Use of culture-independent analysis to reveal alteration of intestinal microflora by heat-killed Lactobacillus pentosus in a mouse model of endogenous sepsis

• Published in: Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy Vol. 19; no. 4; pp. 673 - 676
• Main Authors: Sugasawa, Yasuyuki, Saga, Tomoo, Kimura, Soichiro, Ishii, Yoshikazu, Yamaguchi, Keizo, Tateda, Kazuhiro
• Format: Journal Article
• Language: English
• Published: Tokyo Elsevier Ltd 01.08.2013; Springer Japan
• Subjects: Animals; Bacteroides; Feces - microbiology; Female; Filamentous bacteria; Hematology, Oncology and Palliative Medicine; Infectious Diseases; Intestinal microbiology; Intestines - microbiology; Lactobacillus; Lactobacillus - growth & development; Lactobacillus - isolation & purification; Lactobacillus pentosus; Medical Microbiology; Medicine; Medicine & Public Health; Mice; Mice, Inbred ICR; Microbiota - genetics; Microbiota - physiology; Original Article; Polymorphism, Restriction Fragment Length; Pseudomonas aeruginosa; Quantitative PCR (qPCR); Real-Time Polymerase Chain Reaction; Sepsis - microbiology; Terminal-restriction fragment length polymorphism (T-RFLP); Virology; Quantitative PCR (qPCR); Filamentous bacteria; Intestinal microbiology; Terminal-restriction fragment length polymorphism (T-RFLP); Lactobacillus
• ISSN: 1341-321X; 1437-7780; 1437-7780
• DOI: 10.1007/s10156-012-0541-1

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.