Bifidobacterium

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; P </= 0.03) . The administration of SIM altered the microflora profiles of TG rats by increasing the diversity and stimulating specifically the growth of Bifidobacterium animalis . The probiotic bacteria added to SIM were below the detection level in cecal stool samples at the end of the study period . The administration of SIM resulted in a measurable impact on the cecal microflora profiles of TG rats with attenuation of colitis . The lack of detection of any added probiotic bacteria in the cecal content suggests that prebiotic inulin is the major effective compound.

Int J Food Microbiol, 2004 Jun 1, 93(2), 249 - 57
Determinations of the antimutagenic activities of several probiotic bifidobacteria under acidic and bile conditions against benzo{a}pyrene by a modified Ames test; Lo PR et al.; Antimutagenic activities of six bifidobacteria, after acidic and bile treatment mimicking gastrointestinal conditions, against benzo{a}pyrene (B{a}P) were determined by a modified Ames test using Salmonella typhimurium TA 100 . Results showed that when bifidobacteria were treated at pH 2.0 for 3 h or 1% bile for 6 h, their antimutagenic activities against B{a}P were increased as compared to controls at pH 7.0 for 0 h . After sequential acidic pH and bile treatments, Bifidobacterium lactis Bb-12 displayed the highest antimutagenic activity (71.5%), although its viable cell number was less than 2.0 log cfu/ml . The antimutagenic activity of B . lactis Bb-12 against B{a}P was increased as pH values were increased from 2.0 to 7.0, and reaction time was extended from 1 to 3 h . However, antimutagenic activity was decreased as bile salt concentration was increased from 0.5% to 2.0% . The antimutagenic activity of B . lactis Bb-12 against B{a}P was increased in the presence of whole milk (WM), semiskimmed milk (SSM) and skimmed milk (SM) . When B . lactis Bb-12 was preincubated with B{a}P and milk substrates, its antimutagenic activity was increased to 99-100% .

Int J Food Microbiol, 2004 May 15, 93(1), 109 - 13
H+-ATPase activity in Bifidobacterium with special reference to acid tolerance; Matsumoto M et al.; The acid tolerance of 17 strains of nine species of bifidobacteria was compared using brief exposures to acidic conditions (pH 2-5) . In addition, because it has been hypothesized that the acid tolerance of bifidobacteria depends on H+-ATPase activity, the activity of this enzyme in various strains and species was compared . In general, the acid tolerance of bifidobacteria was found to be weak, with the exception of Bifidobacterium lactis and Bifidobacterium animalis . High numbers of all strains of B . lactis and B . animalis survived exposure to pH 3-5 for 3 h . The H+-ATPase activity of the acid-tolerant strains B . lactis LKM512 and JCM 10602T, and B . animalis JCM 1190T, JCM 1253, JCM 7117, and JCM 7124 was higher at pH 4 than at pH 5 . In contrast, the H+-ATPase activity of nonacid-tolerant strains was lower at pH 4 than at pH 5 .

J Physiol, 2004 Jul 1, 558(Pt 1), 263 - 75 Epub 2004 May 07.
Postnatal microbial colonization programs the hypothalamic-pituitary-adrenal system for stress response in mice; Sudo N et al.; Indigenous microbiota have several beneficial effects on host physiological functions; however, little is known about whether or not postnatal microbial colonization can affect the development of brain plasticity and a subsequent physiological system response . To test the idea that such microbes may affect the development of neural systems that govern the endocrine response to stress, we investigated hypothalamic-pituitary-adrenal (HPA) reaction to stress by comparing germfree (GF), specific pathogen free (SPF) and gnotobiotic mice . Plasma ACTH and corticosterone elevation in response to restraint stress was substantially higher in GF mice than in SPF mice, but not in response to stimulation with ether . Moreover, GF mice also exhibited reduced brain-derived neurotrophic factor expression levels in the cortex and hippocampus relative to SPF mice . The exaggerated HPA stress response by GF mice was reversed by reconstitution with Bifidobacterium infantis . In contrast, monoassociation with enteropathogenic Escherichia coli, but not with its mutant strain devoid of the translocated intimin receptor gene, enhanced the response to stress . Importantly, the enhanced HPA response of GF mice was partly corrected by reconstitution with SPF faeces at an early stage, but not by any reconstitution exerted at a later stage, which therefore indicates that exposure to microbes at an early developmental stage is required for the HPA system to become fully susceptible to inhibitory neural regulation . These results suggest that commensal microbiota can affect the postnatal development of the HPA stress response in mice.

Appl Environ Microbiol, 2004 May, 70(5), 3110 - 21
Bifidobacterium lactis DSM 10140: identification of the atp (atpBEFHAGDC) operon and analysis of its genetic structure, characteristics, and phylogeny; Ventura M et al.; The atp operon is highly conserved among eubacteria, and it has been considered a molecular marker as an alternative to the 16S rRNA gene . PCR primers were designed from the consensus sequences of the atpD gene to amplify partial atpD sequences from 12 Bifidobacterium species and nine Lactobacillus species . All PCR products were sequenced and aligned with other atpD sequences retrieved from public databases . Genes encoding the subunits of the F(1)F(0)-ATPase of Bifidobacterium lactis DSM 10140 (atpBEFHAGDC) were cloned and sequenced . The deduced amino acid sequences of these subunits showed significant homology with the sequences of other organisms . We identified specific sequence signatures for the genus Bifidobacterium and for the closely related taxa Bifidobacterium lactis and Bifidobacterium animalis and Lactobacillus gasseri and Lactobacillus johnsonii, which could provide an alternative to current methods for identification of lactic acid bacterial species . Northern blot analysis showed that there was a transcript at approximately 7.3 kb, which corresponded to the size of the atp operon, and a transcript at 4.5 kb, which corresponded to the atpC, atpD, atpG, and atpA genes . The transcription initiation sites of these two mRNAs were mapped by primer extension, and the results revealed no consensus promoter sequences . Phylogenetic analysis of the atpD genes demonstrated that the Lactobacillus atpD gene clustered with the genera Listeria, Lactococcus, Streptococcus, and Enterococcus and that the higher G+C content and highly biased codon usage with respect to the genome average support the hypothesis that there was probably horizontal gene transfer . The acid inducibility of the atp operon of B . lactis DSM 10140 was verified by slot blot hybridization by using RNA isolated from acid-treated cultures of B . lactis DSM 10140 . The rapid increase in the level of atp operon transcripts upon exposure to low pH suggested that the ATPase complex of B . lactis DSM 10140 was regulated at the level of transcription and not at the enzyme assembly step.

Biotechnol Lett, 2004 Mar, 26(6), 539 - 42
Light sensitivity of Bifidobacterium longum in bioreactor cultivations; Kiviharju K et al.; Bifidobacteria are gaining commercial significance due to their probiotic properties . However, little is still known about the production of these bacteria and their behavior in bioreactors . Two Bifidobacterium longum strains were sensitive to light when grown in a transparent (glass) bioreactor under microaerophilic growth conditions (i.e . no gases added and slow mixing) . The sensitivity was less clear the more anaerobic the initial conditions were . In a darkened bioreactor in microaerophilic conditions, the two strains grew with maximum specific growth rates of 0.36 h(-1) and 0.48 h(-1) . In an illuminated bioreactor neither strain grew . In comparison, Lactobacillus reuteri was not sensitive to light under the same conditions.

Best Pract Res Clin Gastroenterol, 2004 Apr, 18(2), 299 - 313
Microbial-gut interactions in health and disease . Probiotics; Isolauri E et al.; The definition of probiotics has evolved from a live active culture which improves the balance of the gut microbiota composition to specific effects, in particular, the immunomodulatory potential of clearly defined strains . The strains with beneficial properties, potential sources of probiotics, most frequently belong to the genera Bifidobacterium and Lactobacillus, and some of these strains exhibit powerful anti-inflammatory properties . Indeed, probiotic therapy has attracted research interest in human infectious, inflammatory and allergic disease . The most fully documented disease altering the gut microbiota is acute infectious diarrhoea in childhood . Current probiotic research aims to provide safe but sufficient bacterial stimulus in order to avert deviant immune responsiveness related to allergic and inflammatory diseases . However, further rigorous scientific efforts are required to characterize the immunomodulatory potential of specific probiotic strains for these targets.

Best Pract Res Clin Gastroenterol, 2004 Apr, 18(2), 287 - 98
Microbial-gut interactions in health and disease . Prebiotics; Manning TS et al.; In nutritional sciences there is much interest in dietary modulation of the human gut . The gastrointestinal tract, particularly the colon, is very heavily populated with bacteria . Most bacteria are benign; however, certain gut species are pathogenic and may be involved in the onset of acute and chronic disorders . Bifidobacteria and lactobacilli are thought to be beneficial and are common targets for dietary intervention . Prebiotic is a non-viable food ingredient selectively metabolized by beneficial intestinal bacteria . Dietary modulation of the gut microflora by prebiotics is designed to improve health by stimulating numbers and/or activities of the bifidobacteria and lactobacilli . Having an 'optimal' gut microflora can increase resistance to pathogenic bacteria, lower blood ammonia, increase stimulation of the immune response and reduce the risk of cancer . This chapter examines how prebiotics are being applied to the improvement of human health and reviews the scientific evidence behind their use.

Southeast Asian J Trop Med Public Health, 2003 Dec, 34(4), 845 - 8
Management of infant diarrhea with high-lactose probiotic-containing formula; Sudarmo SM et al.; To study the influence of high-lactose probiotic-containing formula on the course of acute diarrhea, an experiment using a randomized controlled clinical trial with patients having acute diarrhea for 3 days was conducted . One hundred patients were allocated into two groups that were comparable for age, sex, and nutritional status . The test group was administered high-lactose Bifidobacterium bifidum-containing formula, while the control group had no high-lactose probiotic until the end of the experiment . The degree of subsequent diarrhea and recovery were monitored in both groups . The results for the test and control groups were analyzed and compared using the chi-square test and Fisher exact test with a significance level (alpha) of 0.05 . The study results revealed that there was no significant difference between the test and control groups (p>0.05) as well as at positive clinical test (13%) and positive floating test (65%) . However, the patients receiving probiotic-containing formula had significantly less frequency of stools, when compared with the control group (p<0.05).

Dig Dis Sci, 2004 Feb, 49(2), 320 - 7
Modulation of the effect of dextran sulfate sodium-induced acute colitis by the administration of different probiotic strains of Lactobacillus and Bifidobacterium; Osman N et al.; The pathogenic mechanism of inflammatory bowel diseases is not fully understood but colonic microflora including Lactobacillus and Bifidobacterium species may affect the induction of colonic inflammation . In this study the relative efficacy of different probiotic organisms in the prevention of colitis was compared in an induced rat colitis model . Three Lactobacillus strains and two Bifidobacterium strains were fed to Sprague-Dawley rats for 7 days prior to offering the rats 5% dextran sulfate sodium (DSS) in their drinking water to induce colitis and the administration of the probiotics continued for 7 days with the DSS . Colitis severity was assessed daily using a disease activity index (DAI) . Samples were collected 7 days after colitis induction for intestinal bacterial flora and bacterial translocation . The DAI decreased significantly on days 4, 5, 6, and 7 in the Lactobacillus plantarum DSM 9843, Bifidobacterium sp . 3B1, and Bifidobacterium infantis DSM 15158 groups compared to the colitis control . It decreased significantly on days 5, 6, and 7 in the Bifidobacterium infantis DSM 15158 group compared to the Lactobacillus paracaesi DSM 13434 and Lactobacillus gasseri 5B3 groups . It also decreased significantly on day 7 in the L . plantarum DSM 9843 group compared to the L . gasseri 5B3 group . Bacterial translocation to the mesenteric lymph nodes decreased significantly in all treatment groups compared to the colitis control . Enterobacteriaceae bacterial translocation to the liver decreased in all treatment groups compared to the colitis control . Administration of certain strains of Lactobacillus and Bifidobacterium significantly improves the DAI and reduces bacterial translocation, and L . plantarum DSM 9843, Bifidobacterium sp . 3B1, and Bifidobacterium infantis DSM 15158 seem to have the best effect.

Infect Immun, 2004 May, 72(5), 2671 - 8
Pattern of cytokine responses to gram-positive and gram-negative commensal bacteria is profoundly changed when monocytes differentiate into dendritic cells; Karlsson H et al.; The normal gastrointestinal bacterial flora is crucial for the maturation of acquired immunity via effects on antigen-presenting cells (APCs) . Here we investigated how two types of APCs, monocytes and dendritic cells (DCs), react to different bacterial strains typical of the commensal intestinal microflora . Purified human monocytes and monocyte-derived DCs were stimulated with UV-inactivated gram-positive (Lactobacillus plantarum and Bifidobacterium adolescentis) and gram-negative (Escherichia coli and Veillonella parvula) bacterial strains . Monocytes produced higher levels of interleukin 12p70 (IL-12p70) and tumor necrosis factor (TNF), as detected by an enzyme-linked immunosorbent assay, in response to L . plantarum than in response to E . coli and V . parvula . In contrast, DCs secreted large amounts of IL-12p70, TNF, IL-6, and IL-10 in response to E . coli and V . parvula but were practically unresponsive to L . plantarum and B . adolescentis . The lack of a response to the gram-positive strains correlated with lower surface expression of Toll-like receptor 2 (TLR2) on DCs than on monocytes . The surface expression of TLR4 on DCs was undetectable when it was analyzed by flow cytometry, but blocking this receptor decreased the TNF production in response to V . parvula, indicating that TLR4 is expressed at a low density on DCs . Gamma interferon increased the expression of TLR4 on DCs and also potentiated the cytokine response to the gram-negative strains . Our results indicate that when monocytes differentiate into DCs, their ability to respond to different commensal bacteria dramatically changes, and they become unresponsive to probiotic gram-positive bacteria . These results may have important implications for the abilities of different groups of commensal bacteria to regulate mucosal and systemic immunity.

Vestn Ross Akad Med Nauk, 2004, (2), 13 - 7
{The microecological disorders in clinical pathology and their correction by bifidum-containing probiotics}; Vorob'ev AA et al.; On the basis of independent research and published data surveyed in the paper is the prevalence and nature of disorders of the intestinal microflora observed in adults and children with different pathologies, i.e . chronic diseases of the gastrointestinal tract, acute intestinal infections, acute respiratory viral infections, pneumonias, as well as certain nephrological, gynecological, and surgical diseases etc . The deficit of Bifidobacteria was found to be the most frequent chain in intestinal microflora pathologies . The significance of Bifido-containing drugs as well as current research of Russian scientists, including new probiotics with Bifidobacteria immobilized in coal, is elucidated . The advantages of intestinal dysbacteriosis correction by sorbated Bifido-containing probiotics, like bifidumbacterin forte and probifor, are demonstrated, versus bifidumbacterin, in different pathologies; their related clinical-and-morphological effect is defined . Prospects for designing the combined and polyvalent drugs with immobilized Bifidobacteria are described.

Appl Microbiol Biotechnol, 2004 Oct, 65(5), 606 - 10 Epub 2004 Apr 14.
Growth and final product formation by Bifidobacterium infantis in aerated fermentations; Gonzalez R et al.; Fermentation conditions were developed to allow Bifidobacterium infantis to grow in the presence of air . Batch fermentations in TPYG medium, starting from anoxic conditions followed by the application of low airflow rates {0.02-0.1 air volume, per liquid media volume, per minute (vvm)}, were analyzed for growth, oxygen uptake, and product formation by the bacterium . Under all aerated fermentations, B . infantis showed high aerotolerance, with a maximum oxygen-specific consumption rate of 0.34 mmol oxygen per gram dry cell weight per hour in the presence of 0.06 vvm . Similar growth yields were obtained under oxic and anoxic conditions (0.11-0.13 and 0.11 g dry cell weight per mmol glucose, respectively) . Oxygen also influenced metabolite formation since lactate production and its molar relation to acetate increased and formate decreased with aeration rate . Under anoxic conditions, a maximum concentration of 8.1 mM lactate and an acetate/lactate ratio of 3.5:1 were obtained, while under oxic conditions the lactate concentration increased more than two-fold and the acetate/lactate molar ratio decreased to 1.5:1 . The possibility of balancing acetate/lactate molar ratios for organoleptic purposes as well as for obtaining good growth under microaerated conditions was demonstrated.

J Pediatr Gastroenterol Nutr, 2004 Mar, 38(3), 288 - 92
Acidified milk formula supplemented with bifidobacterium lactis: impact on infant diarrhea in residential care settings; Chouraqui JP et al.; OBJECTIVES: Probiotics may be useful in preventing acute infectious diarrhea . Bifidobacteria are particularly attractive as probiotics agent because they constitute the predominant colonic flora of breastfed infants and are thought to play a role in the decreased incidence of diarrhea in breastfed infants . METHODS: This was a multicenter, double-blind, controlled study to evaluate the efficacy of a milk formula supplemented with viable Bifidobacterium lactis strain Bb 12 (BbF) in the prevention of acute diarrhea in infants younger than 8 months living in residential nurseries or foster care centers . RESULTS: Ninety healthy children received either the BbF or a conventional formula (CF) daily . The mean duration of the stay in the residential center was similar (137 v 148 days) . At enrollment, there were no differences between the two groups with respect to age (3.7 +/- 2.1 months), gender, anthropometric data, history of allergy or gastrointestinal disease, frequency of breast-feeding in the neonatal period or timing of introduction of solid food . Altogether, 28.3% of the BbF infants had diarrhea during the study compared with 38.7% of controls (NS) . There was a statistically insignificant trend for shorter episodes of diarrhea in the BbF group (5.1 +/- 3.3 days v 7 +/- 5.5 days, NS) . The number of days with diarrhea was 1.15 +/- 2.5 in the BbF group with a daily probability of diarrhea of 0.84 versus 2.3 +/- 4.5 days and 1.55, respectively, in the CF group (P = 0.0002 and 0.0014) . Feeding infants with the BbF reduced their risk of getting diarrhea by a factor of 1.9 (range, 1.33-2.6) . Analysis of the cumulative incidence of diarrheal episodes showed a trend that the first onset of diarrhea occurred later in the BbF group . CONCLUSION: These results provide some evidence that viable Bifidobacterium lactis strain Bb 12, added to an acidified infant formula, has some protective effect against acute diarrhea in healthy children.

Appl Environ Microbiol, 2004 Apr, 70(4), 1923 - 30
Short fractions of oligofructose are preferentially metabolized by Bifidobacterium animalis DN-173 010; Van der Meulen R et al.; The growth of Bifidobacterium animalis DN-173 010 on different energy sources was studied through small- and large-scale fermentations . Growth on both more common energy sources (glucose, fructose, galactose, lactose, and sucrose) and inulin-type fructans was examined . High-performance liquid chromatography analysis was used to investigate the kinetics . Gas chromatography was used to determine the fructan degradation during the fermentation process . B . animalis DN-173 010 was unable to grow on a medium containing glucose as the sole energy source . In general, monosaccharides were poor growth substrates for the B . animalis strain . The fermentations with the inulin-type fructans resulted in changes in both growth and metabolite production due to the preferential metabolism of certain fructans, especially the short-chain oligomers . Only after depletion of the shorter chains were the larger fractions also metabolized, although to a lesser extent . Acetic acid was the major metabolite produced during all fermentation experiments . At the beginning of the fermentation, high levels of lactic acid were produced, which were partially replaced by formic acid at later stages . This suggests a shift in sugar metabolism to gain additional ATP that is necessary for growth on oligofructose, which is metabolized more slowly.

Adv Drug Deliv Rev, 2004 Apr 19, 56(6), 809 - 18
Probiotics in inflammatory bowel disease--therapeutic rationale and role; Shanahan F; The intestinal flora has a conditioning effect on intestinal homeostasis, delivering regulatory signals to the epithelium, the mucosal immune system and to the neuromuscular activity of the gut . Beneficial metabolic activities of the enteric flora include nutrient production, metabolism of dietary carcinogens, conversion of prodrugs to active drugs . However, increasing evidence suggests that some components of the enteric flora are essential ingredients in the pathogenesis of inflammatory bowel disease (IBD); this has prompted interest in therapeutic manipulation of the flora with probiotics . Probiotics are biologic control agents-described as live microbial food supplements which confer a health benefit beyond inherent basic nutrition . Multiple potential beneficial effects have been attributed to the probiotic use of lactic acid bacteria, bifidobacteria and other non-pathogenic commensals . At present, much of the promise of probiotics remains outside the realm of evidence-based medicine and awaits the results of prospective trials, now underway . No reliable in vitro predictors of in vivo efficacy of putative probiotics have been identified . Rigorous comparisons of probiotic performance have not been performed and the suitability of a given probiotic for different individuals is largely unexplored . Notwithstanding, an improved understanding of the normal commensal flora and host-flora interactions has the potential to open up new therapeutic strategies for inflammatory disorders of the gut.

Curr Issues Intest Microbiol, 2004 Mar, 5(1), 1 - 8
The role of oxygen in the viability of probiotic bacteria with reference to L . acidophilus and Bifidobacterium spp; Talwalkar A et al.; The various therapeutic benefits of Lactobacillus acidophilus and Bifidobacterium spp . have resulted in their increased incorporation into dairy foods such as yoghurts . Currently however, the efficacy of these probiotic bacteria is limited by their poor survival during the shelf life of yoghurt . Oxygen toxicity is widely considered to be responsible for the cell deaths of these bacteria . The intestinal origins and the microaerophilic and anaerobic characteristics of L . acidophilus and Bifidobacterium spp . respectively, can render them susceptible to oxygen contained in the food products . This review discusses the influence of the dissolved oxygen in yogurt on the viability of these bacteria . Suggested techniques to protect these probiotic bacteria from oxygen toxicity are evaluated . Although the problem of oxygen toxicity in probiotic bacteria is regarded as significant, little is known however about the cellular interaction of these bacteria with oxygen . This review summarizes what is known about the biochemistry of oxygen toxicity in these bacteria . The various metabolic and biochemical responses of L . acidophilus and Bifidobacterium to oxygen are examined . Additionally, the importance of NADH oxidase and NADH peroxidase in the oxygen tolerance of these bacteria is evaluated and assays used to measure their cellular concentrations are discussed.

Di Yi Jun Yi Da Xue Xue Bao, 2004 Mar, 24(3), 264 - 8
{Effect of Bifidobacterial adhesin on lipopolysaccharide- and H2O2-induced proliferation and apoptosis of intestinal epithelial cells in vitro}; Zhong SS et al.; OBJECTIVE: To study the effect of Bifidobacterial adhesin on proliferation and apoptosis of intestinal epithelial cell induced by lypopolysaccharide (LPS) and H(2)O(2) in vitro . METHODS: With (3)H-TdR incorporation method, flow cytometry and fluorochrome staining, the proliferation and apoptosis of intestinal epithelial cells induced by LPS and H(2)O(2) in vitro were studied . RESULTS: LPS at the dose of 100 microg/L effectively stimulated the proliferation and apoptosis of cells, whereas H(2)O(2) at the dose of 200 micromol/L obviously restrained the proliferative ability while enhanced the apoptosis of the cells . Fluorochrome staining showed cell shrinkage, nuclear condensation and fragmentation under microscope . After treatment with Bifidobacterial adhesin, the cell proliferation and apoptosis decreased significantly in LPS group, and in H(2)O(2) group, cell apoptosis was significantly decreased . CONCLUSIONS: Bifidobacterial adhesin can protect intestinal epithelial cells from the damage by LPS and H(2)O(2), and maintain the balance between the proliferation and apoptosis of the cells.

Infect Immun, 2004 Apr, 72(4), 2240 - 7
Probiotic bifidobacteria protect mice from lethal infection with Shiga toxin-producing Escherichia coli O157:H7; Asahara T et al.; The anti-infectious activity of probiotic Bifidobacteria against Shiga toxin-producing Escherichia coli (STEC) O157:H7 was examined in a fatal mouse STEC infection model . Stable colonization of the murine intestines was achieved by the oral administration of Bifidobacterium breve strain Yakult (naturally resistant to streptomycin sulfate) as long as the mice were treated with streptomycin in their drinking water (5 mg/ml) . The pathogenicity of STEC infection, characterized by marked body weight loss and subsequent death, observed in the infected controls was dramatically inhibited in the B . breve-colonized group . Moreover, Stx production by STEC cells in the intestine was almost completely inhibited in the B . breve-colonized group . A comparison of anti-STEC activity among several Bifidobacterium strains with natural resistance to streptomycin revealed that strains such as Bifidobacterium bifidum ATCC 15696 and Bifidobacterium catenulatum ATCC 27539(T) did not confer an anti-infectious activity, despite achieving high population levels similar to those of effective strains, such as B . breve strain Yakult and Bifidobacterium pseudocatenulatum DSM 20439 . The effective strains produced a high concentration of acetic acid (56 mM) and lowered the pH of the intestine (to pH 6.75) compared to the infected control group (acetic acid concentration, 28 mM; pH, 7.15); these effects were thought to be related to the anti-infectious activity of these strains because the combination of a high concentration of acetic acid and a low pH was found to inhibit Stx production during STEC growth in vitro.

Hepatol Res, 2004 Mar, 28(3), 155 - 160
Endotoxinemia and benzodiazepine-like substances in compensated cirrhotic patients: a randomized study comparing the effect of rifaximine alone and in association with a symbiotic preparation; Lighthouse J et al.; Aim: The aim of the present investigation was to test study benzodiazepines (BZDs) profile in patients with viral cirrhosis under different combinations of rifaximine and of a novel symbiotic . Methods: Our study groups consisted of 30 patients with a confirmed diagnosis of HCV-related Child B liver cirrhosis . Patients were randomly allocated into three groups: rifaximine 400mg t.i.d . for 2 weeks; (B) SCM-III (Lactobacillus acidophilus, Lactobacillus helveticus and Bifidobacteria in a ion- and vitamin-enriched medium, Named srl, Italy) 10ml t.i.d . for 2 weeks; (C) rifaximine 400mg t.i.d . for 1 week followed by SCM-III 10ml t.i.d . for 5 weeks . At weekly interval, blood samples were withdrawn to test BZD-like substances, ammonia and endotoxin . Results: Rifaximine treatment brought about a significant early drop of BZDs ( {Formula: see text} versus pre-treatment and versus control) till fourth week of observation when a gradual increase took place with return to pre-treatment values at the sixth week . Symbiotic treatment was comparably effective while given to patients but significantly elevated BZDs level were noted starting from the third week . Similar phenomena were noted for endotoxin and ammonia although symbiotic seemed more effective against endotoxin and rifaximine against ammonia increase . However, the sequential treatment rifaximine-symbiotic brought about a sustained normalization of BZDs, ammonia and endotoxin throughout the 6-week study . Conclusion: The present pilot study suggests that a rifaximine-symbiotic regimen could be an effective tool in compensated liver cirrhosis to limit some triggering factors of hepatic encephalopathy while being amenable to long-term use and devoid of significant side effects.

Int J Food Microbiol, 2004 Apr 1, 92(1), 69 - 78
In vitro inhibition of Escherichia coli O157:H7 by bifidobacterial strains of human origin; Gagnon M et al.; The ability of bifidobacteria isolated from infant feces to inhibit enterohemorrhagic Escherichia coli serotype O157:H7 in vitro and reduce its adhesion to human enterocyte-like Caco-2 cells was evaluated in comparison to American Type Culture Collection bifidobacterial reference strains . Five Bifidobacterium isolates from infant feces were identified and characterized by morphology, fructose-6-phosphate phosphoketolase (F6PPK) assay, polymerase chain reaction using bifidobacterial 16S rDNA specific primers, carbohydrate fermentation patterns, resistance to lysozyme, acid, bile and hydrogen peroxide as well as their ability to inhibit E . coli O157:H7 using the agar spot technique . Infant isolates showed greater resistance to bile, acid, lysozyme and more antimicrobial activity against E . coli O157:H7 than ATCC strains . Two infant isolates identified as B . bifidum RBL 71 and B . bifidum RBL 460 showed good adhesion and significant potential for reducing adhesion of E . coli O157:H7 to Caco-2 cells . This effect was dependent on bifidobacterial cell concentration . These results show that bifidobacteria isolated from infants may be useful for improving probiotic formulae with respect to protection against E . coli O157:H7 infection.

J Vet Sci, 2004 Mar, 5(1), 41 - 8
Immunomodulatory and antitumor effects in vivo by the cytoplasmic fraction of Lactobacillus casei and Bifidobacterium longum; Lee JW et al.; The immunomodulatory and antitumor effects of lactic acid bacteria (LABs) were investigated . Cytoplasmic fraction of Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium longum were tested for the antiproliferative activity in vitro to SNUC2A, SNU1, NIH/3T3 and Jurkat cell lines by crystal violet assay . All cytoplasmic fraction suppressed proliferation of tumor cells, though L . casei and B . longum were more effective . From these results, cytoplasmic fraction of L . casei and B . longum with Y400 as a control were administered as dietary supplements to Balb/c mice for 2, and 4 consecutive wks . Administration for 4 wks enhanced the number of total T cells, NK cells and MHC class II+ cells, and CD4-CD8+ T cells in flow cytometry analysis . To determine of antitumor activity of LABs preparation in vivo, F9 teratocarcinoma cells were inoculated on mice at 14th day . Body weight was decreased with increased survival rate in all groups with the cytoplasm of LABs . Our results showed that cytoplasmic fraction of LABs had direct antiproliferative effects on tumor cell lines in vitro, effects on immune cells in vivo, and antitumor effects on tumor-bearing mice with prolonged survival periods.

Zh Mikrobiol Epidemiol Immunobiol, 2004 Jan-Feb, (1), 62 - 7
{Qualitative and quantitative composition of intestinal microflora in healthy young children}; Postnikova EA et al.; The results of the qualitative and quantitative analysis of the microflora of the large intestine in 45 healthy children aged 6, 8, 10 months and 1 year, living in Moscow, are presented . During the first year of life high concentrations and detection rate of not only bifidobacteria, enterococci, nonpathogenic Escherichia, but also bacteria, commonly regarded as opportunistic were typical . The latter include lecithinase positive Clostridium, citrate assimilating enterobacteria, as well as Escherichia with low biochemical activity and capacity for hemolysin production . In addition, this group also includes coagulase-positive staphylococci . Later on, as these children become older, opportunistic bacteria are partially or completely eliminated under normal conditions . Suggestion is made that the composition of intestinal microflora depends on protective and other systems of the host at different periods of life, as well as on the character of nutrition and the microbial contamination of the environment.

Int J Syst Evol Microbiol, 2004 Mar, 54(Pt 2), 401 - 6
Bifidobacterium psychraerophilum sp . nov . and Aeriscardovia aeriphila gen . nov., sp . nov., isolated from a porcine caecum; Simpson PJ et al.; In a previous study that was based primarily on 16S rDNA sequencing, two groups of bifidobacteria that had been recovered from a pig caecum were proposed to belong to two novel species, termed 'Bifidobacterium pyschroaerophilum' and 'Bifidobacterium aerophilum' . In this study, based on DNA G+C content and partial heat-shock protein 60 (HSP60) gene sequences, the assignment of 'B . pyschroaerophilum', corrected to Bifidobacterium pyschraerophilum, to the genus Bifidobacterium was confirmed . The DNA G+C content of 'B . aerophilum' was relatively low, which was consistent with its segregation into subcluster II of the 16S rDNA phylogenetic tree . Based on partial 16S rDNA and HSP60 gene sequences, the species was transferred to a novel genus and reclassified as Aeriscardovia aeriphila gen . nov., sp . nov . Biochemical profiles and growth parameters were established for both novel species . Interestingly, each had a high tolerance to oxygen and grew on agar media under aerobic conditions, a trait that may relate to their caecal habitat . Under aerobic growth conditions, the short-rod morphology of A . aeriphila lengthened considerably . This appeared to arise from incomplete cell division . In addition, B . pyschraerophilum was unusual in that it grew at temperatures as low as 4 degrees C . On the basis of genetic, phylogenetic and phenotypic data, the identities of Bifidobacterium pyschraerophilum sp . nov . (type strain, T16(T)=LMG 21775(T)=NCIMB 13940(T)) and Aeriscardovia aeriphila gen . nov., sp . nov . (type strain, T6(T)=LMG 21773(T)=NCIMB 13939(T)) are confirmed.

Asia Pac J Clin Nutr . 2003;12 Suppl:S60.
Effect of experimental oligosaccharide on brain and body weight; Wang B et al.; Background - The biological function of human milk oligosaccharides (HMO) is not fully understood . In addition to their ability to stimulate growth of Bifidobacteria and inhibit pathogens, there may be beneficial effects on brain development . Our hypothesis is that oral supplementation of certain oligosaccharides favourably influences brain growth and learning ability in an animal model . The piglet is the most appropriate as its brain growth closely parallels that of the human infant . Objective - To investigate the effects of an experimental oligosaccharide found in human milk, on brain weight, total brain cortex cell number (estimated by DNA content) and body weight development in formula-fed piglets . Design - Twelve 3-day-old male piglets were randomly allocated into a treatment (n =6) or control group (n=6) . Piglets were fed a diet of soy/whey/casein sow's milk replacer (55:9:36) for 32 days . The treatment group were fed the standard diet with 215 mg/kg oligosaccharide per day . Body weight was determined daily using electronic scales prior to the morning feed . The piglets were euthanased on day 36 and brain weight and frontal cortex DNA content were determined . Outcomes - The rate of body weight gain (mean +/- SD) did not differ between the groups: 217 +/- 34 g/d in the treatment group and 216 +/- 39 g/d in the controls . Brain weights were similar 54 +/- 6 g vs 57 +/- 4 g respectively, and cerebrum and cerebellum were 71% and 11% vs 69% and 11% respectively of total brain weight . Brain frontal cortex cell number was 24% higher (110.7 +/- 14.2 x 10(3) cells/mg tissue) in treatment than controls (89.3 +/- 15.0 x 10(3) cells/mg tissue) but the difference did not reach statistical significance (P=0.326) . Conclusions - This study showed that dietary supplementation of the experimental oligosaccharide had no significant effect on body weight gain, brain weight or frontal cortex cell number.

Plasmid, 2004 Mar, 51(2), 87 - 100
Molecular characterization of three plasmids from Bifidobacterium longum; Corneau N et al.; The complete nucleotide sequences for pNAC1 (3538bp) from strain RW048 as well as for pNAC2 (3684bp) and pNAC3 (10,224bp) from strain RW041 of Bifidobacterium longum were determined . The largest ORF (repB) of pNAC1 encodes a putative protein similar to those involved in a rolling-circle (RC) replication mechanism, which was confirmed by demonstration of single-strand intermediates in the host cell . The putative RepB gene product of pNAC2 is most similar to the replication protein of pDOJH10L and pKJ36 . A second gene (mob) is similar to mobilization proteins involved in conjugation . Plasmid pNAC3 is the largest bifidobacterial plasmid to be sequenced to date . Of the eight putative gene products coded by pNAC3, one is similar to replication proteins (RepB), and another (Orf2) to putative transfer proteins (Tra) . Bifidobacterial plasmids were divided into five groups based on Rep amino acid sequence homology and the results suggest a new plasmid family for B . longum.

J Microbiol Methods, 2004 Apr, 57(1), 9 - 16
The evaluation of a mupirocin-based selective medium for the enumeration of bifidobacteria from probiotic animal feed; Simpson PJ et al.; In this study, MRS medium supplemented with cysteine hydrochloride and mupirocin, termed Bifidobacterium selective medium (BSM) was found to be elective for bifidobacteria but inhibitory to a wide range of non-bifidobacteria strains commonly included in probiotic animal feed . Bacilli, lactobacilli, lactococci and streptococci failed to form colonies on BSM and enterococci, pediococci and propionibacteria formed colonies <0.5 mm in diameter . Bifidobacteria formed colonies >1 mm in size and could be readily distinguished . The addition of nystatin to BSM further inhibited Saccharomyces cerevisiae . BSM was successfully used to enumerate the bifidobacteria components, confirmed through fructose-6-phophate-phosphoketolase detection, present in two commercial probiotic feeds . The medium is recommended for the enumeration of bifidobacteria from animal feeds especially when not a numerically dominant component.

J Agric Food Chem, 2004 Mar 10, 52(5), 1338 - 43
Characterization of a new potential functional ingredient: coffee silverskin; Borrelli RC et al.; Dietary fiber (DF) is one of the main dietary factors contributing to consumers' well-being . In this work the possibility of using the roasted coffee silverskin (CS), a byproduct of roasted coffee beans, as a DF-rich ingredient has been evaluated . The results of our investigation showed that this material has 60% total DF, with a relevant component (14%) of soluble DF . Although a small amount of free phenol compounds is present in CS, it has a marked antioxidative activity, which can be attributed to the huge amount of Maillard reaction products, the melanoidins . Static batch culture fermentation experiments showed that CS induces preferential growth of bifidobacteria rather than clostridia and Bacteroides spp . CS can be proposed as a new potential functional ingredient in consideration of the high content of soluble DF, the marked antioxidant activity, and the potential prebiotic activity.

Int J Food Microbiol, 2004 Mar 15, 91(3), 253 - 60
In vitro assessment of the upper gastrointestinal tolerance of potential probiotic dairy propionibacteria; Huang Y et al.; This study aimed to assess the transit tolerance of potential probiotic dairy propionibacteria strains in human upper gastrointestinal tract in vitro, and to evaluate the effect of food addition on viability of these strains in simulated pH 2.0 gastric juices . The transit tolerance of 13 dairy propionibacteria strains was determined at 37 degrees C by exposing washed cell suspensions to simulated gastric juices at pH values at 2.0, 3.0, and 4.0, and simulated small intestinal juices (pH 8.0) with or without 0.3% bile salts . The viability of dairy propionibacteria in pH 2.0 simulated gastric juice with So-Good original soymilk or Up & Go liquid breakfast was also determined . The simulated gastric transit tolerance of dairy propionibacteria was strain-dependent and pH-dependent . All tested strains were tolerant to simulated small intestinal transit . The addition of So-Good original soymilk or Up & Go liquid breakfast greatly enhanced the survival of dairy propionibacteria strains in pH 2.0 simulated gastric juices . Dairy propionibacteria strains demonstrate high tolerance to simulated human upper gastrointestinal tract conditions and offer a relatively overlooked, yet alternative source for novel probiotics besides Lactobacillus and Bifidobacterium.

Poult Sci, 2004 Feb, 83(2), 175 - 82
Effects of mushroom and herb polysaccharides, as alternatives for an antibiotic, on the cecal microbial ecosystem in broiler chickens; Guo FC et al.; An in vivo experiment was conducted to study the potential prebiotic effects of mushroom and herb polysaccharide extracts, Lentinus edodes extract (LenE), Tremella fuciformis extract, and Astragalus membranaceus Radix extract, on chicken growth and the cecal microbial ecosystem, as compared with the antibiotic Apramycin (APR) . This investigation was carried out in terms of a dose-response study . The chickens were naturally infected with avian Mycoplasma gallisepticum prior to the experiment . The BW gain, cecal pH, viscosity, and predominant microbial populations were measured 1 wk after the extract and APR treatments . The extracts and APR significantly stimulated growth of the chickens infected with avian Mycoplasma gallisepticum . The average BW gain of the groups fed with the extracts was significantly lower than that of the antibiotic group . The extracts had no significant effect on cecal pH . However, cecal viscosity and microbial populations were significantly affected by feeding extracts and antibiotic . In contrast to APR, the extracts stimulated the number of the potentially beneficial bacteria (bifidobacteria and lactobacilli), while reducing the number of the potentially harmful bacteria (Bacteroides spp . and Escherichia coli) . Of the 3 extracts, LenE was associated with the most cecal bifidobacteria and lactobacilli . With each increase in the LenE dose, birds tended to have higher BW gain and total aerobe and anaerobe counts . Numbers of predominant cecal bacteria, in particular, E . coli, bifidobacteria, and lactobacilli, were significantly increased with increases in the LenE dose . It would seem that these specific mushroom and herb polysaccharide extracts hold some promise as potential modifiers of intestinal microbiota in diseased chickens.

Lett Appl Microbiol, 2004, 38(3), 229 - 32
Evaluation of the rrn operon copy number in Bifidobacterium using real-time PCR; Candela M et al.; AIMS: A real-time PCR-based method was developed to evaluate the Bifidobacterium rRNA operon copy number . As a result of their repetitive nature, rRNA operons are very suitable targets for chromosomal integration of heterologous genes . METHODS AND RESULTS: The rrn operon multiplicity per chromosome was determined by real-time PCR quantification of the 16S rRNA amplicons obtained from genomic DNA . The values obtained in several bifidobacterial strains of human origin ranged from 1 to 5 . The reliability of the method developed was confirmed by Southern hybridization technique . CONCLUSIONS: In the Bifidobacterium genus the rrn operon copies showed variability at species and strain level . The identification of Bifidobacterium strains with high rRNA multiplicity allowed the selection of potential hosts for chromosomal integration . SIGNIFICANCE AND IMPACT OF THE STUDY: The methodology here proposed represents a rapid, reliable and sensitive new tool for the quantification of rrn operon copy number in bacteria.

Food Chem Toxicol, 2004 Mar, 42(3), 351 - 7
In vitro digestion of sinigrin and glucotropaeolin by single strains of Bifidobacterium and identification of the digestive products; Cheng DL et al.; Three strains of Bifidobacterium sp., B . pseudocatenulatum, B . adolescentis, and B . longum were studied for their ability to digest glucosinolates, sinigrin (SNG) and glucotropaeolin (GTL), in vitro . All strains digested both glucosinolates during 24-48 h cultivation, accompanied by a decline in the medium pH from 7.1 to 5.2 . The digestion of glucosinolates by a cell-free extract prepared from sonicated cells of B . adolescentis, but not cultivated broth, increased in the presence of 0.5 mM l-ascorbic acid . Also, a time-dependent formation of allyl isothiocyanate (AITC) was observed when the cell-free extract was incubated with 0.25 mM SNG for 120 min at pH 7.0 . These reaction features suggest that the digestive activity may have been due to an enzyme similar to myrosinase, an enzyme of plant origin . GC-MS analysis of the Bifidobacterial cultured broth showed that the major products were 3-butenenitrile (BCN) and phenylacetonitrile (PhACN), from SNG and GTL, respectively and nitriles, probably due to a decrease in the pH of the media . AITC and benzyl isothiocyanate (BzITC) were barely detectable in the broth . It was concluded that the three species of Bifidobacteria could be involved in digestive degradation of glucosinolates in the human intestinal tract.

Biotechnol Prog, 2004 Jan-Feb, 20(1), 145 - 50
Continuous production of mixed lactic starters containing probiotics using immobilized cell technology; Doleyres Y et al.; The production of a mixed lactic culture containing Lactococcus lactis subsp . lactis biovar . diacetylactis MD and Bifidobacterium longum ATCC 15707 was studied during a 17-day continuous immobilized-cell culture at different temperatures between 32 and 37 degrees C . The two-stage fermentation system was composed of a first reactor (R1) containing cells of the two strains separately immobilized in kappa-carrageenan/locust bean gum gel beads and a second reactor (R2) operated with free cells released from the first reactor . The system allowed continuous production of a concentrated mixed culture with a strain ratio whose composition depended on temperature and fermentation time . A stable mixed culture (with a 22:1 ratio of L . diacetylactis and B . longum) was produced at 35 degrees C in the effluent of R2, whereas the mixed culture was rapidly unbalanced in favor of B . longum at a higher temperature (37 degrees C) or L . diacetylactis at a lower temperature (32 degrees C) . Strain redistribution in beads originally immobilizing pure cultures of L . diacetylactis or B . longum was observed . At the end of culture, the strain ratio (7:1 L . diacetylactis/B . longum) in bulk bead samples was similar to that of individual beads . The determination of the spatial distribution of the two strains in gel beads by immunofluorescence and confocal laser-scanning microscopy showed that bead cross-contamination was limited to a 100 microm peripheral layer . Data from this study validate a previous model for population dynamics and cell release in gel beads during mixed immobilized-cell cultures.

J Dairy Sci, 2004 Feb, 87(2), 258 - 66
Purification and characterization of three different types of bile salt hydrolases from Bifidobacterium strains; Kim GB et al.; Bile salt hydrolases were purified to electrophoretic homogeneity from Bifidobacterium bifidum ATCC 11863, Bifidobacterium infantis KL412, Bifidobacterium longum ATCC 15708, Bifidobacterium longum KL507, and Bifidobacterium longum KL515 . Three different types (A, B, and C) of bile salt hydrolase (BSH) were revealed during the purification study, exhibiting the type-specific characteristics in their electrophoretic migration and elution profiles from anion exchange and hydrophobic interaction chromatographic columns . The subunit molecular mass estimated by sodium dodecylsulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) was around 35 kDa, and the native molecular mass in all five Bifidobacterium strains was estimated to be between 130 and 150 kDa by gel filtration chromatography, indicating that all BSH enzymes have tetrameric structure . From the isoelectric focusing, an isoelectric point value of 4.45 was obtained with BSH (type B) from B . bifidum ATCC 11863 and the other BSH (types A and C) showed the similar pI values around 4.65 . N-Terminal amino acid sequencing for the proteins of types A and C revealed that 6 out of 20 amino acid residues were different, and highly conserved residues were identified in both N-terminal sequences of types A and C . All BSH enzymes from five strains hydrolyzed six major human bile salts, and they showed a better deconjugation rate on glycine-conjugated bile salts than on taurine-conjugated forms.

Poult Sci, 2004 Jan, 83(1), 84 - 8
The effects of supplementation of humate and probiotic on egg production and quality parameters during the late laying period in hens; Yoruk MA et al.; This study was designed to investigate whether inclusions of humate and probiotic into diets of hens during the late laying period increases egg production and improves egg quality . Hisex Brown layers (n = 300), 54 wk of age, were fed a control diet, 0.1% humate, 0.2% humate, 0.1% probiotic, or 0.2% probiotic for 75 d . Active ingredients of humate and probiotic were polymeric polyhydroxy acids (humic, fulvic, ulmic, and humatomelanic acids) and bacterial cultures (Lactobacillus, Bifidobacterium, Streptococcus, and Enterococcus spp.), respectively . Egg production and feed intake were measured daily, and egg weight was measured biweekly . Also, a sample of 12 eggs from each group was collected randomly to determine egg quality every 25 d . The data were analyzed as repeated measures with time as subplot . There were no effects of dietary treatments on feed intake and egg weight . Egg production for hens supplemented with humate and probiotic was not different but was greater than for control hens . Egg production increased linearly and mortality and feed conversion efficiency (weight of feed/weight of eggs) decreased linearly with increasing levels of supplemental humate and probiotic . There were no effects of treatments on egg quality . In conclusion, supplementation of humate and probiotic during the late laying period increased egg production, reduced mortality, and improved feed conversion efficiency but did not improve egg quality.

Biochemistry, 2004 Feb 10, 43(5), 1156 - 62
Crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis; Sprogoe D et al.; Around 80 enzymes are implicated in the generic starch and sucrose pathways . One of these enzymes is sucrose phosphorylase, which reversibly catalyzes the conversion of sucrose and orthophosphate to d-Fructose and alpha-d-glucose 1-phosphate . Here, we present the crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis (BiSP) refined at 1.77 A resolution . It represents the first 3D structure of a sucrose phosphorylase and is the first structure of a phosphate-dependent enzyme from the glycoside hydrolase family 13 . The structure of BiSP is composed of the four domains A, B, B', and C . Domain A comprises the (beta/alpha)(8)-barrel common to family 13 . The catalytic active-site residues (Asp192 and Glu232) are located at the tips of beta-sheets 4 and 5 in the (beta/alpha)(8)-barrel, as required for family 13 members . The topology of the B' domain disfavors oligosaccharide binding and reduces the size of the substrate access channel compared to other family 13 members, underlining the role of this domain in modulating the function of these enzymes . It is remarkable that the fold of the C domain is not observed in any other known hydrolases of family 13 . BiSP was found as a homodimer in the crystal, and a dimer contact surface area of 960 A(2) per monomer was calculated . The majority of the interactions are confined to the two B domains, but interactions between the loop 8 regions of the two barrels are also observed . This results in a large cavity in the dimer, including the entrance to the two active sites.

World J Gastroenterol, 2004 Feb 1, 10(3), 455 - 7
Probiotics inhibit TNF-alpha-induced interleukin-8 secretion of HT29 cells; Bai AP et al.; AIM: To study the effect of probiotics on interleukin-8 secretion in intestinal epithelia when stimulated by proinflammatory cytokines . METHODS: Colonic adenocarcinoma HT29 cells were cultured and divided into four groups: control, TNF-alpha (group T in short), bifidobacterium (group B), lactobacillus (group L) . B . Longum and L . bulgaricus were suspended in culture medium with a concentration of 1 x 10(8) cfu/ml and added into 24 wells respectively . One hour later TNF-alpha (10 ng/ml) was added into each well of groups T, B, L . The supernatants were collected and measured for IL-8 after 3 hours, nuclear factor-kappaB (NF-kappaB) p65 was also examined by Western blotting . RESULTS: There was less interleukin-8 secretion in HT29 cells when preincubated with B . Longum or L . bulgaricus compared with group T . Less p65 appeared in nuclei in groups B and L compared with group T, as detected by Western blot . CONCLUSION: Probiotics can suppress interleukin-8 secretion in intestinal epithelia when stimulated by proinflammatory cytokines, which is most likely mediated by NF-kappaB.

Zentralbl Gynakol, 2003 Dec, 125(12), 475 - 9
{Probiotics}; Wisker E; The colonic flora is thought to play a key role in human health . Gut bacteria produce desirable as well as undesirable metabolites from fermentation and sometimes they may act as pathogens . Composition as well as activities of the gut bacteria can be influenced by nutrition . Prebiotics are food ingredients that selectively stimulate the growth of colonic bacteria regarded as beneficial, i . e . bifidobacteria and lactobacilli . Fructose polymers (fructo-oligosaccharides, inulin), galacto-oligosaccharides and soya oligosaccharides were shown to exert prebiotic activities . In adults, the growth of bifidobacteria was stimulated especially by inulin and fructo-oligosaccharides . Infant formulas containing galacto-oligosaccharides and inulin promoted the growth of bifidobacteria in faeces of bottle-fed infants and resulted in stool characteristics similar to those found in breast-fed infants . However, at present there is only limited knowledge on the long-term health consequences of increased counts of bifidobacteria . In adults, prebiotics can enhance the absorption of calcium from the colon . They can increase stool frequency and are mildly laxative . Prebiotic consumption is usually accompanied by mild flatulence and in sensitive persons by more severe gastrointestinal symptoms . In contrast to the results of animal experiments, a significant lipid-lowering action of fructo-oligosaccharides has not been found in humans . In animals, fructo-oligosaccharides have shown a favourable effect on experimental carcinogenesis . Whether or not these findings may apply to humans will be subject of future studies.

Lancet, 2004 Jan 24, 363(9405), 325 - 7
Bifidobacterium lipoteichoic acid and false ELISA reactivity in aspergillus antigen detection; Mennink-Kersten MA et al.; A major difficulty with the detection of circulating galactomannan, a cell-wall polysaccharide released by Aspergillus sp during growth, in the serodiagnosis of invasive aspergillosis is the occurrence of false-positive ELISA results, especially in neonates and infants . On the basis of molecule similarity, we postulate that a lipoteichoic acid of Bifidobacterium sp can act as epitope for the monoclonal antibody used in the ELISA . The neonatal gut is heavily colonised with Bifidobacterium sp and these bacteria or their lipoteichoic acid might cause ELISA reactivity with serum after translocation because of immaturity of the intestinal mucosa . If our hypothesis is correct, we might find a method to discriminate between false-positive and true-positive ELISA results and thereby prevent unnecessary pre-emptive treatment of patients.

Int J Food Microbiol, 2004 Feb 1, 90(3), 357 - 62
Binding of Bifidobacterium bifidum and Lactobacillus reuteri to the carbohydrate moieties of intestinal glycolipids recognized by peanut agglutinin; Mukai T et al.; We examined binding of Bifidobacterium bifidum and Lactobacillus reuteri to the carbohydrate moieties of glycolipids extracted from human enterocyte-like Caco-2 cells in this study . In binding assays to reference glycolipids of different carbohydrate compositions, B . bifidum EB102 bound strongly to gangliotetraosylceramide (asialo-GM1) and less strongly to gangliotriaosylceramide (asialo-GM2), lactosylceramide and sulfatide . The binding profile of B . bifidum EB102 was almost identical to that of L . reuteri JCM1081 described previously {Lett . Appl . Microbiol . 27 (1998) 130} . When we examined binding to neutral glycolipids extracted from Caco-2 cells, the binding profiles of B . bifidum EB102 and L . reuteri JCM1081 were very similar to that shown by peanut agglutinin (PNA) . Binding of both strains to periodate-treated intestinal glycolipids was completely abolished, suggesting that the bacterial cells bind to carbohydrate moieties of the glycolipids . Furthermore, B . bifidum EB102 was found to express multiple glycolipid-binding proteinaceaous components on the cell surface . These results strongly suggested involvement of cell-surface proteinaceous components of B . bifidum in binding to the carbohydrate moieties of intestinal glycolipids recognized by PNA . Binding ability of B . bifidum and L . reuteri to intestinal glycolipids may play a crucial role for colonization on the mucosal surface of the intestine.

Appl Microbiol Biotechnol, 2004 Aug, 65(2), 219 - 27 Epub 2004 Jan 22.
Physico-chemical and transglucosylation properties of recombinant sucrose phosphorylase from Bifidobacterium adolescentis DSM20083; van den Broek LA et al.; Clones of a genomic library of Bifidobacterium adolescentis were grown in minimal medium with sucrose as sole carbon source . An enzymatic fructose dehydrogenase assay was used to identify sucrose-degrading enzymes . Plasmids were isolated from the positive colonies and sequence analysis revealed that two types of insert were present, which only differed with respect to their orientation in the plasmid . An open reading frame of 1,515 nucleotides with high homology for sucrose phosphorylases was detected on these inserts . The gene was designated SucP and encoded a protein of 56,189 Da . SucP was heterologously expressed in Escherichia coli, purified, and characterized . The molecular mass of SucP was 58 kDa, as estimated by SDS-PAGE, while 129 kDa was found with gel permeation, suggesting that the native enzyme was a dimer . The enzyme showed high activity towards sucrose and a lower extent towards alpha-glucose-1-phosphate . The transglucosylation properties were investigated using a broad range of monomeric sugars as acceptor substrate for the recombinant enzyme, while alpha-glucose-1-phosphate served as donor . D- and L-arabinose, D- and L-arabitol, and xylitol showed the highest production of transglucosylation products . The investigated disaccharides and trisaccharides were not suitable as acceptors . The structure of the transglucosylation product obtained with D-arabinose as acceptor was elucidated by NMR . The structure of the synthesized non-reducing dimer was alpha-Glcp(1-->1)beta-Araf.

Nippon Jinzo Gakkai Shi, 2003, 45(8), 759 - 64
{Effect of oral intake of an enteric capsule preparation containing Bifidobacterium longum on the progression of chronic renal failure}; Ando Y et al.; Since the accumulation of intestinal putrefactive products, such as indole and phenol, is known to play a role in the exacerbation of chronic renal failure, reduction of these intestinal putrefactive products can be expected to retard the progression of renal failure . In the present study, an enteric capsule preparation containing Bifidobacterium longum(Bifidus HD) was administered orally to 27 patients with chronic renal failure(CRF) for 6 months . Though no significant effect was found in the whole group, a significant retardation of the progression of renal failure was found in patients with an initial serum creatinine level > or = 4.0 mg/dl or those with an initial serum inorganic phosphate level > or = 4.0 mg/dl . There was no adverse effect observed in any case . Bifidus HD is considered a useful tool for suppressing the progression of chronic renal failure(CRF) in the conservative period.

Asian Pac J Cancer Prev, 2003 Aug-Dec, 4(4), 369 - 72
Probiotics and health; Otles S et al.; Probiotics are described as live microbial food ingredients that are beneficial to health of the of host, especially by improving intestinal microbial balance . The major consumption of probiotics is in dairy-based foods form, which is containing intestinal species of Lactobacillus and Bifidobacterium . A number of potential benefits of probiotics have been proposed, including: adherence to cells; exclusion or reduction of pathogenic adherence; production of acids, hydrogen peroxide, and bacteriocins antagonistic to pathogen growth; safe, noninvasive, noncarcinogenic and nonpathogenic characteristics; and congregation to form a normal balanced flora . The interrelation between probiotics and health are reviewed in this article.

Appl Environ Microbiol, 2004 Jan, 70(1), 167 - 73
Quantitative PCR with 16S rRNA-gene-targeted species-specific primers for analysis of human intestinal bifidobacteria; Matsuki T et al.; A highly sensitive quantitative PCR detection method has been developed and applied to the distribution analysis of human intestinal bifidobacteria by combining real-time PCR with Bifidobacterium genus- and species-specific primers . Real-time PCR detection of serially diluted DNA extracted from cultured bifidobacteria was linear for cell counts ranging from 10(6) to 10 cells per PCR assay . It was also found that the method was applicable to the detection of Bifidobacterium in feces when it was present at concentrations of >10(6) cells per g of feces . Concerning the distribution of Bifidobacterium species in intestinal flora, the Bifidobacterium adolescentis group, the Bifidobacterium catenulatum group, and Bifidobacterium longum were found to be the three predominant species by examination of DNA extracted from the feces of 46 healthy adults . We also examined changes in the population and composition of Bifidobacterium species in human intestinal flora of six healthy adults over an 8-month period . The results showed that the composition of bifidobacterial flora was basically stable throughout the test period.

J Nutr, 2004 Jan, 134(1), 153 - 6
Prebiotic inulin enriched with oligofructose in combination with the probiotics Lactobacillus rhamnosus and Bifidobacterium lactis modulates intestinal immune functions in rats; Roller M et al.; Probiotics (PRO) modulate systemic immunity in animals and humans . In contrast, the effects of prebiotics (PRE) on systemic and intestinal immunity have not been investigated . Whether the combined application of PRO and PRE {synbiotics (SYN)} has synergistic or additive effects is presently unknown . Therefore, PRO (Lactobacillus rhamnosus GG and Bifidobacterium lactis Bb12), PRE (inulin enriched with oligofructose), and SYN (combination of PRO and PRE) were fed to F344 rats for 4 wk as supplements to a high fat diet . Functions of immune cells isolated from peripheral blood mononuclear cells (PBMC), spleen, mesenterial lymph nodes and Peyer's patches (PP) were investigated . The SYN supplement increased secretory immunoglobulin A (sIgA) production in the ileum compared with controls fed the high fat diet alone (P<0.05), and decreased the oxidative burst activity of blood neutrophils (P<0.05) compared with rats fed PRO . The PRE supplement enhanced the production of interleukin-10 (P<0.05) in PP as well as the production of sIgA in the cecum (P<0.05), compared with controls . The PRO supplement modestly affected immune functions, whereas systemic immunomodulatory effects were observed in rats fed SYN . The PRE supplement primarily acted at the level of the gut-associated lymphoid tissue . The combined application of PRO and PRE has different effects from those of the individual supplements, but does not simply result in additive or synergistic effects.

Med Dosw Mikrobiol, 2003, 55(3), 245 - 52
{Phenotype and genotype of Clostridium sp . strains producing botulism neurotoxin}; Augustynowicz E et al.; Neurotoxins produced by strains of Clostridium sp . are belonging to the most toxic biological substances . In the study phenotypes and genotypes of C . botulinum strains in animal studies in vivo and on the DNA level were evaluated, respectively . Additionally, the presence of genes encoding BoNT toxins of A, B, and E types among strains of Clostridium sp . were identified . In case of C . botulinum DNA was isolated from vegetative bacterial cells and from spores . Two different genes encoding two different neurotoxins harboured by three strains of Ae biotype/ae genotype, and by two strains of B biotype/be genotype were detected . Additionally, above E type C . botulinum strains, the presence of gene encoding E type neurotoxin, was found in genome of two C . baratii, two C . butyricum, and C . bifidobacterium, and C . oedematicum strains . C . bifidobacterium and C . oedematicum strains positive for presence of gene encoding E type neurotoxin, were found negative for E neurotoxin production in vivo in TN test . The study indicates that genes encoding BoNT/E neurotoxins are very common among Clostridium species . Phenotype and genotype analysis indicated co-presence of B phenotype together with be genotype and A phenotype together with ae genotype among C . botulinum strains.

J Chromatogr B Analyt Technol Biomed Life Sci, 2004 Feb 5, 800(1-2), 27 - 32
Magnetic hydrophilic methacrylate-based polymer microspheres designed for polymerase chain reactions applications; Spanova A et al.; Magnetic hydrophilic non-porous P(HEMA-co-EDMA), P(HEMA-co-GMA) and PGMA microspheres were prepared by dispersion (co)polymerization of 2-hydroxyethyl methacrylate (HEMA) and ethylene dimethacrylate (EDMA) or glycidyl methacrylate (GMA) in the presence of several kinds of magnetite . It was found that some components used in the preparation of magnetic carriers interfered with polymerase chain reaction (PCR) . Influence of non-magnetic and magnetic microspheres, including magnetite nanoparticles and various components used in their synthesis, on the PCR course was thus investigated . DNA isolated from bacterial cells of Bifidobacterium longum was used in PCR evaluation of non-interfering magnetic microspheres . The method enabled verification of the incorporation of magnetite nanoparticles in the particular methacrylate-based polymer microspheres and evaluation of suitability of their application in PCR . Preferably, electrostatically stabilized colloidal magnetite (ferrofluid) should be used in the design of new magnetic methacrylate-based microspheres by dispersion polymerization.

Microbiol Immunol, 2003, 47(12), 911 - 4
In vivo safety assessment of two Bifidobacterium longum strains; Makelainen H et al.; Bifidobacteria are important members of the intestinal microbiota and are considered to contribute to maintaining health . However, the level of bifidobacteria colonising the intestine of elderly subjects tends to be lower than in younger adults . Therefore, two Bifidobacterium longum strains, isolated from healthy elderly, were chosen for supplementation of the endogenous Bifidobacterium microbiota in the elderly . Bifidobacteria are generally regarded safe for human consumption . However, since the strains are intended for consumption by the elderly, whom may be more prone to disease, it is important to ascertain their safety . For this purpose, the strains were given to healthy adult volunteers . No side effects were reported and no undesirable changes observed in the immune parameters measured . Based on this study it appears that the two strains are well tolerated by human subjects and there are no reservations about their food use.

Eur J Biochem, 2004 Jan, 271(1), 87 - 95
Expression of mitochondrial HMGCoA synthase and glutaminase in the colonic mucosa is modulated by bacterial species; Cherbuy C et al.; The expression of the colonic mitochondrial 3-hydroxy 3-methyl glutaryl CoA (mHMGCoA) synthase, a key control site of ketogenesis from butyrate, is lower in germ-free (GF) than in conventional (CV) rats . In contrast, the activity of glutaminase is higher . The objective of this study was to investigate whether the intestinal flora can affect gene expression through short chain fatty acid (SCFA) and butyrate production . GF rats were inoculated with a conventional flora (Ino-CV) or with a bacterial strain producing butyrate (Clostridium paraputrificum, Ino-Cp) or not (Bifidobacterium breve, Ino-Bb) . In the Ino-CV rats, mHMGCoA synthase expression was restored to the CV values 2 days after the inoculation, i.e . concomitantly with SCFA production . In the Ino-Cp group, but not in the Ino-Bb group, mHMGCoA synthase and glutaminase were expressed at the level observed in the CV rats . These data suggest that the intestinal flora, through butyrate production, could control the expression of colonic mHMGCoA synthase and glutaminase . These modifications in gene expression by butyrate in vivo seem unrelated to a modification of histone acetylation.

J Pediatr (Rio J), 2000 Jul, 76(Suppl 1), S209 - 17
{Up-to-date clinical and experimental basis for the use of probiotics}; Penna FJ et al.; OBJECTIVE: To evaluate the potential of probiotics or biotherapeutic agents for the prevention and/or treatment of selected intestinal infections . METHODS: Medline database was searched for all relevant articles between 1990 and February 1998 . Bibliographies of articles were also used . All animal experiments and placebo-controlled human studies were reviewed in order to provide information on the mechanisms of action, potential efficacy, or adverse effects of these biotherapeutic agents . RESULTS: In the first part of this review, the different mechanisms of action that are effective in the treatment of diarrhea were discussed, and they were well demonstrated in laboratory animals . The most important are: enzymatic induction of disaccharidase activity, trophic effects on the intestinal mucosa, action in blocking bacterial toxins, and also induction of the immunologic response . Therapeutic effects of probiotics in humans, mainly in the gastrointestinal tract, were reported in the second part . Placebo-controlled studies have shown that biotherapeutic agents have been used successfully in the treatment of acute diarrhea in infants, traveler s diarrhea, antibiotic-associated diarrhea, with or without Clostridium difficile-associated enterocolitis (pseudomembranous colitis), and in immunosuppression-associated diarrhea, including AIDS . Lactobacillus, Bifidobacterium and Saccharomyces boulardii were the most important biotherapeutic agents to be considered . CONCLUSIONS: Currently, there is evidence that the administration of selected microorganisms is benefic in the prevention and treatment of certain intestinal infections . According to the literature, Saccharomyces boulardii is the most important probiotic . Possible future indications were discussed, such as the probable synergic effect of many probiotics due to their different and complementary mechanisms of action . The importance of new experimental and clinical studies for the better understanding of actions and the use of probiotics in other clinical situations was emphasized.

Syst Appl Microbiol, 2003 Nov, 26(4), 572 - 84
Molecular approaches for the detection and identification of bifidobacteria and lactobacilli in the human gastrointestinal tract; Satokari RM et al.; In this review an overview of various molecular techniques and their application for the detection and identification of bifidobacteria and lactobacilli in the gastrointestinal (GI) tract is presented . The techniques include molecular typing techniques such as amplified ribosomal DNA restriction analysis (ARDRA), randomly amplified polymorphic DNA (RAPD), pulsed field gel electrophoresis (PFGE), ribotyping and community profiling techniques such as PCR coupled to temperature and denaturing gradient gel electrophoresis (PCR-TGGE and PCR-DGGE, respectively) . Special attention is given to oligonucleotide probes and primers that target the ribosomal RNA (rRNA) sequences and their use in PCR and different hybridisation techniques such as DNA microarrays and fluorescent in situ hybridisation (FISH) . In addition, recent findings based on the molecular studies of bifidobacteria and lactobacilli in the GI-tract are reviewed.

Appl Environ Microbiol, 2003 Dec, 69(12), 7545 - 8
Case study of the distribution of mucosa-associated Bifidobacterium species, Lactobacillus species, and other lactic acid bacteria in the human colon; Nielsen DS et al.; The distribution of mucosa-associated bacteria, bifidobacteria and lactobacilli and closely related lactic acid bacteria, in biopsy samples from the ascending, transverse, and descending parts of the colon from four individuals was investigated by denaturing gradient gel electrophoresis (DGGE) . Bifidobacterial genus-specific, Lactobacillus group-specific, and universal bacterial primers were used in a nested PCR approach to amplify a fragment of the 16S rRNA gene . DGGE profiles of the bifidobacterial community were relatively simple, with one or two amplicons detected at most sampling sites in the colon . DGGE profiles obtained with Lactobacillus group-specific primers were complex and varied with host and sampling site in the colon . The overall bacterial community varied with host but not sampling site.

Appl Environ Microbiol, 2003 Dec, 69(12), 7517 - 22
Comparative sequence analysis of the tuf and recA genes and restriction fragment length polymorphism of the internal transcribed spacer region sequences supply additional tools for discriminating Bifidobacterium lactis from Bifidobacterium animalis; Ventura M et al.; The relationship between Bifidobacterium lactis and Bifidobacterium animalis was examined by comparative analysis of tuf and recA gene sequences and by restriction fragment length polymorphism analysis of their internal 16S-23S transcribed spacer region sequences . The bifidobacterial strains investigated could be divided into two distinct groups within a single species based on the tuf, recA, and 16S-23S spacer region sequence analysis . Therefore, all strains of B . lactis and B . animalis could be unified as the species B . animalis and divided into two subspecies, Bifidobacterium animalis subsp . lactis and Bifidobacterium animalis subsp . animalis.

Appl Environ Microbiol, 2003 Dec, 69(12), 7116 - 23
Purification and characterization of alpha-L-arabinopyranosidase and alpha-L-arabinofuranosidase from Bifidobacterium breve K-110, a human intestinal anaerobic bacterium metabolizing ginsenoside Rb2 and Rc; Shin HY et al.; Two arabinosidases, alpha-L-arabinopyranosidase (no EC number) and alpha-L-arabinofuranosidase (EC 3.2.1.55), were purified from ginsenoside-metabolizing Bifidobacterium breve K-110, which was isolated from human intestinal microflora . alpha-L-Arabinopyranosidase was purified to apparent homogeneity, using a combination of ammonium sulfate fractionation, DEAE-cellulose, butyl Toyopearl, hydroxyapatite Ultrogel, QAE-cellulose, and Sephacryl S-300 HR column chromatography, with a final specific activity of 8.81 micro mol/min/mg . alpha-L-Arabinofuranosidase was purified to apparent homogeneity, using a combination of ammonium sulfate fractionation, DEAE-cellulose, butyl Toyopearl, hydroxyapatite Ultrogel, Q-Sepharose, and Sephacryl S-300 column chromatography, with a final specific activity of 6.46 micro mol/min/mg . The molecular mass of alpha-L-arabinopyranosidase was found to be 310 kDa by gel filtration, consisting of four identical subunits (77 kDa each, measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis {SDS-PAGE}), and that of alpha-L-arabinofuranosidase was found to be 60 kDa by gel filtration and SDS-PAGE . alpha-L-Arabinopyranosidase and alpha-L-arabinofuranosidase showed optimal activity at pH 5.5 to 6.0 and 40 degrees C and pH 4.5 and 45 degrees C, respectively . Both purified enzymes were potently inhibited by Cu(2+) and p-chlormercuryphenylsulfonic acid . alpha-L-Arabinopyranosidase acted to the greatest extent on p-nitrophenyl-alpha-L-arabinopyranoside, followed by ginsenoside Rb2 . alpha-L-Arabinofuranosidase acted to the greatest extent on p-nitrophenyl-alpha-L-arabinofuranoside, followed by ginsenoside Rc . Neither enzyme acted on p-nitrophenyl-beta-galactopyranoside or p-nitrophenyl-beta-D-fucopyranoside . These findings suggest that the biochemical properties and substrate specificities of these purified enzymes are different from those of previously purified alpha-L-arabinosidases . This is the first reported purification of alpha-L-arabinopyranosidase from an anaerobic Bifidobacterium sp.

Appl Environ Microbiol, 2003 Dec, 69(12), 6994 - 7001
Investigation of protein export in Bifidobacterium breve UCC2003; MacConaill LE et al.; The molecular interactions between the bifidobacterial cell and its natural environment, namely, the gastrointestinal tract of its host, are particularly important in understanding the presumed positive effects of Bifidobacterium on the health status of the host . In this study an export-specific reporter system, designed for use in gram-positive organisms and based on the use of the staphylococcal nuclease (Nuc) as a reporter, was employed to identify exported proteins in Bifidobacterium breve UCC2003 . A B . breve genomic library of translational fusions to the Nuc-encoding gene devoid of its own export signal was established in the shuttle vector pFUN (I . Poquet, S . D . Ehrlich, and A . Gruss, J . Bacteriol . 180:1904-1912, 1998) and screened for bifidobacterial export signals . Sequence analysis of the fusion proteins obtained that displayed a nuclease-producing phenotype in both Lactococcus lactis and B . breve predicted the presence of a classical signal peptide and/or single or multiple transmembrane domains, thus indicating that some of the export signals in B . breve are comparable to those used in L . lactis . Cell fractionation studies, zymograms, nuclease assays, and Western blotting were employed to confirm the function of the predicted signals and to determine the location and activity of the exported fusion proteins in B . breve and/or L . lactis.

Eksp Klin Gastroenterol, 2003, (4), 59 - 67, 115
{Clinical significance of studies of microorganisms of the intestinal mucosa by culture biochemical methods and mass fragmentography}; Osipov GA et al.; This research is a continuation of the series of studies of the parietal microbiota of the bowel tissue samplings by the gas chromatography and mass spectrometry (GCMS) method {G.A . Ossipov et al.//Journal of the Society of Russian Gastroenterologists, 2001, 1:54-69} . The purpose was to study a number of new microorganisms in view of new data on the composition of their fatty acids (FA) and aldehydes; to confirm the presence of a number of bacteria, fungi and aerobic actinomycetes revealed earlier by FA markers in the composition of the bowel parietal microbiota by their isolation in a pure culture; to amend the estimation of the clinical value of changes in the composition of the human bowel parietal microflora in case of irritable bowel syndrome (IBS) and antibiotics-associated diarrhea (AAD) . We examined 31 patients with IBS with predominating diarrhea, 18 patients with AAD and 3 volunteers (a control group) . We studied the blood samples, tissue samplings of the mucous coat of the jejunum, ileum and colon and composition of healthy people's feces . The GCMS method was applied . Morphology of defined strains was controlled by methods of light and scanning electron microscopy . We found a substantial portion of eubacteria among the bowel microorganisms and specific changes of their species in case of IBS and AAD . Taking into account their physiological and biochemical activity, when regulating their concentration one can expect at least the same effect as when regulating the number of bifidobacteria and lactobacilli in treatment of intestinal pathologies and other diseases related to bowel dysbacteriosis . The analysis of the feces microbiota using the GCMS method by FA of parietal microorganisms provides reliable data on their number both in feces and in tissue samplings . We found a substantial portion of eubacteria among other bowel microorganisms (27% in the jejunum and 16% in the colon) and specific changes of their species in case of IBS and AAD . The concentration of streptomycetes, rhodococci and other members of the Actinomycetales order becomes dozens times more and/or reduces in pathological states . The tenfold concentration of markers of lactobacilli and bifidobacteria in some diseases stimulates the differentiated application of widespread probiotics based on these bacteria.

J Appl Microbiol, 2003, 95(5), 1058 - 69
Production of antibacterial substances by bifidobacterial isolates from infant stool active against Listeria monocytogenes; Toure R et al.; AIMS: This study aimed to characterize new isolates of human bifidobacteria, evaluate some of their probiotic potential and to screen these isolates for their effectiveness at inhibiting Listeria monocytogenes in vitro . METHODS AND RESULTS: Thirty-four Bifidobacterium isolates from infant faeces were identified by fructose-6-phosphate phosphoketolase and PCR . Six isolates, coded RBL67, RBL68, RBL69, RBL70, RBL85 and RBL86, showed higher antagonistic activity against L . monocytogenes . Neutralized culture supernatants of these strains did not inhibit L . monocytogenes when tested by agar diffusion method . However, the concentration of supernatant by speed-vac resulted in the formation of an inhibitory effect with supernatants from strains RBL67, RBL68 and RBL70 . This effect was shown to be related to heat-stable proteinaceous compound(s) which were resistant to heating at 100 degrees C for 5 min but not to pronase-E, proteinase-K or trypsin . The extraction of the inhibitory compounds by methanol-acetone extraction procedure indicated that four strains (RBL67, RBL68, RBL69 and RBL70) were mostly soluble in acetone . However, strain RBL85 produced inhibitory substances that were soluble in methanol . CONCLUSION: Infant bifidobacterial isolates produce heat-stable proteinaceous compounds active against L . monocytogenes . SIGNIFICANCE AND IMPACT OF THE STUDY: Production of antibacterial substances by bifidobacteria would improve intestinal bacterial ecology and inhibit intestinal pathogens.

Int J Food Sci Nutr, 2004 Feb, 55(1), 53 - 60
Survival of Lactobacillus acidophilus LA-5 and Bifidobacterium bifidum BB-02 in white-brined cheese; Yilmaztekin M et al.; The viability of probiotic cultures used as adjunct cultures including Bifidobacterium bifidum BB-02 and Lactobacillus acidophilus LA-5 in white-brined cheese was investigated . The microbiological and biochemical properties of cheeses were assessed throughout 90 days of storage . Two inoculum rates for probiotic starters (2.5% and 5.0%, v/v, corresponding to (1.0-1.3)x10(9) cfu/g and (2.0-2.1)x10(9) cfu/g, respectively) were compared with the control cheese . Results obtained showed that a higher inoculum rate resulted in faster proteolysis . The cheese inoculated with 5% probiotic strains had higher water-soluble nitrogen, non-protein nitrogen, proteose-peptone nitrogen and tyrosine values than the other experimental cheeses . At the end of storage, the cheese supplemented with 5% probiotic culture had a ripening coefficient of 28.3%, followed by 2.5% probiotic strains containing the sample being 24.9%, and the control being 23.6%, respectively . It was also found that the colony counts of both probiotic microorganisms declined during storage . After 90 days of storage, the number of probiotic colonies were still around the threshold for a minimum probiotic effect (10(6) cfu/g).

J Food Prot, 2003 Nov, 66(11), 2076 - 84
Protection of bifidobacteria encapsulated in polysaccharide-protein gel beads against gastric juice and bile; Guerin D et al.; Bifidobacterium cells were encapsulated in a mixed gel composed of alginate, pectin, and whey proteins . Two kinds of capsules were obtained: gel beads without membranes and gel beads with two membranes formed by the transacylation reaction . In vitro studies were carried out to determine the effects of simulated gastric pH and bile salts on the survival of free and encapsulated Bifidobacterium bifidum . The protective effects of gel beads without membranes and gel beads coated with two membranes formed by the transacylation reaction were evaluated . After 1 h in an acidic solution (pH 2.5), the free-cell counts decreased by 4.75 log units, compared with a <1-log decrease for entrapped cells . The free cells did not survive after 2 h of incubation at pH 2.5, while immobilized-cell counts decreased by about 2 log units . After incubation (1 or 3 h) in 2 and 4% bile salt solutions, the bifidobacterium mortality level for membrane-free gel beads (4 to 7 log units) was higher than that for free cells (2 to 3 log units) . However, counts of bifidobacteria immobilized in membrane-coated gel beads decreased by <2 log units . Cell encapsulation in membrane-coated protein-polysaccharide gel beads could be used to increase the survival of healthy probiotic bacteria during their transit through the gastrointestinal tract.

Appl Microbiol Biotechnol, 2004 Apr, 64(3), 396 - 402 Epub 2003 Nov 18.
Diversity of L-leucine catabolism in various microorganisms involved in dairy fermentations, and identification of the rate-controlling step in the formation of the potent flavour component 3-methylbutanal; Smit BA et al.; Various microorganisms, belonging to the genera Lactococcus, Lactobacillus, Streptococcus, Leuconostoc, Bifidobacterium, Propionibacterium, Brevibacterium, Corynebacterium and Arthrobacter, used in dairy fermentations such as cheese making, were analysed for their potential to convert leucine into flavour components, most notably 3-methylbutanal . A large variation between and within species was observed for various enzyme activities involved in the conversion pathway, e.g . transaminases, alpha-hydroxy acid dehydrogenase and alpha-keto acid decarboxylase . In particular, alpha-keto acid decarboxylase activity-leading to 3-methylbutanal-was found to be present in only two of the strains tested . It is proposed that this activity is rate-controlling in the conversion pathway leading to the flavour compound 3-methylbutanal.

Sichuan Da Xue Xue Bao Yi Xue Ban, 2003 Oct, 34(4), 622 - 4
{Bifidobacterium cell wall proteins induced beta-defensin 2 mRNA expression in human intestinal epithelial cells}; Wang G et al.; OBJECTIVE: To examine Bifidobacterium mediated induction of human defensin 2 mRNA expression in human intestinal epithelial cells and define the bioactive components of Bifidobacterium . METHODS: Bifidobacterium was cultured in MRS medium in anaerobic incubator and its cell wall was isolated by sonication and centrifugation . The cell wall proteins of Bifidobacteria were extracted with 2% SDS . The level of hBD-2 mRNA in HT-29 cells was detected by reverse-transcription PCR and Northern blot analysis . RESULTS: There was no detectable signal of hBD-2 mRNA in the unstimulated HT-29 cells . However, the heat-killed Bifidobacteria, cell wall of Bifidobacteria, and cell wall proteins remarkably induced hBD-2 mRNA expression in HT-29 . CONCLUSION: Bifidobacterium or its cell wall proteins can induce hBD-2 gene expression in human intestinal epithelial cells, which may play a role in the mechanisms of innate defense against pathogens in the intestine.

J Invertebr Pathol, 2003 Oct, 84(2), 96 - 103
Bacterial diversity in worker adults of Apis mellifera capensis and Apis mellifera scutellata (Insecta: Hymenoptera) assessed using 16S rRNA sequences; Jeyaprakash A et al.; High-fidelity PCR of 16S rRNA sequences was used to identify bacteria associated with worker adults of the honeybee subspecies Apis mellifera capensis and Apis mellifera scutellata . An expected approximately 1.5-kb DNA band, representing almost the entire length of the 16S rRNA gene, was amplified from both subspecies and cloned . Ten unique sequences were obtained: one sequence each clustered with Bifidobacterium (Gram-positive eubacteria), Lactobacillus (Gram-positive eubacteria), and Gluconacetobacter (Gram-negative alpha-proteobacteria); two sequences each clustered with Simonsiella (beta-proteobacteria) and Serratia (gamma-proteobacteria); and three sequences each clustered with Bartonella (alpha-proteobacteria) . Although the sequences relating to these six bacterial genera initially were obtained from either A . m . capensis or A . m . scutellata or both, newly designed honeybee-specific 16S rRNA primers subsequently amplified all sequences from all individual workers of both subspecies . Attempts to amplify these sequences from eggs have failed . However, the wsp primers designed to amplify Wolbachia DNA from arthropods, including these bees, consistently produced a 0.6-kb DNA band from individual eggs, indicating that amplifiable bacterial DNA was present . Hence, the 10 bacteria could have been acquired orally from workers or from other substrates . This screening of 16S rRNA sequences from A . m . capensis and A . m . scutellata found sequences related to Lactobacillus and Bifidobacterium which previously had been identified from other honeybee subspecies, as well as sequences related to Bartonella, Gluconacetobacter, Simonsiella/Neisseria, and Serratia, which have not been identified previously from honeybees.

Appl Environ Microbiol, 2003 Nov, 69(11), 6908 - 22
Analysis, characterization, and loci of the tuf genes in lactobacillus and bifidobacterium species and their direct application for species identification; Ventura M et al.; We analyzed the tuf gene, encoding elongation factor Tu, from 33 strains representing 17 Lactobacillus species and 8 Bifidobacterium species . The tuf sequences were aligned and used to infer phylogenesis among species of lactobacilli and bifidobacteria . We demonstrated that the synonymous substitution affecting this gene renders elongation factor Tu a reliable molecular clock for investigating evolutionary distances of lactobacilli and bifidobacteria . In fact, the phylogeny generated by these tuf sequences is consistent with that derived from 16S rRNA analysis . The investigation of a multiple alignment of tuf sequences revealed regions conserved among strains belonging to the same species but distinct from those of other species . PCR primers complementary to these regions allowed species-specific identification of closely related species, such as Lactobacillus casei group members . These tuf gene-based assays developed in this study provide an alternative to present methods for the identification for lactic acid bacterial species . Since a variable number of tuf genes have been described for bacteria, the presence of multiple genes was examined . Southern analysis revealed one tuf gene in the genomes of lactobacilli and bifidobacteria, but the tuf gene was arranged differently in the genomes of these two taxa . Our results revealed that the tuf gene in bifidobacteria is flanked by the same gene constellation as the str operon, as originally reported for Escherichia coli . In contrast, bioinformatic and transcriptional analyses of the DNA region flanking the tuf gene in four Lactobacillus species indicated the same four-gene unit and suggested a novel tuf operon specific for the genus Lactobacillus.

Acta Paediatr Suppl, 2003 Sep, 91(441), 68 - 76
Prebiotics in infant milk formulas: new perspectives; Miniello VL et al.; In recent years it has become accepted that healthy human intestinal microflora may play an important part in priming the infants' systemic and mucosal immunity . Dietary modulation of the gut microbiota is a topical area of nutritional sciences and the main focus of many current functional foods such as non-digestible oligosaccharides (NDOs) . Fructo-oligosaccharides (FOS) and trans-beta-galacto-oligosaccharides (TOS) have been claimed to benefit the health of the colon by selectively stimulating the growth of bifidobacteria and lactobacilli (prebiotic effect) . It could be of clinical interest to manipulate colonic flora because it is supposed that specific bacteria in the gut microbial microflora could promote potentially antiallergenic processes and play a key part in atopic disease prevention . Supporting this view is the finding that analysis of the composition of the intestinal bacterial populations showed different microbial patterns between healthy and allergic individuals . Assuming that non-digestible TOS and FOS can affect the intestinal ecosystem beneficially, the opportunity for gut flora manipulation arises in bottle-fed infants . New preterm and term infant milk formulas, supplemented with a mixture of TOS and FOS as prebiotic ingredients induced a significantly higher colonization of bifidobacteria and lactobacilli . In the future, selective manipulation of the intestinal microbiota might be an approach to novel prophylactic and therapeutic intervention strategies of atopy, by redirecting allergic Th-2 responses in favour of Th-1 responses.

Acta Paediatr Suppl, 2003 Sep, 91(441), 64 - 7
Prebiotic concept for infant nutrition; Boehm G et al.; In the neonatal period, the intestine is colonised in a stepwise process that depends on mode of delivery, environmental factors, bacterial interactions, and the host itself resulting in a colonisation with a complex heterogeneous bacterial flora . Oligosaccharides have been identified as an important prebiotic factor of human milk As long as analogues of human milk oligosaccharides are not available now and in the near future it is aimed to resemble the prebiotic effect of human milk by oligosaccharides from available sources . In the present study in preterm infants, a mixture of 90% galacto-oligosaccharides and 10% fructo-oligosaccharides has been tested . The mixture of GOS/FOS was composed to mimic the molecule size distribution of human milk oligosaccharides . Microbiological analysis of the faces was performed before and 7, 14, and 28 days after start of supplementation and stool characteristics have been recorded . Maltodextrin was used as placebo and infants fed human milk have been used as reference . After a 28 days feeding period, the number of bifidobacteria of the group fed the oligosaccharide supplemented formula was in the upper range of the reference group whereas the numbers of the group fed the formula supplemented with the placebo were in the lower range of the reference group (placebo: 7.9 +/- 0.83 and GOS/FOS mixture: 10,0 +/- 2.05 log 10 CFU/g wet stool; reference (M +/- SD): 7.14-10.7 log 10 CFU/g wet stool) . Stool characteristics in the group fed the supplemented formula were close to those found in the human milk fed infants . In summary, supplementation of a preterm formula with a mixture of galacto- and fructo-oligosaccharides has a stimulating effect on the growth of bifidobacteria in the intestine and results in more frequent produced and softer stools . Thus, prebiotic mixtures such like the studied oligosaccharide mixture might help in improving intestinal tolerance to enteral feeding in preterm infants.

Acta Paediatr Suppl, 2003 Sep, 91(441), 56 - 63
Advances in the modulation of the microbial ecology of the gut in early infancy; Chierici R et al.; It is now generally accepted that the microbiota of the human gut may influence health and well-being . Lactic acid bacteria are the most important microorganisms associated with these beneficial effects and the elevated bifidobacterial count may be one of the greatest advantages that breastfed infants have over infants fed with milk formulas . Several studies relative to the selective growth stimulation of bifidobacteria, both in vitro and in vivo, are reported in this review . Over the years, diverse human milk components have been identified as the specific factors able to modulate the growth of bifidobacteria . Even if there is a certain agreement that the bifidogenic activity of human milk may be based not on single growth substances, but on a complex set of interacting factors, the present state of knowledge indicates that the use of non-digestible but fermentable carbohydrates may be an easy and reliable method to influence the growth of lactic acid bacteria . In this context, some of the characteristics of the major physiological effects of inulin-type fructans, of galacto-oligosaccharides, but also of lactoferrin, a milk whey protein fraction with purported bifidogenic activity, are briefly examined.

Antimicrob Agents Chemother, 2003 Nov, 47(11), 3667 - 71
In vitro activities of a new des-fluoro(6) quinolone, garenoxacin, against clinical anaerobic bacteria; Liebetrau A et al.; The antimicrobial activities of garenoxacin and eight other antibiotics against 641 anaerobic isolates were evaluated with the NCCLS agar dilution method . Overall, the MICs of garenoxacin for 50 and 90% of the strains tested (in micrograms per milliliter) were as follows: Bacteroides fragilis group, 0.5 and 2; Prevotella spp., 0.25 and 2; Fusobacterium spp., 0.25 and 0.5; Porphyromonas spp., 0.125 and 0.25; Bilophila wadsworthia, 0.5 and 1; Veillonella spp., 0.25 and 0.5; Clostridium spp., 0.25 and 1; Clostridium difficile, 2 and >64; Bifidobacterium spp., 1 and 2; Eggerthella lenta, 0.25 and 1; Propionibacterium spp., 0.5 and 0.5; gram-positive cocci, 0.125 and 0.25.

J Nutr Biochem, 2003 Oct, 14(10), 598 - 605
Inulin stimulates NO synthesis via activation of PKC-alpha and protein tyrosine kinase, resulting in the activation of NF-kappaB by IFN-gamma-primed RAW 264.7 cells; Koo HN et al.; Inulin, an active component of Chicorium intybus root, has been shown to stimulate the growth of bifidobacteria, and inhibit colon carcinogenesis . NO mediates a number of the host-defense functions of activated macrophages, including antimicrobial and tumoricidal activity . We examined the effect of inulin on the synthesis of NO in RAW 264.7 cells . Inulin alone had no effect, whereas inulin with IFN-gamma synergistically increased the NO production and inducible NO synthase (iNOS) expression in RAW 264.7 cells . Synergy between IFN-gamma and inulin was mainly dependent on inulin-induced TNF-alpha secretion . Also, protein kinase C (PKC)-alpha was involved in the inulin-induced NO production . Inulin-mediated NO production was inhibited by the protein tyrosine kinase (PTK) inhibitor, tyrphostin AG126 . Since iNOS gene transcriptions have been shown to be under the control of the NF-kappaB/Rel family of transcription factors, we assessed the effect of inulin on NF-kappaB/Rel using an EMSA . Inulin produced strong induction of NF-kappaB/Rel binding, whereas AP-1 binding was slightly induced in RAW 264.7 cells . Inulin stimulated phosphorylation and degradation of IkappaB-alpha . These results suggest that in IFN-gamma-primed RAW 264.7 cells inulin might stimulate NO synthesis via activation of PKC-alpha and PTK, resulting in the activation of NF-kappaB.

Int J Antimicrob Agents, 2003 Oct, 22(4), 360 - 6
The role of probiotics in the treatment and prevention of Helicobacter pylori infection; Hamilton-Miller JM; Literature reporting activity of probiotics in infections due to Helicobacter pylori has been reviewed to assess their value in combating such infections . Several in vitro studies show that lactobacilli or their cell-free cultures inhibit or kill H . pylori, prevent its adhesion to mammalian epithelial cells and prevent IL8 release . In vivo models demonstrate that pre-treatment with a probiotic can prevent H . pylori infections and/or that administration of probiotics markedly reduced an existing infection . Thirteen clinical trials have been published . In six (180 patients), a probiotic was used alone; five of these had an encouraging result-in three there were significantly reduced breath test readings and in two others some patients were cleared of infection . In seven further trials (682 patients), probiotics were added to a therapeutic regimen of antibiotics, resulting in an increased cure rate in two studies, and reduced side-effects in four . Trials in which fermented milk products or whole cultures of lactobacilli were used tended to show better results than when the probiotic was taken in the form of bacteria alone . Not all the studies were randomised, double-blind and placebo controlled, and some involved only small numbers of patients . The results suggest that probiotics may have a place as adjunctive treatment in H . pylori infections and possibly in prophylaxis . Future trials should address in particular the type of patient (asymptomatic volunteers, symptomatic patients), choice of probiotic strain(s), a wide range of probiotic strains (Lactobacillus acidophilus, L . johnsonii, L . gasseri, lactobacillus GG, Bifidobacterium longum, and bioyoghurts) have been used-some non-viable, regimens (doses and duration) and criteria of success (breath test, histology, culture, serology).

Biotechnol Lett, 2003 Aug, 25(15), 1211 - 7
Cloning and expression of sucrose phosphorylase gene from Bifidobacterium longum in E . coli and characterization of the recombinant enzyme; Kim M et al.; A DNA fragment, which complemented the growth of E . coli both on M9 medium containing raffinose and on LB medium containing ampicillin, IPTG and 5-bromo-4-chloro-3-indoxyl-alpha-D-galactoside, was isolated from the genomic library of Bifidobacterium longum SJ32, which had been digested with EcoRI . In the cloned DNA fragment, a gene encoding a sucrose phosphorylase (splP) and a partially cloned putative sucrose regulator gene (splR) were identified using the deletion analysis and sequence analysis . A 56 kDa protein was synthesized in E . coli and partially purified by DEAE-ion exchange chromatography . The partially purified enzyme did not react with melibiose, melezitoze and raffinose but did with sucrose . It had transglucosylation activity in addition to hydrolytic activity.

Best Pract Res Clin Gastroenterol, 2003 Oct, 17(5), 775 - 83
Probiotics, antibiotic-associated diarrhoea and Clostridium difficile diarrhoea in humans; Surawicz CM; Probiotics are living organisms which, when ingested, have a beneficial therapeutic effect . Examples are bacteria, especially Lactobacillus rhamnosus GG, and the yeast Saccharomyces boulardii . Controlled trials indicate a benefit of both of these in the prevention of antibiotic-associated diarrhoea . Other less effective probiotics are Lactinex, Enterococcus faecium and bifidobacteria . In the difficult clinical problem of recurrent Clostridium difficile disease, S . boulardii as an adjunct to antibiotics has shown benefit in controlled trials . There is, however, less convincing evidence for the efficacy of Lactobacillus GG in this disease . Additional controlled trials and safety studies are needed before there can be a widespread endorsement of probiotics for these two conditions.

Best Pract Res Clin Gastroenterol, 2003 Oct, 17(5), 741 - 54
Adhesion of probiotic strains to the intestinal mucosa and interaction with pathogens; Servin AL et al.; Probiotic lactic acid strains are live micro-organisms that, when consumed in adequate amounts as part of food, confer a health benefit on the host . The scientific basis for the use of selected probiotic strains has only recently been firmly established, and appropriate and well-conducted experimental in vitro and in vivo studies, as well as clinical studies, are now beginning to be published, especially with regard to the effectiveness of probiotic strains in antagonizing pathogens . In particular, experimental data have allowed new insights into selected probiotic strains that express strain-specific probiotic properties and into the mechanism of action of these strains . The objective of this review is to analyse the in vitro or in vivo experimental studies in which the antimicrobial activity of selected Lactobacillus and Bifidobacterium strains has been documented.

Curr Issues Intest Microbiol, 2003 Sep, 4(2), 71 - 5
Carbohydrate preferences of Bifidobacterium species isolated from the human gut; Palframan RJ et al.; The growth of nine species of Bifidobacterium on media containing glucose, xylose, xylooligosaccharides (XOS), xylan or fructooligosaccharides (FOS) as the sole carbon source were compared in pure culture . The bifidobacteria differed in fermentation profiles when tested on different carbohydrates . All species grew to their highest final optical density (OD) on a glucose containing medium, with the exception of B . catenulatum which demonstrated a preference for xylose over glucose, and XOS over FOS . B . bifidum grew to the highest OD on XOS compared to xylose suggesting a specific transport system for the oligosaccharide over the monomer . This is consistent with a lack of beta-xylosidase activity present in the culture medium . Lactate, formate and acetate levels were determined and the ratios of these metabolites altered between and within species growing on different carbohydrates . In general, high lactate production correlated with low formate production and low lactate concentrations were obtained at higher levels of formate . Bifidobacteria may alter their metabolic pathways based upon the carbohydrates that are available for their use.

Curr Issues Intest Microbiol, 2003 Sep, 4(2), 53 - 60
Intestinal microflora and homeostasis of the mucosal immune response: implications for probiotic bacteria?
Blum S, Schiffrin EJ.
The intestinal microflora can be considered a postnatally acquired organ that is composed of a large diversity of bacteria that perform important functions for the host and can be modulated by environmental factors, such as nutrition . Specific components of the intestinal microflora, including lactobacilli and bifidobacteria, have been associated with beneficial effects on the host, such as promotion of gut maturation and integrity, antagonisms against pathogens and immune modulation . Beyond this, the microflora seems to play a significant role in the maintenance of intestinal immune homeostasis and prevention of inflammation . The contribution of the intestinal epithelial cell in the first line of defense against pathogenic bacteria and microbial antigens has been recognized . However, the interactions of intestinal epithelial cells with indigenous bacteria are less well understood . This review will summarize the increasing scientific attention to mechanisms of the innate immune response of the host towards different components of the microflora, and suggest a potential role for selected probiotic bacteria in the regulation of intestinal inflammation.

Appl Microbiol Biotechnol, 2004 Mar, 64(1), 106 - 11 Epub 2003 Sep 06.
In vitro evaluation of the fermentation properties of galactooligosaccharides synthesised by alpha-galactosidase from Lactobacillus reuteri; Tzortzis G et al.; Stirred, pH-controlled anaerobic batch cultures were used to evaluate the in vitro utilisation by canine gut microflora of novel alpha-galactooligosaccharides synthesised with an enzyme extract from a canine Lactobacillus reuteri strain . Fructooligosaccharides (FOS), melibiose and raffinose were used as reference carbohydrates for the prebiotic properties of the synthesised oligosaccharide (galactosyl melibiose mixture-GMM) . Addition of Lactobacillus acidophilus was used as control for the evaluation of the synbiotic properties of the oligosaccharide with L . reuteri . Populations of predominant gut bacterial groups were monitored over 48 h of batch culture by fluorescent in situ hybridisation, and short-chain fatty acid (SCFA) production was measured . GMM showed a higher increase in bifidobacteria and lactobacilli population number and size as well as a higher decrease in clostridia population number and size compared to the commercial prebiotics (FOS, melibiose, raffinose) . This prebiotic effect was further increased by the addition of L . reuteri followed by a change in the SCFA production pattern compared to GMM alone or GMM with L . acidophilus . The observed change in SCFA production was in accordance with the fermentation properties of L . reuteri, suggesting that the novel synbiotic had a significant effect on the canine gut microflora fermentation.

FEMS Immunol Med Microbiol, 2003 Sep 22, 38(2), 165 - 72
Immunomodulatory effects of probiotic bacteria DNA: IL-1 and IL-10 response in human peripheral blood mononuclear cells; Lammers KM et al.; A new therapeutic approach for inflammatory bowel diseases is based on the administration of probiotic bacteria . Prokaryotic DNA contains unmethylated CpG motifs which can activate immune responses, but it is unknown whether bacterial DNA is involved in the beneficial effects obtained by probiotic treatment . Peripheral blood mononuclear cells (PBMC) from healthy donors were incubated with pure DNA of eight probiotic strains and with total bacterial DNA from human feces collected before and after probiotic ingestion . Cytokine production was analyzed in culture supernatants . Modification of human microflora after probiotic administration was proven by polymerase chain reaction analysis . Here we show that Bifidobacterium genomic DNA induced secretion of the antiinflammatory interleukin-10 by PBMC . Total bacterial DNA from feces collected after probiotic administration modulated the immune response by a decrease of interleukin-1 beta and an increase of interleukin-10.

Ann Rheum Dis, 2003 Oct, 62(10), 976 - 82
Role of peptidoglycan subtypes in the pathogenesis of bacterial cell wall arthritis; Simelyte E et al.; BACKGROUND: Bacterial cell wall (CW) arthritis develops in susceptible strains of rats after a single intraperitoneal injection of the CW from certain bacterial species, both pathogenic and non-pathogenic . For the development of chronic bacterial CW arthritis, the structure of the bacterial peptidoglycan (PG) has been found to be decisive . OBJECTIVE: To define the role of PG subtypes in the pathogenesis of chronic bacterial CW arthritis . METHOD: Arthritis was induced with CWs of Lactobacillus plantarum, L casei B, L casei C, and L fermentum . Gas chromatography-mass spectrometry was used to measure the presence of CW derived muramic acid in the liver and to determine PG subtypes . CWs were also tested for their resistance to lysozyme in vitro . RESULTS: These results and those published previously indicate that PGs of CWs which induce chronic arthritis, no matter whether they were derived from strains of Streptococcus, Bifidobacterium, Collinsella, or Lactobacillus, all have lysine as the third amino acid of the PG stem peptide, representing PG subtypes A3alpha and A4alpha . Those strains which induce only transient acute arthritis or no arthritis at all do not have lysine in this position, resulting in different PG subtypes . CONCLUSIONS: In vivo degradation of only those PGs with the subtypes A3alpha and A4alpha leads to the occurrence of large CW fragments, which persist in tissue and have good proinflammatory ability . CWs with other PG subtypes, even if they are lysozyme resistant, do not cause chronic arthritis, because the released fragments are not phlogistic . It is emphasised that a variety of microbial components not causing inflammation have been found in animal and human synovial tissue.

Lett Appl Microbiol, 2003, 37(4), 281 - 4
Development of a quantitative tool for the comparison of the prebiotic effect of dietary oligosaccharides; Palframan R et al.; AIMS: To develop a quantitative equation {prebiotic index (PI)} to aid the analysis of prebiotic fermentation of commercially available and novel prebiotic carbohydrates in vitro, using previously published fermentation data . METHODS: The PI equation is based on the changes in key bacterial groups during fermentation . The bacterial groups incorporated into this PI equation were bifidobacteria, lactobacilli, clostridia and bacteroides . The changes in these bacterial groups from previous studies were entered into the PI equation in order to determine a quantitative PI score . PI scores were than compared with the qualitative conclusions made in these publications . In general the PI scores agreed with the qualitative conclusions drawn and provided a quantitative measure . CONCLUSIONS: The PI allows the magnitude of prebiotic effects to be quantified rather than evaluations being solely qualitative . SIGNIFICANCE AND IMPACT OF THE STUDY: The PI equation may be of great use in quantifying prebiotic effects in vitro . It is expected that this will facilitate more rational food product development and the development of more potent prebiotics with activity at lower doses.

Antibiot Khimioter, 2003, 48(5), 11 - 4
{Effect of medicinal plants on the antilysozyme activity of microorganisms}; Bukharin OV et al.; Medicinal plants effect on antilysozyme activity of E . coli, Klebsiella, S . aureus, Candida, Bifidobacterium, Bacteroides, Clostridium was evaluated in vitro . It was demonstrated that the effects differed substancially according to the microorganism taxonomy . The experimental data on the plants inhibitory activity may be used for elaboration of effective regimes of infections and disbiosis treatment.

Zh Mikrobiol Epidemiol Immunobiol, 2003 Jul-Aug, (4), 85 - 8
{Evaluation of the effect of medicines on biological properties of Clostridium difficile}; Valyshev AV et al.; The presence of the persistence factors (anti-lysozyme and anti-complement activity) in the vegetative forms of C . difficile was experimentally proved . The effect of different medicines (vitamins B1, B6 and C, prebiotic inulin, probiotics Bifidumbacterin and Enterol) on the persistence factors of C . difficile and microbial resistance to vancomycin, thienam, lincomycin, clindamycin was evaluated . The anti-lysozyme and anti-complement activity of C . difficile was found to decrease under the influence of vitamins B1, B6, C, inulin, exometabolites of bifidobacteria . Under the impact of the preparations used in this study changes in the sensitivity of C . difficile to antibiotics of the lincoamide, carbapenem, glycopeptide groups were found to occur . The data obtained reveal one of the possible mechanisms of the corrective action of the medicines under study on the intestinal microbiocenosis in patients with antibiotic-associated colitis.

Biotechnol Bioeng, 2003 Oct 20, 84(2), 145 - 50
Stability of recombinant plasmids on the continuous culture of Bifidobacterium animalis ATCC 27536; Gonzalez Vara A et al.; Bifidobacterium animalis ATCC 27536 represents among bifidobacteria a host-model for cloning experiments . The segregational and structural stabilities of a family of cloning vectors with different molecular weights but sharing a common core were studied in continuous fermentation of the hosting B . animalis without selective pressure . The rate of plasmid loss (R) and the specific growth rate difference (delta mu) between plasmid-free and plasmid-carrying cells were calculated for each plasmid and their relationship with plasmid size was studied . It was observed that both R and the numerical value of delta mu increased exponentially with plasmid size . The exponential functions correlating the specific growth rate difference and the rate of plasmid loss with the plasmid molecular weight were determined . Furthermore, the smallest of the plasmids studied, pLAV (4.3-kb) was thoroughly characterized by means of its complete nucleotide sequence . It was found that it contained an extra DNA fragment, the first bifidobacterial insertion sequence characterised, named IS 1999 .

Clin Diagn Lab Immunol, 2003 Sep, 10(5), 787 - 92
Effect of probiotic bacteria on induction and maintenance of oral tolerance to beta-lactoglobulin in gnotobiotic mice; Prioult G et al.; In this study, the effect of Lactobacillus paracasei (NCC 2461), Lactobacillus johnsonii (NCC 533) and Bifidobacterium lactis Bb12 (NCC 362) on the induction and maintenance of oral tolerance to bovine beta-lactoglobulin (BLG) was investigated in mice . Germfree mice were monocolonized with one of the three strains before oral administration of whey protein to induce tolerance . Mice were then injected with BLG and sacrificed 28 or 50 days after whey protein feeding for humoral and cellular response measurement . Conventional and germfree mice were used as controls . Both humoral and cellular responses were better suppressed in conventional mice than in germfree and monoassociated mice throughout the experiment and better suppressed in L . paracasei-associated mice than in mice colonized with B . lactis or L . johnsonii . The latter two mono-associations suppressed humoral responses only partially and cellular responses not at all . This study provides evidence that probiotics modulate the oral tolerance response to BLG in mice . The mono-colonization effect is strain-dependant, the best result having been obtained with L . paracasei.

J Pediatr Gastroenterol Nutr, 2003 Sep, 37(3), 273 - 80
rRNA probes used to quantify the effects of glycomacropeptide and alpha-lactalbumin supplementation on the predominant groups of intestinal bacteria of infant rhesus monkeys challenged with enteropathogenic Escherichia coli; Bruck WM et al.; OBJECTIVES: Certain milk factors may help to promote the growth of a host-friendly colonic microflora (e.g . bifidobacteria, lactobacilli) and explain why breast-fed infants experience fewer and milder intestinal infections than those who are formula-fed . The effects of supplementation of formula with two such milk factors was investigated in this study . MATERIALS AND METHODS: Infant rhesus macaques were breast-fed, fed control formula, or formula supplemented with glycomacropeptide (GMP) or alpha-lactalbumin (alpha-LA) from birth to 5 months of age . Blood was drawn monthly and rectal swabs were collected weekly . At 4.5 months of age, 10(8) colony-forming units of enteropathogenic E.coli O127, strain 2349/68 (EPEC) was given orally and the response to infection assessed . The bacteriology of rectal swabs pre- and post-infection was determined by culture independent fluorescence in situ hybridization . RESULTS: Post-challenge, breast-fed infants and infants fed alpha-LA-supplemented formula had no diarrhea, whilst those infants fed GMP-supplemented formula had intermittent diarrhea . In infants fed control formula the diarrhea was acute . CONCLUSIONS: Supplementation of infant formula with appropriate milk proteins may be useful for improving the infant's ability to resist acute infection caused by E.coli.

Appl Environ Microbiol, 2003 Sep, 69(9), 5731 - 5
Selective plating underestimates abundance and shows differential recovery of bifidobacterial species from human feces; Apajalahti JH et al.; The aim of the present work was to compare the efficacies and levels of selectivity of different culture-dependent and -independent methods for analyzing bifidobacteria in human stool samples . The three different culture media used here significantly differed from each other, particularly with regard to the recovery of Bifidobacterium adolescentis . Bifidobacterium medium failed to recover B . adolescentis; Beerens medium recovered some B . adolescentis organisms (17% of total bifidobacteria), whereas tomato-Eugon medium recovered mainly B . adolescentis organisms (58% of total bifidobacteria) . A culture-independent method that combines GC fractionation of bacterial community DNA and 16S rRNA sequencing indicated that B . adolescentis organisms accounted for 85% of all bifidobacteria . Methodological biases, such as those described in this paper, should be taken into account in interpreting earlier studies and designing future experiments.

Appl Environ Microbiol, 2003 Sep, 69(9), 5096 - 103
Purification and functional characterization of a novel alpha-L-arabinofuranosidase from Bifidobacterium longum B667; Margolles A et al.; The gene encoding a novel alpha-L-arabinofuranosidase from Bifidobacterium longum B667, abfB, was cloned and sequenced . The deduced protein had a molecular mass of about 61 kDa, and analysis of its amino acid sequence revealed significant homology and conservation of different catalytic residues with alpha-L-arabinofuranosidases belonging to family 51 of the glycoside hydrolases . Regions flanking the gene comprised two divergently transcribed open reading frames coding for hypothetical proteins involved in sugar metabolism . A histidine tag was introduced at the C terminus of AbfB, and the recombinant protein was overexpressed in Lactococcus lactis under control of the tightly regulated, nisin-inducible nisA promoter . The enzyme was purified by nickel affinity chromatography . The molecular mass of the native protein, as determined by gel filtration, was about 260 kDa, suggesting a homotetrameric structure . AbfB was active at a broad pH range (pH 4.5 to 7.5) and at a broad temperature range (20 to 70 degrees C), and it had an optimum pH of 6.0 and an optimum temperature of 45 degrees C . The enzyme seemed to be less thermostable than most previously described arabinofuranosidases and had a half-life of about 3 h at 55 degrees C . Chelating and reducing agents did not have any effect on its activity, but the presence of Cu(2+), Hg(2+), and Zn(2+) markedly reduced enzymatic activity . The protein exhibited a high level of activity with p-nitrophenyl alpha-L-arabinofuranoside, with apparent K(m) and V(max) values of 0.295 mM and 417 U/mg, respectively . AbfB released L-arabinose from arabinan, arabinoxylan, arabinobiose, arabinotriose, arabinotetraose, and arabinopentaose . No endoarabinanase activity was detected . These findings suggest that AbfB is an exo-acting enzyme and may play a role, together with other glycosidases, in the degradation of L-arabinose-containing polysaccharides.

Poult Sci, 2003 Aug, 82(8), 1242 - 9
Composition of microbiota in content and mucus from cecae of broiler chickens as measured by fluorescent in situ hybridization with group-specific, 16S rRNA-targeted oligonucleotide probes; Zhu XY et al.; Six group-specific 16S rRNA-targeted oligonucleotide probes were used to investigate the composition of the microbiota of cecal content and mucus from broiler chickens . Together, the probes hybridized to as many as 94.7% of the bacteria detectable with the universal probe Bact338 in the content of the cecum of 2-d-old chicks . Fewer bacteria gave signals with these probes as the birds aged, and coverage was as low as 76% for the bacteria in cecal content of a 6-wk-old chicken . In the cecal content of 2-d-old chicks, approximately 56, 34, and 3% of the bacteria detectable with the universal probe reacted with the probes Enter1432 (enterics), Lacto722 (Lactobacillus/Streptococcus/Enterococcus), and Bif164 (bifidobacteria), respectively . Probes Clept1240 (Clostridium leptum subgroup), Erec482 (Clostridium coccoides-Eubacterium rectale), and Bacto1080 (Bacteroides groups) did not produce signals . In cecal content from 1-wk-old chicks, all six probes gave signals, and in samples from 6-wk-old birds approximately 3, 9, 6, 32, 22, and 8% of the bacteria detectable with the universal probe hybridized with the probes Enter1432, Lacto722, Bif164, Clept1240, Erec482, and Bacto1080, respectively . At this age, the six probes detected the phylogenetic groups in similar proportions in the microbiota of cecal content and cecal mucus . The exception was the enterics probe because more bacteria from the mucus fraction than from cecal content gave signals with this probe (13.4 vs . 4.4%, P<0.001).

J Dairy Sci, 2003 Aug, 86(8), 2537 - 46
Metabolic and biochemical responses of probiotic bacteria to oxygen; Talwalkar A et al.; The interaction between oxygen and probiotic bacteria was studied by growing Lactobacillus acidophilus and Bifidobacterium spp . in 0, 5, 10, 15, and 21% oxygen in a hypoxic glove box . The metabolic responses of each probiotic strain in the different oxygen environments were monitored by measuring the levels of lactic acid and determining the lactate-to-acetate ratio . Biochemical changes induced by oxygen were examined by monitoring the specific activities of NADH oxidase, NADH peroxidase, and superoxide dismutase . In addition, the ability to decompose hydrogen peroxide and the sensitivity of each strain to hydrogen peroxide was also determined . With an increase in oxygen percentage, levels of lactic acid in L . acidophilus strains decreased, whereas the lactate-to-acetate ratio reduced in all the bifidobacteria tested . At 21% oxygen, the specific activities of NADH oxidase and NADH peroxidase, and the hydrogen peroxide decomposing ability of five probiotic strains was significantly higher than at 0% oxygen . The sensitivity of the probiotic strains to hydrogen peroxide however, remained unaffected in all the different oxygen percentages . Superoxide dismutase levels did not reveal any conclusive trend . In both L . acidophilus and Bifidobacterium spp., NADH oxidase and NADH peroxidase functioned optimally at pH 5 . Growth in the various oxygen environments did not change this optimum pH.

Int J Food Microbiol, 2003 Oct 15, 87(1-2), 17 - 27
Growth and metabolism of selected strains of probiotic bacteria in milk; Ostlie HM et al.; Growth and metabolism of five probiotic strains with well-documented health effects were studied in ultra-high temperature (UHT) treated milk, supplemented with 0.5% (w/v) tryptone or 0.75% (w/v) fructose . The probiotic strains were Lactobacillus acidophilus La5, Lb . acidophilus 1748, Lactobacillus rhamnosus GG, Lactobacillus reuteri SD 2112 and Bifidobacterium animalis BB12 . Fermentation was followed for 72 h at 37 degrees C and the samples were analysed for pH, log cfu ml(-1), volatile compounds, organic acids and carbon dioxide . The strains reduced pH from 6.7 to between 3.9 and 4.4 after 24 h of incubation . All strains attained viable cell counts above 8.7-9.18 log cfu ml(-1) after 6-16 h of incubation . The two Lb . acidophilus strains showed a stable level of viable cells during 12-72 h of incubation but the three other strains showed a reduction of 0.4-1.1 log cfu ml(-1) from 24 to 72 h of incubation . However, all strains showed cell levels between 7.8 and 8.7 log cfu ml(-1) after 72 h of incubation . After 48 h of incubation, the amount of lactic acid produced varied according to strain from 6949 to 14,000 mg kg(-1) and acetic acid produced varied from 0 to 6901 mg kg(-1) . Three of the strains metabolised citrate but only low amounts of diacetyl and acetoin were detected within strains, 0.2-0.8 and 6.5-10 mg kg(-1), respectively . Carbon dioxide produced varied from 221 to 3942 mg kg(-1) and was connected to the citrate-fermenting ability of the strain used and their carbohydrate fermentation pathway . Three of the strains produced detectable levels of acetaldehyde and the concentration varied from 9.4 to 12.6 mg kg(-1) after 24 h of incubation . All five probiotic strains showed very different profiles of metabolites during fermentation; however, the two Lb . acidophilus strains were the most alike . Our findings show the importance of controlling the fermentation time since the probiotic strains produced different amounts of metabolic products according to fermentation time.

Drug Discov Today, 2003 Aug 1, 8(15), 692 - 700
Using probiotics and prebiotics to improve gut health; Tuohy KM et al.; Recent molecular-based investigations have confirmed the species diversity and metabolic complexity of the human gut microbiota . It is also increasingly clear that the human gut microbiota plays a crucial role in host health, both as a source of infection and environmental insult and, conversely, in protection against disease and maintenance of gut function . Although little is known about the health impact of the dominant groups of gut bacteria it is generally accepted that bifidobacteria and lactobacilli are important components of what might be termed the beneficial gut microbiota . The microbiota management tools of probiotics, prebiotics and synbiotics have been developed and, indeed, commercialized over the past few decades with the expressed purpose of increasing numbers of bifidobacteria and/or lactobacilli within the gastrointestinal tract.

J Agric Food Chem, 2003 Aug 27, 51(18), 5278 - 84
Andean yacon root (Smallanthus sonchifolius Poepp . Endl) fructooligosaccharides as a potential novel source of prebiotics; Pedreschi R et al.; The ability of three known probiotic strains (two lactobacilli and one bifidobacterium) to ferment fructooligosaccharides (FOS) from yacon roots (Smallanthus sonchifolius Poepp . Endl) was compared to commercial FOS in this study . Results indicate that Lactobacillus acidophilus NRRL-1910, Lactobacillus plantarum NRRL B-4496, and Bifidobacterium bifidum ATCC 15696 were able to ferment yacon root FOS . FOS consumption apparently depended on the degree of polymerization and the initial FOS composition . L . plantarum NRRL B-4496 and L . acidophilus NRRL B-1910 completely utilized 1-kestose molecules, while B . bifidum was able to utilize 1-kestose molecules as well as molecules with a higher degree of polymerization.

Int J Food Microbiol, 2003 Sep 15, 86(3), 293 - 301
Viability of microencapsulated bifidobacteria in simulated gastric juice and bile solution; Lian WC et al.; Microencapsulated cells of Bifidobacterium longum B6 and Bifidobacterium infantis CCRC 14633 were prepared by spray drying the cell suspension containing the test organism and 10% (w/w) of the carrier material of either gelatin, soluble starch, skim milk or gum arabic . Survival of these microencapsulated and free cells of bifidobacteria in simulated gastric juice (pH 2.0 and 3.0) and bile solution (0.5% and 2.0%) was then examined . B . infantis CCRC 14633 was more susceptible than B . longum B6 to the simulated gastric environment and bile solution tested . Microencapsulated bifidobacteria exhibited a lower population reduction than free cells during exposure to simulated gastric environment and bile solution . This phenomenon was most pronounced when the test organism was exposed to gastric juice at pH 2.0 or 2.0% bile solution . Moreover, it was also observed that the protective effect exerted by encapsulation with spray drying varied with the carriers used and the strains of bifidobacteria.

Biol Pharm Bull, 2003 Aug, 26(8), 1170 - 3
Purification and characterization of ginsenoside Ra-hydrolyzing beta-D-xylosidase from Bifidobacterium breve K-110, a human intestinal anaerobic bacterium; Shin HY et al.; Beta-D-Xylosidase (EC 3.2.1.37) has been purified from ginsenoside Ra-metabolizing Bifidobacterium breve K-110, which was isolated from human intestinal microflora . beta-D-Xylosidase was purified to apparent homogeneity by a combination of ammonium sulfate precipitation, QAE-cellulose, butyl-toyopearl, hydroxyapatit and Q-Sepharose column chromatographies with the final specific activity of 51.8 micromol/min/mg . Molecular weight of beta-D-xylosidase is 49 kDa by SDS-PAGE and gel filtration, which consisted of a single subunit . beta-D-Xylosidase showed optimal activity at pH 5.0 and 37 degrees C . The purified enzyme was potently inhibited by PCMS . beta-D-Xylosidase acted to the greatest extent on p-nitrophenyl-beta-D-xylopyranoside, followed by ginsenoside Ra1 and ginsenoside Ra2 . This enzyme hydrolyzed xylan to xylose, but did not act on p-nitrophenyl-beta-glucopyranoside, p-nitrophenyl-beta-galactopyranoside or p-nitrophenyl-beta-D-fucopyranoside . These findings suggest that this is the first reported purification of ginsenoside-hydrolyzing beta-D-xylosidase from an anaerobic Bifidobacterium sp.

Sichuan Da Xue Xue Bao Yi Xue Ban, 2003 Jul, 34(3), 435 - 8
{Investigation on tumor-targeting characteristics of bifidobacterium infantis to melanoma in mice}; Wu Y et al.; OBJECTIVE: To investigate the tumor-targeting of bifidobacterium infantis to melanoma . METHODS: After bolus administration of bifidobacterium infantis with 3H-TdR tagged, the values of radioactivity in tumor and organs were examined at days 1, 2, 3, 4 and 7 . Anaerobic culture and histological observation of tumor and normal organs were taken for the examination of tumor-targeting characteristics of bifidobacterium infantis . RESULTS: The radioactivity in melanoma tissue increased progressively, while the radioactivity in normal organs became attenuated with time . The anaerobic culture showed an obvious proliferation of bifidobacterium infantis in tumor tissue . A large part of area was Gram positive in the tumor tissue section, whereas the normal tissue was Gram negative . CONCLUSION: Bifidobacterium infantis has good tumor-targeting characteristics in mice melanoma.

Microbiol Immunol, 2003, 47(6), 371 - 8
The suppressive effect of bifidobacteria on Bacteroides vulgatus, a putative pathogenic microbe in inflammatory bowel disease; Shiba T et al.; Bacteroides, a predominant commensal bacteria in the gut, are thought to be responsible for the development of inflammatory bowel disease (IBD) . In the present study, we examined whether or not bifidobacteria suppress B . vulgatus, a representative pathogenic Bacteroides species, in both the coculture system and the gnotobiotic murine model . As a result, Bifidobacterium infantis 1222 highly inhibited the growth of B . vulgatus in the coculture and also significantly suppressed the systemic antibody response raised by B . vulgatus colonizing the gut in gnotobiotic mice . Colonization of the mice by B . vulgatus increased the number of Peyer's patch (PP) cells bearing PNA (peanut agglutinin)+/anti-kappa+ phenotype, which represents plasma cell-like B cells . Moreover, treatment of those B . vulgatus-implanted mice with B . infantis 1222 abrogated such increase in the number of PNA+/anti-kappa+ cells . These results thus suggested that B . infantis 1222 protected the gut epithelial layer including the PP from being invaded by Bacteroides, thereby suppressing the systemic antibody response raised by Bacteroides.

Clin Infect Dis, 2003 Aug 15, 37(4), 490 - 7 Epub 2003 Jul 30.
Human cervicofacial actinomycoses: microbiological data for 1997 cases; Pulverer G et al.; Actinomycoses are sporadically occurring endogenous polymicrobial inflammatory processes, in which fermentative actinomycetes of the genera Actinomyces, Propionibacterium, or Bifidobacterium act as the principal pathogens . Difficulties in diagnosing the disease in a timely and reliable fashion have led clinicians and microbiologists to grossly underestimate its medical importance . Therefore, we evaluated microbiological and selected clinical data derived from 1997 culture-positive cases of human cervicofacial actinomycoses examined in our laboratories during 1972-1999 . The causative actinomycetes belonged to at least 9 different species, among which Actinomyces israelii and Actinomyces gerencseriae predominated . The well-known predisposition of male patients to acquire the disease varied with age and appeared to be especially pronounced in patients aged 20-60 years, the highest incidence being found in female patients aged 11-40 years and in male patients aged 21-50 years . The relevant procedures necessary for diagnosing human actinomycoses reliably, as well as details of their complex etiology, are discussed.

Microbiology, 2003 Aug, 149(Pt 8), 2031 - 7
Cholic acid accumulation and its diminution by short-chain fatty acids in bifidobacteria; Kurdi P et al.; Cholic acid (CA) transport was investigated in nine intestinal Bifidobacterium strains . Upon energization with glucose, all of the bifidobacteria accumulated CA . The driving force behind CA accumulation was found to be the transmembrane proton gradient (Delta pH, alkaline interior) . The levels of accumulated CA generally coincided with the theoretical values, which were calculated by the Henderson-Hasselbalch equation using the measured internal pH values of the bifidobacteria, and a pK(a) value of 6.4 for CA . These results suggest that the mechanism of CA accumulation is based on the diffusion of a hydrophobic weak acid across the bacterial cell membrane, and its dissociation according to the Delta pH value . A mixture of short-chain fatty acids (acetate, propionate and butyrate) at the appropriate colonic concentration (117 mM in total) reduced CA accumulation in Bifidobacterium breve JCM 1192(T) . These short-chain fatty acids, which are weak acids, reduced the Delta pH, thereby decreasing CA accumulation in a dose-dependent manner . The bifidobacteria did not alter or modify the CA molecule . The probiotic potential of CA accumulation in vivo is discussed in relation to human bile acid metabolism.

Appl Environ Microbiol, 2003 Aug, 69(8), 4743 - 52
An in vitro study of the probiotic potential of a bile-salt-hydrolyzing Lactobacillus fermentum strain, and determination of its cholesterol-lowering properties; Pereira DI et al.; This study evaluated the use of a bile-salt-hydrolyzing Lactobacillus fermentum strain as a probiotic with potential hypocholesterolemic properties . The effect of L . fermentum on representative microbial populations and overall metabolic activity of the human intestinal microbiota was investigated using a three-stage continuous culture system . Also, the use of galactooligosaccharides as a prebiotic to enhance growth and/or activity of the Lactobacillus strain was evaluated . Administration of L . fermentum resulted in a decrease in the overall bifidobacterial population (ca . 1 log unit) . In the in vitro system, no significant changes were observed in the total bacterial, Lactobacillus, Bacteroides, and clostridial populations through L . fermentum supplementation . Acetate production decreased by 9 to 27%, while the propionate and butyrate concentrations increased considerably (50 to 90% and 52 to 157%, respectively) . A general, although lesser, increase in the production of lactate was observed with the administration of the L . fermentum strain . Supplementation of the prebiotic to the culture medium did not cause statistically significant changes in either the numbers or the activity of the microbiota, although an increase in the butyrate production was seen (29 to 39%) . Results from this in vitro study suggest that L . fermentum KC5b is a candidate probiotic which may affect cholesterol metabolism . The short-chain fatty acid concentrations, specifically the molar proportion of propionate and/or bile salt deconjugation, are probably the major mechanism involved in the purported cholesterol-lowering properties of this strain.

Zhonghua Wai Ke Za Zhi, 2003 May, 41(5), 385 - 8
{Bifidobacterial supplement enhances the expression and excretion of intestinal sIgA in severely burned rats}; Wang ZT et al.; OBJECTIVE: To investigate the potential effect of bifidobacterial supplement on intestinal mucosal immunity associated with severe burns . METHODS: Wistar rats were randomly divided into burn control group (BC group, n = 30), treatment group (BT group, n = 30), and sham-burn group (NC group, n = 10) . Rats in BT group were fed bifidobacterial preparation (5 x 10(9) CFU/ml) after 30% total body surface area full-thickness burns, 1.5 ml, twice daily . Rats in BC group and NC group were fed normal saline, 1.5 ml, twice daily . Samples were taken on post-burn 1-, 3-, and 5-day . The incidence of bacterial translocation and bifidobacteria counts in the cecum mucosa were determined with standard methods . The sIgA levels in the mucus of the small intestine were measured by RIA . The positive sIgA expression in the lamina propria was detected by immunohistochemical staining . RESULTS: The incidence of bacterial translocation was 42% and 16% in BC and BT groups on post-burn day 3 (P = 0.004), 30% and 8% on day 5 (P = 0.002), respectively . Plasma endotoxin levels were markedly higher in BC and BT groups than in NC group at the early stage post-burn . There was a significant decrease between BT group and BC group on post-burn day 1 (P = 0.0412) . Bifidobacteria counts in cecum mucosa were reduced by 10- to 60-fold after thermal injury, but there was a remarkable increase in bifidobacteria counts in animals fed with bifidobacteria . sIgA levels in the intestinal mucus were significantly decreased in group BC, but they returned to normal range in BT group on post-burn day 5 . Similarly, sIgA expression in the lamina propria was also weakened after burns, and had a tendency to recover after prescription of a 5-day bifidobacteria-supplemented formula . A strong positive correlation was observed between the counts of bifidobacteria in the cecal mucosa and the levels of sIgA in the intestinal mucus (r = 0.7534, P = 0.0000) . CONCLUSIONS: The expression and excretion of sIgA in the intestine appear to be markedly inhibited following a severe thermal injury . The supplement of exogenous bifidobacteria could improve sIgA formation in the small intestine, thereby reducing the incidence of bacterial/endotoxin translocation secondary to major burns.

Zh Mikrobiol Epidemiol Immunobiol, 2003 May-Jun, (3), 57 - 62
{Evaluation on monkeys of reactogenicity and effectiveness of the complex immunoglobulin preparation formulation}; Lapin BA et al.; Experiments on 75 monkeys, 20--healthy and 75--with diarrhea, showed that ready medicinal forms of the complex immunoglobulin preparation in tablets, capsules or in a dried form (vials) were safe, well tolerated, clinically and bacteriologically effective . When used simultaneously with antiparasitic treatment, this preparation, irrespective of the kind of its ready medicinal form, showed high curative effect (80-90%) . The diarrhea ceasing was accompanied by the gradual normalization of intestinal microbiocenosis (an increased concentrations of Escherichia coli and Bifidobacterium, a decreased amount of opportunistic bacteria) . The complex method of the treatment of diarrhea in monkeys (antibiotics, trichopol, phage and probiotics) made it possible to achieve curative effect only in 60% of cases, which required the additional course of treatment, namely the use of rehydration therapy.

Dig Dis Sci, 2003 Jul, 48(7), 1324 - 32
In vitro growth responses of bifidobacteria and enteropathogens to bovine and human lactoferrin; Griffiths EA et al.; A series of in vitro experiments was performed to test the ability of bovine and human lactoferrin to influence the growth of the gram-positive probiotic bacteria, Bifidobacterium bifidum, Bifidobacterium infantis, and Lactobacillus acidophilus, as well as the gram-negative enteric bacteria, E . coli O157:H7 and Salmonella typhimurium . None of the lactoferrin preparations stimulated the growth of the tested strains . However, iron-free apo-lactoferrin (bovine and human) and 66% iron-saturated bovine lactoferrin dramatically slowed the growth of E . coli O157:H7 in single culture experiments, while 98% iron-saturated preparations had no effect . In coculture experiments of B . infantis and E . coli, the iron-limited preparations of lactoferrin also slowed the growth of the latter without inhibiting the bifidobacteria . These results suggest that lactoferrin in iron-limited forms may have the potential to be combined with probiotic bacteria in biotherapeutic products, which could help balance human gut microflora and limit the overgrowth of certain enteric microorganisms.

Curr Opin Infect Dis, 2003 Aug, 16(4), 349 - 55
Neonatal necrotizing enterocolitis: an overview; Kafetzis DA et al.; PURPOSE OF REVIEW: Necrotizing enterocolitis represents a disease entity that remains quite challenging for neonatologists all around the world, in that its aetiology has yet to be revealed, but it is the cause of death for many premature infants each year, affecting up to 28% of very low birthweight infants . This is an attempt to improve the management of affected babies and stimulate more research concerning new diagnostic tools . RECENT FINDINGS: Current trends in the field of (early) diagnosis, such as: (1) imaging techniques, e.g . contrast radiography, portal vein ultrasonography, magnetic resonance, radionuclide scanning; (2) gastrointestinal tonometry; (3) the detection of biochemical markers, cytokines, growth factors; and (4) the determination of the mean peak hydrogen: carbon dioxide ratio excreted in breath, are only some of those mentioned . Various novel preventive techniques are also presented, among which platelet-activating factor acetyl hydrolase activity enhancement, platelet-activating factor receptor antagonists and probiotics, such as Bifidobacterium infantis and Lactobacillus acidophilus, seem quite promising . Regarding treatment, the use of oxygenated perfluorocarbon has added to the limited alternatives available . These data along with other recent discoveries concerning the risk factors and pathogenesis of this disease create a full picture of the current opinion on this topic . SUMMARY: Keeping in mind that the key to confronting such a devastating disorder as necrotizing enterocolitis is early diagnosis and prevention, both clinically applicable and experimental advances are presented with the hope of improving the survival rates of patients affected.

FEMS Microbiol Lett, 2003 Jul 15, 224(1), 143 - 9
A PCR-based method for identification of bifidobacteria from the human alimentary tract at the species level; Venema K et al.; A polymerase chain reaction (PCR)-based method was developed for the identification of isolates of Bifidobacterium at the species level . Using two Bifidobacterium-specific primers directed against the 16S ribosomal gene (Bif164 and Bif662), a PCR product was obtained from the type strains of 12 different bifidobacterial species that have been found in the human alimentary tract . After purification of the PCR products, the DNA was restricted with five different restriction enzymes . The size of the different restriction fragments was used as a fingerprint for the identification of individual bifidobacterial species . The amplified ribosomal DNA restriction analysis method was subsequently used to speciate bifidobacterial isolates from child's feces and from an in vitro model of the large intestine.

Biocell, 2003 Apr, 27(1), 1 - 9
Interaction of bifidobacteria with the gut and their influence in the immune function; Perdigon G et al.; Bifidobacteria are predominant in the lumen of the large intestine and confer various health benefits on the host . They are also used in the preparation of new fermented milks (bioyogurts) or added to conventional yogurt to generate probiotic effects . The colonization of the gut by bacteria tends to be host specific due partly to the way in which bacteria adhere to the intestinal wall . Using a homologous strain of Bifidobacterium animalis in an experimental mouse model, we analyzed by immunofluorescence labelled-bacteria and transmission electronic microscopy the importance of the bacterial interaction with epithelial an immune cells associated to the gut, and the effect of feeding of B . animalis in the immune response . It was able to adhere and interact with both small and large intestine . In spite of this interaction with the gut, no modifications in the immune state (secretory or systemic response) were observed . A heterologous strain of Bifidobacterium adolescentis from human faeces, was neither incapable of binding to the intestine, nor influence the immune system activation, when it was administered during 2, 5 or 7 consecutive days; we believe that using a homologous strain, oral tolerance is developed even when the microorganism interacts with the immune cells associated with the intestine . However, we cannot ignore the beneficial effect of these microorganisms, especially in the prevention of intestinal infections . We think that this property exerted by bifidobacteria is more related to other mechanisms such as competitive inhibition, acid production or others, than enhancement of the immune state.

Br J Nutr, 2003 Jul, 90(1), 119 - 25
Plasma enterolactone or intestinal Bifidobacterium levels do not explain adenoma formation in multiple intestinal neoplasia (Min) mice fed with two different types of rye-bran fractions; Oikarinen S et al.; The study was designed to evaluate whether two types of rye-bran fractions result in distinct bifidogenic effect or enterolactone production in multiple intestinal neoplasia (Min) mice and whether these parameters are associated with intestinal tumorigenesis in this animal model . The experimental diets were a non-fibre diet (control), a rye-bran diet, and diets containing either the soluble extract or the insoluble fraction prepared from rye bran . The main result on adenoma formation in these experiments was the observation that the soluble extract increased number (P=0.012) and size (P=0.008) of adenomas in the distal small intestine when compared with the non-fibre group . All rye-supplemented diets supported similarly the in vivo growth of Bifidobacterium (10(8)-10(9) colony forming units/g) in Min mice, whereas the non-fibre diet lowered intestinal Bifidobacterium below the level of detection . The results show that water solubility does not affect the bifidogenicity of rye bran . Mean plasma enterolactone concentration was highest in the rye-bran group (30.0 nmol/l; P=0.002), which along with the soluble-extract group (16.2 nmol/l; P=0.024) differed significantly from the non-fibre diet group (7.5 nmol/l) . Thus, the mice fed with the rye bran were the best enterolactone producers . In conclusion, rye bran and rye fractions influence adenoma formation in Min mice to a varying degree but plasma enterolactone levels or the production of bifidogenic bacteria do not mediate the effect.

Ann Pharm Fr, 2003 Jul, 61(4), 276 - 81
{Intestinal flora and Crohn's disease}; Desreumaux P et al.; The pathogenesis of inflammatory bowel diseases (IBD) proceeds through stages of initiation, amplification and healing . Abundant clinical and experimental data incriminate luminal bacteria or bacterial products in both the initiation and perpetuation of chronic intestinal inflammation . Macrophage and T-cell activation with accompanying inflammatory cytokine production appears to be an early event . Studies of lymphocyte responsiveness to autologous and heterologous intestinal bacteria have suggested that this activation may result from a breakdown in tolerance to the enteric flora in IBD . This lack of tolerance might be due to an imbalance between protective and aggressive commensal luminal bacterial species (dysbiosis), a decreased barrier function and/or an impaired mucosal clearance allowing the access of bacteria to the mucosal immune system and lack of regulatory mediators or cells . There is still controversy over whether the virulence traits of bacteria are expressed broadly or just in a small subset of bacteria . Individual bacterial species within the indigenous flora vary in their capacity to drive intestinal inflammation . In experimental models, some bacteria such as Bacteroides vulgatus can cause colitis alone when monoassociated in the HLA-B27 transgenic rat model . Others, including Lactobacillus and Bifidobacterium species have no proinflammatory capacity and have been used as probiotics . In patients with IBD, systematic approach to this issue is hampered by the limited knowledge of intestinal flora . Adherent-invasive Escherichia coli are a possible candidate for the onset and/or persistence of intestinal inflammation in patients with Crohn's disease, since they possess all the virulence factors that allow the bacteria to cross the intestinal barrier, to move to deep tissues, and to continuously activate macrophages . The recent identification of NOD2/CARD15 as a susceptibility gene for Crohn's disease has provided another link between the immune response to enteric bacteria and the development of mucosal inflammation . NOD2/CARD15 is composed of two caspase recruitment domain (CARD), a nucleotide-binding domain (NBD) and a leucin-rich-repeat (LRR) region . The LRR domain of NOD2/CARD15 has binding activity for bacterial peptidoglycans and its deletion stimulates the NF-kappaB pathway . The most frequent variants of NOD2/CARD15 observed in Crohn's disease tend to cluster in the LRR and its adjacent regions . This suggests that the LRR domain of CD-associated variants is likely to be impaired in its recognition of microbial components . Continuing studies are investigating the pathophysiological mechanisms induced by NOD2/CARD15 variants in the intestinal mucosa.

Appl Environ Microbiol, 2003 Jul, 69(7), 4296 - 301
Identification and tracing of Bifidobacterium species by use of enterobacterial repetitive intergenic consensus sequences; Ventura M et al.; Eighty-nine Bifidobacterium strains from 26 species were identified and classified to the species level with an enterobacterial repetitive intergenic consensus (ERIC)-PCR approach . We demonstrated that ERIC-PCR is useful for a phylogenetic and taxonomical analysis but as well as for a species composition analysis of mixed bifidobacterial cultures isolated from dairy products and other environments.

Appl Environ Microbiol, 2003 Jul, 69(7), 4219 - 26
Identification of two-component regulatory systems in Bifidobacterium infantis by functional complementation and degenerate PCR approaches; MacConaill LE et al.; Two-component signal transduction systems (2CSs) are widely used by bacteria to sense and adapt to changing environmental conditions . With two separate approaches, three different 2CSs were identified on the chromosome of the probiotic bacterium Bifidobacterium infantis UCC 35624 . One locus was identified by means of functional complementation of an Escherichia coli mutant . Another two were identified by PCR with degenerate primers corresponding to conserved regions of one protein component of the 2CS . The complete coding regions for each gene cluster were obtained, which showed that each 2CS-encoding locus specified a histidine protein kinase and an assumed cognate response regulator . Transcriptional analysis of the 2CSs by Northern blotting and primer extension identified a number of putative promoter sequences for this organism while revealing that the expression of each 2CS was growth phase dependent . Analysis of the genetic elements involved revealed significant homology with several distinct regulatory families from other high-G+C-content bacteria . The conservation of the genetic organization of these three 2CSs in other bacteria, including a number of recently published Bifidobacterium genomes, was investigated.

Biosci Biotechnol Biochem, 2003 May, 67(5), 951 - 7
Decrease in ovalbumin specific IgE of mice serum after oral uptake of lactic acid bacteria; Ishida Y et al.; Different kinds of lactobacilli and Bifidobacteria fermented milk were fed to ovalbumin-specific IgE-elevated mice for 3 days, and after the final administration, changes in the ovalbumin-specific IgE values for each sample were compared to the value for non-fermented milk . Seven of the Lactobacillus-fermented milks caused a significant decrease in the serum ovalbumin-specific IgE levels . Above all, Lactobacillus acidophilus L92, Lactobacillus acidophilus CP1613, and Lactobacillus fermentum CP34 fermented milk had the most significant effects of decreasing the serum ovalbumin-specific IgE levels compared to a control group . The L . acidophilus L92 and L . fermentum CP34 cells also showed significant ovalbumin-specific IgE lowering activities . From these results, an active component seems to exist in the cells of L . acidophilus L92 and L . fermentum CP34 strains . Recovery of the radiolabeled L . acidophilus L92 and L . fermentum CP34 cells from the small intestine and the large intestine of the mouse 13 h after oral administration were higher than the recovery of any other strain.

Zhongguo Wei Zhong Bing Ji Jiu Yi Xue, 2003 Mar, 15(3), 154 - 8
{Improvement of bifidobacterial supplement on the barrier function of intestinal mucosa and microbe flora induced by thermal injury in rats}; Wang ZT et al.; OBJECTIVE: To investigate the potential effect of bifidobacterial supplement on intestinal mucosal and biological barrier following severe burns . METHODS: Seventy Wistar rats were randomly divided into burn control group (BC, n=30), burn + treatment group (BT, n=30), and sham-burn group (NC, n=10) . Animals in BT group were fed bifidobacterial preparation (5x10(12) CFU/L) after 30% total body surface area (TBSA) full-thickness thermal injury, 1.5 ml, twice daily . Samples were taken on postburn days 1,3,and 5 . The incidence of bacteria/endotoxin translocation and the counts of bifidobacteria, E.coli, and fungi in ileo-cecum mucosa were determined with standard methods . Ileum mucosal injuries were evaluated light and electron microscopically by blinded examiners . RESULTS: (1) The incidence of bacterial translocation were 42% and 16% in BC and BT groups on postburn day 3 (P=0.004 0), 30% and 6% on day 5 (P=0.002 0), respectively . Plasma endotoxin levels were markedly higher in BC and BT groups than that in NC group at the early stage postburn, while there was a significant decrease in BT group compared with BC group on postburn day 1 (P=0.04) . (2) The bifidobacteria counts in ileo-cecum mucosa were reduced by 10- to 60-fold after thermal injury, while there was a remarkable increase in animals fed with bifidobacteria . The E.coli counts in ileo -cecum mucosa were increased by 20- to 30-fold on postburn days 1 and 3, whereas those markedly reduced after 3-day bifidobacteria-supplemented formula, tending to normal range.(3)There were obviously ileum mucosal lesions on postburn days 1 and 3,the ileum lesion scores remained significant higher in B C group than that in NC group on postburn 5 day (P<0.05).However,the damage d ileum mucosa was markedly repaired after 3-day bifidobacteria-supplemented formula, and almost restored 5 days later . CONCLUSION: The intestinal mucosal and biological barrier appear to be markedly damaged after severe burns . The supplement of exogenous bifidobacteria can facilitate the improvement of mucosal as well as biological barrier function, thereby reducing the incidence of bacteria/endotoxin translocation secondary to major burns.

Poult Sci, 2003 Jun, 82(6), 1030 - 6
Effects of dietary fructooligosaccharide on digestive enzyme activities, intestinal microflora and morphology of male broilers; Xu ZR et al.; Two hundred forty male Avian Farms broiler chicks, 1 d of age, were randomly allocated to four treatments, each of which had five pens of 12 chicks per pen . The chicks were used to investigate the effects of fructooligosaccharide (FOS) on digestive enzyme activities and intestinal microflora and morphology . The chicks received the same basal diet based on corn-soybean meal, and FOS was added to the basal diet at 0, 2.0, 4.0, and 8.0 g/kg diet at the expense of corn . Addition of 4.0 g/kg FOS to the basal diet significantly increased average daily gain of broilers . The feed-to-gain ratios were significantly decreased for the birds fed diets with 2.0 and 4.0 g/kg FOS versus the control . Addition of 4.0 g/kg FOS enhanced the growth of Bifidobacterium and Lactobacillus, but inhibited Escherichia coli in the small intestinal and cecal digesta . Supplementation of 2.0 or 4.0 g/kg FOS to chicks significantly improved the activities of amylase compared to the control (12.80 or 14.75 vs . 8.42 Somogyi units) . A significant increase in the activities of total protease was observed in 4.0 g/kg FOS-treated birds versus controls (83.91 vs . 65.97 units) . Morphology data for the duodenum, jejunum, and ileum showed no significant differences for villus height, crypt depth, or microvillus height at the duodenum . By contrast, addition of 4.0 g/kg FOS significantly increased ileal villus height, jejunal and ileal microvillus height, and villus-height-to-crypt-depth ratios at the jejunum and ileum and decreased crypt depth at the jejunum and ileum . However, addition of 8.0 g/kg FOS had no significant effect on growth performance, digestive enzyme activities, intestinal microflora, or morphology.

Am J Clin Nutr, 2003 Jun, 77(6), 1537S - 1543S
Nutritional and physiologic significance of human milk proteins; Lonnerdal B; Human milk contains a wide variety of proteins that contribute to its unique qualities . Many of these proteins are digested and provide a well-balanced source of amino acids to rapidly growing infants . Some proteins, such as bile salt-stimulated lipase, amylase, beta-casein, lactoferrin, haptocorrin, and alpha1-antitrypsin, assist in the digestion and utilization of micronutrients and macronutrients from the milk . Several proteins with antimicrobial activity, such as immunoglobulins, kappa-casein, lysozyme, lactoferrin, haptocorrin, alpha-lactalbumin, and lactoperoxidase, are relatively resistant against proteolysis in the gastrointestinal tract and may, in intact or partially digested form, contribute to the defense of breastfed infants against pathogenic bacteria and viruses . Prebiotic activity, such as the promotion of the growth of beneficial bacteria such as Lactobacilli and Bifidobacteria, may also be provided by human milk proteins . This type of activity can limit the growth of several pathogens by decreasing intestinal pH . Some proteins and peptides have immunomodulatory activities (eg, cytokines and lactoferrin), whereas others (eg, insulin-like growth factor, epidermal growth factor, and lactoferrin) are likely to be involved in the development of the intestinal mucosa and other organs of newborns . In combination, breast-milk proteins assist in providing adequate nutrition to breastfed infants while simultaneously aiding in the defense against infection and facilitating optimal development of important physiologic functions in newborns.

J Appl Microbiol, 2003, 95(1), 146 - 54
Employment of stressful conditions during culture production to enhance subsequent cold- and acid-tolerance of bifidobacteria; Maus JE et al.; AIMS: This study examined whether exposure of early stationary phase Bifidobacterium longum and B . lactis cells to various combinations of reduced temperature, reduced pH and starvation would enhance the cells' subsequent cold- and/or acid-tolerance . METHODS AND RESULTS: Survival of B . longum in growth medium at 6 degrees C significantly (P < 0.05) increased as a result of starving cells for 30 or 60 min without any simultaneous decrease in temperature or pH . Acid-tolerance of B . lactis (at pH 3.5 in synthetic gastric fluid) increased significantly when the growth medium pH was decreased from 6.0 to 5.2 and cells experienced 30 or 60 min of starvation . Enhanced B . lactis acid-tolerance persisted through 8-11 weeks of -80 degrees C storage in the pH 5.2 growth medium . Upon addition to milk during yogurt manufacture, these cells initially had enhanced acid-tolerance relative to untreated cells but untreated cells became equally acid-tolerant during the first 2.5 h of yogurt manufacture . CONCLUSIONS: The cold- and acid-tolerance of bifidobacteria vary widely, but may be significantly increased by application of sub-lethal stress to early stationary phase cells during culture production . SIGNIFICANCE AND IMPACT OF THE STUDY: The enhancement of B . lactis acid-tolerance observed in this study may be of potential importance in the production of effective ready-to-consume probiotic dietary supplements.

J Appl Microbiol, 2003, 95(1), 7 - 12
Growth-inhibiting effects of seco-tanapartholides identified in Artemisia princeps var . orientalis whole plant on human intestinal bacteria; Cho SH et al.; AIMS: The present work aimed at isolating antibacterial constituents from the whole plant of Artemisia princeps var . orientalis active towards nine human intestinal bacteria . METHODS AND RESULTS: The growth-inhibiting activities of materials derived from the Artemisia whole plant towards test bacteria were examined using an impregnated paper disc method . The biologically active constituents of the Artemisia whole plant were characterized as the sesquiterpene lactones seco-tanapartholides A and B by spectroscopic analysis . In a test using 1 mg per disc, seco-tanapartholides A and B produced a clear inhibitory effect against Clostridium perfringens, Bacteroides fragilis and Staphylococcus aureus . These compounds did not affect the growth of test lactic acid-producing bacteria (Bifidobacterium adolescentis, Bif . breve, Lactobacillus acidophilus and Lact . casei) and Escherichia coli, whereas weak growth inhibition towards Bif . bifidum was observed . At 0.5 mg per disc, seco-tanapartholides A and B exhibited moderate growth inhibition towards Cl . perfringens but weak growth inhibition towards Bact . fragilis and Staph . aureus . CONCLUSIONS: Inhibitory action of seco-tanapartholides A and B towards specific bacteria without any adverse effects on lactic acid-producing bacteria may be an indication of at least one of the pharmacological actions of A . princeps var . orientalis whole plant . SIGNIFICANCE AND IMPACT OF THE STUDY: These naturally occurring Artemisia whole plant-derived materials could be useful as a new preventive agent against various diseases caused by harmful intestinal bacteria such as clostridia.

Arch Tierernahr, 2003 Feb, 57(1), 1 - 10
Effects of stachyose on performance, diarrhoea incidence and intestinal bacteria in weanling pigs; Liying Z et al.; A three-week trial was conducted to evaluate the effects of the oligosaccharide stachyose on performance, diarrhoea incidence and intestinal bacterial populations in weaned pigs (7.96 +/- 0.2kg) . A total of 144 crossbred (Landrace x Large White x Duroc) pigs weaned at 28 days were allotted to one of four treatments with six pens per treatment and six pigs per pen . The experimental diets were based on maize, dried whey and whole-fat milk and were supplemented with 0, 1 or 2% stachyose . A fourth diet contained no milk but instead contained 20% soybean meal to provide 0.78% stachyose and 0.21% raffinose . Inclusion of 1 or 2% stachyose in the diet depressed growth compared with pigs fed the control diet . Pigs fed the soybean meal diet gained weight at a rate similar to pigs fed the diet containing 1% added stachyose . Diarrhoea incidence was highest for pigs fed the soybean meal diet and lowest for pigs fed the control diet, with pigs fed the diets containing stachyose being intermediate . Pigs fed 1% stachyose had more lactobacilli in the ileum as well as more bifidobacteria in the caecum and colon than control pigs . They also had fewer enterobacteria in the colon . In contrast, pigs fed the diet containing 2% stachyose had fewer lactobacilli and bifidobacteria in the jejunum, ileum and caecum than did control pigs . Volatile fatty acids in the ileum, caecum and colon were highest for pigs fed 1% stachyose and lowest for pigs fed 2% stachyose . Volatile fatty acid concentrations were not significantly different between pigs fed the soybean meal diet and those fed the control . The overall results of this experiment indicate that the oligosaccharide stachyose had a negative effect on pig performance and its presence may partially explain the poorer performance observed when soybean meal is used as the sole source of supplemental protein in cereal-based diets fed to weaned pigs.

Int J Immunopathol Pharmacol, 1999 Jan-Apr, 12(1), 31 - 6
Intestinal mucosa-associated bacteria modulate rat mast cell reactivity; Brzezinska-Blaszczyk E et al.; It is well known that in some conditions bacteria of physiological flora of gastrointestinal tract may become pathogenic . Each complaint which causes injury of gastrointestinal wall integrity permits bacteria to penetrate the tissues and affect the tissue cells . Since mast cells represent one of very important and numerous cellular elements of the gastrointestinal tract walls, bacteria can exert the effects on them . Therefore, the aim of our study was to examine the influence of four strains of intestinal mucosa-associated bacteria--Bacteroides thetaiotaomicron, Bacteroides fragilis, Bifidobacterium adolescentis and Escherichia coli on the mast cell reactivity . Our experiments were performed in vitro on isolated rat peritoneal mast cells and the reactivity of these cells was estimated on the basis of histamine release . We used the suspensions of whole bacteria, killed by heating at 65 degrees C . We have noticed that the magnitude of bacteria-induced histamine release from mast cells was very low (up to 6.0%) when compared with histamine release induced with Con A, compound 48/80 and TNF-alpha . However, all studied bacteria changed the reactivity of mast cells in anaphylactic (with ConA) and anaphylactoid (with compound 48/80) reactions . After 40 min preincubation with B . thetaioataomicron, B . fragilis, B . adolescentis or E . coli ConA-induced histamine release was diminished up to 25%, 71%, 58% and and 68% of maximal histamine release, respectively . Preincubation of rat mast cells with B . thetaioataomicron, B . fragilis, B . adolescentis or E . coli also changed their reactivity in anaphylactoid reaction with compound 48/80 (histamine release was diminished up to 70%, 63%, 63% and 60%, respectively).

Nutrition, 2003 Jun, 19(6), 549 - 52
Partially hydrolyzed guar gum: clinical nutrition uses; Slavin JL et al.; OBJECTIVE: This paper provides a review of research on partially hydrolyzed guar gum that is relevant to clinical nutrition practice . METHODS: All relevant papers published on partially hydrolyzed guar gum were reviewed and the results summarized . RESULTS: Partially hydrolyzed guar gum (PHGG) is a water-soluble dietary fiber with a wide range of uses in clinical nutrition . Its low viscosity allows its use in enteral products and beverages . PHGG can be added to enteral formulas and food products as a dietary fiber source . PHGG provides the benefits associated with dietary fiber ingestion . Addition of PHGG to the diet reduced laxative dependence in a nursing home population . PHGG also reduced the incidence of diarrhea in septic patients receiving total enteral nutrition and reduced symptoms of irritable bowel syndrome . PHGG also increased production of Bifidobacterium in the gut . CONCLUSION: The ease of use of PHGG and its clinical effectiveness make it a good choice in clinical nutrition practice.

J Antimicrob Chemother, 2003 Jul, 52(1), 96 - 102 Epub 2003 May 29.
Effects of cefotaxime and desacetylcefotaxime upon Clostridium difficile proliferation and toxin production in a triple-stage chemostat model of the human gut; Freeman J et al.; Clostridium difficile is recognized as an important nosocomial pathogen . C . difficile infection (CDI) is thought to arise as a result of depletion of the normal gut flora by antimicrobial agents . Cefotaxime (CTX) is well-known for its propensity to cause CDI, but the reasons behind its particular predisposition to the disease remain unclear . Previous investigations have so far relied upon the hamster model of CDI or human volunteers . We have used a triple-stage chemostat model of the human gut to investigate the behaviour of C . difficile and components of the normal gut flora, in response to exposure to CTX alone, and in combination with its active metabolite desacetylcefotaxime (dCTX) . C . difficile remained in a steady state during non-antibiotic exposed periods, with no detectable cytotoxin . During both antibiotic exposure regimens, proliferation of C . difficile and elevated cytotoxin levels were observed . Cessation of antibiotic instillation produced a reduction in cytotoxin levels and viable counts . Decreases in bacterial counts were observed in response to both antibiotic exposure regimens, notably for bifidobacteria and bacteroides . Numbers of bacteroides were profoundly affected by exposure to the CTX/dCTX combination, and this may indicate a possible role for bacteroides in colonization resistance . We believe that the gut model is a promising method for studying C . difficile pathogenesis in conditions analogous to the in vivo situation.

Prikl Biokhim Mikrobiol, 2003 May-Jun, 39(3), 307 - 12
{Effect of antibacterial factors and cell metabolism biostimulants on bacterial luminescence}; Pshenichnov RA et al.; Bioluminescence was used as an index of effects of a series of factors (differing in origin and structure) on cell metabolism . Luminescence of intact cells of Escherichia coli lum+ (a genetically modified strain) was measured on exposure to antibiotics, probiotics, phages, and biostimulators . The sensitivity of E . coli lum+ to antibiotics correlated with the antibiotic-induced luminescence quenching . Bioluminescence-based assessment of the antagonistic activity of the main probiotics (bificoll, bifidobacterin, acylact, colibacterin, and a composite probiotic preparation), each taken at a concentration of 1 dose/ml demonstrated that bacterial luminescence was inhibited by 75-99.9% (exposure time, 30 min) . Bioluminescence changes reflected cell damage associated with phage infection . It was shown that bioluminescence stimulation could be used as an index of the effect of immunomodulators (olexin and vermin) on cell metabolism in bacterial cultures.

Cancer Res, 2003 May 15, 63(10), 2388 - 92
Identification of mucin-depleted foci in the unsectioned colon of azoxymethane-treated rats: correlation with carcinogenesis; Caderni G et al.; We tested the association between aberrant crypt foci (ACF) and tumor induction by feeding azoxymethane-induced rats (15 mg/kg x 2, s.c.) with synbiotics (Raftilose Synergy 1, a derivative of inulin, 10% of the diet, along with lactobacilli and bifidobacteria) . After 16 weeks of feeding, synbiotics significantly increased ACF multiplicity . On the contrary, after 32 weeks, synbiotics significantly decreased intestinal tumors . When the same unsectioned colon used for ACF determination was stained with high-iron diamine Alcian blue, foci of crypts with scarce or absent mucins were identified . We defined these lesions as mucin-depleted foci (MDF), and they were visible in all azoxymethane-treated rats and correlated with tumor induction (MDF/colon: 8.2 +/- 0.9 and 3.8 +/- 0.9 in controls and synbiotic-fed rats, respectively, P < 0.01; crypts/MDF: 12.2 +/- 2 and 6.4 +/- 1 in controls and synbiotic-fed rats, respectively, P < 0.05, means +/- SE, n = 7) . There were fewer MDF/colon than ACF, and they were histologically more dysplastic than mucinous lesions identified as ACF in high-iron diamine Alcian blue-stained colon . In conclusion, MDF may be premalignant lesions that predict colon carcinogenesis.

Proc Nutr Soc, 2003 Feb, 62(1), 177 - 82
Oligosaccharides: state of the art; Delzenne NM; Oligosaccharides, consisting of a mixture of hexose oligomers with a variable extent of polymerisation, are food products with interesting nutritional properties . They may be naturally present in food, mostly in fruits, vegetables or grains, or produced by biosynthesis from natural sugars or polysaccharides and added to food products because of their nutritional properties or organoleptic characteristics . The dietary intake of oligosaccharides is difficult to estimate, but it may reach 3-13 g/d per person (for fructo-oligosaccharides), depending on the population . The extent of resistance to enzymic reactions occurring in the upper part of the gastrointestinal tract allows oligosaccharides to become 'colonic nutrients', as some intestinal bacterial species express specific hydrolases and are able to convert oligosaccharides into short-chain fatty acids (acetate, lactate, propionate, butyrate) and/or gases by fermentation . Oligosaccharides that selectively promote some interesting bacterial species (e.g . lactobacilli, bifidobacteria), and thus equilibrate intestinal microflora, are now termed prebiotics . The pattern of short-chain fatty acid production in the caeco-colon, as well as the prebiotic effect, if demonstrated, are dynamic processes that vary with the type of oligosaccharide (e.g . extent of polymerisation, nature of hexose moieties), the duration of the treatment, the initial composition of flora or the diet in which they are incorporated . Experimental data obtained in vitro and in vivo in animals, and also recent data obtained in human subjects, support the involvement of dietary oligosaccharides in physiological processes in the different intestinal cell types (e.g . mucins production, cell division, immune cells function, ionic transport) and also outside the gastrointestinal tract (e.g . hormone production, lipid and carbohydrates metabolism) . The present paper gives an overview of the future development of oligosaccharides, newly recognised as dietary fibre.

Proc Nutr Soc, 2003 Feb, 62(1), 67 - 72
Regulation of short-chain fatty acid production; Macfarlane S et al.; Short-chain fatty acid (SCFA) formation by intestinal bacteria is regulated by many different host, environmental, dietary and microbiological factors . In broad terms, however, substrate availability, bacterial species composition of the microbiota and intestinal transit time largely determine the amounts and types of SCFA that are produced in healthy individuals . The majority of SCFA in the gut are derived from bacterial breakdown of complex carbohydrates, especially in the proximal bowel, but digestion of proteins and peptides makes an increasing contribution to SCFA production as food residues pass through the bowel . Bacterial hydrogen metabolism also affects the way in which SCFA are made . This outcome can be seen through the effects of inorganic electron acceptors (nitrate, sulfate) on fermentation processes, where they facilitate the formation of more oxidised SCFA such as acetate, at the expense of more reduced fatty acids, such as butyrate . Chemostat studies using pure cultures of saccharolytic gut micro-organisms demonstrate that C availability and growth rate strongly affect the outcome of fermentation . For example, acetate and formate are the major bifidobacterial fermentation products formed during growth under C limitation, whereas acetate and lactate are produced when carbohydrate is in excess . Lactate is also used as an electron sink in Clostridium perfringens and, to a lesser extent, in Bacteroides fragilis . In the latter organism acetate and succinate are the major fermentation products when substrate is abundant, whereas succinate is decarboxylated to produce propionate when C and energy sources are limiting.

Curr Microbiol, 2003 Jun, 46(6), 391 - 7
Identification of the gene for beta-fructofuranosidase of Bifidobacterium lactis DSM10140(T) and characterization of the enzyme expressed in Escherichia coli; Ehrmann MA et al.; Bifidobacterium lactis is a moderately oxygen-tolerant, saccharolytic bacterium often used in combination with fructooligosaccharides (FOS) as a probiotic supplement in diverse dairy products . This is the first report describing the gene structure and enzymatic properties of a beta-fructofuranosidase {EC 3.2.1.26} from Bifidobacteria . BfrA was identified in Bifidobacterium lactis DSM 10140(T) and heterologously expressed in Escherichia coli . The G+C content was identical with the G+C content as determined for the total genomic DNA (61.9 mol %) . The gene codes for a 532-aa residue polypeptide of 59.4 kDa . Surprisingly, the deduced aa sequence revealed only minor similarity to other fructofuranosidases (18% to E . coli cscA) . The enzyme was purified to homogeneity after incorporation of a C-terminal 6 x HIS affinity tag . It hydrolased sucrose, 1-kestose, Raftilose, Actilight, inulin, and raffinose (100%, 91%, 84%, 80%, 37%, 4%) . Fructose moieties were released in an exo-type fashion . Substrates with alpha-glycosidic linkages or residues other than fructose were not attacked . The kinetic parameters K(m) and V(max) for sucrose hydrolysis were 10.3 m M and 0.031 microM/min (pH 7.6; 37 degrees C) . The activity was abolished by Zn(2+) (1 m M) and significantly inhibited by Fe(2+) and Ni(2+) (10 m M) . The enzyme showed its maximal activity at 40 degrees C.

Wei Sheng Yan Jiu, 2003 Jan, 32(1), 54 - 5
{Study on the regulative effect of isomaltooligosaccharides on human intestinal flora}; Gu Q et al.; OBJECTIVE: In order to study the effect of Isomaltooligosaccharides on Human and mouse intestinal flora in vivo . METHODS: 1 . 30 health subjects (15 male, 15 female) were randomly selected from the clinic of Tianjin CDC . They were provide 15 g Isomaltooligosaccharides once a day for 7 days; 2 . 40 BABL/c mice were divided into 4 groups, 3 experimental groups and 1 control group . These 3 experimental groups were fed different dose of Isomaltooligosaccharides by gavage for 7 days . Feces of both human and mice were determined before and at the end of the experiment . RESULTS: The results showed that the reproduction of Bifidobacteria and Lactobacillus greatly increased, and the growth of Clostridium perfriengenes was significantly inhibited both in mice and human intestinal tracts . CONCLUSION: Isomaltooligosaccharides could regulate and improve the intestinal flora in both mice and human intestine.

Zh Mikrobiol Epidemiol Immunobiol, 2000 Jul-Aug, (4 Suppl), 77 - 9
{Effect of bifidobacteria on the anti-lysozyme activity of enterobacteria}; Valyshev AV et al.; The influence of the subinhibitory concentrations of bifidobacterial exometabolites on the capacity for inactivating lysozyme in pathogenic and opportunistic enterobacteria was determined . The supernatant of B . bifidum 1 suppressed antilysozyme activity (ALA) in most Klebsiella strains and in some pathogenic Escherichia coli strains, but increased ALA in some E . coli of indigenous flora and failed to influence ALA in Salmonellae . The stimulating effect of the vital activity products of bifidobacteria with respect to the antilysozyme sign in some S . enteritidis isolates and diarrheagenic E . coli was found . These properties should be taken into consideration while selecting strains for the construction of probiotics.

Poult Sci, 2003 Apr, 82(4), 627 - 31
Application of prebiotics and probiotics in poultry production; Patterson JA et al.; The intestinal microbiota, epithelium, and immune system provide resistance to enteric pathogens . Recent data suggest that resistance is not solely due to the sum of the components, but that cross-talk between these components is also involved in modulating this resistance . Inhibition of pathogens by the intestinal microbiota has been called bacterial antagonism, bacterial interference, barrier effect, colonization resistance, and competitive exclusion . Mechanisms by which the indigenous intestinal bacteria inhibit pathogens include competition for colonization sites, competition for nutrients, production of toxic compounds, or stimulation of the immune system . These mechanisms are not mutually exclusive, and inhibition may comprise one, several, or all of these mechanisms . Consumption of fermented foods has been associated with improved health, and lactic acid bacteria (lactobacilli and bifidobacteria) have been implicated as the causative agents for this improved health . Research over the last century has shown that lactic acid bacteria and certain other microorganisms can increase resistance to disease and that lactic acid bacteria can be enriched in the intestinal tract by feeding specific carbohydrates . Increased bacterial resistance to antibiotics in humans has caused an increase in public and governmental interest in eliminating sub-therapeutic use of antibiotics in livestock . An alternative approach to sub-therapeutic antibiotics in livestock is the use of probiotic microorganisms, prebiotic substrates that enrich certain bacterial populations, or synbiotic combinations of prebiotics and probiotics . Research is focused on identifying beneficial bacterial strains and substrates along with the conditions under which they are effective.

Int J Food Microbiol, 2003 Jun 15, 83(2), 161 - 70
A collaborative study of a method for the enumeration of probiotic bifidobacteria in animal feed; Leuschner RG et al.; An enumeration method to be used as an official control method in the framework of Council Directive 70/524/EEC for probiotic bifidobacteria used as feed additives was validated . Seventeen laboratories in 11 European Countries carried out a collaborative study . A spread plate method following BS ISO 15214:1998 using four different agars, Man Rogosa Sharpe (MRS), acidified MRS, MRS with triphenyl tetrazolium chloride (TTC) and a selective bifidobacteria medium, was validated . Precision data in terms of repeatability (r) and reproducibility (R) of the method for each medium using different feeding stuffs with a high and a low inoculation level were determined . Bifidobacteria were present in the samples as a single component or in mixtures with other probiotics . The enumeration of bifidobacteria on all agars showed a relative standard deviation of repeatability (RSD(r)) between 1.2% and 6.3% and a relative standard deviation of reproducibility (RSD(R)) between 2.6% and 8.7% . MRS agar was preferred, followed by acidified MRS and MRS+TTC agar . The selective bifidobacteria medium gave similar counts as the MRS media . For routine analysis, the use of MRS agar with supplementation of cysteine hydrochloride (the selective bifidobacteria medium without antibiotics) is recommended . Depending on the presence and concentration of other probiotics such as enterococci, lactobacilli and pediococci, acidified MRS or MRS+TTC agar is recommended . The selective bifidobacteria medium was selective for bifidobacteria.An official control method for enumeration of probiotic bifidobacteria as a single component and in mixtures with other probiotic microorganisms in feeding stuffs was validated . The methodology is not applicable to mineral feed . The results are intended for consideration for adaptation as CEN and ISO standards.

Med Hypotheses, 2003 Jun, 60(6), 915 - 23
Chronic fatigue syndrome: lactic acid bacteria may be of therapeutic value; Logan AC et al.; Chronic fatigue syndrome (CFS) is complex illness with unknown aetiology . Recent research shows that patients with CFS have marked alterations in microbial flora, including lowered levels of bifidobacteria and small intestinal bacterial overgrowth (SIBO) . Research also indicates that CFS patients are under increased oxidative stress, have a type 2 helper cell dominate cytokine profile, frequently report allergies, have altered essential fatty acid (EFA) status and may have malabsorption of certain micronutrients . Lactic acid bacteria (LAB) have the potential to influence the immune system in CFS patients by supporting T helper cell 1 driven cellular immunity and may decrease allergies . In addition LAB are strong antioxidants, may improve EFA status, can enhance absorption of micronutrients by protecting the intestinal epithelial barrier, and have been used to treat SIBO . It is our contention that LAB may have a therapeutic role in the treatment of CFS.

J Food Prot, 2003 Apr, 66(4), 674 - 8
Inhibition of nonproteolytic Clostridium botulinum with lactic acid bacteria and their bacteriocins at refrigeration temperatures; Rodgers S et al.; Nonproteolytic Clostridium botulinum (strains 17B, Beluga, and 202F) was found to be inhibited by Lactobacillus, Lactococcus, Streptococcus, and Pediococcus species in tests by the spot-on-the-lawn simultaneous-antagonism method at 10, 15, and 25 degrees C . C . botulinum 17B was the most resistant strain . Inhibition zone size increased with decreasing incubation temperature . Six strains of Lactobacillus acidophilus and seven strains of bifidobacteria failed to produce an inhibition zone on buffered reinforced clostridium Prussian blue agar seeded with spores of any of the selected C . botulinum strains . C . botulinum 17B was sensitive to 50 to 100 IU of nisin per ml and to 10 to 20 AU of pediocin A per ml.

Curr Issues Intest Microbiol, 2003 Mar, 4(1), 9 - 20
Intestinal bacteria and ulcerative colitis; Cummings JH et al.; Convincing evidence from both animal models and the study of patients with ulcerative colitis (UC) implicates the intestinal microflora in the initiation and maintenance of the inflammatory processes in this condition . Despite this, no specific pathogen has been identified as causal and the disease is widely believed to occur as the result of a genetically determined, but abnormal immune response to commensal bacteria . When compared with healthy people, UC patients have increased levels of mucosal IgG directed against the normal microflora . Studies of mucosal bacterial populations in UC indicate that there may be increased numbers of organisms, but reduced counts of "protective" bacteria such as lactobacilli and bifidobacteria . In animal models of colitis, antibiotics, particularly metronidazole, clindamycin, ciprofloxacin and the combination of vancomycin/impinemem protect against UC, especially if given before the onset of inflammation . These antibiotics target anaerobes and some Gram-positive organisms such as enterococci . However, antibiotic use in more than a dozen randomised control trials has been very disappointing, probably because we do not know which species to target, when to give the antibiotics, for how long and in what combinations . Surprisingly, therefore, there is a consistent benefit in the small number of studies reported of probiotics to manage UC and pouchitis . There is scope for more work in this area focussing on the mucosal microflora, its interactions with the gut immune system, its metabolic properties and the potential ways of modifying it.

Curr Opin Infect Dis, 2002 Oct, 15(5), 501 - 6
Probiotics, infection and immunity; Macfarlane GT et al.; PURPOSE OF REVIEW: Taking live bacteria by mouth to improve health (probiotics) is not intuitively rational yet it is a practice with a long history . As interest in the effects on health of the intestinal flora has developed, along with major advances in the technology for studying it, so has come a new interest in establishing the true benefits of probiotic therapy . This review summarizes the most recent contributions to this rapidly developing area . RECENT FINDINGS: Probiotic bacteria, mainly bifidobacteria and lactobacilli for historical reasons, can prevent or ameliorate some diseases . Many empirical studies have been done, but work to develop the ideal characteristics of probiotics lags behind . Current literature covers survival of probiotics in the gut, mucosal adherence, antibacterial/pathogen mechanisms, effects on immune function and clinical studies . SUMMARY: Probiotic bacteria are effective in preventing and reducing the severity of acute diarrhoea in children . They are also useful in antibiotic associated diarrhoea but not for elimination of Helicobacter pylori . In inflammatory bowel disease, especially ulcerative colitis, probiotics offer a safe alternative to current therapy . Probiotics have been used to prevent urogenital tract infection with benefit and, perhaps more intriguingly, to reduce atopy in children . Probiotics do not invariably work and study of mechanisms is urgently needed.

Biotechnol Prog, 2003 Mar-Apr, 19(2), 672 - 5
Quantitative comparison of lactose and glucose utilization in Bifidobacterium longum cultures; Kim TB et al.; In batch cultures, Bifidobacterium longum SH2 has a higher final cell concentration and greater substrate consumption when grown on lactose versus glucose . Continuous cultures were used to compare lactose and glucose utilization by B . longum quantitatively . In the continuous culture, the estimated maintenance coefficients (m) were similar when on lactose and glucose; the maximum cell yield coefficient (Y(X/S)(max)) was higher on lactose; and the specific consumption rate of lactose (q(S)) was lower than that of glucose . Assuming that cell growth followed the Monod model, the maximum specific growth rates (mu(max)) and saturation constants (K(S)) in lactose and glucose media were determined using the Hanes-Woolf plots . The respective values were 0.40 h(-)(1) and 78 mg/L for lactose and 0.46 h(-)(1) and 697 mg/L for glucose . The kinetic parameters of the continuous cultures showed that B . longum preferred lactose to glucose, although the specific consumption rate of glucose was higher than that of lactose.

Int J Food Microbiol, 2003 May 25, 83(1), 105 - 9
Performance of bifidobacteria in oat-based media; Laine R et al.; Thirteen Bifidobacterium strains were isolated form the faeces of healthy elderly subjects . The ability of the isolated strains to acidify and survive in an oat-based medium was assessed and compared to a commercial starter Bifidobacterium strain . Since adhesion to the intestinal mucosa is one of the main selection criteria for potential probiotics, the influence of the oat medium on the ex vivo adhesion of the isolates to both human colonic tissue and to mucus from elderly subjects was assessed . Seven of the fourteen strains survived the 24-h fermentation, six of these acidified the medium to below pH 5 . Supplementation of the medium with soy protein had a negative effect on survival and the acidification . The strains isolated from the elderly subjects were found to adhere in high levels to immobilised intestinal mucus from elderly . However, their adhesion to colonic tissue was significantly lower . Prior exposure to the oat-based medium increased the adhesion of two of the strains.

Int J Antimicrob Agents, 2003 Apr, 21(4), 340 - 6
Identification and screening for antimicrobial activity against Clostridium difficile of Bifidobacterium and Lactobacillus species isolated from healthy infant faeces; Lee YJ et al.; The antimicrobial activity against Clostridium difficile of 109 lactic acid bacteria (LAB) isolated from 32 healthy Korean infants was measured . The ability to show similar activity against Escherichia coli O157:H7 and Staphylococcus aureus was also looked for . Twelve of the 109 LAB showed activity against C . difficile and 19 strains were active against E . coli O157:H7, but none against S . aureus . Four strains had antimicrobial activity against both C . difficile and E . coli O157:H7 . Of the 12 strains that had activity against C . difficile, four strains were excluded as Streptococcus species, while the other eight were identified using polymerase chain reaction (PCR) assays using group-specific primers designed from the nucleotide sequences of the 16S rDNA and internal transcribed spacer (ITS) regions of the Bifidobacterium and Lactobacillus species . Based on the sequencing results, the eight strains screened were identified as Bifidobacterium infantis and Lactobacillus salivarius.

J Bacteriol, 2003 Apr, 185(8), 2571 - 81
Genomic diversity and relatedness of bifidobacteria isolated from a porcine cecum; Simpson PJ et al.; This study initially involved the isolation of a number of bifidobacteria from either the lumen or the epithelium of a porcine cecum . A total of 160 isolates were selected at random on MRS plates containing cysteine hydrochloride (0.5 g/liter) and mupirocin (50 mg/liter) . All were identified as bifidobacteria based on fructose-6-phosphate phosphoketolase activity . Following genomic digestion with the restriction enzyme XbaI and pulsed-field gel electrophoresis (PFGE), the isolates produced 15 distinct macro-restriction patterns . Several of the PFGE patterns differed by only 1, 2, or 3 DNA fragments and were grouped as related patterns into seven PFGE types, termed A through G . The related patterns appeared to show genomic plasticity within the isolates arising from chromosomal mutations or possibly horizontal transfer of plasmids . The relative frequency of each PFGE type was maintained within each cecal sample, with PFGE type E representing approximately 50% of the isolates . Randomly amplified polymorphic DNA PCR, cell morphology, whole-cell protein profiling, 16S ribosomal DNA sequencing, and DNA-DNA hybridization were used to determine if the seven apparently unrelated PFGE types represented genetically distinct isolates . Four groups were identified: PFGE types A, C/D/G, B/E, and F, and these appeared to represent Bifidobacterium minimum, Bifidobacterium pseudolongum subsp . pseudolongum, and Bifidobacterium pseudolongum subsp . globosum and two new species, respectively . The data demonstrate the presence of considerable genomic diversity within a relatively simple bifidobacteria population, consisting of 15 distinct strains representing four groups, which was maintained throughout the porcine cecal contents and epithelial layer.

Am J Clin Nutr, 2003 Apr, 77(4), 937 - 42
Oligofructose-supplemented infant cereal: 2 randomized, blinded, community-based trials in Peruvian infants; Duggan C et al.; BACKGROUND: Prebiotics are nondigestible food ingredients that stimulate the growth of Bifidobacterium and other bacteria in the gastrointestinal tract . Improved gastrointestinal and other health effects have been attributed to them . OBJECTIVE: The objective of this study was to evaluate the effects of dietary supplementation with the prebiotic oligofructose with and without zinc on the prevalence of diarrhea in a community with a high burden of gastrointestinal and other infections . DESIGN: Two consecutive randomized, blinded, controlled clinical trials were performed in a shantytown community near Lima, Peru . The first trial compared an infant cereal supplemented with oligofructose (0.55 g/15 g cereal) with nonsupplemented cereal . During the second trial, zinc (1 mg/15 g cereal) was added to both oligofructose-supplemented and control cereals . RESULTS: We enrolled 282 infants in the first trial and 349 in the second . In the first trial, mean (+/- SD) days of diarrhea were 10.3 +/- 9.6 in the nonsupplemented cereal group and 9.8 +/- 11.0 in the prebiotic-supplemented cereal group (P = 0.66) . In the second trial, mean days of diarrhea were 10.3 +/- 8.9 in the group consuming cereal fortified only with zinc and 9.5 +/- 8.9 in the group consuming cereal containing both zinc and prebiotics (P = 0.35) . Postimmunization titers of antibody to Haemophilus influenzae type B were similar in all groups, as were gains in height, visits to clinic, hospitalizations, and use of antibiotics . CONCLUSIONS: Cereal supplemented with prebiotics was not associated with any change in diarrhea prevalence, use of health care resources, or response to H . influenzae type B immunization . Infants and young children who continue to breast-feed may not benefit from prebiotic supplementation.

Appl Microbiol Biotechnol, 2003 Mar, 61(1), 55 - 60 Epub 2003 Jan 25.
Cloning and characterization of two alpha-glucosidases from Bifidobacterium adolescentis DSM20083; van den Broek LA et al.; Two alpha-glucosidase encoding genes (aglA and aglB) from Bifidobacterium adolescentis DSM 20083 were isolated and characterized . Both alpha-glucosidases belong to family 13 of the glycosyl hydrolases . Recombinant AglA (EC 3.2.1.10) and AglB (EC 3.2.1.20), expressed in Escherichia coli, showed high hydrolytic activity towards isomaltose and pnp-alpha-glucoside . The K(m) for pnp-alpha-glucoside was 1.05 and 0.47 mM and the V(max) was 228 and 113 U mg(-1) for AglA and AglB, respectively . Using pnp-alpha-glucoside as substrate, the pH optimum for AglA was 6.6 and the temperature optimum was 37 degrees C . For AglB, values of pH 6.8 and 47 degrees C were found . AglA also showed high hydrolytic activity towards isomaltotriose and, to a lesser extent, towards trehalose . AglB has a high preference for maltose and less activity towards sucrose; minor activity was observed towards melizitose, low molecular weight dextrin, maltitol, and maltotriose . The recombinant alpha-glucosidases were tested for their transglucosylation activity . AglA was able to synthesize oligosaccharides from trehalose and sucrose . AglB formed oligosaccharides from sucrose, maltose, and melizitose.

Microbes Infect, 2003 Feb, 5(2), 115 - 22
Prevention of gut inflammation by Bifidobacterium in dextran sulfate-treated gnotobiotic mice associated with Bacteroides strains isolated from ulcerative colitis patients; Setoyama H et al.; Indigenous Bacteroides strains are closely associated with the occurrence and exacerbation of ulcerative colitis (UC) . In this study, we aimed to clarify the effect of Bifidobacterium strains, another major member of colonic bacteria, on the development of gut inflammation using gnotobiotic mouse models associated with Bacteroides strains isolated from UC patients . Dextran sulfate (DSS) administration induced inflammation in the large intestine, in particular of the cecum, in the gnotobiotic mice associated with three strains of Bacteroides vulgatus, judging from the myeloperoxidase activity, occult blood score, and IgG leakage into the intestinal contents . However, the severity of the inflammation was greatly reduced in the gnotobiotic mice associated with both B . vulgatus and Bifidobacterium strains . The severity of the cecal inflammation was well correlated with the concentration of succinic acid in the cecum . Bacteriologically, the density of B . vulgatus strain A (BV-A) greatly decreased and the predominant strain changed from BV-A to BV-B on additional association with Bifidobacterium strains . Among gnotobiotic mice associated with a single B . vulgatus strain, the severity of cecal inflammation in BV-A-associated mice was greater than that in BV-B-associated mice . Each Bifidobacterium strain produced compound(s) more effectively inhibiting the growth of BV-A than BV-B in in vitro culture . Taken together, these results suggest that the severity of DSS-induced gut inflammation is closely associated with a particular B . vulgatus strain, and that Bifidobacterium strains may repress exacerbation of intestinal inflammation through growth inhibition of the B . vulgatus strain.

Toxicol In Vitro, 2003 Apr, 17(2), 145 - 52
Growth inhibition of intestinal bacteria and mutagenicity of 2-, 3-, 4-aminobiphenyls, benzidine, and biphenyl; Chung KT et al.; 2-Aminobiphenyl (2-ABP), 3-aminobiphenyl (3-ABP) and 4-aminobiphenyl (4-ABP), but not benzidine (Bz) and biphenyl (Bp), were found to be inhibitory to the growth of human intestinal bacteria Bifidobacterium infantis ATCC 15697, B . bifidium ATCC 11863, Clostridium perfringens ATCC 13124, Escherichia coli ATCC 25922, E . coli ATCC 35218, Enterobacter cloacae ATCC 13047 and Salmonella typhimurium TA98, TA100, YG1041 at 10-200 microg/ml in culture broth . Bacteroides distasonis ATCC 8503, B . fragilis ATCC 25285, B . theataiotaomicron ATCC 29741, C . paraputrificum ATCC 26780, C . clostridiiforme ATCC 25537, Lactobacillus acidophilus ATCC 4356 and Enterococcus faecium ATCC 19434 were not inhibited by the above mentioned compounds in concentrations up to 200 microg/ml . The Ames Salmonella/microsome assay was employed to test the mutagenicity of the above-mentioned compounds using strains TA98 and TA100 in the presence and absence of Aroclor 1254-induced rat S9 mix . It was found that 4-ABP was mutagenic to both TA98 and TA100, and Bz was mutagenic to TA98 in the presence of rat S9 mix . 2-Aminobiphenyl, 3-ABP, and Bp were not mutagenic to both strains tested . 2-Aminobiphenyl and 3-ABP are chemical isomers of 4-ABP and are as strong as 4-ABP in inhibiting the growth of intestinal bacteria but not as mutagenic as 4-ABP . Evidence suggested that the mechanism of growth inhibition is not involved with the interaction of DNA that causes mutations, but rather on the electron transport system of these organisms.

Zhonghua Shao Shang Za Zhi, 2002 Dec, 18(6), 365 - 8
{The relationship between intestinal bifidobacteria and bacteria/endotoxin translocation in scalded rats}; Wang Z et al.; OBJECTIVE: To investigate the potential role of intestinal bifidobacteria in the pathogenesis of gut-origin bacteria/endotoxin translocation in scalded rats . METHODS: Wistar rats inflicted with 30% III degree scalding on the back were employed as the model with the rats undergoing sham injury as the control . The intestinal bacteria/endotoxin translocation and the changes in cecal mucosal microflora were determined by routine methods . And the plasma IL-6 concentration was measured with ELISA . RESULTS: The incident of bacterial translocation into internal organs increased markedly in scalded rats (P = 0.001) . The plasma LPS levels on 1, 3 and 5 postburn days (PBDs) in scalded rat group were much higher than those in sham injury group . The number of bifidobacteria decreased sharply 20 - 250 fold, the fungi increased 5 - 60 fold and E . coli increased 0.5 - 30 fold in the caecal mucosal microflora in the scalding group . The ratio of bifidobacteria to E . coli in the scalding group (4 - 800:1) was much lower than that in the sham injury group (25000:1) . Furthermore, the plasma IL-6 level increased evidently in the scalding group . It was indicated by further analysis that compared with the rats without bacterial translocation, the bifidobacteria decreased 120 fold, the fungal number increased 50 fold and the E . coli number increased 30 fold in the scalded rats . The bifidobacterial number in the caecal mucosal microflora was negatively correlated with the plasma concentrations of IL-6 and LPS (P < 0.01) in the scalding rat group, and the plasma concentration of IL-6 was significantly and positively correlated with that of LPS . CONCLUSION: Severe scalding injury could lead to an the imbalance of intestinal microflora and the increased intestinal translocation of bacteria and LPS . The decrease of the ratio and number of bifidobacteria in the caecal mucosal microflora might be a contribute to the occurrence of postburn intestinal bacteria/endotoxin translocation.

Lett Appl Microbiol, 2003, 36(4), 213 - 6
An in vitro model for investigating intestinal adhesion of potential dairy propionibacteria probiotic strains using cell line C2BBe1; Huang Y et al.; AIMS: The purposes of this study were to screen the adhesion properties of dairy propionibacteria strains and evaluate whether C2BBe1 could be used in the screening of potential probiotic strains . METHODS AND RESULTS: Thirteen dairy propionibacteria strains and two control strains, Lactobacillus acidophilus MJLA1 and Bifidobacterium lactis BDBB2, were tested for adhesion to C2BBe1 . Electron microscopic observations demonstrated that the control strains, L . acidophilus MJLA1 and B . lactis BDBB2, had similar adhesive ability to C2BBe1 as had been previously shown to Caco-2 . Only one of the 13 strains of dairy propionibacteria, strain P . jensenii 702, demonstrated adhesion to C2BBe1 . CONCLUSIONS: C2BBe1 can provide an alternative to Caco-2 for assessing in vitro adhesion properties of probiotic strains . Adhesion properties of dairy propionibacteria were strain-dependent . SIGNIFICANCE AND IMPACT OF THE STUDY: C2BBe1 is highly suitable for application in bacterial adhesion studies, and was used successfully to select a new potential probiotic.

Skin Pharmacol Appl Skin Physiol, 2003 Mar-Apr, 16(2), 108 - 16
Bifidobacterium-fermented soy milk extract stimulates hyaluronic acid production in human skin cells and hairless mouse skin; Miyazaki K et al.; We examined the effects of Bifidobasterium-fermented (BE) and nonfermented (SME) soy milk extracts on the production of hyaluronic acid (HA) in vitro and in vivo . BE, but not SME, significantly enhanced the production of HA in monolayer and organotypic cultures of human keratinocytes, in cultures of human skin fibroblasts, and in hairless mouse skin following topical application for 2 weeks . In the organotypic cultures formed by a similar structure to human epidermis, BE also extended the distribution of HA . Genistein and daidzein, known to stimulate HA production, were detected in BE at a concentration of 0.18 and 0.07 mM, respectively, but not in SME . Therefore, BE has the potential to enhance HA production in the epidermis and dermis, mainly due to genistein released from its glycoside during fermentation . BE is expected to prevent the age-dependent loss of cutaneous HA .

Med Dosw Mikrobiol, 2002, 54(3), 259 - 64
{Bacterial flora from bronchoalveolar lavage and occurrence of class IgG and IgA antibodies to Chlamydia pneumoniae in patients with chronic obstructive pulmonary disease (COPD)}; Chylak J et al.; Bronchoalveolar lavage taken from 46 patients (ranging in age from 21 to 71 years, mean 50.6 +/- 13.9) was examined for aerobic and anaerobic bacterial flora . Sera taken from 39 of patients as well as sera taken from 25 healthy blood donors of similar age (P = 0.99) were examined to determine IgG and IgA antibodies to C . pneumoniae . Bacterial flora was routinely cultured and determined using ATB computer system (bioMerieux,) . IgG and IgA antibodies were tested by the enzyme immunoassays (Labsystems, Finland, Helsinki) . Sera containing anti -C . pneumoniae IgG antibodies with titers of 45 EIU or higher and IgA with titers of 12 EIU or higher were considered positive . 143 of aerobic and 74 of anaerobic bacterial strains were cultured . Streptococci group viridans, pneumococci, enteric bacilli, Haemophilus spp., Prevotella spp., Actinomyces spp., Bifidobacterium spp . and Veilonella spp . were most often cultured . 66.6% of patients had IgG or IgA antibodies, in contrast, to the control group in which 60.0% and 44.0% of examined blood donors had IgG and IgA antibodies respectively . COPD patients were more frequently positive for specific anti-C . pneumoniae antibodies than the healthy donors (p = 0.003) . The difference in a seropositivity rate of specific IgA and IgG antibodies was significant (p = 0.00002 and p = 0.003 respectively) . Bronchoalveolar lavage of patients suffering from COPD can be contaminated with high number of aerobic and anaerobic bacterial species, and immunological status of the patients indicated persistent infection caused by C . pneumoniae more often than in controls.

Clin Infect Dis, 2003 Mar 15, 36(6), 775 - 80 Epub 2003 Mar 05.
Safety of probiotics that contain lactobacilli or bifidobacteria; Borriello SP et al.; Lactobacilli and bifidobacteria are extremely rare causes of infection in humans, as are probiotics based on these organisms . This lack of pathogenicity extends across all age groups and to immunocompromised individuals . Strains used for new probiotics should be chosen from the commensal flora of humans and should not carry intrinsic resistance to antibiotics that would prevent treatment of a rare probiotic infection . Vigilance regarding the detection of possible rare cases of infection due to probiotics should be maintained, and isolates should be sent to reference centers for molecular characterization and confirmation.

Anal Chem, 2003 Feb 15, 75(4), 818 - 24
Mechanism of microbial aggregation during capillary electrophoresis; Zheng J et al.; We studied the aggregation of a rod-shaped bacteria, Bifidobacterium infantis, during capillary electrophoresis (CE) . A microscope with an intensified CCD camera was employed to monitor the migration and aggregation of bacteria, which are labeled with fluorescent dye Syto 9 and excited with a 488-nm argon ion laser . A collision-based aggregation mechanism is proposed, in which collisions between microbes result from different mobilities and migration directions in the electric field . Individual microbes are aligned differently with respect to the direction of the electric field and exhibit different drag coefficients . The long-range forces include van der Waals attraction and electrostatic repulsion as qualitatively described by DLVO theory . Collisions in CE produce sufficient energy to overcome electrostatic repulsion, thus improving the efficiency of aggregation . This is supported by the fact that higher electric fields always resulted in faster aggregation . Also, when sodium phosphate buffer was used, increasing the ionic strength resulted in faster aggregation . However, when Tris-boric acid-EDTA (TBE, pH 9.1) buffer was used, the aggregation speed decreased when the ionic strength increased . We attribute this to the change of the surface of the bacteria at high borate and EDTA concentration, such as the loss of polysaccharides or the presence of complexation . This reduces the hydrophobicity of the surface and, thus, the short-range attractive forces . The addition of 0.05% poly(ethylene oxide) (PEO) into high ionic strength TBE buffer increased the aggregation rate . This can be attributed to the bridging effect of PEO between microbes . Further increase in the concentration of polymer reduced the aggregation rate, especially when the electric field was low, due in part to the increase in viscosity . The decrease in migration velocity produced lower collision energies and lower aggregation efficiencies as well.

Nutrition, 2003 Mar, 19(3), 270 - 4
Oral administration of Bifidobacterium longum culture condensate in a diet-restricted murine peritonitis model enhances polymorphonuclear neutrophil recruitment into the local inflammatory site; Hidemura A et al.; Dietary restriction impairs polymorphonuclear neutrophil (PMN) recruitment into the local inflammatory site, resulting in susceptibility to infection . Probiotics enhance host immunity via conditioning host intestinal microflora . Oral administration of Bifidobacterium longum culture condensate (BCC) in a diet-restricted murine peritonitis model may enhance PMN recruitment into the inflammatory site.Male ICR mice (n = 40) were assigned in equal numbers to control or BCC groups and subjected to 75% restricted food intake for 7 d . During dietary restriction, controls received only standard mouse chow, whereas the BCC group received standard mouse chow containing 1% BCC . Mice were killed before (0 h) or after (2 or 4 h) intraperitoneal glycogen injection . Peritoneal lavage fluid and exudative cells were recovered by peritoneal lavage . Peritoneal exudative cell number was counted . Tumor necrosis factor-alpha, interleukin-6, macrophage inflammatory protein-2, and interleukin-10 concentrations in peritoneal lavage fluid were determined by enzyme-linked immunosorbent assay . CD11b, CD18, CD31, and CD62L expressions on circulating PMNs were measured by flow cytometry.Oral BCC administration upregulated PMN recruitment into the peritoneal cavity and increased peritoneal fluid cytokine concentrations as well as CD18 and CD62L expressions on circulating PMNs during glycogen-induced peritonitis.Oral BCC administration in a diet-restricted murine peritonitis model augmented PMN recruitment into the inflammatory site by upregulating cytokine concentrations in the local inflammatory site and adhesion molecule expression on circulating PMNs . Oral BCC administration may be a favorable modality for improving dietary restriction-induced host immunosuppression.

Am J Kidney Dis, 2003 Mar, 41(3 Suppl 1), S142 - 5
Bifidobacterium in gastro-resistant seamless capsule reduces serum levels of indoxyl sulfate in patients on hemodialysis; Takayama F et al.; BACKGROUND: Intestinal microflora is deranged in hemodialysis (HD) patients as increased aerobacteria such as Escherichia coli, and decreased anaerobacteria such as bifidobacteria . Indole, a precursor of indoxyl sulfate, is produced by E coli but not by bifidobacteria . The serum levels of indoxyl sulfate are increased markedly in HD patients and cannot be reduced efficiently by HD because of its albumin binding . METHODS: To compare the effect of oral administration of Bifidobacterium longum in gastro-resistant seamless capsule (Bifina) on indoxyl sulfate levels with that of Bifidobacteria in powder formulation (Lac B), Bifina was administered to 11 HD patients for 5 weeks, and Lac B to another group of 11 HD patients for 5 weeks . The authors measured the serum level of indoxyl sulfate by using high-performance liquid chromatography . RESULTS: The pre-HD serum levels of indoxyl sulfate significantly decreased in Bifina-treated patients (before, 4.9 +/- 1.7 mg/dL, 4.5 mg/dL, mean +/- SD, median, after 5 weeks, 3.5 +/- 1.3 mg/dL, 3.8 mg/dL; P < 0.005) . However, they did not decrease in the Lac B-treated patients (before, 4.8 +/- 1.4 mg/dL, 4.5 mg/dL, after 5 weeks, 5.2 +/- 2.0 mg/dL, 5.1 mg/dL) . CONCLUSION: Oral administration of Bifina to HD patients is effective in reducing the serum levels of indoxyl sulfate by correcting the intestinal microflora . Gastro-resistant seamless capsule prevents bifidobacteria from its inactivation in acidic gastric juice, and allows it to be actived in the intestine.

J Pediatr Gastroenterol Nutr, 2003 Mar, 36(3), 343 - 51
Randomized double-blind study of the nutritional efficacy and bifidogenicity of a new infant formula containing partially hydrolyzed protein, a high beta-palmitic acid level, and nondigestible oligosaccharides; Schmelzle H et al.; OBJECTIVES: The aim of this study was to evaluate the nutritional efficacy and bifidogenic characteristics of a new infant formula containing partially hydrolyzed whey protein, modified vegetable oil with a high beta-palmitic acid content, prebiotic oligosaccharides, and starch . METHODS: In a double-blind study, healthy formula-fed term infants aged younger than 2 weeks were randomized to receive either the new infant formula (NF) or a standard formula (SF) until the age of 12 weeks . Anthropometric measurements were taken at enrollment, 6 weeks, and 12 weeks . In a subsample of infants, blood samples were taken at 6 weeks and stool samples were taken at enrollment and 6 weeks . Blood samples were analyzed for biochemical measures of protein status and amino acids, and stools were analyzed for total bacteria and bifidobacteria . Mothers completed a feeding diary and questionnaire at 6 and 10 weeks . RESULTS: One hundred fifty-four infants were enrolled in the study; 102 completed the trial . The growth of infants in both formula groups was in line with published growth curves . During the first 6 weeks, NF girls gained more weight and head circumference than the SF girls . These velocity differences were not maintained throughout the 12-week study period . The NF stools had a higher proportion of bifidobacteria at 6 weeks compared with the SF stools, and they were softer . There were no clinically significant differences in the blood biochemical and amino acid values between groups . Both formulas were well tolerated by the infants . CONCLUSIONS: When compared with a standard infant formula, the new formula supported satisfactory growth, led to higher counts of bifidobacteria in the feces, produced blood bio-chemical values typical of formula-fed infants, and was well tolerated.

Cir Pediatr, 2002 Oct, 15(4), 162 - 5
{Beneficial effects of Bifidobacterium lactis in the prevention of bacterial translocation in experimental short bowel syndrome}; Garcia-Urkia N et al.; BACKGROUND: Probiotics are live organisms that survive passage through the gastrointestinal tract and have beneficial effects on the host . Lactobacillus and Bifidobacterium have been recommended in cholesterol lowering, acute diarrhea, prevention of cancer or inflammatory bowel disease . On the other hand, after massive bowel resection bacterial overgrowth is frequent and favours the occurrence of bacterial translocation (BT) . The possible beneficial effects of Bifidobacterium lactis (BL) administration on BT in experimental short bowel syndrome (SBS), have not been investigated . AIM: To test the hypothesis that BL administration decreases BT in SBS in animals fed orally . METHODS: Thirty-two adult Wistar rats fed orally with standard rat chow and tap water "ad libitum" were maintained individual metabolic cages for ten days after 80% gut resection from the duodeno-jejunal angle to 10 cm above the cecum and divided in two groups: -Group A (n = 14): served as control . -Group B (n = 18): daily 7.8 x 109 CFU Bifidobacterium lactis administration, after orgastric intubation . At the end of the experiment they were sacrificed and mesenteric lymph nodes (MLN), and peripheral and portal blood specimens were recovered and cultured . Bacterial identification in blood was made by conventional methods and MLN culture was considered positive with a growth over 100 CFU/g . RESULTS: Bacterial translocation was detected in 93% of Group A rats . The incidence of BT in Group B was 44% . The relative risk reduction (RRR) was 0.52 (95% confidence interval 0.23-0.81) and the number needed to treat (NNT) was 2 (95% confidence interval between 1-5) . CONCLUSION: Administration of Bifidobacterium lactis reduces the incidence of BT in adult Wister rats, after 80% gut resection.

Syst Appl Microbiol, 2002 Dec, 25(4), 483 - 90
Lactic acid bacteria isolated from dairy products inhibit genotoxic effect of 4-nitroquinoline-1-oxide in SOS-chromotest; Cenci G et al.; Antigenotoxic activity against 4-nitroquinoline-1-oxide (4-NQO) of lactic acid bacteria isolated from commercial dairy products was studied using SOS-Chromotest . The supernatants from bacteria-genotoxin co-incubations in general exhibited a strong suppression on SOS-induction produced by 4-NQO on the tester organism Escherichia coli PQ37 (sfiA::lacZ) . High genotoxicity inhibition (>75%) was found for 31/67 of the examined bacteria and the maximum values of some strains within the species were as follows: Lactobacillus casei, 99.1%; L . plantarum, 93.3%; L . rhamnosus, 93.4%; L . acidophilus, 90.9%; L . delbrueckii subsp . bulgaricus, 85.7% and Bifidobacterium bifidum, 89.6%; Strains with low antigenotoxicity (5-60%) were evidenced in both L . acidophilus and L . delbrueckii subsp . bulgaricus, whereas some inactive strains were found only in L . casei and L . delbrueckii subsp . bulgaricus . Cell exposure to 100 degrees C for 15 min prevented antigenotoxicity and no effect was evidenced for cell-free spent media . The active strains survived at 0.1 mM 4-NQO exposure and generally presented some relevant functional properties, such as tolerance to bile (0.5%) or acid environment (pH 2.0) and adherence to Caco-2 enterocytes . Antigenotoxicity was always associated with modification of the 4-NQO absorbance profile.

Curr Pharm Des, 2003, 9(4), 333 - 46
Modification of intestinal flora in the treatment of inflammatory bowel disease; Kanauchi O et al.; Because the intestinal microflora play an important role in the development of inflammatory bowel disease (IBD), there is currently some interest in the manipulation of the composition of the microflora towards a potentially more remedial community . This review summarizes the clinical and experimental efficacy of the manipulation of microflora by the use of prebiotics, probiotics, synbiotics, and antibiotics in IBD . Prebiotics, defined as nondigestible food ingredients that beneficially affect the host by selectively stimulating the growth or activity of one or a limited number of bacterial species already resident in the colon, can modulate the colonic microbiota by increasing the number of specific bacteria and thus changing the composition of the microbiota . Prebiotics for IBD include lactosucrose, oligofructose, inulin, bran, psyllium, and germinated barley foodstuff (GBF) . GBF, which mainly consists of dietary fiber and glutamine-rich protein, is a prebiotic foodstuff for ulcerative colitis . GBF has shown to be converted into a preferential nutrient for colonocytes through Eubacterium and Bifidobacterium and also inactivate nuclear factor kappa B (NFkB) . Moreover, it exhibits a potent water-holding capacity and bile-acid binding capacity . Probiotics, which are microbial food supplements that beneficially affect the host by improving the intestinal microbial balance, have been used to change the composition of colonic microbiota . The approaches for IBD include VSL#3, Nissle1917, Clostridium butyricum and Bifidobacterium-fermented milk . Use of Lactococci secreting IL-10 provides excellent results . The combination of prebiotics and probiotics in a synbiotic has not been studied in IBD but is promising . The use of antibiotics continues to be of interest . Although these strategies hold great promise and appear to be useful in some settings, more clinical study is needed to firmly establish the relevance of these therapies.

Int J Food Microbiol, 2003 Apr 25, 82(2), 191 - 8
Characterisation of a Bifidobacterium strain with acquired resistance to cholate--a preliminary study; Margolles A et al.; The susceptibility levels against bile salts (ox gall) and sodium cholate of 19 Bifidobacterium strains from different origins ranged between 0.125% and 2.0% and 0.05% and 0.4%, respectively . Seven of these strains were subsequently selected for the isolation of sodium cholate resistant derivatives by exposure to gradually increasing concentrations of this compound . Derivatives resistant to at least 1.2% of cholate were obtained from Bifidobacterium bifidum CECT 4549 and B . bifidum M6 . Further analysis of the CECT 4549 derivative indicated that the phenotype induced was stable; the enzymatic activities analysed remained unchanged and no major rearrangements of chromosomal DNA were produced . However, the cells of the resistant derivative were smaller and more regular in average size than those of the original strain . In addition, the derivative had lost the ability to use lactose, fructose and galactose, and showed reduced expression levels of two membrane proteins of 78 and 114 kDa . Finally, the resistant strain displayed higher survival and lower growth inhibition in the presence of cholate than the original strain.

Wei Sheng Wu Xue Bao, 2002 Feb, 42(1), 56 - 61
{Amplification of bacterial heat shock protein 60 gene using inverse PCR method}; Jian W et al.; A method is presented for rapid in vitro amplification of DNA sequences of bacterial HSP60 gene . In our previous work, using degenerate oligonucleotide primers for conserved regions of HSP60 gene, a 600 bp fragment was amplified by PCR, cloned and sequenced . An inverse PCR method, with the primers oriented in the reversed direction of the usual orientation, is used to amplify the DNA sequences that flank the 600 bp known region . The feasibility of this method is shown by amplifying the complete HSP60 gene sequences of 4 bacterial strains, namely Bifidobacterium denticolens, Bifidobacterium inopinatum, Bifidobacterium adolescentis and Gardnerella vaginalis.

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