Proteins were transferred to a (polyvinylidene difluoride (PVDF) membrane (Millipore, Bedford,

Massachusetts, USA). The membranes were blocked and epitopes detected with monoclonal antibodies against gp340 (mAb143) [34] or LUM7-2 [35]. Membranes were washed with TBS (gp340) or PBS (MUC7) and incubated with HRP-conjugated anti-mouse (SAB-100, Stressgen, Victoria, Canada) for gp340 or HRP-conjugated anti-rabbit MRT67307 (P0448, DAKO, Glostrup, Denmark) for MUC7 and detected using Super Signal west Dura Extended Duration Substrate (Thermo Scientific, Rockford, IL, USA). Data processing and statistical analyses The power calculation for the parent study was based on body weight as main outcome [36] with a statistical power of 80% and a level of significance of 0.05% (unpublished data, Timby N, Hernell O, Lönnerdal B and Domellöf M). Based on previous investigations [37], IWP-2 the number of infants included in this study was sufficient to detect a difference in bacterial colonization pattern. Data handling and statistical analyses were performed using PASW Statistics

20 (IBM Corporation Route 100, Somers, New York, USA). Anthropometric measures for infants were averaged, and means with 95% CI reported. Differences between means were tested using analysis of variance (ANOVA) followed by a Bonferroni post hoc test. Differences between means for lactobacilli Amino acid detected in saliva and swabs were tested using generalized linear modeling adjusted for delivery method and exposure to probiotic drops at 4 months. L. gasseri detected in swabs was additionally adjusted for amount

of DNA. Categorical data are presented as proportions (%) and differences between groups were tested with a Chi2 test. A p-value <0.05 was considered statistically significant. Multivariate partial least squares analysis (PLS) was performed (SIMCA P+, version 12.0, Umetrics AB, Umeå, Sweden) as previously described [38, 39]. Cross-validation (Q2 values) was performed by a systematic prediction of 1/7th of the data by the remaining 6/7th of the data. The importance of each variable in the model was displayed in a loading scatter plot. R2- and Q2-values give the capacity of the x-variables to explain (R2) and predict (Q2) the outcome. Results Among the 133 infants, the proportions of boys and girls, infants delivered vaginally, mean body weight and length at birth and at 4 months of age (screening age) did not differ significantly between infants fed breast milk, the standard formula or the MFGM-enriched formula (Table 1). This observation was not affected by exclusion of infants given antibiotics or probiotic drops.