Structure and function of non-digestible carbohydrates in the gut microbiome.

Together with proteins and fats, carbohydrates are one of the macronutrients in the human diet. Digestible carbohydrates, such as starch, starch-based products, sucrose, lactose, glucose and some sugar alcohols and unusual (and fairly rare) α-linked glucans, directly provide us with energy while other carbohydrates including high molecular weight polysaccharides, mainly from plant cell walls, provide us with dietary fibre. Carbohydrates which are efficiently digested in the small intestine are not available in appreciable quantities to act as substrates for gut bacteria. Some oligo- and polysaccharides, many of which are also dietary fibres, are resistant to digestion in the small intestines and enter the colon where they provide substrates for the complex bacterial ecosystem that resides there. This review will focus on these non-digestible carbohydrates (NDC) and examine their impact on the gut microbiota and their physiological impact. Of particular focus will be the potential of non-digestible carbohydrates to act as prebiotics, but the review will also evaluate direct effects of NDC on human cells and systems.

Simulating colonic survival of probiotics in single-strain products compared to multi-strain products.

Background: Probiotic formulations can be single- or multi-strain. Commercially, multi-strain preparations have been suggested to have improved functionality over single-strain cultures. Probiotics are often tested as single-strain preparations but may subsequently be commercially formulated as multi-strain products. Objective: The aim of this study was to determine what happens at the site of action, the intestine, with probiotics as single- compared to multi-strain preparations. The human gastrointestinal tract contains a broad mixture of different microbes which may affect the survival of different probiotics in different ways. Design: The current study was performed to evaluate, in an in vitro colon simulation, whether probiotics influence each other's survival when they are taken as a combination of several strains (HOWARU Restore; Lactobacillus acidophilus NCFM, Lactobacillus paracasei Lpc-37, Bifidobacterium lactis Bl-04 and B. lactis Bi-07) compared to the strains as single preparations. Results: All strains could be detected after the colon simulations and there were no substantial differences in levels of the same strain when comparing single- and multi-strain products. Conclusions: It can be concluded that probiotics do not have an antagonistic effect on each other's survival when used in a multi-strain product compared to a single-strain product, at least within a microbiota in a simulated colonic environment.

Importance of the gastrointestinal life cycle of Bacillus for probiotic functionality.

Bacillus spp. are widely used in animal production for their probiotic properties. In many animal species, feed supplementation with specific Bacillus strains can provide numerous benefits including improvement in digestibility, the gut microbiota and immune modulation, and growth performance. Bacilli are fed to animals as spores that can sustain the harsh feed processing and long storage. However, the spores are metabolically quiescent and it is widely accepted that probiotics should be in a metabolically active state to perform certain probiotic functions like secretion of antimicrobial compounds and enzymes, synthesis of short chain fatty acids, and competition for essential nutrients. These functions should become active in the host gastrointestinal tract (GIT) soon after digestion of spores in order to contribute to microbiota and host metabolism. Considering that bacterial spores are metabolically dormant and many health benefits are provided by vegetative cells, it is of particular interest to discuss the life cycle of Bacillus in animal GIT. This review aims to capture the main characteristics of spores and vegetative cells and to discuss the latest knowledge in the life cycle of beneficial Bacillus in various intestinal environments. Furthermore, we review how the life cycle may influence probiotic functions of Bacillus and their benefits for human and animal health.

Influence of sucrose and xylitol on an early Streptococcus mutans biofilm in a dental simulator.

OBJECTIVES: In vitro methods to study dental biofilms are useful in finding ways to support a healthy microbial balance in the oral cavity. The effects of sucrose, xylitol, and their combination on three strains of Streptococcus mutans and one strain of Streptococcus sobrinus were studied using a dental simulator. METHODS: A simulator was used to mimic the oral cavity environment. It provided a continuous-flow system using artificial saliva (AS), constant temperature, mixing, and hydroxyapatite (HA) surface in which the influence of xylitol was studied. The quantities of planktonic and adhered bacteria were measured by real-time qPCR. RESULTS: Compared against the untreated AS, adding 1% sucrose increased the bacterial colonization of HA (p<0.0001) whereas 2% xylitol decreased it (p<0.05), with the exception of clinical S. mutans isolate 117. The combination of xylitol and sucrose decreased the bacterial quantities within the AS and the colonization on the HA by clinical S. mutans isolate 2366 was reduced (p<0.05). Increasing the concentration (2%-5%) of xylitol caused a reduction in bacterial counts even in the presence of sucrose. CONCLUSIONS: The continuous-culture biofilm model showed that within a young biofilm, sucrose significantly promotes whereas xylitol reduces bacterial colonization and proliferation. The results indicate that xylitol affects the ability of certain S. mutans strains to adhere to the HA. Clinical studies have also shown that xylitol consumption decreases caries incidence and reduces the amount of plaque. This study contributes to the understanding of the mechanism behind these clinical observations.

Lactobacillus acidophilus supplementation in human subjects and their resistance to enterotoxigenic Escherichia coli infection.

To assess the effect of Lactobacillus acidophilus (American Type Culture Collection (ATCC) 700396) on enterotoxigenic Escherichia coli (ETEC) infection, in the present study, a parallel, double-blind, placebo-controlled 4-week intervention was performed in healthy males. The subjects largely consumed their habitual diet, but had to abstain from consuming dairy foods generally high in Ca. The subjects were randomised into the L. acidophilus (dose 109 colony-forming units twice daily; n 20) or the placebo (n 19) group. After an adaptation period of 2 weeks, the subjects were orally infected with a live, but attenuated, ETEC vaccine, able to induce mild, short-lived symptoms. Before and after the challenge, the subjects recorded stool consistency, bowel habits, and frequency and severity of gastrointestinal complaints. The ETEC challenge led to a significant increase in faecal output on the 2nd day and a concomitant increase in Bristol stool scale scores. Likewise, abdominal pain, bloating, flatulence, fever, headache and nausea peaked 1 d after the oral challenge. The concentrations of faecal calprotectin and IgA peaked 2 d after and that of serum IgM peaked 9 and 15 d after the oral challenge. The concentrations of serum IgA and IgG were unaffected. The ETEC challenge led to a reduction in the number of Bacteroides-Prevotella, Bifidobacterium, Clostridium cluster XIVab and total faecal bacteria. Probiotic treatment was associated with a larger increase in Bristol stool scale scores and more fever, headache and nausea after the ETEC challenge compared with the placebo treatment. These differences were, however, small and with substantial variation within the groups. Oral application of an attenuated live ETEC vaccine provides a useful model for food-borne infections. Supplementation with L. acidophilus ATCC 700396, however, was ineffective in reducing ETEC infection symptoms in healthy men.

The effect of probiotics on faecal microbiota and genotoxic activity of faecal water in patients with atopic dermatitis: a randomized, placebo-controlled study.

BACKGROUND: Colonic microbiota is involved in the etiology of colon cancer according to several reports. Studies also indicate that the microbiota differs between atopic patients and healthy subjects. OBJECTIVE: To evaluate whether a probiotic mix containing Lactobacillus paracasei Lpc-37, Lactobacillus acidophilus 74-2, and Bifidobacterium animalis subsp. lactis DGCC 420 can affect the microbiota and its genotoxic activity in healthy subjects and patients with atopic dermatitis (AD). METHODS: A placebo-controlled cross-over study was conducted. Fifteen healthy adults and 15 adult AD patients consumed 2×100 ml/d of either a probiotic or a placebo drink for 8 weeks followed by a wash out period of 2 weeks before crossing the intervention. Faecal water was isolated from stool samples collected at the end of each period. HT29c19a cells incubated with faecal water were measured for DNA damage using single-cell gel electrophoresis ("comet assay"). Bacterial species were determined by qPCR and concentrations of short-chain fatty acids were measured by means of gas chromatography. RESULTS: Probiotic supplementation resulted in a significant increase in lactobacilli, whereas numbers of Bifidobacteria and Bacteroidetes remained unchanged. Clostridium perfringens cluster I-II was significantly reduced in healthy subjects. Genotoxic potential (expressed as tail intensity) of faecal water, was not affected. However, tail intensity decreased significantly in the probiotic period compared to placebo (23.5 vs. 16.7%) in AD patients. Although faecal concentrations of short-chain fatty acids were not affected, faecal pH was significantly reduced (7.0 vs. 6.6) in AD patients after probiotics. CONCLUSION: The results indicate that probiotics lower the genotoxic potential of faecal water in AD patients. The faecal C. perfringens cluster I-II levels remained unaffected suggesting either a change in their activity, or the fact that other bacterial species are responsible for the reduced genotoxic activity of faecal water.

Emergence of extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae during the years 2000 and 2004 in Helsinki, Finland.

The molecular epidemiology of 33 Escherichia coli and 81 Klebsiella pneumoniae extended-spectrum beta-lactamase-producing healthcare-associated and community-acquired isolates collected in the Helsinki district during 2000-2004 was investigated. Clonality studies, antimicrobial susceptibility and genotyping of the isolates were performed. Newly emerging CTX-M-producing E. coli and bla(SHV-12)-producing K. pneumoniae isolates were detected. Clonal clusters of both species persisted throughout the study period.