Fecal samples were collected from 67 healthy Japanese subjects who had not taken any antibiotics for 1 month. Thirty-six healthy breast-fed infants (18 male and 18 female babies, average age = 2.9 ± 1.2 months, age range = 1.4 to 5.7 months) who were born at full term were recruited from Nagao Midwife Clinic (Kyoto, Japan). Breast milk was donated from 32 mothers at the same day when their infant stools were collected. Thirty-one healthy adults (16 males and 15 nonpregnant females who are not kin of the recruited infants, average age = 42.3 ± 16.8 years, age range = 21 to 75 years) were recruited from Ishikawa Prefectural University (Ishikawa, Japan). All samples were frozen immediately after collection and transferred to the laboratories. The samples were subjected to quantitative PCR (qPCR)–based microbial abundance analysis (infant and adult stools) and HMO analysis (breast milk and infant stools) as described later. All samples were blinded before data evaluation.

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