Clin Exp Immunol, 2005 Feb, 139(2), 245 - 56 Production of interleukin (IL)-10 and IL-12 by murine colonic dendritic cells in response to microbial stimuli; Rigby RJ et al.; Summary Intestinal dendritic cells (DC) are likely to regulate immunity to gut microflora, but little is known about their responses to bacterial antigens . Therefore, DC from normal murine colon were characterized and their cytokine responses to components of Gram-negative and/or Gram-positive bacteria assessed . Cells were obtained by digestion of colonic tissue and contained DC that were identified by flow cytometry as CD11c(+) major histocompatibility complex (MHC) class II(+) cells . Purified DC were obtained by immunomagnetic separation plus cell sorting . DC had the morphology of immature myeloid cells, were endocytically active, expressed low levels of co-stimulatory molecules and stimulated a weak allogeneic mixed leucocyte reaction . Analysis of flow cytometry data by a sensitive subtraction method allowed measurement of production of interleukin (IL)-12 and IL-10 by small numbers of gut DC by intracellular staining . Fewer than 5% of unstimulated DC produced either IL-10 or IL-12 . IL-10 production was significantly up-regulated following stimulation with Bifidobacteria longum, but not after exposure to lipopolysaccharide (LPS) or Streptococcus faecium . In contrast, colonic DC produced IL-12 in response to both LPS and B.longum . Thus, colonic DC can produce both IL-12 and IL-10 following bacterial stimulation . Cell wall components from different bacteria stimulate distinct responses and may direct immune responses differentially in the gut. Appl Microbiol Biotechnol . 2005 Jan 14; {Epub ahead of print} Cloning and characterization of arabinoxylan arabinofuranohydrolase-D3 (AXHd3) from Bifidobacterium adolescentis DSM20083; van den Broek LA et al.; Arabinoxylan arabinofuranohydrolase-D3 (AXHd3) from Bifidobacterium adolescentis releases only C3-linked arabinose residues from double-substituted xylose residues . A genomic library of B . adolescentis DSM20083 was screened for the presence of the axhD3 gene . Two plasmids were identified containing part of the axhD3 gene . The nucleotide sequences were combined and three open reading frames (ORFs) were found . The first ORF showed high homology with xylanases belonging to family 8 of the glycoside hydrolases and this gene was designated xylA . The second ORF was the axhD3 gene belonging to glycoside hydrolase family 43 . The third (partial) ORF coded for a putative carboxylesterase . The axhD3 gene was cloned and expressed in Escherichia coli . Several substrates were employed in the biochemical characterization of recombinant AXHd3 . The enzyme showed the highest activity toward wheat arabinoxylan oligosaccharides . In addition, beta-xylanase from Trichoderma sp . was able to degrade soluble wheat arabinoxylan polymer to a higher extent, after pretreatment with recombinant AXHd3 . Arabinoxylan oligosaccharides incubated with a combination of recombinant AXHd3 and an alpha-L: -arabinofuranosidase from Aspergillus niger did not result in a higher maximal release of arabinose than incubation with these enzymes separately. J Ren Nutr, 2005 Jan, 15(1), 77 - 80 Beneficial effects of Bifidobacteria in a gastroresistant seamless capsule on hyperhomocysteinemia in hemodialysis patients; Taki K et al.; Intestinal microflora is deranged in hemodialysis (HD) patients as an increase in aerobic bacteria such as Escherichia coli and a decrease in anaerobic bacteria such as Bifidobacterium . Bifidobacteria ferment carbohydrates to produce acetic acid and lactic acid, which inhibit the intestinal putrefaction . Thus, intake of Bifidobacteria effectively restores the disturbed microflora to normal . However, Bifidobacteria in most medical products and healthy foods cannot usually survive because of exposure to gastric juices before it reaches the intestines . A gastroresistant seamless capsule prevents Bifidobacteria from inactivation by acidic gastric juice and allows it to be active in the intestines . We showed that the oral administration of Bifidobacterium longum in a gastroresistant seamless capsule to HD patients is effective in decreasing the pre-HD serum levels of homocysteine, indoxyl sulfate, and triglyceride . The reduction in the serum level of homocysteine is mainly attributable to the supply of folate produced by Bifidobacterium longum in the human intestines. Gut, 2005 Feb, 54(2), 242 - 9 Synbiotic therapy (Bifidobacterium longum/Synergy 1) initiates resolution of inflammation in patients with active ulcerative colitis: a randomised controlled pilot trial; Furrie E et al.; BACKGROUND AND AIMS: Ulcerative colitis (UC) is an acute and chronic inflammatory disease of the large bowel with unknown aetiology . The immune response against normal commensal microorganisms is believed to drive inflammatory processes associated with UC . Therefore, modulation of bacterial communities on the gut mucosa, through the use of probiotics and prebiotics, may be used to modify the disease state . METHODS: A synbiotic was developed for use in UC patients combining a probiotic, Bifidobacterium longum, isolated from healthy rectal epithelium, and a prebiotic (Synergy 1), a preferential inulin-oligofructose growth substrate for the probiotic strain . Treatment was employed in a double blinded randomised controlled trial using 18 patients with active UC for a period of one month . Clinical status was scored and rectal biopsies were collected before and after treatment, and transcription levels of epithelium related immune markers were measured . RESULTS: Sigmoidoscopy scores (scale 0-6) were reduced in the test group (start 4.5 (1.4), end 3.1 (2.5)) compared with placebo (start 2.6 (2.1), end 3.2 (2.2)) (p = 0.06) . mRNA levels for human beta defensins 2, 3, and 4, which are strongly upregulated in active UC, were significantly reduced in the test group after treatment (p = 0.016, 0.038, and 0.008, respectively) . Tumour necrosis factor alpha and interleukin 1alpha, which are inflammatory cytokines that drive inflammation and induce defensin expression, were also significantly reduced after treatment (p = 0.018 and 0.023, respectively) . Biopsies in the test group had reduced inflammation and regeneration of epithelial tissue . CONCLUSIONS: Short term synbiotic treatment of active UC resulted in improvement of the full clinical appearance of chronic inflammation in patients receiving this therapy. Appl Environ Microbiol, 2005 Jan, 71(1), 487 - 500 Gene Structure and Transcriptional Organization of the dnaK Operon of Bifidobacterium breve UCC 2003 and Application of the Operon in Bifidobacterial Tracing; Ventura M et al.; The incorporation and delivery of bifidobacterial strains as probiotic components in many food preparations expose these microorganisms to a multitude of environmental insults, including heat and osmotic stresses . We characterized the dnaK gene region of Bifidobacterium breve UCC 2003 . Sequence analysis of the dnaK locus revealed four genes with the organization dnaK-grpE-dnaJ-ORF1, whose deduced protein products display significant similarity to corresponding chaperones found in other bacteria . Northern hybridization and real-time LightCycler PCR analysis revealed that the transcription of the dnaK operon was strongly induced by osmotic shock but was not induced significantly by heat stress . A 4.4-kb polycistronic mRNA, which represented the transcript of the complete dnaK gene region, was detected . Many other small transcripts, which were assumed to have resulted from intensive processing or degradation of this polycistronic mRNA, were identified . The transcription start site of the dnaK operon was determined by primer extension . Phylogenetic analysis of the available bifidobacterial grpE and dnaK genes suggested that the evolutionary development of these genes has been similar . The phylogeny derived from the various bifidobacterial grpE and dnaK sequences is consistent with that derived from 16S rRNA . The use of these genes in bifidobacterial species as an alternative or complement to the 16S rRNA gene marker provides sequence signatures that allow a high level of discrimination between closely related species of this genus. Curr Pharm Des, 2005, 11(1), 75 - 90 Modulation of the human gut microflora towards improved health using prebiotics - assessment of efficacy; Tuohy KM et al.; There is increasing awareness that the human gut microflora plays a critical role in maintaining host health, both within the gastrointestinal tract and, through the absorption of metabolites, systemically . An 'optimal' gut microflora establishes an efficient barrier to the invasion and colonisation of the gut by pathogenic bacteria, produces a range of metabolic substrates which in turn are utilized by the host (e.g . vitamins and short chain fatty acids) and stimulates the immune system in a non-inflammatory manner . Although little is known about the individual species of bacteria responsible for these beneficial activities, it is generally accepted that the bifidobacteria and lactobacilli constitute important components of the beneficial gut microflora . A number of diet-based microflora management tools have been developed and refined over recent decades including probiotic, prebiotic and synbiotic approaches . Each aims to stimulate numbers and/or activities of the bifidobacteria and lactobacilli within the gut microflora . The aim of this article is to examine how prebiotics are being applied to the improvement of human health and to review the scientific evidence supporting their use. Curr Pharm Des, 2005, 11(1), 17 - 23 Regulating the safety of probiotics - the European approach; von Wright A; This article reviews pertinent legislation regulating the safety of probiotics within the European Union (EU) . Currently available probiotic organisms and known issues regarding their safety are briefly summarised . While most of the species and genera, particularly lactobacilli and bifidobacteria are apparently safe, certain micro-organisms may be problematic; particularly the enterococci, which are associated with nosocomial infections and harbour transmissible antibiotic resistance determinants . At present, probiotic human foods are not governed under specific EU regulatory frameworks, although the Novel Food Regulation EU 258/97, could be relevant in some specific cases . However, microbial feed additives (regulated by Council Directive 70/254/EEC and in accordance with guidelines of the Scientific Committee on Animal Nutrition (SCAN)) are subjected to detailed safety assessment with the intention of ensuring that they are innocuous to target animals, users and consumers . Particular attention is focused on the presence of transmissible antibiotic resistance markers, and to the potential for production of harmful metabolites . The guidelines do not differentiate between species and strains with long histories of safe use and other micro-organisms . This has caused some concern regarding overregulation, if the same principles are to be applied to probiotics or starter cultures intended for human food use . Accordingly, SCAN has launched an initiative towards a "Qualified Perception of Safety" (QPS) concept, which would allow strains with established safety status to enter the market without extensive testing requirements . It is likely that the European Food Safety Authority (EFSA) will play a central role in the regulation of both human and animal probiotics. Biofactors, 2004, 21(1-4), 123 - 5 Production of nitric oxide, tumor necrosis factor-alpha and interleukin-6 by RAW264.7 macrophage cells treated with lactic acid bacteria isolated from kimchi; Hur HJ et al.; The present study investigated immunopotentiating activities of lactic acid bacteria isolated from kimchi (KLAB) in vitro . A RAW264.7 macrophage cell line was stimulated with four strains of KLAB and two strains of bifidobacteria, and then the production of nitric oxide (NO) and cytokines - tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) - was determined . Lactobacillus plantarum, a lactic acid bacteria involved in the latent fermentation of kimchi, was the most effective for the generation of NO, TNF-alpha, and IL-6 in macrophage . All strains generated NO, while for the TNF-alpha and IL-6, only L . plantarum had significant activity . Our results indicate that L . plantarum plays an important role in the immunopotentiating activity of kimchi. Pediatrics, 2005 Jan, 115(1), 5 - 9 Effect of a probiotic infant formula on infections in child care centers: comparison of two probiotic agents; Weizman Z et al.; OBJECTIVE: To investigate the effect of 2 different species of probiotics in preventing infections in infants attending child care centers . METHODS: A double-blind, placebo-controlled, randomized trial was conducted from December 1, 2000, to September 30, 2002, at 14 child care centers in the Beer-Sheva area of Israel in healthy term infants 4 to 10 months old . Infants were assigned randomly to formula supplemented with Bifidobacterium lactis (BB-12), Lactobacillus reuteri (American Type Culture Collection 55730), or no probiotics . Duration of feeding, including follow-up, for each participant was 12 weeks . All infants were fed only the assigned formula and were not breastfed due to parental decision before recruitment to the study . Probiotic or prebiotic food products or supplements were not allowed . Main outcome measures were number of days and number of episodes with fever (>38 degrees C) and number of days and number of episodes with diarrhea or respiratory illness . RESULTS: Participants (n = 201) were similar regarding gestational age, birth weight, gender, and previous breastfeeding . The controls (n = 60), compared with those fed B lactis (n = 73) or L reuteri (n = 68), had significantly more febrile episodes (mean {95% confidence interval}: 0.41 {0.28-0.54} vs 0.27 {0.17-0.37} vs 0.11 {0.04-0.18}, respectively) . The controls also had more diarrhea episodes (0.31 {0.22-0.40} vs 0.13 {0.05-0.21} vs 0.02 {0.01-0.05}, respectively) and episodes of longer duration (0.59 {0.34-0.84} vs 0.37 {0.08-0.66} vs 0.15 {0.12-0.18} days, respectively) . The L reuteri group, compared with BB-12 or controls, had a significant decrease of number of days with fever, clinic visits, child care absences, and antibiotic prescriptions . Rate and duration of respiratory illnesses did not differ significantly between groups . CONCLUSIONS: Child care infants fed a formula supplemented with L reuteri or B lactis had fewer and shorter episodes of diarrhea, with no effect on respiratory illnesses . These effects were more prominent with L reuteri, which was also the only supplement to improve additional morbidity parameters. Pediatrics, 2005 Jan, 115(1), 1 - 4 Oral probiotics reduce the incidence and severity of necrotizing enterocolitis in very low birth weight infants; Lin HC et al.; OBJECTIVE: We evaluated the efficacy of probiotics in reducing the incidence and severity of necrotizing enterocolitis (NEC) in very low birth weight (VLBW) infants . PATIENTS AND METHODS: A prospective, masked, randomized control trial was conducted to evaluate the beneficial effects of probiotics in reducing the incidence and severity of NEC among VLBW (<1500 g) infants . VLBW infants who started to fed enterally and survived beyond the seventh day after birth were eligible for the trial . They were randomized into 2 groups after parental informed consents were obtained . The infants in the study group were fed with Infloran (Lactobacillus acidophilus and Bifidobacterium infantis) with breast milk twice daily until discharged . Infants in the control group were fed with breast milk alone . The clinicians caring for the infants were blinded to the group assignment . The primary outcome was death or NEC (>or= stage 2) . RESULTS: Three hundred sixty-seven infants were enrolled: 180 in the study group and 187 in the control group . The demographic and clinical variables were similar in both groups . The incidence of death or NEC (>or= stage 2) was significantly lower in the study group (9 of 180 vs 24 of 187) . The incidence of NEC (>or= stage 2) was also significantly lower in the study when compared with the control group (2 of 180 vs 10 of 187) . There were 6 cases of severe NEC (Bell stage 3) in the control group and none in the study group . None of the positive blood culture grew Lactobacillus or Bifidobacterium species . CONCLUSION: Infloran as probiotics fed enterally with breast milk reduces the incidence and severity of NEC in VLBW infants. Proteomics . 2004 Dec 23; {Epub ahead of print} A proteomic view of Bifidobacterium infantis generated by multi-dimensional chromatography coupled with tandem mass spectrometry; Vitali B et al.; Bifidobacteria are Gram-positive prokaryotes that naturally colonize the human gut where they exert several health-promoting effects . The present paper reports the use of a strong cation exchange-reversed-phase-tandem mass spectrometry strategy to catalogue the most abundantly expressed proteins of a probiotic Bifidobacterium infantis strain . A global view of the B . infantis proteome was obtained . The bimodal representation of the proteins identified by mass spectrometry provides the first theoretical two-dimensional map of protein distribution for this organism . Among the 136 proteins identified by multidimensional protein identification technology (MudPIT) analysis, 118 showed the highest similarity with the translated sequences of B . longum genome, two proteins were similar to other Bifidobacterium species and the remaining 16 were similar to different genera . Specific biological activities have been assigned to 115 identified proteins, whereas 21 have been referred to the group of hypothetical proteins . The MudPIT approach allowed us to identify high mass and basic isoelectric point proteins that are generally challenging to visualize using the traditional two-dimensional electrophoresis technique . Redundancy in peptide and protein identification using the double chromatography technique was also evaluated. Med Hypotheses, 2005, 64(3), 533 - 8 Major depressive disorder: probiotics may be an adjuvant therapy; Logan AC et al.; Major depressive disorder (MDD) is an extremely complex and heterogeneous condition . Emerging research suggests that nutritional influences on MDD are currently underestimated . MDD patients have been shown to have elevated levels of pro-inflammatory cytokines, increased oxidative stress, altered gastrointestinal (GI) function, and lowered micronutrient and omega-3 fatty acid status . Small intestinal bacterial overgrowth (SIBO) is likely contributing to the limited nutrient absorption in MDD . Stress, a significant factor in MDD, is known to alter GI microflora, lowering levels of lactobacilli and bifidobacterium . Research suggests that bacteria in the GI tract can communicate with the central nervous system, even in the absence of an immune response . Probiotics have the potential to lower systemic inflammatory cytokines, decrease oxidative stress, improve nutritional status, and correct SIBO . The effect of probiotics on systemic inflammatory cytokines and oxidative stress may ultimately lead to increased brain derived neurotrophic factor (BDNF) . It is our contention that probiotics may be an adjuvant to standard care in MDD. J Vet Sci, 2004 Dec, 5(4), 391 - 5 Isolation of cholesterol-lowering lactic acid bacteria from human intestine for probiotic use; Lim HJ et al.; Cholesterol-lowering effect of lactic acid bacteria (LAB: Streptococcus, Lactobacillus and Bifidobacterium) is well-known . Thus, we investigated LAB isolated from human intestine on the cholesterol-lowering effect in vitro . Seven Streptococcus (61.1%), 11 Lactobacillus (71.8%) and 7 Bifidobacterium (27.9%) were isolated as acid (pH 2.5 and 3.0) and bile (0.3% oxgall) tolerant strains . Streptococcus HJS-1, Lactobacillus HJL-37 and Bifidobacterium HJB-4 were finally selected as probiotic strains to use through the bile salt hydrolase (BSH) activity assay by using MRS media added taurodeoxycholic acid (TDCA) and the cholesterol-lowering test by using soluble cholesterol containing MRS broth . These studies suggested that the isolated LAB had an excellent hypocholesterolemic effect. Br J Nutr, 2004 Dec, 92(6), 931 - 8 Intestinal immunity of rats with colon cancer is modulated by oligofructose-enriched inulin combined with Lactobacillus rhamnosus and Bifidobacterium lactis; Roller M et al.; Probiotics (PRO) are known to modulate immunity in animals and human subjects and to inhibit colon carcinogenesis in experimental models, but the effects of synbiotics (SYN) are not well understood . Therefore, the effects of PRO (Lactobacillus rhamnosus GG and Bifidobacterium lactis Bb12), PRE (inulin-based enriched with oligofructose, 100 g/kg) and SYN (combination of PRO and PRE) on the immune system of rats were investigated in the azoxymethane (AOM)-induced colon cancer model . After 33 weeks, rats with and without AOM treatment were killed and immune cells were isolated from spleen, mesenterial lymph nodes (MLN) and Peyer's patches (PP) . AOM treatment significantly reduced natural killer (NK) cell-like cytotoxicity in control rats and in PRO- and PRE-supplemented rats . SYN supplementation prevented the AOM-induced suppression of NK cell-like cytotoxicity in PP compared with control rats (P<0.01) . SYN and PRE supplementation stimulated IL-10 production in PP in these rats (P<0.01) and in MLN of rats not treated with AOM (P<0.05) . Interferon-gamma production in PP was decreased by PRO supplementation (PRO and SYN groups combined; P<0.05) . Proliferative responsiveness of lymphocytes (PP) from AOM-treated rats was suppressed in SYN-supplemented rats (P<0.01) . Overall, SYN supplementation in carcinogen-treated rats primarily modulated immune functions in the PP, coinciding with a reduced number of colon tumours . PRE and PRO provided in combination as SYN may contribute to the suppression of colon carcinogenesis by modulating the gut-associated lymphoid tissue. J Cosmet Sci, 2004 Sep-Oct, 55(5), 473 - 9 Topical application of Bifidobacterium-fermented soy milk extract containing genistein and daidzein improves rheological and physiological properties of skin; Miyazaki K et al.; The authors examined the effects of Bifidobacterium-fermented soy milk extract (BE) containing genistein and daidzein on the hyaluronic acid (HA) content and rheological and physiological properties of hairless mouse and/or human skin . Topical application of BE for six weeks significantly restored changes in the elasticity and viscoelasticity of mouse skin, increased the HA content, and hydrated and thickened mouse skin . Also, topical application of a gel formula containing 10% BE to the human forearm for three months significantly lessened the decrease in skin elasticity . Therefore, BE is expected to become a new cosmetic ingredient to prevent the loss of skin elasticity through enhancement of HA production. FEMS Immunol Med Microbiol, 2005 Jan 1, 43(1), 59 - 65 Similar bifidogenic effects of prebiotic-supplemented partially hydrolyzed infant formula and breastfeeding on infant gut microbiota; Rinne MM et al.; The aim of the study was to assess the quantitative and qualitative differences of the gut microbiota in infants . We evaluated gut microbiota at the age of 6 months in 32 infants who were either exclusively breast-fed, formula-fed, nursed by a formula supplemented with prebiotics (a mixture of fructo- and galacto-oligosaccharides) or breast-fed by mothers who had been given probiotics . The Bifidobacterium, Bacteroides, Clostridium and Lactobacillus/Enterococcus microbiota were assessed by the fluorescence in situ hybridization, and Bifidobacterium species were further characterized by PCR . Total number of bifidobacteria was lower among the formula-fed group than in other groups (P=0.044) . Total amounts of the other bacteria were comparable between the groups . The specific Bifidobacterium microbiota composition of the breast-fed infants was achieved in infants receiving prebiotic supplemented formula . This would suggest that early gut Bifidobacterium microbiota can be modified by special diets up to the age of 6 months. Arch Pharm Res, 2004 Nov, 27(11), 1136 - 40 Protective effect of fermented red ginseng on a transient focal ischemic rats; Bae EA et al.; Red ginseng and fermented red ginseng were prepared, and their composition of ginsenosides and antiischemic effect were investigated . When ginseng was steamed at 98-100 degrees C for 4 h and dried for 5 h at 60 degrees C, and extracted with alcohol, its main components were ginsenoside Rg3> ginsenoside Rb1 > ginsenoside Rb2 . When the ginseng was suspended in water and fermented for 5 days by previously cultured Bifidobacterium H-1 and freeze-dried (fermented red ginseng), its main components were compound K > ginsenoside Rg3 > or = ginsenoside Rh2 . Orally administered red ginseng extract did not protect ischemia-reperfusion brain injury . However, fermented red ginseng significantly protected ischemica-reperfusion brain injury . These results suggest that ginsenoside Rh2 and compound K, which was found to be at a higher content in fermented red ginseng than red ginseng, may improve ischemic brain injury. J Zhejiang Univ Sci, 2005 Jan, 6(1), 14 - 21 Intestinal microflora in rats with ischemia/reperfusion liver injury; Xing HC et al.; Objectives: To investigate the intestinal microflora status related to ischemia/reperfusion (I/R) liver injury and explore the possible mechanism . Methods: Specific pathogen free grade Sprague-Dawley rats were randomized into three groups: Control group (n=8), sham group (n=6) and I/R group (n=10) . Rats in the control group did not receive any treatment, rats in the I/R group were subjected to 20 min of liver ischemia, and rats in the sham group were only subjected to sham operation . Twenty-two hours later, the rats were sacrificed and liver enzymes and malondialdehyde (MDA), superoxide dismutase (SOD), serum endotoxin, intestinal bacterial count, intestinal mucosal histology, bacterial translocation to mesenteric lymph nodes, liver, spleen, and kidney were studied . Results: Ischemia/reperfusion increased liver enzymes, MDA, decreased SOD, and was associated with plasma endotoxin elevation in I/R group campared to those in the sham group . Intestinal Bifidobacterium and Lactobacillus decreased and intestinal Enterobacteria and Enterococci, bacterial translocation to kidney increased in the I/R group compared to the sham group . Intestinal microvilli were lost, disrupted and the interspace between cells became wider in the I/R group . Conclusion: I/R liver injury may lead to disturbance of intestinal microflora and impairment of intestinal mucosal barrier function, which contributes to endotoxemia and bacterial translocation to kidney. Curr Cancer Drug Targets, 2004 Dec, 4(8), 689 - 702 Bacteria and bacterial toxins as therapeutic agents for solid tumors; Michl P et al.; Patients with advanced solid tumors frequently relapse and succumb to their metastatic disease after developing resistance to conventional treatment modalities such as chemotherapy and radiotherapy . In these patients, novel strategies of targeting widespread tumors are urgently needed . The increasing knowledge of the underlying pathogenetic mechanisms has led to the identification of numerous molecules that are overexpressed in various tumors and accumulate at the cell surface . The use of genetically modified bacteria and their toxins targeting these surface molecules has emerged as a promising new treatment strategy in refractory cancers . This review focuses on bacterial toxins such as Diphtheria toxin (DT), Pseudomonas exotoxin A (PE) and Clostridium perfringens enterotoxin (CPE) . In addition, the use of anaerobic bacteria such as Clostridium, Salmonella and Bifidobacterium spp . as drug-delivery systems targeting hypoxic tumor areas will be discussed as a new therapeutic modality of advanced solid tumors. Appl Environ Microbiol, 2004 Dec, 70(12), 7220 - 8 Use of 16S rRNA gene-targeted group-specific primers for real-time PCR analysis of predominant bacteria in human feces; Matsuki T et al.; 16S rRNA gene-targeted group-specific primers were designed and validated for specific detection and quantification of the Clostridium leptum subgroup and the Atopobium cluster . To monitor the predominant bacteria in human feces by real-time PCR, we used these specific primers together with four sets of group-specific primers for the Clostridium coccoides group, the Bacteroides fragilis group, Bifidobacterium, and Prevotella developed in a previous study (T . Matsuki, K . Watanabe, J . Fujimoto, Y . Miyamoto, T . Takada, K . Matsumoto, H . Oyaizu, and R . Tanaka, Appl . Environ . Microbiol . 68:5445-5451, 2002) . Examination of DNA extracted from the feces of 46 healthy adults showed that the C . coccoides group was present in the greatest numbers (log10 10.3 +/- 0.3 cells per g {wet weight} {average +/- standard deviation}), followed by the C . leptum subgroup (log10 9.9 +/- 0.7 cells per g {wet weight}), the B . fragilis group (log10 9.9 +/- 0.3 cells per g {wet weight}), Bifidobacterium (log10 9.4 +/- 0.7 cells per g {wet weight}), and the Atopobium cluster (log10 9.3 +/- 0.7 cells per g {wet weight}) . These five bacterial groups were detected in all 46 volunteers . Prevotella was found in only 46% of the subjects at a level of log10 9.7 +/- 0.8 cells per g (wet weight) . Examination of changes in the population and the composition of the intestinal flora for six healthy adults over an 8-month period revealed that the composition of the flora of each volunteer remained stable throughout the test period. J Antimicrob Chemother, 2005 Jan, 55(1), 38 - 44 Epub 2004 Dec 1. Antimicrobial susceptibility of bifidobacteria; Moubareck C et al.; OBJECTIVES: The aim of our study was to analyse the antibiotic susceptibility of various strains of Bifidobacterium spp . to a wide range of antimicrobial agents . METHODS: Fifty strains belonging to eight species of bifidobacteria, isolated from humans, animals or probiotic products, were tested for susceptibility to 30 antibiotics by disc diffusion on Brucella agar supplemented with 5% laked sheep blood and vitamin K1 (1 mg/L) . MICs of nine anti-anaerobe agents, including three new molecules (telithromycin, linezolid and gatifloxacin), were determined using the reference agar-dilution method . RESULTS: All strains of bifidobacteria, whatever the species, were sensitive to penicillins: penicillin G, amoxicillin (MIC(50) 0.06 mg/L), piperacillin, ticarcillin, imipenem and usually anti-Gram-positive antibiotics (macrolides, clindamycin, pristinamycin, vancomycin and teicoplanin) . Susceptibility to cefalothin and cefotetan was variable . Most isolates (70%) were resistant to fusidic acid . As expected, high resistance rates were observed for aminoglycosides . Metronidazole, an agent known for its anti-anaerobe activity, was ineffective against 38% of the strains . The newly commercialized molecules, telithromycin, linezolid and gatifloxacin, were active with MIC(50)S of 1 mg/L . The only variation in susceptibility observed among the different species concerned Bifidobacterium breve, which appeared to be generally more resistant . Potentially acquired resistance was only observed against tetracycline and minocycline, in 14% of the strains . CONCLUSIONS: With regard to a general concern about the safety of probiotics, such as potential transferability of resistance determinants, bifidobacteria, with their low natural and acquired resistance to 30 antibiotics, appear risk-free. J Pediatr Gastroenterol Nutr, 2004 Nov, 39(5), 465 - 73 Prebiotic oligosaccharides in dietetic products for infants: a commentary by the ESPGHAN Committee on Nutrition; Agostoni C et al.; This article by the ESPGHAN Committee on Nutrition summarizes available information on the effects of adding prebiotic oligosaccharides to infant and follow-on formulae . Currently there are only limited studies evaluating prebiotic substances in dietetic products for infants . Although administration of prebiotic oligosaccharides has the potential to increase the total number of bifidobacteria in feces and may also soften stools, there is no published evidence of clinical benefits of adding prebiotic oligosaccharides to dietetic products for infants . Data on oligosaccharide mixtures in infant formulae do not demonstrate adverse effects, but further evaluation is recommended . Combinations and dosages in addition to those so far studied need to be fully evaluated with respect to both safety and efficacy before their use in commercial infant food products . Well-designed and carefully conducted randomized controlled trials with relevant inclusion/exclusion criteria, adequate sample sizes and validated clinical outcome measures are needed both in preterm and term infants . Future trials should define optimal quantity and types of oligosaccharides with prebiotic function, optimal dosages and duration of intake, short and long term benefits and safety . At the present time, therefore, the Committee takes the view that no general recommendation on the use of oligosaccharide supplementation in infancy as a prophylactic or therapeutic measure can be made. J Nutr, 2004 Dec, 134(12), 3277 - 83 Probiotic consumption does not enhance the cholesterol-lowering effect of soy in postmenopausal women; Greany KA et al.; Numerous studies report that soy lowers cholesterol . Probiotic bacteria were also reported to lower total cholesterol (TC) and LDL cholesterol (LDL-C) . We hypothesized that by altering intestinal microflora, probiotic consumption may also change phytoestrogen metabolism and enhance the effects of soy . To evaluate the independent and interactive effects of probiotic bacteria and soy on plasma TC, LDL-C, HDL cholesterol (HDL-C), and triglycerides (TG), 37 women with a baseline TC of 5.24 mmol/L were given the following 4 treatments for 6 wk each in a randomized crossover design: soy protein isolate (26 +/- 5 g soy protein containing 44 +/- 8 mg isoflavones/d); soy protein isolate + probiotic capsules (10(9) colony-forming units Lactobacillus acidophilus DDS-1 and Bifidobacterium longum); milk protein isolate (26 +/- 5 g milk protein/d); and milk protein isolate + probiotic . Soy consumption decreased plasma TC by 2.2% (P = 0.02) and LDL-C by 3.5% (P = 0.005), increased HDL-C by 4.2% (P = 0.006) and tended to decrease TG (P = 0.07) compared with milk protein intake . When divided according to initial TC concentration, soy effects were observed only in hypercholesterolemic women (TC > 5.17 mmol/L) . In this subgroup, soy treatments decreased plasma TC by 3.3% (P = 0.01), LDL-C by 4.5% (P = 0.004), and TG by 10.6% (P = 0.02), and increased HDL-C by 4.2% (P = 0.02) . When subjects were divided on the basis of plasma and urine concentrations of the isoflavone metabolite, equol, equol producers and nonproducers did not differ in baseline lipids or in the effects of soy . Probiotics did not lower cholesterol or enhance the effects of soy . These results confirm a beneficial effect of soy on plasma cholesterol in mildly hypercholesterolemic postmenopausal women independent of equol production status, but do not support an independent or additive effect of these particular probiotic bacteria. Fitoterapia, 2004 Dec, 75(7-8), 760 - 3 The bifidogenic effect of Taraxacum officinale root; Trojanova I et al.; The infusion of dandelion root (Taraxacum officinale) stimulated in vitro the growth of 14 strains of bifidobacteria . The utilization of oligofructans, glucose, fructose and total saccharides was determined by enzymatic and phenol-sulfuric methods . Dandelion oligofructans were important source of carbon and energy for bifidobacteria tested. Cancer Gene Ther . 2004 Nov 26; {Epub ahead of print} Bifidobacterium longum as an oral delivery system of endostatin for gene therapy on solid liver cancer; Fu GF et al.; To overcome difficulties that hampered widespread application of a specific delivery system in cancer gene therapy and to inhibit the growth of solid liver cancer, we utilized a strain of Bifidobacterium longum as a delivery system to transport an endostatin gene that can inhibit growth of tumor . The B . longum strain with the endostatin gene (B . longum-En) was taken orally by tumor-bearing nude mice through drencher preparation . The results showed that B . longum-En could strongly inhibit the growth of solid liver tumor in nude mice and prolong the survival time of tumor-bearing nude mice . Furthermore, tumor growth was inhibited more efficiently when the B . longum-En treatment included selenium . Enriching the B . longum-En treatment with selenium improves the activity of NK and T cells and stimulates the activity of IL-2 and TNF-alpha in BALB/c mice . These results suggest that B . longum may be a highly specific and efficient vector for transporting anticancer genes in cancer gene therapy.Cancer Gene Therapy advance online publication, 26 November 2004; doi:10.1038/sj.cgt.7700758. Eksp Klin Gastroenterol, 2004, (3), 83 - 7, 105 Probiotics and mechanisms of their curative action. {Intestinal bacterial colonisation and translocation in C3H/HeJ conventional mice fed a unique dose of live or dead Bifidobacterium breve C50.} Mullie C, Yazourh A, Parser-Andrada A, Romond MB, Romond Ch. Faculte de pharmacie, universite de Picardie Jules Verne, 1, rue des Louvels, F 80037 Amiens Cedex 1, France . catherine.mullie@u-picardie.fr The efficiency and safety of use of Bifidobacterium breve C50 (BbC50), a potential probiotic, was assessed as regards intestinal microbial colonisation and bacterial translocation . A suspension of BbC50, containing 1-5 to 107-108 live bacteria, was fed to C3H/HeJ mice . The passage of live BbC50 was not demonstrated by culture either in the intestine or extra-intestinal organs . However, mice receiving the highest dose of live bacteria harbored more lactobacilli and less Bacteroides fragilis group in the cecum and colon when compared to control mice . Translocation of lactobacilli observed in the control group was not regulated by Bb50 feeding . Indeed, the spleen was significantly more frequently contaminated in mice fed BbC50, whatever the dose of live bacteria . The kidneys were also significantly more contaminated with lactobacilli in mice fed the highest dose of live Bb50 . Moreover, higher dose of live BbC50 was associated with greater number of extra-intestinal contaminated organs . To conclude, BbC50 feeding induced a favorable balance in the mouse intestinal flora and was never found translocating, demonstrating its efficiency and safety of use . However, BbC50 seemed to interfere with the ability of lymphoid organs (e.g . the spleen) to eliminate translocating lactobacilli. J Pediatr Surg, 2004 Nov, 39(11), 1686 - 92 Experience of long-term synbiotic therapy in seven short bowel patients with refractory enterocolitis; Kanamori Y et al.; BACKGROUND/PURPOSE: Probiotic and prebiotic therapies are potent new strategies to treat various intestinal diseases, including inflammatory bowel disease and viral and bacterial infections . Synbiotics is defined as the combined use of probiotics and prebiotics and is expected to have a stronger effect on intestinal diseases than probiotics or prebiotics alone, but there has been no report of its clinical application . The authors designed a protocol for synbiotic therapy composed of Bifidobacterium breve, Lactobacillus casei, and galactooligosaccharides and preliminarily ascertained its clinical effects in humans . METHODS: This protocol of synbiotic therapy was applied for more than 1 year to 7 malnourished patients with short bowels who suffered from refractory enterocolitis . RESULTS: The therapeutic protocol improved the intestinal bacterial flora (inducing the domination by anaerobic bacteria and suppressing the residence of pathogenic bacteria) and increased short chain fatty acids in the feces (from 27.8 to 65.09 micromol/g wet feces) . All patients but 1 accelerated their body weight gain, and 5 patients showed increased serum rapid turnover proteins . CONCLUSIONS: This protocol for synbiotic therapy might be a potent modulator of intestinal flora and a promising strategy to treat short bowel patients with refractory enterocolitis. J Dairy Sci, 2004 Dec, 87(12), 4073 - 9 Suppressive effect of functional drinking yogurt containing specific egg yolk immunoglobulin on Helicobacter pylori in humans; Horie K et al.; Helicobacter pylori is a human pathogen that infects over 50% of the population worldwide . It is the most important etiologic agent of gastroduodenal ulcers and malignancies . Helicobacter pylori urease enzyme is considered the main factor for the organism's colonization in the gastroduodenal mucosa . Hens immunized with the purified urease produce a highly specific anti-H . pylori urease immunoglobulin (IgY-urease) in their egg yolks . Immunoglobulin Y-urease was stable at 60 to 65 degrees C for 30 min and at pH 4.0 for 7 h . Its activity was lost at 80 degrees C for 20 min and at pH 2 for 4 h . Specially designed functional drinking yogurt containing Lactobacillus acidophilus and Bifidobacterium spp . with 1% egg yolk IgY-urease was produced commercially . Immunoglobulin Y-urease activity showed stability in the product up to 7 d, and then decreased to 85% after 3 wk of storage . A clinical study was conducted to determine the effectiveness of IgY-urease yogurt to suppress infection in humans . Forty-two volunteers who tested positive for H . pylori using a 13C-urea breath test were recruited . A total of 450 mL of IgY-urease (test group) or IgY-urease-free yogurt (control group) was consumed in 150-mL portions 3 times daily for 4 wk . Volunteers were tested after 2 and 4 wk; urea breath test values significantly decreased in the test group compared with the control group . The results indicate that suppression of H . pylori infection in humans could be achieved by consumption of drinking yogurt fortified with IgY-urease. J Nutr, 2004 Sep, 134(9), 2359 - 64 Maternal adaptive immunity influences the intestinal microflora of suckling mice; Diaz RL et al.; The microflorae in the intestine of breast-fed infants are distinct from those that typically populate the intestine of formula-fed infants . Although the acquisition of passive immunity through breast-feeding may play a critical role in influencing the pattern of bacterial colonization of the gut, the precise mechanisms underlying the differences in the commensal microflorae of breast and formula-fed children have not been established . We hypothesized that the assemblage of commensal microflorae in suckling and weaned mice may be influenced by the maternal adaptive immune system . To test this hypothesis, we analyzed the intestinal microflorae of mice reared in the presence (wild-type) or absence of an intact maternal immune system (T- and B-cell deficient) . Several types of bacteria (Lactobacillus, Enterococcus, Clostridium perfringens, Bifidobacterium, and Bacteroides) were isolated and enumerated from both the small and large intestine of 10-, 18-, 25- and 40- to 60-d old mice using selective media . The densities of bacteria were significantly lower in the small intestine of weaned mice that were reared by wild-type (WT) compared with immunodeficient (ID) dams . However, the microflorae were generally more abundant in the large intestine of suckling pups reared by WT compared with ID dams . Our results indicate that intestinal microflorae change throughout the suckling phase of development and that the maternal adaptive immune system influences the pattern and abundance of bacteria within the gut in an age- and site-specific manner. Med Sci Monit, 2004 Sep, 10(9), BR346 - 50 Epub 2004 Aug 20. Microbiological and genetic analysis of probiotic products licensed for medicinal purposes; Szajewska H et al.; BACKGROUND: The purpose of our study was to investigate both qualitatively and quantitatively the microbial content of probiotic products licensed for medicinal purposes . MATERIAL/METHODS: Microbiological analysis was performed on five different brands of probiotic products that claimed to contain lactobacilli and/or bifidobacteria . The species were determined based on phenotypic characters, using API 50CH, API 20A, and API rapid ID 32A kits . Bacterial strains belonging to the Bifidobacterium genus were further identified using genotypic methods (amplification of specific DNA fragments by PCR and analysis of their nucleotide sequences) . The products were also analyzed for pathogenic bacteria . The number of microorganisms contained in four of the products was determined using the plate-count method and the most-probable-number method . The actual and claimed content of probiotic products was analyzed statistically . RESULTS: Microbiological and genetic analysis showed that, in terms of quality, only three of the five products contained the bacterial strains claimed on the label . None of the tested products contained pathogens . Quantitative analysis demonstrated that 57 of 64 samples (89% {95% CI: 81-97}) contained bacterial counts at the cell densities (doses) claimed on the label . CONCLUSIONS: Our study demonstrates unsatisfactory qualitative microbiological specification in the tested products . However, there was good quantitative agreement with the labeling . Our findings indicate that regulations governing the labeling of probiotic products are urgently required. Am J Clin Nutr, 2004 Sep, 80(3), 737 - 41 Effects of ingesting Lactobacillus- and Bifidobacterium-containing yogurt in subjects with colonized Helicobacter pylori; Wang KY et al.; BACKGROUND: Evidence suggests that ingesting lactic acid bacteria exerts a suppressive effect on Helicobacter pylori infection in both animals and humans . Supplementing with Lactobacillus- and Bifidobacterium-containing yogurt (AB-yogurt) was shown to improve the rates of eradication of H . pylori in humans . OBJECTIVE: We administered AB-yogurt to subjects with asymptomatic H . pylori to test whether the yogurt could inhibit H . pylori growth . DESIGN: The in vitro inhibition of H . pylori growth was determined by inoculating Lactobacillus acidophilus La5 or Bifidobacterium lactis Bb12 on plates that were inoculated with H . pylori . Assessment of the viability of H . pylori was performed by the mixed culture method with La5 or Bb12 . In an intervention study, 59 adult volunteers infected with H . pylori were given AB-yogurt (10(7) colony-forming units of both La5 and Bb12/mL) twice daily after a meal for 6 wk . Eleven subjects positive for H . pylori infection were treated with milk placebo as control subjects . H . pylori bacterial loads were determined with use of the (13)C-urea breath test, which was performed before and 4 and 8 wk after the start of AB-yogurt supplementation . RESULTS: Bb12 exerted an in vitro inhibitory effect against H . pylori, whereas La5 did not show an effect . Administration of AB-yogurt decreased the urease activity of H . pylori after 6 wk of therapy (P < 0.0001) . CONCLUSION: Regular intake of yogurt containing Bb12 and La5 effectively suppressed H . pylori infection in humans. Int Endod J, 2004 Sep, 37(9), 579 - 87 Gram-positive rods prevailing in teeth with apical periodontitis undergoing root canal treatment; Chavez de Paz LE et al.; AIMS: To identify Gram-positive rods from root canals of teeth with apical periodontitis and to examine their associations with other species . METHODOLOGY: Consecutive root canal samples (RCSs) from 139 teeth undergoing root canal treatment were analyzed prospectively for cultivable microbes . Gram-positive rods in the first RCS submitted after chemo-mechanical preparation were categorised to genus level by selective media and gas-liquid chromatography (GLC), and identified to species level by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) . Associations between organisms were measured by odds ratios (OR) . RESULTS: In the first samples submitted a total of 158 Gram-positive rods, 115 Gram-positive cocci, 26 Gram-negative rods and 9 Gram-negative cocci, were identified . At genus levels Gram-positive rods were classified into: Lactobacillus spp . (38%), Olsenella spp . (18%), Propionibacterium spp . (13%), Actinomyces spp . (12%), Bifidobacterium spp . (13%) and Eubacterium spp . (6%) . The most frequent species were Olsenella uli, Lactobacillus paracasei and Propionibacterium propionicum . In subsequent samples taken during treatment, Gram-positive rods were also identified, although the number of strains was considerably reduced . Positive associations were observed between members of the genus lactobacilli and Gram-positive cocci (OR>2) . CONCLUSIONS: Olsenella uli and Lactobacillus spp . predominated over other Gram-positive rods . A possible association exists between Lactobacillus spp . and Gram-positive cocci in root canals of teeth with apical periodontitis receiving treatment. Postgrad Med J, 2004 Aug, 80(946), 447 - 51 Probiotics and prebiotics in the elderly; Hamilton-Miller JM; Probiotics (usually lactobacilli and bifidobacteria) and prebiotics (non-digestible oligosaccharides) have been shown to be useful in preventing certain disease conditions as well as possibly promoting specific aspects of health . In the present review, the evidence from clinical trials for benefits from probiotics and prebiotics to elderly populations is presented and discussed, specifically in respect of three common conditions found in the elderly . Both probiotics and prebiotics may be helpful in malnutrition, particularly in lactose intolerance and calcium absorption, and in constipation . Probiotics have been shown clearly to boost immunity in the elderly, but the clinical significance of this remains to be clarified . These results are encouraging, and further large scale studies seem justified to establish the place of probiotic and prebiotic supplements in elderly subjects. Asia Pac J Clin Nutr . 2004;13(Suppl):S28. Efficacy of milk fortified with a probiotic Bifidobacterium lactis (DR-10TM) and prebiotic galacto-oligosaccharides in prevention of morbidity and on nutritional status; Sazawal S et al.; Background - Diarrhoea is still the major cause of childhood morbidity and mortality; estimated to cause more than 3 million deaths among children globally . Any non-invasive easily administrable intervention that may affect these morbidities would have very important implications for child health and survival . In recent years there has been increasing evidence for the role of probiotics in treatment of diarrhoea . There are very sparse data on prophylactic probiotic therapy and its effects . Objective - To evaluate the impact of milk fortified with pre- and probiotics in prevention of diarrhoea and on iron status and growth . Design - The study was conducted in Sangam Vihar, a peri-urban population in south Delhi . Healthy children aged 1- 3 years, permanent residents in the area and without any chronic illnesses or severe malnutrition were invited to participate . After informed consent 634 children were enrolled and randomly allocated to either receive a milk formulation containing Bifidobacterium lactis HN019 (DR-10 trade mark ) minimum of 10(7)-10(8) CFU/100 g and galacto-oligosaccharides 2.5 g/100 g (PP Milk), or the same milk without these two . The milk was provided in sachets of 32 g and children were advised to consume up to 3 sachets per day for 12 months . A blood sample was taken to measure the detailed haemogram, ZPP and retic count at baseline and end study . Twice weekly home visitations were done to collect data on morbidity and compliance . At baseline, six months and one year anthropometric measurements (weight, height) were made . Outcomes - Baseline characteristics of subjects in the two groups were comparable . Compliance was above 80%, with most children consuming at least two serves per day . Supplementation with PP milk resulted in a significant reduction in the incidence of dysentery OR 0.78 (95% CI 0.61, 1.00) as well as prevalence of dysentery OR 0.85 (95% CI 0.71, 1.01) . Reduction in incidence of diarrhoea was 10%, but was not statistically significant . Consistent with improved immunity PP milk supplementation caused a significant reduction in the prevalence of severe illness days OR 0.84 (95%CI 0.74-0.95, p<0.001), days with fever OR 0.68 (95%CI 0.54-0.84) and prevalence of ear infections OR 0.93 (95%CI 0.87-1.00) . Data on haematological parameters indicate a significant increase in the proportion of children with Hb levels above 10 g/L in the PP milk group . There was also a 35 % reduction in the proportion of iron deficient children and a significant reduction in stress of bone marrow as observed by a increase in the proportion of children with normal retic count, in the PP milk group . Children fed PP milk had significantly better growth at six month and one year assessments . After 1 year the children in the PP milk group had significantly better z scores for WAZ (mean diff 0.22, 95%CI 0.02-0.41; p=0.03) and WHZ (mean diff 0.18, p=0.05), and higher weight gain (mean diff: 130g, 95%CI 30-230; p=0.02) . Conclusions - Bifidobacterium lactis HN019 (DR-10 trade mark ) and galacto-oligosaccharide fortified milk resulted in better iron status even when both groups were receiving iso-caloric diets with the same iron content . This effect could be either due to better absorption due to effects on gut flora or more likely secondary to morbidity prevention effects . There was a significant reduction in bloody diarrhoea and a non significant 10% reduction in all diarrhoea . The magnitude of significant reduction in non diarrheal morbidity suggests effects are most likely on both viral and bacterial infections . This study was funded by New Zealand Milk Ltd, who also provided the milk products. Lett Appl Microbiol, 2004, 39(3), 301 - 5 A fluorescence-based method for the detection of adhesive properties of lactic acid bacteria to Caco-2 cells; Bianchi MA et al.; AIMS: The ability of probiotic micro-organisms to adhere to the intestinal surface is regarded as a substantial advantage in terms of bacteria persistence in the gastrointestinal tract . The aim of the present study was the development of a method based on fluorescent staining of bacteria and subsequent spectrofluorimetric detection to quantify the adhesion of several strains of Lactobacillus and Bifidobacterium to Caco-2 cells . METHODS AND RESULTS: Lactic acid bacteria strains were subjected to fluorescent staining using the viable probe carboxyfluorescein diacetate and subsequently incubated on Caco-2 monolayers . The adhesion of the micro-organisms was determined by spectrofluorimetry following the lysis of the attached bacterial cells and expressed as adhesion percentage . The values obtained for the micro-organisms tested ranged from 4% for Bifidobacterium infantis Bi1 to 10% for a Bifidobacterium mixture containing three different strains . CONCLUSIONS: In the present study we successfully applied fluorescent labelling and fluorimetric detection to investigate the adhesive properties of some Lactobacillus and Bifidobacterium strains and a Bifidobacterium mixture to Caco-2 cells . SIGNIFICANCE AND IMPACT OF THE STUDY: The results proved that fluorescent labelling is suitable for adhesion studies and provides a reliable and safer alternative to radioactive labelling. Poult Sci, 2004 Jul, 83(7), 1148 - 54 Effects of dietary mannan oligosaccharide, bacitracin methylene disalicylate, or both on the live performance and intestinal microbiology of turkeys; Sims MD et al.; Hybrid male turkeys were fed to 18 wk of age in a completely randomized design with 10 replicate pens (18 birds each) per treatment to compare growth promoters . Four dietary treatments were used: negative control (CON), bacitracin methylene disalicyate (BMD) at 55 mg/kg to 6 wk and 27.5 mg/kg thereafter, mannan oligosaccharide (MOS) at 0.1% to 6 wk and 0.05% thereafter, and BMD and MOS at concentrations listed above . There were 3 toms/m2 (3.59 ft2/tom) on fresh pine shavings inoculated with used litter . A 6-phase feeding program was used, with crumbles the first 3 wk and pellets thereafter . At wk 6 and 18, one bird per pen was killed, and the large intestinal microbial populations, after being frozen, were enumerated (i.e., bifidobacteria, Clostridium perfringens, coliforms, enterococci, Escherichia coli, lactobacilli, and total anerobes) . Body weights at wk 18 were as follows: CON, 11.87 kg; BMD, 12.46 kg; MOS: 12.56 kg; and BMD + MOS, 12.79 kg . The BMD and MOS turkeys were (P < 0.05) heavier than CON birds, and those fed the combination were significantly heavier than all other treatments . At wk 18, BMD + MOS feed conversion ratio of 2.66 was significantly lower than CON at 3.00 with BMD and MOS being intermediate (2.83 and 2.79, respectively) . Mortality was not affected by treatment . The BMD and MOS each reduced large intestinal concentrations of Clostridium perfringens (log transformed data analysis) at wk 6 but not at wk 18 . The BMD or MOS each improved turkey performance, and when used together, exhibited further beneficial effects. Poult Sci, 2004 Jul, 83(7), 1093 - 8 Microflora ecology of the chicken intestine using 16S ribosomal DNA primers; Amit-Romach E et al.; The microflora in the gastrointestinal tract of broiler chickens influences digestion, health, and wellbeing . Analysis of chicken gut microflora has been mainly by culture-based methods . Studies using these techniques have been useful for identification and analysis of specific groups of bacteria, however, the use of enrichment medium precludes even relative quantitation of bacterial species . Recent advances in ribosomal DNA-based molecular techniques make it possible to identify different bacterial populations in environmental samples without cultivation . In this study, the intestinal microflora was examined using 16S ribosomal DNA (rDNA) targeted probes from bacterial DNA isolated from intestinal and cecal contents of chickens at 4, 14, and 25 d of age . The ribosomal gene sequence was amplified using PCR with universal primers to determine total bacterial DNA and specific primers directed at 6 bacterial species: Lactobacillus, Bifidobacterium, Salmonella, Campylobacter, Escherichia coli, and Clostridium . The use of universal primers extends these methods to allow determination of relative proportions of different bacterial species . The results indicated that in young chicks the major species present in the small intestines and ceca was Lactobacilli, with a Bifidobacteria population becoming more dominant in the ceca at older age . Clostridium was detected in some segments of the small intestine in young chicks . In older chickens, Salmonella, Campylobacter, and E . coli species were found in the ceca . This study has demonstrated the use of molecular techniques for determining relative proportions of bacterial species and monitoring pathogens in the chick gastrointestinal tract. J Nutr, 2004 Aug, 134(8), 1998 - 2003 Plasma phytoestrogens are not altered by probiotic consumption in postmenopausal women with and without a history of breast cancer; Nettleton JA et al.; Soy phytoestrogens were suggested to reduce the risk of a number of diseases including breast cancer . Given that these compounds are metabolized by bacteria, alteration of intestinal bacteria and enzymes may affect phytoestrogen metabolism . We hypothesized that probiotics, when consumed with soy protein, would increase plasma isoflavones, as well as equol producer frequency, in postmenopausal women . We further hypothesized that these effects would differ between women who have had breast cancer and women who have not . To test these hypotheses, 20 breast cancer survivors and 20 controls completed four 6-wk treatments in a randomized, crossover design: supplementation with soy protein (S) (26.6 +/- 4.5 g protein, 44.4 +/- 7.5 mg isoflavones/d); soy + probiotics (S+P) (10(9) colony-forming units Lactobacillus acidophilus DDS+1 and Bifidobacterium longum, 15-30 mg fructooligosaccharide/d); milk protein (M) (26.6 +/- 4.5 g protein/d); and milk + probiotics (M+P) . Plasma phytoestrogen concentrations did not differ between controls and survivors, although genistein tended to be lower in survivors at baseline (P = 0.15), and during soy (P = 0.16) and milk protein (P = 0.16) consumption . As expected, soy consumption increased plasma phytoestrogen concentrations (P < 0.0001) . Plasma phytoestrogen concentrations and the number of equol producers did not differ between the S and S+P diets . At the same time, plasma equol concentrations as well as urinary equol excretion in 2 subjects were more than 7-fold different between the 2 diets . These results indicate that this particular probiotic supplement does not generally affect plasma isoflavones, although the large differences between plasma and urinary equol in some subjects suggest that equol producer status may be modifiable in some individuals. Prikl Biokhim Mikrobiol, 2004 May-Jun, 40(3), 301 - 6 {Antibacterial effects of water-soluble low-molecular-weight chitosans on different microorganisms}; Gerasimenko DV et al.; Low-molecular-weight chitosans with a viscosity-average molecular weight (Mv) of 5 to 27 kDa and equal degree of deacetylation (DD, 85%) were highly active against Pseudomonas aureofaciens, Enterobacter agglomerans, Bacillus subtilis, and Bifidobacterium bifidum 791, causing death of 80 to 100% of cells . An exception to this tendency was Escherichia coli, for which the rate of cell death, induced by the 5-kDa chitosan, was 38% . The antibacterial effect was manifested as early as 10 min after incubation of 12-kDa chitosan with B . subtilis or E . coli cells . Candida krusei was almost insensitive to the above crab chitosans . However, Candida krusei was highly sensitive to chitosans with Mv 5, 6, 12, 15.7, and 27 kDa: the minimum inhibitory concentration (MIC) varied from 0.06 to 0.005% . Chitosans with M, 5, 12, and 15.7 kDa exerted an antibacterial effect on Staphylococcus aureus . Chitosans with Mv 5, 15.7, and 27 kDa had no effect on Bifidobacterium bifidum ATCC 14893 . The antibacterial effect of the 4-kDa chitosan on E . coli and B . bifidum 791 increased with DD in the range 55-85%. J Appl Microbiol, 2004, 97(3), 527 - 39 Increased stress tolerance of Bifidobacterium longum and Lactococcus lactis produced during continuous mixed-strain immobilized-cell fermentation; Doleyres Y et al.; AIMS: The effect of immobilization and long-term continuous culture was studied on probiotic and technological characteristics of lactic acid and probiotic bacteria . METHODS AND RESULTS: A continuous culture in a two-stage system was carried out for 17 days at different temperatures ranging from 32 to 37 degrees C, with a first reactor containing Bifidobacterium longum ATCC 15707 and Lactococcus lactis subsp . lactis biovar . diacetylactis MD immobilized separately in gel beads, and a second reactor operated with free cells released from the first reactor . The tolerance of free cells from both strains produced in the effluent medium of both reactors to hydrogen peroxide, simulated gastric and intestinal juices, antibiotics and nisin, and freeze-drying markedly increased with culture time and was generally higher after 6 days than that of stationary-phase cells produced during free-cell batch fermentations . The reversibility of the acquired tolerance of B . longum, but not L . diacetylactis, to antibiotics was shown during successive free-cell batch cultures . CONCLUSIONS: Free cells produced from continuous immobilized-cell culture exhibited altered physiology and increased tolerance to various chemical and physico-chemical stresses . SIGNIFICANCE AND IMPACT OF THE STUDY: Continuous culture with immobilized cells could be used to produce probiotic and lactic acid bacteria with enhanced technological and probiotic characteristics . J Appl Microbiol, 2004, 97(3), 459 - 70 Genetic heterogeneity and functional properties of intestinal bifidobacteria; Matto J et al.; AIMS: The aim of the present study was to compare several molecular methods for the identification and genotyping of bifidobacteria, and further to investigate genetic heterogeneity and functional properties of bifidobacterial isolates from intestinal samples of Finnish adult subjects . METHODS AND RESULTS: A total of 153 intestinal bifidobacterial isolates were included in initial screening and 34 isolates were further characterized . Identification results obtained with PCR-ELISA and ribotyping were well in accordance with each other, while randomly amplified polymorphic DNA (RAPD) gave tentative identification only to Bifidobacterium bifidum and to 65% of the B . longum isolates . The most commonly detected species were B . longum biotype longum followed by B . adolescentis and B . bifidum . In addition, B . animalis (lactis), B . angulatum and B . pseudocatenulatum were found . Ribotyping and pulsed-field gel electrophoresis (PFGE) proved to be discriminatory methods for bifidobacteria, but also RAPD revealed intraspecies heterogeneity . Besides two B . animalis (lactis) isolates with very close similarity to a commercially available probiotic strain, none of the intestinal isolates showed optimal survival in all tolerance (acid, bile and oxygen) or growth performance tests . CONCLUSIONS: Several species/strains of bifidobacteria simultaneously colonize the gastrointestinal tract of healthy Finnish adults and intestinal Bifidobacterium isolates were genetically heterogeneous . Functional properties of bifidobacteria were strain-dependent . SIGNIFICANCE AND IMPACT OF THE STUDY: Applicability of ribotyping with the automated RiboPrinter System for identification and genotyping of bifidobacteria was shown in the present study . Int J Syst Evol Microbiol, 2004 Jul, 54(Pt 4), 1137 - 43 Polyphasic taxonomic analysis of Bifidobacterium animalis and Bifidobacterium lactis reveals relatedness at the subspecies level: reclassification of Bifidobacterium animalis as Bifidobacterium animalis subsp . animalis subsp . nov . and Bifidobacterium lactis as Bifidobacterium animalis subsp . lactis subsp . nov; Masco L et al.; The taxonomic standing of Bifidobacterium lactis and Bifidobacterium animalis was investigated using a polyphasic approach . Sixteen representatives of both taxa were found to be phenotypically similar and shared more than 70% DNA-DNA relatedness (76-100%), which reinforces the conclusions of previous studies in which B . lactis and B . animalis were considered to be one single species . However, the results of protein profiling, BOX-PCR fingerprinting, Fluorescent Amplified Fragment Length Polymorphism (FAFLP), and atpD and groEL gene sequence analysis demonstrate that representatives of B . animalis and B . lactis constitute two clearly separated subgroups; this subdivision was also phenotypically supported based on the ability to grow in milk . Given the fact that B . lactis Meile et al . 1997 has to be considered as a junior synonym of B . animalis (Mitsuoka 1969) Scardovi and Trovatelli 1974, our data indicate that the latter species should be split into two new subspecies, i.e . Bifidobacterium animalis subsp . animalis subsp . nov . (type strain R101-8T=LMG 10508T=ATCC 25527T=DSM 20104T=JCM 1190T) and Bifidobacterium animalis subsp . lactis subsp . nov . (type strain UR1T=LMG 18314T=DSM 10140T=JCM 10602T). J Microbiol Methods, 2004 Sep, 58(3), 413 - 21 Development of multi-color FISH method for analysis of seven Bifidobacterium species in human feces; Takada T et al.; We have developed a multi-color fluorescence in situ hybridization (FISH) method which detects, by a single reaction, all seven species of Bifidobacterium (B . adolescentis, B . angulatum, B . bifidum, B . breve, B . catenulatum, B . dentium, and B . longum), the dominant bacteria in human feces . First, eight new types of oligonucleotide probe were designed, complementary with the 16S rRNA sequence specific to genus Bifidobacterium and each bifidobacterial species described above . Using whole cell hybridization, the fluorescent intensity was measured against the bacterial species targeted by each probe, to show that each probe is specific to the targeted bacteria and that the relative fluorescent intensity (RFI) as an indicator of probe accessibility is high at 61-117% . Then, bacterial species-specific probes were labeled with fluorochromes (FITC, TAMRA and Cy5) in seven different ways, singly or in combination . Using these probes, seven species of Bifidobacterium were differentially stained in mixed samples of cultured bacteria and feces from adult volunteers, proving the efficacy of this technique. J Am Soc Mass Spectrom, 2004 Aug, 15(8), 1222 - 7 Rapid identification of stress-related fingerprint from whole bacterial cells of Bifidobacterium lactis using matrix assisted laser desorption/ionization mass spectrometry; Marvin-Guy LF et al.; Whole cells of Bifidobacterium lactis were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) . Characteristic and reproducible mass spectra were obtained in the mass range from 6 to 19 kDa . After several days of bacterial cell storage at 4 degrees C (D0, D2, and D6), only minor signal differences were observed . Under identical and reproducible conditions, fourteen relevant diagnostic ions were identified . Moreover, control- and stress-related fingerprints were rapidly obtained using MALDI-TOFMS by comparison of protein patterns obtained from non-stressed (control) versus stressed cells (addition of bile salts during growth) . After quantitative validation of the MALDI-MS data by a statistical approach, two and eight signals were assigned as control- and stress-specific ions, respectively . This work provides the evidence that MALDI-TOFMS can be used for the identification of stress-related fingerprint of B . lactis bacterial cells and could have a high potential for the assessment of the physiological status of the cells. J Infect Dis, 2004 Aug 15, 190(4), 826 - 34 Epub 2004 Jul 21. Viral and bacterial pathogens at the maternal-fetal interface; McDonagh S et al.; We studied the incidence of pathogenic bacteria and concurrent infections with human cytomegalovirus (CMV) and herpes simplex virus (HSV) type 1 and 2 in biopsy samples from the placenta and decidua of women with healthy pregnancies . By polymerase chain reaction analysis, we found that 38% of placental samples were positive for selected bacteria and viruses . CMV, HSV-1, and HSV-2 were detected in isolation or with bacteria in first- and second-trimester samples . Certain bacteria were detected more often during the second trimester than during the first--Ureaplasma urealyticum, Mycoplasma hominis, and Gardnerella/Bifidobacterium species . In paired samples from first-trimester tissues, the detection rate for viruses, compared with most bacteria, was higher in the decidua than in the adjacent placenta . In contrast, bacteria were more frequently detected in placenta . Analyses of immunoglobulin G isolated from the placenta support the hypothesis that immune responses suppress CMV reactivation in the presence of pathogenic bacteria at the maternal-fetal interface. J Bacteriol, 2004 Aug, 186(15), 4885 - 93 Molecular cloning and characterization of Bifidobacterium bifidum 1,2-alpha-L-fucosidase (AfcA), a novel inverting glycosidase (glycoside hydrolase family 95); Katayama T et al.; A genomic library of Bifidobacterium bifidum constructed in Escherichia coli was screened for the ability to hydrolyze the alpha-(1-->2) linkage of 2'-fucosyllactose, and a gene encoding 1,2-alpha-l-fucosidase (AfcA) was isolated . The afcA gene was found to comprise 1,959 amino acid residues with a predicted molecular mass of 205 kDa and containing a signal peptide and a membrane anchor at the N and C termini, respectively . A domain responsible for fucosidase activity (the Fuc domain; amino acid residues 577 to 1474) was localized by deletion analysis and then purified as a hexahistidine-tagged protein . The recombinant Fuc domain specifically hydrolyzed the terminal alpha-(1-->2)-fucosidic linkages of various oligosaccharides and a sugar chain of a glycoprotein . The stereochemical course of the hydrolysis of 2'-fucosyllactose was determined to be inversion by using (1)H nuclear magnetic resonance . The primary structure of the Fuc domain exhibited no similarity to those of any glycoside hydrolases (GHs) but showed high similarity to those of several hypothetical proteins in a database . Thus, it was revealed that the AfcA protein constitutes a novel inverting GH family (GH family 95). Fitoterapia, 2004 Jul, 75(5), 505 - 9 Growth responses of Cassia obtusifolia toward human intestinal bacteria; Sung BK et al.; 1,2-Dihydroxyanthraquinone (1) was isolated from the seed of Cassia obtusifolia through bioassay-guided fractionation . 1,2-Dihydroxyanthraquinone strongly inhibited the growth of Clostridium perfringens and Escherichia coli . Structure-activity relationship revealed that 1,4-dihydroxyanthraquinone (2) and 1,8-dihydroxyanthraquinone (3) had strong growth-inhibition against C . perfringens . In growth-promoting activity, 1,2-, 1,4-, and 1,8-dihydroxyanthraquinones exhibited strong growth-promoting activity to Bifidobacterium bifidum. Int J Food Microbiol, 2004 Aug 15, 95(1), 103 - 6 Use of a probiotic Bifidobacterium in a dry food matrix, an in vivo study; Ouwehand AC et al.; Probiotics are commonly included in dairy products . These products require cold storage and transportation, which limits their use . Here, we describe the inclusion of the probiotic strain Bifidobacterium lactis Bb-12 in a dry food matrix, an oat-based cereal bar, and its detection in faeces after consumption of this product . One week after cessation of B . lactis Bb-12 feeding, it could be identified in the faeces of five of the nine subjects. Br J Nutr, 2004 Jul, 92(1), 81 - 6 Inhibitory effect of yoghurt and soya yoghurt containing bifidobacteria on the proliferation of Ehrlich ascites tumour cells in vitro and in vivo in a mouse tumour model; Abd el-Gawad IA et al.; The effect of yoghurt and soya yoghurt containing Bifidobacterium lactis Bb-12 or B . longum Bb-46 on Ehrlich ascites tumour cell proliferation was investigated in vitro and in vivo . Tumour cells were incubated with B . lactis Bb-12 or B . longum Bb-46 cultivated in de Mann Rogosa Sharpe (MRS) broth medium, or with their centrifuged supernatant fractions or sediments, for 2 h at 37 degrees C . Treatment resulted in the inhibition of tumour cell proliferation by 85.42 (SD 0.78) and 85.10 (SD 1.28) % by intact micro-organisms, 77.61 (SD 0.29) and 71.43 (SD 1.75) % by their supernatant fractions, but only 4.00 (SD 0.19) and 9.09 (SD 1.24) % by the two sedimented bacteria, respectively . The incubation of tumour cells with yoghurt and soya yoghurt containing Bb-12 for 2 h resulted in 83.01 (SD 0.11) and 88.23 (SD 0.06) % inhibition, respectively, while it was 83.82 (SD 0.24) and 86.36 (SD 0.06) %, respectively for the same products containing Bb-46 . Corresponding values for plain yoghurt and soya milk (without bifidobacteria) were 32.81 (SD 0.14) and 5.55 (SD 0.12) %, respectively . The differences between yoghurt or soya yoghurt containing Bb-12 or Bb-46 and plain yoghurt, soya milk or control treatments were statistically significant (n 3; P<0.05) . Female Swiss albino mice were injected intraperitoneally with the same tumour cells . The lifespan of mice fed diets supplemented with yoghurt or soya yoghurt containing Bb-12 or Bb-46 was prolonged by 16, 23, 34 and 39 %, respectively compared with that of the positive control group (n 6; P<0.05) . The lifespan of groups fed plain yoghurt or soya milk was prolonged by 15 and 8 %, respectively . Prolongation of lifespan was positively correlated with faeces bifidobacterial count in the groups fed yoghurt or soya yoghurt containing bifidobacteria (r 0.917; P<0.05). Folia Microbiol (Praha), 2004, 49(2), 175 - 7 Detection of cellulolytic bacteria from the human colon; Kopecny J et al.; The main representatives of bacteria in the human colon were investigated by specific PCR and denaturing gradient gel electrophoresis (DGGE) . Prevalent in both cases were species of Bifidobacterium, Clostridium, Bacteroides, Faecalibacterium and Eubacterium . Simultaneously, cellulolytic bacteria were isolated from the human feces . The largest proportion was represented by ruminococcus-like isolates . Their presence was confirmed both by PCR and DGGE methods; the latter one was able to give more comprehensive data about the composition of bacterial population in the human colon chyme. Folia Microbiol (Praha), 2004, 49(2), 143 - 6 Aggregation of Lactobacilli and Bifidobacteria with Escherichia coli O157; Bujnakova D et al.; A total of 5 Bifidobacterium spp . isolated from pig and children' feces and 6 Lactobacillus spp . from chicken feces were examined for expression of aggregation, cell surface hydrophobicity (CSH) and adherence to intestinal mucin . Co-aggregation activity was seen in 3 strains of auto-aggregative bifidobacteria and 4 auto-aggregative strains of Lactobacillus spp . with 2 enterohemorrhagic E . coli (O157) . CSH correlated with Lactobacillus auto-aggregating activity and adherence to mucin but the correlation between Bifidobacterium adherence to mucin and CSH was not confirmed. Biometals, 2004 Jun, 17(3), 279 - 83 Growth-promoting effects of lactoferrin on L . acidophilus and Bifidobacterium spp; Kim WS et al.; We investigated the effects of lactoferrin on the growth of L . acidophilus CH-2, Bifidobacterium breve ATCC 15700, B . longum ATCC 15707, B . infantis ATCC 15697, and B . bifidum ATCC 15696 . The growth of L . acidophilus was stimulated by bovine holo-lactoferrin but not by apo-lactoferrin . With bifidobacteria, bovine lactoferrin stimulated growth of three strains: B . breve, B . infantis and B . bifidum under certain conditions . Both apoprotein and holoprotein had similar effects . However, B . longum growth was not affected by lactoferrin . Thus, the mechanism of stimulating growth of bifidobacteria may be different from that of L . acidophilus . By far-western blotting using biotinylated lactoferrin and horseradish peroxidase-conjugated streptavidin, lactoferrin-binding proteins were detected in the membrane protein fraction of L . acidophilus, B . bifidum, B . infantis and B . breve . The molecular weights of lactoferrin-binding proteins of L . acidophilus were estimated from SDS-polyacrylamide gel electrophoresis to be 27, 41 and 67 kDa, and those of the three bifidobacterial strains were estimated to be 67-69 kDa . However, no such lactoferrin-binding components were detected in the membrane fraction of B . longum . It is interesting that the appearance of lactoferrin-binding proteins in the membrane fraction of these species corresponds to their growth stimulation by lactoferrin. J Clin Gastroenterol, 2004 Jul, 38(6 Suppl), S91 - 3 Microflora in inflammatory bowel diseases: a pediatric perspective; Bruzzese E et al.; Several lines of evidence link inflammatory bowel diseases to modifications of intestinal microflora . Epidemiologic and clinical data suggest a triggering role for select agents in ulcerative colitis and in Crohn disease . Experimental evidence indicates that intestinal microorganisms are needed for developing intestinal inflammation in IL-10 knockout mice, and this is associated with an increased number of adherent clostridia and a decrease of lactobacilli and bifidobacteria . It may be hypothesized that a host-agent-specific relationship leads to an abnormal immune response, which may be genetically driven in select inflammatory bowel diseases . However, different from adults, the pattern of intestinal microflora undergoes profound changes during the early stage of life, contributing to the development of the immune system . A close relationship exists between microbiologic and immunologic imprinting . The microbiologic imprinting in neonates may be modified using bacterial probiotics that colonize the intestine, modify the immune response, and decrease the risk for atopy . Probiotics may decrease the recurrences of inflammatory bowel diseases . Preliminary evidence of intestinal antiinflammatory effects has been detected in children with cystic fibrosis . Overall these data provide the rationale to investigate the interaction between intestinal microflora and the local and general immune response in children with, or at risk for, inflammatory bowel diseases . This approach may be a key for understanding the pathophysiology of intestinal inflammation and may disclose novel strategies to educate better the immune system, particularly during its developmental stage. J Clin Gastroenterol, 2004 Jul, 38(6 Suppl), S80 - 3 The first prebiotics in humans: human milk oligosaccharides; Coppa GV et al.; The development of intestinal microflora in newborns is strictly related to the kind of feeding . Breast-fed infants, unlike the bottle-fed ones, have an intestinal ecosystem characterized by a strong prevalence of bifidobacteria and lactobacilli . Data available so far in the literature show that, among the numerous substances present in human milk, oligosaccharides have a clear prebiotic effect . They are quantitatively one of the main components of human milk and are only partially digested in the small intestine, so they reach the colon, where they stimulate selectively the development of bifidogenic flora . Such results have been recently proved both by characterization of oligosaccharides in breast-fed infant feces and by the study of intestinal microflora using new techniques of molecular analysis, confirming that human milk oligosaccharides represent the first prebiotics in humans. Syst Appl Microbiol, 2004 May, 27(3), 279 - 85 Hydrolysis of oligofructoses by the recombinant beta-fructofuranosidase from Bifidobacterium lactis; Janer C et al.; The ability of the beta-fructofuranosidase (EC 3.2.1.26) from Bifidobacterium lactis DSM 10140T to cleave a variety of fructooligosaccharides was characterised . We identified its gene on a cloned chromosomal DNA fragment by sequence similarity (69% identity) to the putative CscA protein encoded in the Bifidobacterium longum genome . The deduced amino acid sequence of 532 residues (59.4 kDa) appeared to be identical to the beta-fructofuranosidase from the same strain recently described by Ehrmann et al . (Curr . Microbiol . 2003, 46, 391-397) . However, the characterisation of the heterologously expressed enzyme showed several discrepancies to the referred study . First, the B . lactis beta-fructofuranosidase gene was found to have 41% identity with CscA from E . coli in contrast to the 16% reported, therefore it was assigned to as CscA protein instead of BfrA . Second, we observed only low activity of the enzyme towards sucrose (6%) instead of the 100% previously reported . Instead, we measured highest activity (100%) of the enzyme with the oligofructose Raftilose as a substrate compared with the inulin of low degree of polymerisation Raftiline LS (29%) and the highly polymerised Raftiline HP (10%) . Altogether, the enzyme showed high affinity to terminal beta(2-1) glycosyl linkages between fructose moieties . The Km values obtained for Raftilose, Raftiline LS and sucrose were 0.12, 7.08 and 8.37 mM, respectively, and V(max) values for the conversion to fructose were calculated to be 5, 21 and 17 micromol/min per mg of protein, respectively . Growth of B . lactis was supported by fructans of low degree of polymerisation (Raftilose and Raftiline LS), whereas we observed no growth with highly polymerised inulin (Raftiline HP). Eur J Clin Nutr, 2004 Dec, 58(12), 1635 - 42 Consumption of Lactobacillus acidophilus and Bifidobacterium longum do not alter urinary equol excretion and plasma reproductive hormones in premenopausal women; Bonorden MJ et al.; OBJECTIVE: To confirm the results of an earlier study showing premenopausal equol excretors to have hormone profiles associated with reduced breast cancer risk, and to investigate whether equol excretion status and plasma hormone concentrations can be influenced by consumption of probiotics . DESIGN: A randomized, single-blinded, placebo-controlled, parallel-arm trial . SUBJECTS: In all, 34 of the initially enrolled 37 subjects completed all requirements . INTERVENTION: All subjects were followed for two full menstrual cycles and the first seven days of a third cycle . During menstrual cycle 1, plasma concentrations of estradiol (E(2)), estrone (E(1)), estrone-sulfate (E(1)-S), testosterone (T), androstenedione (A), dehydroepiandrosterone-sulfate (DHEA-S), and sex-hormone-binding globulin (SHBG) were measured on cycle day 2, 3, or 4, and urinary equol measured on day 7 after a 4-day soy challenge . Subjects then received either probiotic capsules (containing Lactobacillus acidophilus and Bifidobacterium longum) or placebo capsules through day 7 of menstrual cycle 3, at which time both the plasma hormone concentrations and the post-soy challenge urinary equol measurements were repeated . RESULTS: During menstrual cycle 1, equol excretors and non-excretors were not significantly different with respect to subject characteristics, diet, or hormone concentrations . Significant inverse correlations were found between E(2) and body mass index (BMI) (P=0.02), SHBG and BMI (P=0.01), DHEA-S and dietary fiber (P=0.04), and A and protein:carbohydrate ratio (P=0.02) . Probiotic consumption failed to significantly alter equol excretor status or hormone concentrations during menstrual cycle 3, although there were trends towards decreased concentrations of T (P=0.14) and SHBG (P=0.10) in the probiotic group . CONCLUSIONS: We were unable to verify a previously reported finding of premenopausal equol excretors having plasma hormone concentrations different from those of nonexcretors . Furthermore, a 2-month intervention with probiotic capsules did not significantly alter equol excretion or plasma hormone concentrations. FEMS Microbiol Lett, 2004 Jul 1, 236(1), 153 - 9 Developing a quantitative approach for determining the in vitro prebiotic potential of dietary oligosaccharides; Vulevic J et al.; Prebiotics are nondigestible carbohydrates that beneficially affect the host by selectively stimulating the growth and/or activity of one, or a limited number of, bacteria present in the colon . The selected genera should have the capacity to improve host health (e.g . Bifidobacterium, Lactobacillus) . To help identify preferred types, for inclusion into the diet, a quantitative equation {measure of the prebiotic effect (MPE)} is suggested . This will help evaluate, in vitro, the fermentation of dietary carbohydrates and compare their prebiotic effect . Although the approach is not meant to define health values, it is formulated to better inform the choice of prebiotic . It therefore, compares measurements of bacterial changes through the determination of maximum growth rates of predominant groups present in faeces, rate of substrate assimilation and the production of lactic, acetic, propionic and butyric acids . The equation will allow further in vitro comparisons of MPE, leading towards further studies (e.g . in humans) to determine the success of dietary intervention. Zh Mikrobiol Epidemiol Immunobiol, 2004 Mar-Apr, (2), 76 - 8 {Correction of intestinal dysbacteriosis in elderly patients with the use of bifidobacterin introduced by different routes}; Chulok TA et al.; The results of the correction of disturbances in intestinal microflora in elderly patients with the use of bifidumbacterin in the form of powder or suppositories are presented . The effectiveness of the rectal administration of bifidumbacterin (by microinjections through an enema or in suppositories) is shown in comparison with the oral administration of the preparation . The administration of bifidumbacterin in suppositories for 15 days resulted in decreased content of hemolytic Escherichia below the threshold of determination in all examined patients. Zh Mikrobiol Epidemiol Immunobiol, 2004 Mar-Apr, (2), 70 - 4 {In vitro evaluation of influence of medicinal plant decoctions and antibacterial antibodies to bifidobacteria on bifidobacterium adhesion}; Popkova SM et al.; Natural antibodies to the surface antigens of bifidobacteria contained in blood sera and determined in the passive hemagglutination test blocked the adhesion of bifidobacteria to human red blood cells in vitro . The decoctions of medicinal plants, often used by pediatricians for therapeutic purposes, were found to have a pronounced inhibiting effect on the adhesion of bifidobacteria after the preliminary treatment of both microorganisms and human red blood cells by these decoctions . Suggestion was made on the possible role of natural human antibodies in changing the contents of the indigenous intestinal microflora; the prolonged use of preparations obtained from medicinal plants could probably produce negative influence on the microbiocenosis of the intestine. Zh Mikrobiol Epidemiol Immunobiol, 2004 Mar-Apr, (2), 60 - 4 {Immunomodulating effect of protein fractions isolated from bifidobacteria}; Nikolaeva TN et al.; The screening of the immunomodulating activity (IMA) of different protein fractions isolated from bifidobacteria was carried out and the capacity of these fractions for changing the proliferative activity of immunocompetent cells was evaluated . Soluble proteins were extracted from lyophilized and sonicated bacterial mass of B . bifidum strain 1 in Na2HPO4 (pH 8) in a water bath at 65 degrees C for 30 minutes . After the formation and removal of nucleic acid sediment the resulting supernatant fluid was dialyzed, its adsorption spectra were analyzed and the fluid was fractionated in a specially proposed device for preparative electrophoresis . Protein fractions were tested for IMA on spleen cells of CBA mice in the reaction of lymphocyte blast-transformation by the level of the inclusion 3H thymidine . The analysis of IMA of protein fractions revealed that their high-molecular components produced a pronounced dose-dependent effect on the proliferative activity of spleen cells . The fractions containing low-molecular components were either inactive (fraction 4) or active only in the maximum dose (fraction 5). Dig Dis Sci, 2004 Apr, 49(4), 579 - 89 In vivo effects of bifidobacteria and lactoferrin on gut endotoxin concentration and mucosal immunity in Balb/c mice; Griffiths EA et al.; The aim of the present study was to examine the effects of oral supplementation of newborn Balb/c mice with bifidobacteria (B . infantis, B . bifidum) and iron-free apo-lactoferrin (bovine, human) on gut endotoxin concentration and mucosal immunity . Endotoxin concentration was measured in ileocecal filtrates at 7, 14, 21, and 28 days postdelivery by a quantitative limulus amebocyte lysate test . While endotoxin levels in bifidobacteria-fed mice showed a steady rise over time, they were consistently lower than that observed in control animals . Results of lactoferrin supplementation varied depending on the specific time point, but overall by day 28, all treatment groups showed lower intestinal endotoxin concentrations compared to saline fed animals . Neither bifidobacteria nor lactoferrin stimulated an increase in B or T cells, or in cytokine production (IL-6, TNF-alpha, INF-gamma), in Peyer's patches as measured by flow cytometry . Bifidobacteria and lactoferrin were well tolerated as dietary supplements and showed promising potential to reduce gut endotoxin levels. Antibiot Khimioter, 2003, 48(12), 9 - 13 {Growth and morphological characteristics of industrial strains of Bifidobacterium and Lactobacillus cultivated in hydrolysate-milk and hydrolysate-soybean media}; Tsinberg MB et al.; Growth features of industrial strains of Bifidobacterium adolescentis MC-42, B . bifidum 1, B . longum B-379, Lactobacillus acidophilus, L . delbrueckii subsp . bulgaricus 8-79 and L . plantarum 8PA3 cultivated in hydrolysate-milk or hydrolysate-soybean media (HMM and HSM respectively) were analysed comparatively . The bacterial cells were investigated morphometrically with atom strength microscopy . It was shown that HSM vs HMM provided a higher growth rate of the strains (shortened growth phases and higher growth rates) that was more evident for the bifidobacteria as compared to the lactobacilli . At the same time, the morphological features of the bacterial cells slightly depended on the medium composition and were mainly defined by the genus. J Nutr, 2004 Jun, 134(6), 1523 - 8 Xylooligosaccharides and fructooligosaccharides affect the intestinal microbiota and precancerous colonic lesion development in rats; Hsu CK et al.; Certain nondigestible oligosaccharides can be selectively utilized by probiotics and reduce the risk of colon cancer . However, the inhibitory effects of xylooligosaccharides (XOS) on colon cancer are not well documented . This study evaluated the effects of xylooligosaccharides and fructooligosaccharides (FOS) on the alteration of cecal microbiota, cecal pH, cecal weight, and serum lipid levels, and also their inhibitory effect on precancerous colon lesions in male Sprague-Dawley rats . The rats were randomly assigned to 4 groups: control, treatment with 1,2-dimethylhydrazine (DMH) {15 mg/(kg body wt.wk) for 2 wk}, treatment with DMH + 60 g XOS/kg diet, and treatment with DMH + 60 g FOS/kg diet . Rats were fed the experimental diets for 35 d, beginning 1 wk after the second dose of DMH . Both XOS and FOS markedly decreased the cecal pH and serum triglyceride concentration, and increased the total cecal weight and bifidobacteria population . XOS had a greater effect on the bacterial population than did FOS . Moreover, both XOS and FOS markedly reduced the number of aberrant crypt foci in the colon of DMH-treated rats . These results suggest that XOS and FOS dietary supplementation may be beneficial to gastrointestinal health, and indicate that XOS is more effective than FOS. Int J Food Microbiol, 2004 Jul 1, 94(1), 79 - 86 Effect of the adaptation to high bile salts concentrations on glycosidic activity, survival at low PH and cross-resistance to bile salts in Bifidobacterium; Noriega L et al.; Six derivatives with increased resistance to ox gall (MIC: > or = 1% w/v) and one derivative resistant to sodium cholate (MIC: 0.8% w/v) were obtained from more sensitive original Bifidobacterium strains . These microorganisms, and two additional cholate resistant derivatives obtained in a previous study (Int . J . Food Microbiol . 82 (2003) 191), were partially characterised in this study . Acquisition of resistance against a given bile salt, also conferred cross-resistance to other bile salts, and promoted an increase in the survival of these microorganisms at low pH . Bile resistance levels of derivatives were dependent on the external pH so that the resistance was lower at neutral pH values than in acidic environments . In addition, the acquisition of bile resistance induced changes on glycoside-hydrolysing activities of derivatives obtained from five out of eight original strains, with certain activities such as beta-glucosidase showing more than tenfold increases in some of these microorganisms . These data suggest that the exposure to high bile salts concentrations may have induced a synergic response on Bifidobacterium for the adaptation to the conditions of the gastrointestinal tract . This could have improved the survival at low pH in these microorganisms, the resistance to high bile salts concentrations, and the assimilation of non-digestible carbohydrates by the enhancement of some glycoside-hydrolysing activities. Gastroenterology, 2004 May, 126(6), 1620 - 33 Therapeutic manipulation of the enteric microflora in inflammatory bowel diseases: antibiotics, probiotics, and prebiotics; Sartor RB; Crohn's disease, ulcerative colitis, and pouchitis are caused by overly aggressive immune responses to a subset of commensal (nonpathogenic) enteric bacteria in genetically predisposed individuals . Clinical and experimental studies suggest that the relative balance of aggressive and protective bacterial species is altered in these disorders . Antibiotics can selectively decrease tissue invasion and eliminate aggressive bacterial species or globally decrease luminal and mucosal bacterial concentrations, depending on their spectrum of activity . Alternatively, administration of beneficial bacterial species (probiotics), poorly absorbed dietary oligosaccharides (prebiotics), or combined probiotics and prebiotics (synbiotics) can restore a predominance of beneficial Lactobacillus and Bifidobacterium species . Current clinical trials do not fulfill evidence-based criteria for using these agents in inflammatory bowel diseases (IBD), but multiple nonrigorous studies and widespread clinical experience suggest that metronidazole and/or ciprofloxacin can treat Crohn's colitis and ileocolitis (but not isolated ileal disease), perianal fistulae and pouchitis, whereas selected probiotic preparations prevent relapse of quiescent ulcerative colitis and relapsing pouchitis . These physiologic approaches offer considerable promise for treating IBD, but must be supported by rigorous controlled therapeutic trials that consider clinical disease before their widespread clinical acceptance . These agents likely will become an integral component of treating IBD in combination with traditional anti-inflammatory and immunosuppressive agents. World J Gastroenterol, 2004 Jun 1, 10(11), 1630 - 3 Competitive inhibition of adherence of enterotoxigenic Escherichia coli, enteropathogenic Escherichia coli and Clostridium difficile to intestinal epithelial cell line Lovo by purified adhesin of Bifidobacterium adolescentis 1027; Zhong SS et al.; AIM: To observe competitive inhibition of adherence of enterotoxigenic Escherichia coli (ETEC), enteropathogenic Escherichia coli (EPEC) and Clostridium difficile (C . difficile) to intestinal epithelial cell line Lovo by purified adhesin of Bifidobacterium adolescentis 1027 (B . ado 1027) . METHODS: The binding of bacteria to intestinal epithelial cell line Lovo was counted by adhesion assay . The inhibition of adherence of ETEC, EPEC and C . difficile to intestinal epithelial cell line Lovo by purified adhesin of B . ado 1027 was evaluated quantitatively by flow cytometry . RESULTS: The purified adhesin at the concentration of 10 microg/mL, 20 microg/mL and 30 microg/mL except at 1 microg/mL and 5 microg/mL could inhibit significantly the adhesion of ETEC, EPEC and C . difficile to intestinal epithelial cell line Lovo . Moreover, we observed that a reduction in bacterial adhesion was occurred with increase in the concentration of adhesin, and MFI (Mean fluorescent intensity) was decreased with increase in the concentration of adhesin . CONCLUSION: The purified adhesin of B . ado 1027 can inhibit the adhesion of ETEC, EPEC and C . difficile to intestinal epithelial cell line Lovo in a dose-dependent manner. World J Gastroenterol, 2004 Jun 1, 10(11), 1619 - 24 Risk factors of development of gut-derived bacterial translocation in thermally injured rats; Wang ZT et al.; AIM: Studies have demonstrated that gut-derived bacterial translocation (BT) might play a role in the occurrence of sepsis and multiple organ dysfunction syndrome (MODS) . Yet, no convincing overall analysis of risk factors for BT has been reported . The purpose of this study was to evaluate the related factors for the development of BT in burned rats . METHODS: Wistar rats were subjected to 30% third-degree burns . Then samples were taken on postburn d 1, 3, and 5 . Incidence of BT and counts of mucosal bifidobacteria, fungi and E . coli, mucus sIgA, degree of injury to ileal mucosa, and plasma interleukin-6 were observed . Univariate analysis and multivariate logistic regression analysis were performed . RESULTS: The overall BT rate was 53.9% (69 in 128) . The result of univariate analysis showed that the levels of plasma endotoxin and interleukin-6, the counts of mucosal fungi and E . coli, and the scores of ileum lesion were markedly increased in animals with BT compared with those without (P=0.000-0.005), while the levels of mucus sIgA and the counts of mucosal bifidobacteria were significantly reduced in animals with translocation compared with those without (P=0.000) . There was a significant positive correlation between mucus sIgA and the counts of mucosal bifidobacteria (r=0.74, P=0.001) . Moreover, there were strong negative correlations between scores of ileum-lesion and counts of bifidobacteria (r=-0.67, P=0.001) . Multivariate logistic regression revealed that ileum lesion score (odds ratio {OR} 45.52, 95% confidence interval {CI} 5.25-394.80), and counts of mucosal bifidobacteria (OR 0.039, 95% CI 0.0032-0.48) were independent predictors of BT secondary to severe burns . CONCLUSION: Ileal lesion score and counts of mucosal bifidobacteria can be chosen as independent prognosis factors of the development of BT . Specific interventions targeting these high-risk factors might be implemented to attenuate BT, including strategies for repair of damaged intestinal mucosae and restoration of the balance of gastrointestinal flora. FEMS Microbiol Lett, 2004 Jun 1, 235(1), 35 - 41 Acquired resistance to bile increases fructose-6-phosphate phosphoketolase activity in Bifidobacterium; Sanchez B et al.; Two Bifidobacterium strains with acquired resistance to bile were used in this study . Significant differences on membrane-associated protein profiles were found between the bile resistant derivatives and their corresponding original strains . One of the major species detected in one of the resistant derivatives had an apparent denatured molecular mass of approximately 90 kDa, and was identified as xylulose-5-phosphate/fructose-6-phosphate phosphoketolase, the key enzyme of Bifidobacterium carbohydrate catabolism . Phosphoketolase activity was considerably higher in membrane preparations and cell-free extracts of the two bile resistant derivatives . This correlated to a greater consumption rate of glucose in resistant strains . Fructose-6-phosphate phosphoketolase activity in the strain Bifidobacterium bifidum CECT4549 and its resistant derivative was found to be partially associated with the cytoplasmic membrane through weak interactions. Methods Mol Biol, 2004, 268, 453 - 8 Microtechnique for identification of lactic acid bacteria; Gusils C et al.; Certain gut species are pathogens, but a number of other resident bacteria may be of some benefit to host health . Examples include enterococci, lactobacilli, propionibacteria, and bifidobacteria, which are present in the colon in significant numbers . Identification and classification are not identical . A group can be identified only after it has been classified, based on a pattern of properties shown by all the members of the group that other groups do not possess . The properties used in identification are often different from those used in classification . Biochemical, nutritional, and physiological characterization tests (usually carried out in bottles and tubes of solid and liquid media and on plates) have been developed and modified since the earliest days of bacteriology . Generally, the characteristics chosen for an identification plan should be easily determinable, whereas those used for classification (such as DNA homology) may be quite difficult to determine . Genera and species identification might not be based on only a few tests, but rather on the pattern given by a whole battery of tests . The members of the family Lactobacillaceae represent one example of this . Some probiotic strains can be selected for their benefical properties as active antimicrobial agents against pathogenic microorganisms, hydrophobic ability, presence of substances with a capacity for adherence to epithelium, and so on . After isolation, identification is an important step before selecting probiotic strains.These identifications can be facilitated through microtechniques . To alleviate the need to inoculate large numbers of tubes with media (conventional test), some rapid multitest systems have been devised and are commercially available (such as the API or Biolog systems) . Although they are expensive for large studies and not always sufficiently versatile, these kits do offer the advantages of convenience, miniaturization, rapidity, and, above all, strict standardization . Other methods such as ribotyping or randomly amplified polymorphic DNA do not take into account the phenotypic characteristics (biotyping) . In this chapter, we describe a simple, rapid, and economical microplates technique to identify lactic acid bacteria and enterobacteria based on general metabolic characteristics, adapted for routine study of a large number of strains. Methods Mol Biol, 2004, 268, 423 - 32 Assessing survival of dairy propionibacteria in gastrointestinal conditions and adherence to intestinal epithelia; Zarate G et al.; The genus Propionibacterium consists of two principal groups, cutaneous and classical or dairy . Cutaneous species are predominant members of the microbial population of human skin and have also been isolated from the feces of humans and other vertebrate animals . They are often considered opportunistic organisms and have been occasionally associated with infections in humans.Dairy propionibacteria are microorganisms extensively used in the industry for manufacture of Swiss-type cheeses and biological production of propionic acid and vitamin B12 . They can be isolated from soil, vegetables, silage, raw milk, and dairy products such as kefir and different cheeses with "eyes."In the last decade, several studies have demonstrated probiotic properties for members of the genus Propionibacterium . The effects claimed are based on the production of bacteriocins, vitamins, stimulation of growth of other colonic bacteria like bifidobacteria, beneficial modification of the composition and metabolic activities of the intestinal microflora, immunomodulation, and antimutagenic activity.It is thought that to produce many of these health benefits, the probiotic microorganisms must be able to survive the transit through the hostile conditions of the gastrointestinal tract (GIT) and remain at high levels in the intestine, avoiding removal by peristaltic contractions of the gut . In this sense, microorganisms with a short generation time or the ability to adhere to the intestinal mucosa will survive for prolonged periods in the body of the host . Therefore, two desirable properties for probiotic microorganisms are (1) resistance to gastric acidity, bile, and pancreatic enzymes; and (2) adhesion ability to mucosal surfaces.Dairy bacteria are traditionally not considered to persist as normal inhabitants of the human intestinal tract . Therefore, survival under GIT conditions and adherence are important properties to be considered, and tests to study them would be useful tools.In the present chapter we describe the methods used in our laboratory to assess survival, metabolic activity, and adhesion of dairy propionibacteria to intestinal epithelial cells after gastrointestinal digestion. J Appl Microbiol, 2004, 96(6), 1205 - 14 Stationary-phase acid and heat treatments for improvement of the viability of probiotic lactobacilli and bifidobacteria; Saarela M et al.; AIMS: To investigate whether sublethal treatments of stationary-phase probiotic cultures enhance their survival during lethal treatments and to adapt these treatments to the fermenter-scale production of probiotic cultures . METHODS AND RESULTS: Conditions for acid and heat pretreatments were screened for three Lactobacillus and two Bifidobacterium strains . Strains were sublethally treated both at laboratory scale and at fermenter scale in a strain-specific manner and exposed to a subsequent lethal treatment . At laboratory scale viability improvement was detected in each strain . However, improvement was more pronounced in the Lactobacillus than in the Bifidobacterium strains . At fermenter scale three strains were tested: for the two Lactobacillus strains a marked improvement in viability was obtained whereas for the Bifidobacterium strain the improvement was either minor or not detected . CONCLUSIONS: Development of treatments for viability enhancement of probiotic strains is feasible, but strain-specific optimization is necessary to obtain notable improvements . SIGNIFICANCE AND IMPACT OF THE STUDY: Strain-specific treatments were developed for the viability enhancement of stationary-phase probiotic cells both at laboratory and fermenter scale . These results can be utilised in the production of probiotic cultures with improved viability. Clin Diagn Lab Immunol, 2004 May, 11(3), 581 - 7 Effects of feeding a probiotic preparation (SIM) containing inulin on the severity of colitis and on the composition of the intestinal microflora in HLA-B27 transgenic rats; Schultz M et al.; An overly aggressive immune response to the intestinal microflora in a genetically susceptible host background has been implicated in the pathogenesis of inflammatory bowel diseases . We measured the impact of a probiotic preparation (SIM) containing inulin on the severity of colitis and on intestinal microflora profiles of HLA-B27-beta(2)-microglobulin transgenic (TG) rats . SIM is a mixture of lactobacilli, bifidobacteria, and inulin . Two-month-old TG rats received either SIM or water . Control TG rats received metronidazole, alone or in combination with SIM, for 8 weeks . Nontransgenic rats received SIM or water . The cecal content was removed for analysis of the intestinal microflora by PCR combined with denaturing gradient gel electrophoresis . The colon was scored for histological evidence of inflammation, colonic myeloperoxidase activity and interleukin-1beta RNA levels were measured photometrically or by real-time quantitative PCR . At 4 months, the colonic inflammation of TG rats treated with SIM was histologically diminished compared to that in untreated TG rats (2.2 +/- 0.2 versus 2.9 +/- 0.1;