect its host. This study provides new information on the proportions of commensal bacteria in human milk in Taiwan.