Publications (5) View all
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Article: Quantitative strain-specific detection of Lactobacillus rhamnosus GG in human faecal samples by real-time PCR.
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ABSTRACT: To develop a strain-specific rapid assay for identification and quantification of Lactobacillus rhamnosus GG in human faecal samples. A unique random amplified polymorphic DNA (RAPD) band of the L. rhamnosus GG strain was isolated and sequenced. Strain-specific polymerase chain reaction (PCR) primers and probes were designed based on the sequence. Quantification was performed by the real-time PCR using a fluorescent resonance energy transfer (FRET) system. The specificity of the assay was tested with DNA isolated from a set of known strains and human faecal samples. The analytical sensitivity of the method for L. rhamnosus GG was about 10 CFU per assay, which corresponds to 10(5) CFU g(-1) of wet faeces. Quantitative real-time PCR is a suitable method for strain-specific identification of L. rhamnosus GG in human faecal samples. Lactobacillus rhamnosus GG is one of the most studied probiotic strains in clinical trials but still lacks a DNA-based identification method. This study describes a real-time PCR method for strain-specific identification and quantification of L. rhamnosus GG in human faecal samples.Journal of Applied Microbiology 03/2009; 106(2):506-14. · 2.34 Impact Factor -
Article: Effects of anti-Helicobacter pylori treatment and probiotic supplementation on intestinal microbiota.
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ABSTRACT: The aims of this study were (i) to evaluate the effect of recommended antimicrobial treatment of Helicobacter pylori infection, consisting of clarithromycin, amoxicillin and lansoprazole, on intestinal microbiota and (ii) to determine the ability of a probiotic combination containing Lactobacillus rhamnosus GG, L. rhamnosus LC705, Propionibacterium freudenreichii ssp. shermanii JS and Bifidobacterium breve Bb99 to prevent treatment-induced alterations in the intestinal microbiota. Faecal samples were obtained from 39 H. pylori-infected patients randomised into two treatment groups. In addition, 19 H. pylori-negative volunteers were included in the study as a control group. Samples were collected before, during and after treatment and microbiota were analysed by fluorescence in situ hybridisation and culture. The quantities of the predominant bacterial groups were altered significantly in both groups and disturbances were seen even 9 weeks after treatment was complete. Probiotics slightly counteracted the effects of anti-H. pylori treatment, seen as significantly less alterations in the total numbers of aerobes and lactobacilli/enterococci. At baseline, the composition of the microbiota between H. pylori-positive versus H. pylori-negative control individuals differed with regard to clostridia and the total number of anaerobes. The recommended treatment for H. pylori infection induces long-term disturbances in the intestinal microbiota. The probiotic combination appeared to result in only minor changes in the microbiota.International Journal of Antimicrobial Agents 02/2007; 29(1):66-72. · 4.13 Impact Factor -
Article: Probiotic supplementation improves tolerance to Helicobacter pylori eradication therapy--a placebo-controlled, double-blind randomized pilot study.
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ABSTRACT: H. pylori is the major cause of chronic gastritis, and a risk factor for peptic ulcer and gastric cancer. To investigate the effect of probiotic supplementation on the tolerance and efficacy of H. pylori eradication treatment in a randomized, double-blind, placebo-controlled trial. A total of 338 volunteers were screened for H. pylori infection. The eligibility criteria were met by 47 subjects whose H. pylori infection was verified at the outset and re-evaluated after the treatment by the 13C-urea breath test and by enzyme immunoassay serology. The subjects were randomized to receive probiotic therapy (Lactobacillus rhamnosus GG, L. rhamnosusLC705, Bifidobacterium breve Bb99 and Propionibacterium freudenreichii ssp. shermanii JS) or a placebo during H. pylori eradication and for 3 weeks following the treatment, and recorded their daily symptoms in a standardized diary. When the frequencies of new or aggravated symptoms were evaluated, no significant differences were found between the two groups for individual symptoms. However, the probiotic group showed less treatment-related symptoms as measured by the total symptom score change (P = 0.038) throughout the H. pylori eradication therapy in contrast to the placebo group. The H. pylori eradication rate was non-significantly higher in the group receiving probiotic therapy (91% vs. 79%, P = 0.42). In this group the recovery of probiotic bacteria in the faeces increased significantly (P < 0.001). In this pilot study, probiotic supplementation did not diminish significantly the frequency of new or aggravated symptoms during H. pylori eradication. However, our data suggest an improved tolerance to the eradication treatment when total symptom severity was taken into account. Furthermore, the results show that probiotic bacteria are able to survive in the gastrointestinal tract despite the intensive antimicrobial therapy.Alimentary Pharmacology & Therapeutics 06/2005; 21(10):1263-72. · 3.77 Impact Factor -
Article: The effect of ageing with and without non-steroidal anti-inflammatory drugs on gastrointestinal microbiology and immunology.
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ABSTRACT: Elderly individuals are more susceptible to gastrointestinal problems such as constipation than young adults. Furthermore, the common use of non-steroidal anti-inflammatory drugs (NSAID) among the elderly is known to further increase such gastrointestinal ailments. To describe the specific changes in elderly, intestinal microbes, their metabolites and immune markers were measured from faecal samples obtained from fifty-five elderly individuals (aged 68-88 years), using either NSAID or not, and fourteen young adults (aged 21-39 years). The faecal DM content increased with age but was significantly lower among the elderly NSAID users. The microbial metabolism was especially influenced by NSAID use and/or ageing, although fewer changes were observed in the composition of the microbial community, whilst the level of aerobes was increased in the elderly and the level of Clostridium coccoides-Eubacterium rectale reduced in the elderly NSAID users as compared with young adults. An increase in the concentrations of some branched SCFA and l-lactate but a decrease in some major SCFA concentrations were observed. Evidently, the decreased defecation frequency in the elderly directed colonic fermentation toward an unfavourable microbial metabolism but this was partially offset by the use of NSAID. Irrespective of the use of NSAID, the elderly subjects had significantly lower concentrations of faecal PGE2 than the young adults, reflecting possibly a reduced immune response. According to the present study more attention should be paid to the development of dietary products that seek to enhance bowel function, saccharolytic fermentation and immune stimulation in the elderly population.The British journal of nutrition 08/2008; 100(1):130-7. · 3.45 Impact Factor -
Article: In school-aged children a combination of galacto-oligosaccharides and Lactobacillus GG increases bifidobacteria more than Lactobacillus GG on its own.
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ABSTRACT: The aim of this study was to compare a combination of Lactobacillus GG (LGG) and galacto-oligosaccharides (GOS) with LGG on its own, and their effects on the intestinal microbiota in school-aged children. The randomized, double-blinded, crossover study comprised 30 healthy children. There were two 3-week study periods with a 4-week wash-out period in between. The children ingested daily 65 ml of milk-based fruit juice containing either LGG alone (6.5 x 10(9) CFU) or LGG plus 2 g of GOS. Symptom diaries were filled during the study periods. Fecal samples were collected at the beginning and end of both study periods. At the end of both study periods, the amount of bifidobacteria was significantly greater after the ingestion of LGG + GOS compared with LGG alone (geometric mean 9.33 x 10(9) vs. 4.28 x 10(9) CFU/g, p < 0.001). No significant differences were seen in the amount of lactobacilli or LGG, nor did gastrointestinal symptoms, defecation frequency, consistency of stools or ease of defecation differ between the two study periods. Ingestion of LGG combined with 2 g of GOS increased the bifidobacteria more than LGG on its own and thus GOS clearly has a prebiotic effect in children. The children tolerated well a daily intake of 2 g of GOS.Annals of Nutrition and Metabolism 01/2008; 52(3):204-8. · 2.26 Impact Factor
