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Applied and Environmental Microbiology, January 2004, p. 558-568, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.558-568.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Genotypic and Phenotypic Studies of Murine Intestinal Lactobacilli: Species Differences in Mice with and without Colitis

J. A. Peña,1,2 S. Y. Li,2,3 P. H. Wilson,2,3 S. A. Thibodeau,4 A. J. Szary,5 and J. Versalovic1,2,3*

Departments of Molecular Virology & Microbiology,1 Pathology, Baylor College of Medicine,3 Department of Pathology, Texas Children's Hospital, Houston, Texas 77030,2 Department of Pathology, Massachusetts General Hospital, Charlestown, Massachusetts 02129,4 Division of Health Science and Technology, Harvard Medical School, Boston, Massachusetts 021155

Received 9 July 2003/ Accepted 7 October 2003

Lactobacilli represent components of the commensal mammalian gastrointestinal microbiota and are useful as probiotics, functional foods, and dairy products. This study includes systematic polyphasic analyses of murine intestinal Lactobacillus isolates and correlation of taxonomic findings with data from cytokine production assays. Lactobacilli were recovered from mice with microbiota-dependent colitis (interleukin-10 [IL-10]-deficient C57BL/6 mice) and from mice without colitis (Swiss Webster and inducible nitric oxide synthetase-deficient C57BL/6 mice). Polyphasic analyses were performed to elucidate taxonomic relationships among 88 reference and murine gastrointestinal lactobacilli. Genotypic tests included single-locus analyses (16S ribosomal DNA sequencing and 16S-23S rRNA intergenic spacer region PCR) and genomic DNA profiling (repetitive DNA element-based PCR), and phenotypic analyses encompassed more than 50 tests for carbohydrate utilization, enzyme production, and antimicrobial resistance. From 20 mice without colitis, six Lactobacillus species were recovered; the majority of the mice were colonized with L. reuteri or L. murinus (72% of isolates). In contrast, only, L. johnsonii was isolated from 14 IL-10-deficient mice. Using an in vitro assay, we screened murine isolates for their ability to inhibit tumor necrosis factor alpha (TNF-{alpha}) secretion by lipopolysaccharide-activated macrophages. Interestingly, a subpopulation of lactobacilli recovered from mice without colitis displayed TNF-{alpha} inhibitory properties, whereas none of the L. johnsonii isolates from IL-10-deficient mice exhibited this effect. We propose that differences among intestinal Lactobacillus populations in mammals, combined with host genetic susceptibilities, may account partly for variations in host mucosal responses.

* Corresponding author. Mailing address: Texas Children's Hospital, Department of Pathology, 6621 Fannin St., MC 1-2261, Houston, TX 77030. Phone: (832) 824-2213. Fax: (832) 825-1032. E-mail: jxversal{at}texaschildrenshospital.org.

Applied and Environmental Microbiology, January 2004, p. 558-568, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.558-568.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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