title: Use of culture-independent analysis to reveal alteration of intestinal microflora by heat-killed Lactobacillus pentosus in a mouse model of endogenous sepsis.
abstract: In this study we evaluated alteration of intestinal microflora by terminal-restriction fragment length polymorphism (T-RFLP) analysis and quantitative PCR (qPCR) for specific microbes. The effects of orally administered heat-killed Lactobacillus pentosus strain b240 (HK-b240) in immunosuppressed mice with endogenous Pseudomonas aeruginosa sepsis was estimated. By T-RFLP analysis, 5 dominant operational taxonomic units (OTUs) including Bacteroides spp. (OTU460) and Lactobacillus spp. (OTU657) were consistently observed, irrespective of treatment, at all time points. A significantly higher population of segmented filamentous bacteria (SFB) was observed by qPCR after 3 weeks of HK-b240 administration; thereafter, the difference was not sustained during immunosuppression and progression of sepsis. Although not significant, Lactobacillus spp. accounted for a larger population in the HK-b240-treated group. In conclusion, this study demonstrated successful application of culture-independent assays for evaluating biological agents by detecting changes in microflora even if the protection was not sufficient to result in significant survival change.
authors.affiliation: Department of Microbiology and Infectious Diseases, Toho University School of Medicine, 5-21-16 Omori-nishi, Ota-ku, Tokyo, 143-8540, Japan.
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Sugasawa
authors.forename: Tomoo
authors.initials: T
authors.lastname: Saga
authors.forename: Soichiro
authors.initials: S
authors.lastname: Kimura
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Ishii
authors.forename: Keizo
authors.initials: K
authors.lastname: Yamaguchi
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Tateda
mesh_terms: D000818:Animals; D005243:Feces / Q000382:microbiology; D005260:Female; D007422:Intestines / Q000382:microbiology*; D007778:Lactobacillus / Q000254:growth & development* / Q000302:isolation & purification; D051379:Mice; D008813:Mice, Inbred ICR; D064307:Microbiota / Q000235:genetics / Q000502:physiology*; D012150:Polymorphism, Restriction Fragment Length; D060888:Real-Time Polymerase Chain Reaction; D018805:Sepsis / Q000382:microbiology*