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Applied and Environmental Microbiology, July 2004, p. 3821-3830, Vol. 70, No. 7
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.7.3821-3830.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Specific Response of a Novel and Abundant Lactobacillus amylovorus-Like Phylotype to Dietary Prebiotics in the Guts of Weaning Piglets

Sergey R. Konstantinov,1* Ajay Awati,2 Hauke Smidt,1 Barbara A. Williams,2 Antoon D. L. Akkermans,1 and Willem M. de Vos1

Laboratory of Microbiology, Agrotechnology and Food Sciences Group, Wageningen University,1 Animal Nutrition Group, Wageningen Institute of Animal Sciences, Wageningen, The Netherlands2

Received 29 December 2003/ Accepted 4 March 2004

Using 16S rRNA gene-based approaches, we analyzed the responses of ileal and colonic bacterial communities of weaning piglets to dietary addition of four fermentable carbohydrates (inulin, lactulose, wheat starch, and sugar beet pulp). An enriched diet and a control diet lacking these fermentable carbohydrates were fed to piglets for 4 days (n = 48), and 10 days (n = 48), and the lumen-associated microbiota were compared using denaturing gradient gel electrophoresis (DGGE) analysis of amplified 16S rRNA genes. Bacterial diversities in the ileal and colonic samples were measured by assessing the number of DGGE bands and the Shannon index of diversity. A higher number of DGGE bands in the colon (24.2 ± 5.5) than in the ileum (9.7 ± 4.2) was observed in all samples. In addition, significantly higher diversity, as measured by DGGE fingerprint analysis, was detected in the colonic microbial community of weaning piglets fed the fermentable-carbohydrate-enriched diet for 10 days than in the control. Selected samples from the ileal and colonic lumens were also investigated using fluorescent in situ hybridization (FISH) and cloning and sequencing of the 16S rRNA gene. This revealed a prevalence of Lactobacillus reuteri in the ileum and Lactobacillus amylovorus-like populations in the ileum and the colon in the piglets fed with fermentable carbohydrates. Newly developed oligonucleotide probes targeting these phylotypes allowed their rapid detection and quantification in the ileum and colon by FISH. The results indicate that addition of fermentable carbohydrates supports the growth of specific lactobacilli in the ilea and colons of weaning piglets.

* Corresponding author. Mailing address: Laboratory of Microbiology, Agrotechnology and Food Sciences Group, Wageningen University, Hesselink van Suchtelenweg 4, 6703 CT Wageningen, The Netherlands. Phone: 31 317 483118. Fax: 31 317 483829. E-mail: sergey.konstantinov{at}wur.nl.

Applied and Environmental Microbiology, July 2004, p. 3821-3830, Vol. 70, No. 7
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.7.3821-3830.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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