Microbiological Effects of Consuming a Synbiotic Containing Bifidobacterium bifidum, Bifidobacterium lactis, and Oligofructose in Elderly Persons, Determined by Real-Time Polymerase Chain Reaction and Counting of Viable Bacteria
TL;DR: Synbiotic consumption increased the size and diversity of protective fecal bifidobacterial populations, which are often very much reduced in older people.
Abstract: Background Because of changes in gut physiology, immune system reactivity, and diet, elderly people are more susceptible to gastrointestinal infections than are younger adults. The gut microflora, which provides a natural defense against invading microorganisms, changes in elderly people with the development of potentially damaging bacterial populations, which may lead to alterations in bacterial metabolism and higher levels of infection. Methods A randomized, double-blind, controlled feeding trial was done with 18 healthy elderly volunteers (age, >62 years) using a synbiotic comprising Bifidobacterium bifidum BB-02 and Bifidobacterium lactis BL-01 (probiotics) together with an inulin-based prebiotic (Synergy 1; Orafti). Real-time PCR was employed to quantitate total bifidobacteria, B. bifidum, and B. lactis in fecal DNA before, during, and after synbiotic consumption. Counting all viable anaerobes, bifidobacteria, and lactobacilli and identification of bacterial isolates to species level was also done. Results Throughout feeding, both bifidobacteria species were detected in fecal samples obtained from all subjects receiving the synbiotic, with significant increases in the number of copies of the 16S rRNA genes of B. bifidum, B. lactis, and total bifidobacteria, compared with the control week and the placebo group. At least 1 of these species remained detectable in fecal samples 3 weeks after feeding in individuals that had no fecal B. bifidum and/or B. lactis in the control week, indicating that the probiotics persisted in the volunteers. Counting of viable organisms showed significantly higher total numbers of fecal bifidobacteria, total numbers of lactobacilli, and numbers of B. bifidum during synbiotic feeding. Conclusion Synbiotic consumption increased the size and diversity of protective fecal bifidobacterial populations, which are often very much reduced in older people.
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01 Jan 2007
TL;DR: Post-genomics studies of fecal bifidobacteria and batch cultures of B. longum resulted in enhanced understanding of the life style and generated important leads for further investigation of genes for metabolism and colonization of intestinal bifIDobacteria within the human host.
Abstract: Following birth, the human gastrointestinal tract is rapidly colonized by microorganisms that profoundly impact the physiology and health of the host by contributing to host nutrition and natural defense, amongst other activities. In the large intestine, bifidobacteria are especially predominant in infants, where they can comprise up to 90% of the total microbiota, and in adults still account for a significant several percent. In this thesis, we applied post-genomic technologies, including metaproteomics and bifidobacterial community transcript profiling, to deepen our understanding of the activity of commensal bifidobacteria in the human intestine. Proteins were extracted from total infant fecal microbiota that was dominated by bifidobacteria, and two-dimensional gel electrophoresis was used to visualize the metaproteomes. The succession of the proteins was studied in fecal microbiota of two infants during 40 days. This revealed that the number and intensity of protein spots changed in time, but the patterns remained similar and specific for each infant. In-gel digestion of protein-spots and sequencing of the peptides, revealed the presence of a bifidobacterial transaldolase supporting the future application of this approach. Global transcript profiling of the infant fecal bifidobacteria was also performed. Total RNA from the fecal microbiota of infants that were solely breast-fed or formula-fed was hybridized to a DNA microarray comprising clones covering the genomes of several bifidobacterial species. Significantly hybridizing clones were sequenced and compared with those in the public databases. While some sequences were found to be bifidobacterial ribosomal RNA, the majority showed similarity to protein-encoding genes predicted to be involved in carbohydrate metabolism, processing of information and housekeeping functions. Remarkably, significant similarity was observed to an operon involved in the utilization of specifically human milk oligosaccharides and mucin sugars, supporting the functionality of bifidobacteria in the infant intestine. Overall, transcript profiling revealed significant differences between breast-fed and formula-fed infants, which was also reflected in the diversity of fecal bifidobacterial species measured with quantitative real time PCR. In another study, a specific prebiotic mixture showed a shift in the dominant adult fecal microbiota as well as abundance of the different bifidobacterial species. Subsequent transcriptomics using the same bifidobacterial-targeted microarrays showed the activity of genes in a wide range of functional groups, the majority being involved in metabolism of carbohydrates of plant origin, house keeping functions, and membrane transport of a wide variety of substrates including sugars and metals. Furthermore, the transcriptome of Bifidobacterium longum was studied in vitro in human and formula milk and semi-synthetic medium with galactooligosaccharides which showed quite some differentially expressed genes for sugar utilization. Overall, the transcription of genes involved in carbohydrate metabolism and uptake were specifically induced. In conclusion, post-genomics studies of fecal bifidobacteria and batch cultures of B. longum resulted in enhanced understanding of the life style and generated important leads for further investigation of genes for metabolism and colonization of intestinal bifidobacteria within the human host.
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08 Aug 2012
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TL;DR: It was shown that real-time PCR can be regarded as an alternative to conventional techniques and also as a complement to results obtained from conventional culture method.
Abstract: The aim of the current study was to compare three methods for determining the influence of different feeding strategies on the gut microbiota of piglets. Forty-eight weanling piglets were fed four different diets enriched with insoluble dietary fibre (wheat bran and pollen from Pinus massoniana). Starting from ileal and colonic samples, the total microbial DNA was isolated and bacterial parameters (lactobacilli, bifidobacteria, Bacteroides vulgatus and total bacterial counts) were quantified using real-time polymerase chain reaction (PCR). The results for lactobacilli, bifidobacteria and total bacterial counts were compared with those obtained by fluorescence in situ hybridization (FISH) and cultivation method. No significant differences could be observed between dietary treatments with real-time PCR and FISH for all investigated parameters. Comparing the applied three methods no consistent results were achieved, whereas FISH usually showed lower values. It was shown that real-time PCR can be regarded as an alternative to conventional techniques and also as a complement to results obtained from conventional culture method.
5 citations
Cites background from "Microbiological Effects of Consumin..."
...Higher real-time PCR results may be explained due to multiple 16S rRNA operons in chromosomes and the additional detection of dead and viable but not culturable cells (Huijsdens et al. 2002; Bartosch et al. 2005; Fu et al. 2006)....
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References
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TL;DR: By combining the rationale of pro- and prebiotics, the concept of synbiotics is proposed to characterize some colonic foods with interesting nutritional properties that make these compounds candidates for classification as health-enhancing functional food ingredients.
Abstract: Because the human gut microbiota can play a major role in host health, there is currently some interest in the manipulation of the composition of the gut flora towards a potentially more remedial community. Attempts have been made to increase bacterial groups such as Bifidobacterium and Lactobacillus that are perceived as exerting health-promoting properties. Probiotics, defined as microbial food supplements that beneficially affect the host by improving its intestinal microbial balance, have been used to change the composition of colonic microbiota. However, such changes may be transient, and the implantation of exogenous bacteria therefore becomes limited. In contrast, prebiotics are nondigestible food ingredients that beneficially affect the host by selectively stimulating the growth and/or activity of one or a limited number of bacterial species already resident in the colon, and thus attempt to improve host health. Intake of prebiotics can significantly modulate the colonic microbiota by increasing the number of specific bacteria and thus changing the composition of the microbiota. Nondigestible oligosaccharides in general, and fructooligosaccharides in particular, are prebiotics. They have been shown to stimulate the growth of endogenous bifidobacteria, which, after a short feeding period, become predominant in human feces. Moreover, these prebiotics modulate lipid metabolism, most likely via fermentation products. By combining the rationale of pro- and prebiotics, the concept of synbiotics is proposed to characterize some colonic foods with interesting nutritional properties that make these compounds candidates for classification as health-enhancing functional food ingredients.
7,232 citations
"Microbiological Effects of Consumin..." refers background in this paper
...Inulin-type fructans are commonly used prebiotics, which have been defined as nondigestible dietary components that selectively stimulate the growth and/or activities of bacteria in the large bowel [15]....
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TL;DR: In this paper, the authors used probiotic treatments to re-establish the natural condition which exists in the wild animal but which has been disrupted by modern trends in conditions used for rearing young animals, including human babies, and in modern approaches to nutrition and disease therapy.
Abstract: There is good evidence that the complex microbial flora present in the gastrointestinal tract of all warm-blooded animals is effective in providing resistance to disease. However, the composition of this protective flora can be altered by dietary and environmental influences, making the host animal susceptible to disease and/or reducing its efficiency of food utilization. What we are doing with the probiotic treatments is re-establishing the natural condition which exists in the wild animal but which has been disrupted by modern trends in conditions used for rearing young animals, including human babies, and in modern approaches to nutrition and disease therapy. These are all areas where the gut flora can be altered for the worse and where, by the administration of probiotics, the natural balance of the gut microflora can be restored and the animal returned to its normal nutrition, growth and health status.
4,055 citations
Journal Article•
TL;DR: These are all areas where the gut flora can be altered for the worse and where, by the administration of probiotics, the natural balance of the gut microflora can be restored and the animal returned to its normal nutrition, growth and health status.
Abstract: There is good evidence that the complex microbial flora present in the gastrointestinal tract of all warm-blooded animals is effective in providing resistance to disease. However, the composition of this protective flora can be altered by dietary and environmental influences, making the host animal susceptible to disease and/or reducing its efficiency of food utilization. What we are doing with the probiotic treatments is re-establishing the natural condition which exists in the wild animal but which has been disrupted by modern trends in conditions used for rearing young animals, including human babies, and in modern approaches to nutrition and disease therapy. These are all areas where the gut flora can be altered for the worse and where, by the administration of probiotics, the natural balance of the gut microflora can be restored and the animal returned to its normal nutrition, growth and health status.
3,391 citations
TL;DR: This work has created a phylogenetically arranged report on rRNA gene copy number for a diverse collection of prokaryotic microorganisms in an attempt to understand the evolutionary implications of rRNA operon redundancy.
Abstract: The Ribosomal RNA Operon Copy Number Database (rrndb) is an Internet-accessible database containing annotated information on rRNA operon copy number among prokaryotes. Gene redundancy is uncommon in prokaryotic genomes, yet the rRNA genes can vary from one to as many as 15 copies. Despite the widespread use of 16S rRNA gene sequences for identification of prokaryotes, information on the number and sequence of individual rRNA genes in a genome is not readily accessible. In an attempt to understand the evolutionary implications of rRNA operon redundancy, we have created a phylogenetically arranged report on rRNA gene copy number for a diverse collection of prokaryotic microorganisms. Each entry (organism) in the rrndb contains detailed information linked directly to external websites including the Ribosomal Database Project, GenBank, PubMed and several culture collections. Data contained in the rrndb will be valuable to researchers investigating microbial ecology and evolution using 16S rRNA gene sequences. The rrndb web site is directly accessible on the WWW at http://rrndb.cme.msu.edu.
1,051 citations
Additional excerpts
...The aim of this double-blind, randomized, controlled feeding trial was to study the effects of ingestion of a synbiotic containing 2 bifidobacterial species (Bifidobacterium bifidum BB-02 and Bifidobacterium lactis BL-01) and oligofructose on the composition of intestinal bifidobacteria and Lactobacillus populations in older people....
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...Another reason why rRNA gene copy numbers were higher than cell numbers is that rRNA operons vary widely in bacteria, and between 2 to 5 rRNA operons have been found in different species belonging to the genus Bifidobacterium [34]....
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...A randomized, double-blind, controlled feeding trial was done with 18 healthy elderly volunteers (age, 162 years) using a synbiotic comprising Bifidobacterium bifidum BB-02 and Bifidobacterium lactis BL-01 (probiotics) together with an inulin-based prebiotic (Synergy 1; Orafti)....
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...With use of primers specific for the genus Bifidobacterium, significantly higher copy numbers of target DNA were found in the synbiotic group during the feeding period (weeks 4 and 5) and during the postfeeding period (weeks 6 and 8) (table 4)....
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...Bifidobacterium adolescentis, Bifidobacterium angulatum, and Bi- Synbiotic Feeding Study • CID 2005:40 (1 January) • 33 fidobacterium dentium predominated (table 2); Bifidobacterium breve, Bifidobacterium longum, and Bifidobacterium pullorum were occasionally found; and Bifidobacterium boum and Bifidobacterium catenulatum were detected in only a few stool samples....
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TL;DR: Since the total culturable counts were only a fraction of the total microscopic counts, the contribution of bifidobacteria to the total intestinal microflora was overestimated by almost 10-fold when cultural methods were used as the sole method for enumeration.
Abstract: Three 16S rRNA hybridization probes were developed and tested for genus-specific detection of Bifidobacterium species in the human fecal flora. Variable regions V2, V4, and V8 of the 16S rRNA contained sequences unique to this genus and proved applicable as target sites for oligodeoxynucleotide probes. Determination of the genus specificity of the oligonucleotides was performed by whole-cell hybridization with fluorescein isothiocyanate-labelled probes. To this end, cells were fixed on glass slides, hybridized with the probes, and monitored by videomicroscopy. In combination with image analysis, this allowed quantification of the fluorescence per cell and objective evaluation of hybridization experiments. One of the probes developed was used to determine the population of Bifidobacterium spp. in human fecal samples. A comparison was made with results obtained by cultural methods for enumeration. Since both methods gave similar population estimates, it was concluded that all bifidobacteria in feces were culturable. However, since the total culturable counts were only a fraction of the total microscopic counts, the contribution of bifidobacteria to the total intestinal microflora was overestimated by almost 10-fold when cultural methods were used as the sole method for enumeration.
979 citations
"Microbiological Effects of Consumin..." refers background in this paper
...Bifidobacterium genus Bif164F 5′-GGG TGG TAA TGC CGG ATG-3′ 457 59 [22] Bif601R 5′-TAA GCC ATG GAC TTT CAC ACC-3′ ....
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