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. 2005 Jan;71(1):501-6.
doi: 10.1128/AEM.71.1.501-506.2005.

Monitoring of antibiotic-induced alterations in the human intestinal microflora and detection of probiotic strains by use of terminal restriction fragment length polymorphism

Cecilia Jernberg  1 , Asa Sullivan, Charlotta Edlund, Janet K Jansson
Affiliations

Monitoring of antibiotic-induced alterations in the human intestinal microflora and detection of probiotic strains by use of terminal restriction fragment length polymorphism

Cecilia Jernberg et al. Appl Environ Microbiol. 2005 Jan.

Abstract

Terminal restriction fragment length polymorphism (T-RFLP) was investigated as a tool for monitoring the human intestinal microflora during antibiotic treatment and during ingestion of a probiotic product. Fecal samples from eight healthy volunteers were taken before, during, and after administration of clindamycin. During treatment, four subjects were given a probiotic, and four subjects were given a placebo. Changes in the microbial intestinal community composition and relative abundance of specific microbial populations in each subject were monitored by using viable counts and T-RFLP fingerprints. T-RFLP was also used to monitor specific bacterial populations that were either positively or negatively affected by clindamycin. Some dominant bacterial groups, such as Eubacterium spp., were easily monitored by T-RFLP, while they were hard to recover by cultivation. Furthermore, the two probiotic Lactobacillus strains were easily tracked by T-RFLP and were shown to be the dominant Lactobacillus community members in the intestinal microflora of subjects who received the probiotic.

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Figures

FIG. 1.
FIG. 1.
UPGMA dendrograms constructed from Euclidean pairwise distances between time points and corresponding PCA analyses of T-RFLP data from days 0, 7, and 21 for three combined restriction fragment profiles. Subjects A, B, C, and D received a probiotic during antibiotic administration, and subjects E, F, G, and H received ordinary yogurt during antibiotic administration. PCA1, principal component 1; PCA2, principal component 2.
FIG. 2.
FIG. 2.
TRFs from the MspI restriction digest representing members of the C. coccoides subgroup that were negatively affected by clindamycin. Each bar represents pooled relative abundance values for a group of TRFs whose sizes range from 222 to 227 bp (typical for this subgroup). Subjects A, B, C, and D (upper panel) received the probiotic, and subjects E, F, G, and H (lower panel) received ordinary yogurt.
FIG. 3.
FIG. 3.
Relative abundance values for TRFs obtained by using a Lactobacillus-specific reverse primer and HaeIII restriction digestion. Blue cones, day 0; yellow cones, day 7; red cones, day 21. The 247-bp TRF represents L. acidophilus NCFB 1748, and the 328-bp TRF represents L. paracasei F19. The data are for one representative subject who ingested the probiotic (subject A).
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