Effects of alternative dietary substrates on competition between human colonic bacteria in an anaerobic fermentor system

S H Duncan, K P Scott, A G Ramsay, H J M Harmsen, G W Welling, C S Stewart, H J Flint

Research output: Contribution to journalArticle

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Abstract

Duplicate anaerobic fermentor systems were used to examine changes in a community of human fecal bacteria supplied with different carbohydrate energy sources. A panel of group-specific fluorescent in situ hybridization probes targeting 16S rRNA sequences revealed that the fermentors supported growth of a greater proportion of Bacteroides and a lower proportion of gram-positive anaerobes related to Faecalibacterium prausnitzii, Ruminococcus flavefaciens-Ruminococcus bromii, Eubacterium rectale-Clostridium coccoides, and Eubacterium cylindroides than the proportions in the starting fecal inoculum. Nevertheless, certain substrates, such as dahlia inulin, caused a pronounced increase in the number of bacteria related to R. flavefaciens-R. bromii and E. cylindroides. The ability of three strictly anaerobic, gram-positive bacteria to compete with the complete human fecal flora was tested in the same experiment by using selective plating to enumerate the introduced strains. The Roseburia-related strain A2-183(R), was able to grow on all substrates despite the fact that it was unable to utilize complex carbohydrates in pure culture, and it was assumed that this organism survived by cross-feeding. In contrast, Roseburia intestinalis L1-82(R) and Eubacterium sp. strain A2-194(R) survived less well despite the fact that they were able to utilize polysaccharides in pure culture, except that A2-194(R) was stimulated 100-fold by inulin. These results suggest that many low-G+C-content gram-positive obligate anaerobes may be selected against during in vitro incubation, although several groups were stimulated by inulin. Thus, considerable caution is necessary when workers attempt to predict the in vivo effects of probiotics and prebiotics from their effects in vitro.

Original languageEnglish
Pages (from-to)1136-1142
Number of pages7
JournalApplied and Environmental Microbiology
Volume69
Issue number2
DOIs
Publication statusPublished - Feb 2003

Keywords

  • 16S ribosomal-RNA
  • in-situ hybridization
  • human fecal flora
  • targeted oligonucleotide probes
  • semicontinuous culture system
  • green fluorescent proteins
  • human large-intestine
  • lactic-acid bacteria
  • human gut
  • resistant starch

Cite this

Effects of alternative dietary substrates on competition between human colonic bacteria in an anaerobic fermentor system. / Duncan, S H ; Scott, K P ; Ramsay, A G ; Harmsen, H J M ; Welling, G W ; Stewart, C S ; Flint, H J .

In: Applied and Environmental Microbiology, Vol. 69, No. 2, 02.2003, p. 1136-1142.

Research output: Contribution to journalArticle

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AB - Duplicate anaerobic fermentor systems were used to examine changes in a community of human fecal bacteria supplied with different carbohydrate energy sources. A panel of group-specific fluorescent in situ hybridization probes targeting 16S rRNA sequences revealed that the fermentors supported growth of a greater proportion of Bacteroides and a lower proportion of gram-positive anaerobes related to Faecalibacterium prausnitzii, Ruminococcus flavefaciens-Ruminococcus bromii, Eubacterium rectale-Clostridium coccoides, and Eubacterium cylindroides than the proportions in the starting fecal inoculum. Nevertheless, certain substrates, such as dahlia inulin, caused a pronounced increase in the number of bacteria related to R. flavefaciens-R. bromii and E. cylindroides. The ability of three strictly anaerobic, gram-positive bacteria to compete with the complete human fecal flora was tested in the same experiment by using selective plating to enumerate the introduced strains. The Roseburia-related strain A2-183(R), was able to grow on all substrates despite the fact that it was unable to utilize complex carbohydrates in pure culture, and it was assumed that this organism survived by cross-feeding. In contrast, Roseburia intestinalis L1-82(R) and Eubacterium sp. strain A2-194(R) survived less well despite the fact that they were able to utilize polysaccharides in pure culture, except that A2-194(R) was stimulated 100-fold by inulin. These results suggest that many low-G+C-content gram-positive obligate anaerobes may be selected against during in vitro incubation, although several groups were stimulated by inulin. Thus, considerable caution is necessary when workers attempt to predict the in vivo effects of probiotics and prebiotics from their effects in vitro.

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KW - targeted oligonucleotide probes

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KW - lactic-acid bacteria

KW - human gut

KW - resistant starch

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JO - Applied and Environmental Microbiology

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