Analysis of intestinal flora development in breast-fed and formula-fed infants by using molecular identification and detection methods.

By H. Harmsen, A. C. Wildeboer-veloo, G. C. Raangs, Arjen A. Wagendorp, N. Klijn, J. G. Bindels, G. W. Welling. Journal of Pediatric Gastroenterology and Nutrition - JPGN, 2000

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BACKGROUND An obvious difference between breast-fed and formula-fed newborn infants is the development of the intestinal flora, considered to be of importance for protection against harmful micro-organisms and for the maturation of the intestinal immune system. In this study, novel molecular identification methods were used to verify the data obtained by traditional culture methods and to validate the culture independent fluorescent in situ hybridization (FISH) technique. METHODS From each of six breast-fed and six formula-fed newborn infants, six fecal samples were obtained during the first 20 days of life. The microbial compositions of the samples were analyzed by culturing on specific media and by FISH, by using specific 16S rRNA-targeted oligonucleotide probes. The colonies growing on the media were identified by random amplified polymorphic DNA pattern analysis and by polymerase chain reaction amplification and subsequent analysis of the 16S rRNA gene. RESULTS Molecular identification of the colonies showed that the selective media are insufficiently selective and unsuitable for quantitative analyses. Qualitative information from the culturing results combined with the data obtained by the FISH technique revealed initial colonization in all infants of a complex (adult-like) flora. After this initial colonization, a selection of bacterial strains began in all infants, in which Bifidobacterium strains played an important role. In all breast-fed infants, bifidobacteria become dominant, whereas in most formula-fed infants similar amounts of Bacteroides and bifidobacteria (approximately 40%) were found. The minor components of the fecal samples from breast-fed infants were mainly lactobacilli and streptococci; samples from formula-fed infants often contained staphylococci, Escherichia coli, and clostridia. CONCLUSIONS This study confirms the differences in development of intestinal flora between breast-fed and formula-fed infants. The results obtained from the FISH technique were consistent. Although the repertoire of probes for this study was not yet complete, the FISH technique will probably become the method of reference for future studies designed to develop breast-fed-like intestinal flora in formula-fed infants.

Subfields: Endocrinology, Food Science, Nutrition and Dietetics

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Citations in the corpus

The probiotic Bifidobacteria infantis: An assessment of potential antidepressant properties in the rat. (2008, background): “Shortly after birth, up to 90% of the bacteria found in the infant GIT is composed of bifidobacteria (Harmsen et al., 2000), and in the adult they still make up approximately 3–5% of the microflora (Harmsen et al.”
High-throughput diversity and functionality analysis of the gastrointestinal tract microbiota (2008, result): “previous studies which showed that bifidobacteria are the most dominant group in infants.(78) 79”
Bifidobacteria and Their Role as Members of the Human Gut Microbiota (2016, background): “Bifidobacteria dominate the total gut bacterial population in healthy breast-fed infants (Harmsen et al., 2000; Favier et al., 2002; Leahy et al., 2005), although this dominance decreases following weaning (Ventura et al., 2004).”
Bacterial metabolism and health‐related effects of galacto‐oligosaccharides and other prebiotics (2007, background): “These microbiotas are characterized by slightly lower counts of bifidobacteria, with greater numbers of more potentially harmful organisms such as clostridia and enterococci (Harmsen et al. 2000; Macfarlane et al. 2004).”
The Impact of Diet and Lifestyle on Gut Microbiota and Human Health (2014, background): “Oligosaccharides present in breast milk promote the growth of Lactobacillus and Bifidobacterium, which dominate the infant gut, and this can strengthen or promote development of the immune system and may help prevent conditions such as eczema and asthma [28–30].”
Selective increases of bifidobacteria in gut microflora improve high-fat-diet-induced diabetes in mice through a mechanism associated with endotoxaemia (2007, methodology): “The probes used were: EREC 482 [19] for Eubacterium rectale– Clostridium coccoides group, Lab158 [20] for lactobacilli and enterococci and Bif164 [21] for Bifidobacterium spp. (Gram-positive bacteria); and Bac303 [22] for Bacteroides– Prevotella spp., MIB 661 [23] for Bacteroides mouse intestinal ba”
Development of the Human Infant Intestinal Microbiota (2007, background): “to breast-fed infants [20,21,23–25], yet other reports have”
16S rRNA gene-based analysis of fecal microbiota from preterm infants with and without necrotizing enterocolitis (2009, background): “The initial colonization patterns in normal full-term infants are dependent on environmental influences such as mode of delivery and type of feeding (Gronlund et al., 1999; Harmsen et al., 2000; Penders et al., 2006; Palmer et al., 2007).”
Impacts of Gut Bacteria on Human Health and Diseases (2015, neutral): “[30–32].”
Distinct patterns of neonatal gut microflora in infants in whom atopy was and was not developing. (2001, result): “The results demonstrated that the ratio of bifidobacteria to clostridia was reduced in atopic subjects (geometric mean [95% confidence interval]: 19 [3-122]) compared with that in nonatopic subjects (185 [57-604], P = .”