16S rRNA Gene-targeted TTGE in Determining Diversity of Gut Microbiota during Acute Diarrhoea and Convalescence

The human gut microbiota play a vital role in health and nutrition but are greatly modified during severe diarrhoea due to purging and pathogenic colonization. To understand the extent of loss during and after diarrhoea, faecal samples collected from children (n=21) suffering from acute diarrhoea and from their healthy siblings (n=9) were analyzed by 16S rRNA gene-targeted universal primer polymerase chain reaction (PCR), followed by temporal temperature gradient gel electrophoresis (TTGE). The gut microbiota decreased significantly as indicated by the number of TTGE bands at day 0 of acute diarrhoea [patients vs healthy siblings: 11±0.9 vs 21.8±1.1 (mean±standard error), p<0.01]. The number of bands showed a steady increase from day 1 to day 7; however, it remained significantly less than that in healthy siblings (15±0.9, p<0.01). These results suggest that appropriate therapeutic and post-diarrhoeal nutritional intervention might be beneficial for the early microbial restoration and recovery.

[1]  G. Nair,et al.  Time Course of Bacterial Diversity in Stool Samples of Malnourished Children With Cholera Receiving Treatment , 2009, Journal of pediatric gastroenterology and nutrition.

[2]  L. Beaugerie,et al.  Effect of Antibiotic Therapy on Human Fecal Microbiota and the Relation to the Development of Clostridium difficile , 2008, Microbial Ecology.

[3]  P. Pochart,et al.  A longitudinal study of infant faecal microbiota during weaning. , 2006, FEMS microbiology ecology.

[4]  P. Pochart,et al.  Low species diversity and high interindividual variability in faeces of preterm infants as revealed by sequences of 16S rRNA genes and PCR-temporal temperature gradient gel electrophoresis profiles. , 2006, FEMS microbiology ecology.

[5]  A. Murray,et al.  Investigations into the influence of host genetics on the predominant eubacteria in the faecal microflora of children. , 2005, Journal of medical microbiology.

[6]  F. Guarner,et al.  Gut flora in health and disease , 2003, The Lancet.

[7]  B. Deplancke,et al.  Selective growth of mucolytic bacteria including Clostridium perfringens in a neonatal piglet model of total parenteral nutrition. , 2002, The American journal of clinical nutrition.

[8]  G. Tannock.  Molecular methods for exploring the intestinal ecosystem , 2002, British Journal of Nutrition.

[9]  L. Cocolin,et al.  Denaturing Gradient Gel Electrophoresis Analysis of the 16S rRNA Gene V1 Region To Monitor Dynamic Changes in the Bacterial Population during Fermentation of Italian Sausages , 2001, Applied and Environmental Microbiology.

[10]  G. Macfarlane,et al.  Age and disease related changes in intestinal bacterial populations assessed by cell culture, 16S rRNA abundance, and community cellular fatty acid profiles , 2001, Gut.

[11]  J. Doré,et al.  Direct Analysis of Genes Encoding 16S rRNA from Complex Communities Reveals Many Novel Molecular Species within the Human Gut , 1999, Applied and Environmental Microbiology.

[12]  E. Zoetendal,et al.  Temperature Gradient Gel Electrophoresis Analysis of 16S rRNA from Human Fecal Samples Reveals Stable and Host-Specific Communities of Active Bacteria , 1998, Applied and Environmental Microbiology.

[13]  H. Backhaus,et al.  Direct ribosome isolation from soil to extract bacterial rRNA for community analysis , 1996, Applied and environmental microbiology.

[14]  K. Wilson,et al.  Human colonic biota studied by ribosomal DNA sequence analysis , 1996, Applied and environmental microbiology.

[15]  G. Muyzer,et al.  Distribution of sulfate-reducing bacteria in a stratified fjord (Mariager Fjord, Denmark) as evaluated by most-probable-number counts and denaturing gradient gel electrophoresis of PCR-amplified ribosomal DNA fragments , 1996, Applied and environmental microbiology.

[16]  D. M. Ward,et al.  Denaturing Gradient Gel Electrophoresis Profiles of 16 S rRNA-Defined Populations Inhabiting a Hot Spring Microbial Mat Community , 1996 .

[17]  L. Poulsen,et al.  Phylogeny of not-yet-cultured spirochetes from termite guts , 1996, Applied and environmental microbiology.

[18]  A. Uitterlinden,et al.  Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA , 1993, Applied and environmental microbiology.

[19]  S. Saidi,et al.  Ecological studies on intestinal microbial flora of Kenyan children with diarrhoea. , 1990, The Journal of tropical medicine and hygiene.

[20]  S. Finegold,et al.  Intestinal beta-lactamase activity in ampicillin-induced, Clostridium difficile-associated ileocecitis. , 1983, The Journal of infectious diseases.

[21]  R. Berg.  Promotion of the translocation of enteric bacteria from the gastrointestinal tracts of mice by oral treatment with penicillin, clindamycin, or metronidazole , 1981, Infection and immunity.

[22]  S. J. Baker,et al.  Faecal flora of South Indian infants and young children in health and with acute gastroenteritis. , 1978, Journal of medical microbiology.

[23]  R. Sack,et al.  Acute undifferentiated human diarrhea in the tropics. I. Alterations in intestinal micrflora. , 1971, The Journal of clinical investigation.

[24]  R. Dubos,et al.  ALTERATIONS IN THE MOUSE CECUM AND ITS FLORA PRODUCED BY ANTIBACTERIAL DRUGS , 1968, The Journal of experimental medicine.

[25]  D. Ahearn,et al.  Microbial intestinal flora in acute diarrheal disease. , 1967, JAMA.

[26]  M. Blaut,et al.  Long-term stability of the human gut microbiota in two different rat strains. , 2008, Current issues in molecular biology.

[27]  K. Brigham,et al.  Intestinal microflora in Asiatic cholera. I. "Rice-water" stool. , 1970, The Journal of infectious diseases.

[28]  D. Mahalanabis,et al.  Intestinal microflora in Asiatic cholera. 3. Studies in pediatric cholera. , 1970, The Journal of infectious diseases.

[29]  K. Brigham,et al.  Intestinal microflora in Asiatic cholera. II. The small bowel. , 1970, The Journal of infectious diseases.