Breast or bottle feeding information including type of formula given to infants before recruitment and consumption of probiotics products were obtained from infant’s diet records. The current study population of 133 infants, comprised 43 breastfed infants, 43 standard formula-fed infants and 47 infants fed the MFGM enriched formula. Saliva could not be collected
from six infants (2 breastfed, and 4 MFGM formula-fed), and oral swabs were not obtained from five infants (2 breastfed, 3 MFGM formula-fed). One standard formula-fed infant had received antibiotics at birth and one MFGM enriched formula-fed infant received antibiotics Y-27632 supplier at 3 months of age. Twenty-five infants had been given commercially available probiotic oral drops (Semper Magdroppar, BioGaia AB, Lund, Sweden) containing L. reuteri ATCC 17938 (~108 CFU in 5 drops) at 1, 2, 3 or 4 months of age. Infants given
probiotic drops did not differ between the three feeding groups (p≥0.401). The study was approved by the Regional Ethical Review Board in Umeå, Sweden. All caregivers signed informed consent when recruited. Culture of salivary lactobacilli and characterization of isolates Whole saliva was collected from the infants and Lactobacillus cultured using selective medium as previously described . Up to 30 isolates were selected from MI-503 supplier each plate and were identified by comparing 16S rRNA gene sequences to databases HOMD (http://www.homd.org) and NCBI (http://blast.ncbi.nlm.nih.gov/Blast.cgi). qPCR for L. gasseri in mucosal swabs The mucosa of the cheeks, the tongue and alveolar ridges of the infants were swabbed using sterile cotton swabs (Applimed SA, Chatel-St-Denis, Switzerland). Samples storage, DNA purification and L. gasseri level quantification by qPCR were as described previously [13, 27]. Growth inhibition by L. gasseri Cultural conditions and bacterial strains used in growth inhibition tests Lactobacillus isolates were maintained
on de Man, Rogosa, Sharpe Agar Baricitinib (MRS) (Fluka, Buchs, Switzerland) and grown in MRS broth. S. mutans strains Ingbritt, NG8, LT11 and JBP, S. sobrinus strains OMZ176 and 6715, Actinomyces naeslundii genospecies 1 strains ATCC 35334 and ATCC 29952, and Actinomyces oris (previously A. naeslundii genospecies 2) strains T14V and M4366 were maintained on Columbia agar plates (Alpha BioScience, Baltimore, Maryland, USA) supplemented with 5% horse blood (CAB) and grown in Todd-Hewitt broth (Fluka). Fusobacterium nucleatum strains ATCC 25586 and UJA11-a were maintained on Fastidious Anaerobe Agar (FAA, Lab M, Bury, UK) and grown in Peptone yeast extract broth (PY, Sigma-Aldrich Co., St. Louis, Missouri, USA). Bacteria were cultured anaerobically at 37°C for 48–72 h (maintenance) or 24 h (growth).