SUMMARYBackground: H. pylori is the major cause of chronic gastritis, and a risk factor for peptic ulcer and gastric cancer. Aim: 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. Methods: 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 13 C-urea breath test and by enzyme immunoassay

Aims:  To develop a strain‐specific rapid assay for identification and quantification of Lactobacillus rhamnosus GG in human faecal samples. Methods and Results:  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 105 CFU g−1 of wet faeces. Conclusions:  Quantitative real‐time PCR is a suitable method for strain‐specific identification of L. rhamnosus GG in human faecal samples. Significance and Impact of the Study:  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.

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Elderly individuals are more susceptible to gastrointestinal problems such as constipation than young adults. Furthermore, the common use of nonsteroidal 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 PGE 2 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. Intestinal bacteria: Immune responses: Ageing: Non-steroidal anti-inflammatory drugsAgeing is associated with significant changes in gastrointestinal function. Due to changes in gut physiology, immune system reactivity and diet, elderly individuals are more susceptible to gastrointestinal problems and diseases than young adults. Diseases such as peptic ulcer and gastric cancer, irritable bowel syndrome, diverticulosis and colon cancer are relatively common problems of ageing. Constipation, another common concern in the elderly, often results in increased use of laxatives with ageing (1) .Furthermore, the common use of non-steroidal anti-inflammatory drugs (NSAID) among the elderly may affect their intestinal health. NSAID have analgesic, antipyretic and anti-inflammatory effects. The most common adverse effect caused by NSAID is damage to the mucosa in the gastrointestinal tract (2,3) . According to a recent study by Hartikainen et al. (4) , 70 % of the over-75-year-olds in the city of Kuopio (Finland) take at least one analgesic, of which NSAID are the most commonly used.During infancy, the intestinal microbiota changes dramatically (5) after which it remains relatively stable during childhood and throughout adult life (6) . However, during old age, it has been observed that changes in the composition of the microbiota occur again. Using cul...

Propionibacterium freudenreichii is a commercially important bacterium that is essential for the development of the characteristic eyes and flavor of Swiss-type cheeses. These bacteria grow actively and produce large quantities of flavor compounds during cheese ripening at warm temperatures but also appear to contribute to the aroma development during the subsequent cold storage of cheese. Here, we advance our understanding of the role of P. freudenreichii in cheese ripening by presenting the 2.68-Mbp annotated genome sequence of P. freudenreichii ssp. shermanii JS and determining its global transcriptional profiles during industrial cheese-making using transcriptome sequencing. The annotation of the genome identified a total of 2377 protein-coding genes and revealed the presence of enzymes and pathways for formation of several flavor compounds. Based on transcriptome profiling, the expression of 348 protein-coding genes was altered between the warm and cold room ripening of cheese. Several propionate, acetate, and diacetyl/acetoin production related genes had higher expression levels in the warm room, whereas a general slowing down of the metabolism and an activation of mobile genetic elements was seen in the cold room. A few ripening-related and amino acid catabolism involved genes were induced or remained active in cold room, indicating that strain JS contributes to the aroma development also during cold room ripening. In addition, we performed a comparative genomic analysis of strain JS and 29 other Propionibacterium strains of 10 different species, including an isolate of both P. freudenreichii subspecies freudenreichii and shermanii. Ortholog grouping of the predicted protein sequences revealed that close to 86% of the ortholog groups of strain JS, including a variety of ripening-related ortholog groups, were conserved across the P. freudenreichii isolates. Taken together, this study contributes to the understanding of the genomic basis of P. freudenreichii and sheds light on its activities during cheese ripening.

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 × 10⁹ 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 × 10⁹ vs. 4.28 × 10⁹ 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.

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Background: Helicobacter pylori (Hp) is the major cause of chronic gastritis, and a risk factor for peptic ulcer and gastric cancer. Aim: In this study we investigated the effect of probiotic supplementation on the tolerance and efficacy of Hp eradication treatment in a randomised, double-blind, placebo-controlled trial. Methods: A total of 338 volunteers were screened for Hp infection. The eligibility criteria were met by 47 subjects whose Hp infection was verified at the outset and re-evaluated after the treatment by the 13C-urea breath test and by EIA serology. The subjects were randomised to receive probiotic therapy (Lactobacillus rhamnosus GG, Lactobacillus rhamnosus LC750, Bifidobacterium breve Bb99 and Propionibacterium freudenreichii ssp.shermanii JS) or a placebo during Hp eradication and for three weeks following the treatment, and recorded their daily symptoms in a standardised diary. Results: 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 Hp eradication therapy in contrast to the placebo group. The Hp 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). Conclusions: In this pilot study, probiotic supplementation did not diminish significantly the frequency of new or aggravated symptoms during Hp eradication. However, our data suggest an improved tolerance to the eradication treatment when also the 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.