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Published on: November 17, 2018
Characterisation of intestinal bacteria in infant stools using real-time PCR and northern hybridisation analyses
Mark J Hopkins1, George T Macfarlane, Elizabeth Furrie
1Microbiology and Gut Biology Group, University of Dundee, Ninewells Hospital Medical School, UK.
Insights
Infant gut bacteria composition changes with age, with bifidobacteria increasing in breast-fed babies and sulphate-reducing bacteria in bottle-fed infants. These shifts are crucial for infant gut health.
Area of Science:
- Microbiology
- Infant gut microbiome research
Background:
- The infant gut microbiome plays a critical role in health and development.
- Understanding early-life microbial colonization is essential for identifying potential health impacts.
Purpose of the Study:
- To quantify bacterial populations in infant feces.
- To investigate the impact of age and feeding method (breast vs. bottle) on gut microbiota composition.
Main Methods:
- Utilized real-time PCR and northern hybridizations targeting 16S rRNA genes.
- Analyzed fecal samples from 40 infants across three age groups (0-6, 7-12, 13-24 months).
- Developed specific PCR primers for key bacterial groups: Bifidobacteria, Bacteroides, sulphate-reducing bacteria, and Enterococcus faecalis.
Main Results:
- Bacteroides and Desulfovibrio populations increased significantly in infants aged 7-24 months, while Enterococcus faecalis decreased.
- Northern hybridizations confirmed these trends (except for Desulfovibrios) and indicated increased colonization by Clostridium coccoides group and Faecalibacterium prausnitzii after 6 months.
- Breast-fed infants showed higher Bifidobacteria levels, whereas bottle-fed infants had elevated Desulfovibrio levels.
Conclusions:
- Infant gut microbiota undergoes significant compositional changes during the first two years of life.
- Feeding method influences early-life gut bacterial colonization, with distinct patterns observed in breast-fed versus bottle-fed infants.
- These findings contribute to understanding the microbial basis of infant gut health and development.
Abstract:
Real-time PCR and northern hybridisations were used to quantify bacterial populations in the large gut of infants. PCR primers for rapid, sensitive, high throughput detection of bifidobacteria, bacteroides, sulphate-reducing bacteria and Enterococcus faecalis, based on analysis of 16S rRNA genes were used. Bacterial populations were analysed in faeces from 40 infants aged 0-6, 7-12 and 13-24 months. The effects of breast versus bottle feeding was also investigated. Real-time PCR indicated that bacteroides and desulfovibrio numbers increased markedly in the 7-12 and 13-24 month age groups, and that the reverse occurred with Ent. faecalis. With the exception of desulfovibrios, this was seen with northern hybridisations, which also showed increased colonisation by the Clostridium coccoides group and Faecalibacterium prausnitzii after 6 months. Both methodologies indicated increased bifidobacteria in breast-fed babies, and higher levels of desulfovibrios in bottle-fed children.
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