Bifidobacterium

Eur J Clin Nutr, 2002 Dec, 56 Suppl 4, S16 - 20
The role of fermented milk in complementary feeding of young children: lessons from transition countries; Branca F et al.; Probiotic bacteria are used for production of fermented dairy products . The use of probiotic bacteria has the potential to replenish the natural intestinal flora of the body . These bacteria competitively inhibit the growth and colonization of pathogenic bacteria . Breastmilk is the best food for babies, also from a probiotic point of view . Human milk, in fact, contains many substances that stimulate the growth of bifidobacteria in vitro and in the small intestine of infants . Improvement of lactose digestion and avoidance of symptoms of intolerance in lactose malabsorbers are the most profoundly studied health-relevant effects of fermented milk . In fact fermented milks are nutritionally similar to unfermented milk, except that some of lactose is broken down to glucose and galactose . The role of fermented milk in complementary feeding and in particular for the prevention of anaemia is an innovative theme, recently focused . Iron deficiency in infants and young children is widespread and has serious consequences for child health . Prevention of iron deficiency should therefore be given high priority . The too-early introduction of unmodified cow's milk and milk products is an important nutritional risk factors for the development of iron-deficiency anaemia . Fermented milks represent an excellent source of nutrients such as calcium, protein, phosphorus and riboflavin . During the fermentation of milk, lactic acid and other organic acids are produced and these increase the absorption of iron . If fermented milk is consumed at mealtimes, these acids are likely to have a positive effect on the absorption of iron from other foods.

Eur J Clin Nutr, 2002 Dec, 56 Suppl 4, S2 - S15
Fermented milks: a historical food with modern applications--a review; Tamime AY; BACKGROUND:: This paper was presented at the symposium which was organized by Instituto Danone Mexico in May 2001, and it provides an overview of the current scientific knowledge on fermented milks concerning the historical developments, manufacturing stages, classification of such products, and nutritional aspects . Particular attention has been paid to the human health benefits associated with the consumption of these products, the use of probiotic starter cultures and their industrial applications, and the significance of using a trained sensory panel for the evaluation of probiotic fermented milks made with different commercial blends of starter cultures . The paper also highlights the future research areas for the exploitation of starter microflora (Lactobacillus, Bifidobacterium and Enterococcus species) in fermented milk products . CONCLUSION:: This review indicates that the complex metabolism of the starter cultures is well established; however, more information is still needed on specific microbial metabolites such as polymerization of milk sugars for the production of exopolysaccharides and the modification of the milk peptides and secretion of bacteriocins . More clinical studies are still required to establish the 'functional' health benefits of probiotic fermented milks to humans.

Wei Sheng Wu Xue Bao, 1998 Oct, 38(5), 341 - 7
{Numerical taxonomy of bifidobacteria and homology analysis of some representative strains}; Ling D et al.; Fifty five strains of bifidobacteria belonging to Bifidobacterium from various sources were studied by using numerical taxonomic techniques . These organisms included 13 reference strains and 42 newly isolated strains (19 strains of human sources, 25 strains of animal sources and 11 strains obtained from sewage) . Several strains isolated from the animals weren't isolated before . Overall similarities of the strains for 75 characters contained morphological, physiological and biochemical characteristics and antibacterial susceptibility were determined . Dendrogram of numerical analysis based on unweighted pair cluster average linkage were achieved . Five clusters were defined at 70% Sm level . The relationships of these strains among five clusters were analysed . The strains of human sources were basically separated from the animal strains, and the sewage strains were distributed between the animal clusters and the human clusters . On the basis of the numerical analysis, the G + C content of DNA of some representative strains among these clusters were measured . Biotin-labeled DNA segments, complementary to variable regions of 16S rRNA, were amplified using PCR as probes . In the dot-blot hybridization the relationships of DNA segments among some strains were discussed . The test results showed that the homologies among the strains in same species and various species were different . The studies are valuable for further research in the relationships of some species among bifidobacteria of various sources.

Cancer Gene Ther, 2003 Feb, 10(2), 105 - 11
Bifidobacterium adolescentis as a delivery system of endostatin for cancer gene therapy: selective inhibitor of angiogenesis and hypoxic tumor growth; Li X et al.; In order to overcome difficulties that hampered widespread application of antiangiogenesis in cancer therapy, a highly specific delivery system may be engaged in vivo to deliver and express antiangiogenic genes . We selected a strain of Bifidobacterium adolescentis (B . adolescentis) as the delivery system to transport endostatin gene to solid tumors . B . adolescentis with endostatin gene were injected into tumor-bearing mice through the tail vein . After the mice were sacrificed, the tumor and some normal tissues of the mice were examined . B . adolescentis were only found in the tumors and no bacilli were found in other normal tissues . Also, a strong inhibition of angiogenesis had been shown to inhibit local tumor growth in the administrated group . These results suggested that B . adolescentis only germinated and proliferated in solid tumors and might be a highly specific and efficient vector for transporting anticancer genes into target tumor in cancer gene therapy.

Zh Mikrobiol Epidemiol Immunobiol, 2002 Sep-Oct, (5), 98 - 104
{Characteristics of microorganisms colonizing human intestine}; Efimov BA et al.; Introduction of novel methods of microbial diagnostics has considerably broadened our conceptions on the qualitative and quantitative variety of microorganisms inhabiting human gastrointestinal tract . In this review morphological and functional properties of obligate anaerobic bacteria (bifidobacteria, lactobacilli, eubacteria, peptostreptococci, clostridia, bacteroids, fusobacteria) and facultative anaerobic microorganisms (enterobacteria, staphylococci, streptococci, yeast-like fungi of the genus Candida) capable of colonizing human intestine are briefly characterized.

Microbiol Immunol, 2002, 46(11), 781 - 5
Stimulation of the secretion of pro-inflammatory cytokines by Bifidobacterium strains; He F et al.; To characterize the ability of bifidobacteria to affect the production of macrophage-derived cytokines, a murine macrophage-like cell line, J774.1, was cultured in the presence of 27 strains of heat-inactivated bifidobacteria . Bifidobacterium adolescentis and B . longum, known as adult-type bifidobacteria, induced significantly more pro-inflammatory cytokine secretion, IL-12 and TNF-alpha, by J774.1 cells, than did the infant-type bifidobacteria, B . bifidum, B . breve, and B . infantis (P<0.01) . In contrast, B . adolescentis did not stimulate the production of anti-inflammatory IL-10 from J774.1 cells as the other tested bacteria did . The results suggest that the adult-type bifidobacteria, especially B . adolescentis, may be more potent to amplify but less able to down-regulate the inflammatory response.

Appl Environ Microbiol, 2003 Jan, 69(1), 24 - 32
Induction of sucrose utilization genes from Bifidobacterium lactis by sucrose and raffinose; Trindade MI et al.; The probiotic organism Bifidobacterium lactis was isolated from a yoghurt starter culture with the aim of analyzing its use of carbohydrates for the development of prebiotics . A sucrose utilization gene cluster of B . lactis was identified by complementation of a gene library in Escherichia coli . Three genes, encoding a sucrose phosphorylase (ScrP), a GalR-LacI-type transcriptional regulator (ScrR), and a sucrose transporter (ScrT), were identified by sequence analysis . The scrP gene was expressed constitutively from its own promoter in E . coli grown in complete medium, and the strain hydrolyzed sucrose in a reaction that was dependent on the presence of phosphates . Primer extension experiments with scrP performed by using RNA isolated from B . lactis identified the transcriptional start site 102 bp upstream of the ATG start codon, immediately adjacent to a palindromic sequence resembling a regulator binding site . In B . lactis, total sucrase activity was induced by the presence of sucrose, raffinose, or oligofructose in the culture medium and was repressed by glucose . RNA analysis of the scrP, scrR, and scrT genes in B . lactis indicated that expression of these genes was influenced by transcriptional regulation and that all three genes were similarly induced by sucrose and raffinose and repressed by glucose . Analysis of the sucrase activities of deletion constructs in heterologous E . coli indicated that ScrR functions as a positive regulator.

Int J Syst Evol Microbiol, 2002 Nov, 52(Pt 6), 1945 - 51
Unification of Bifidobacterium infantis and Bifidobacterium suis as Bifidobacterium longum; Sakata S et al.; The relationships between Bifidobacterium infantis, Bifidobacterium longum and Bifidobacterium suis were examined by means of carbohydrate fermentation, DNA-DNA hybridization, ribotyping and random amplified polymorphic DNA-PCR (RAPD-PCR) . The levels of DNA-DNA hybridization among the strains of B . infantis, B . longum and B . suis used in this study were 67-81% under optimal conditions (42 degrees C) and 63-85% under stringent conditions (52 degrees C) . Although the strains showed varied carbohydrate-fermentation patterns, the three species were divided into three types, namely the infantis type, the longum type and the suis type, by ribotyping and RAPD-PCR . On the basis of these results, strains of B . infantis, B . longum and B . suis were recognized as distinct groups within a single species . It is concluded that B . infantis and B . suis should be unified as B . longum, the latter species being divided into three biotypes, the infantis type, the longum type and the suis type, by molecular methods.

Biosci Biotechnol Biochem, 2002 Nov, 66(11), 2362 - 6
Cloned cytosine deaminase gene expression of Bifidobacterium longum and application to enzyme/pro-drug therapy of hypoxic solid tumors; Nakamura T et al.; Bifidobacterium longum is a nonpathogenic anaerobic bacterium among normal bacterial flora . Recently, it was reported that B . longum accumulated in hypoxic solid tumors . The gene of interest was expressed in transfected B . longum by the shuttle vector pBLES100 in solid tumors . In this report, we constructed pBLES100-S-eCD, which included the cytosine deaminase gene . We confirmed by western blotting that transfected B . longum produced cytosine deaminase . In addition, transfected B . longum produced cytosine deaminase that converted 5-fluorocytosine into 5-fluorouracil . B . longum could be useful for enzyme/pro-drug therapy of hypoxic solid tumors.

Int J Food Microbiol, 2003 Jan 26, 82(1), 59 - 70
Bacterial composition of commercial probiotic products as evaluated by PCR-DGGE analysis; Fasoli S et al.; The use of Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) technique in identifying the microorganisms present in commercial probiotic yoghurts and lyophilised products was evaluated . Two reference ladders were assembled constituted by PCR-amplified V2-V3 regions of 16S rDNA from bacterial species generally used as probiotics . Identification was achieved comparing the PCR-DGGE patterns obtained from the analysed products with the ladder bands . Bands from members of the same species showed the same migration distance in denaturing gel, hence supporting the identificative value of the method . The validity of the technique was also proven confirming the PCR-DGGE identification results by sequence data analysis and by species-specific PCR . General congruence between microorganisms declared on the label and those revealed by PCR-DGGE was found for probiotic yoghurts . Conversely, some discrepancies were observed for probiotic lyophilised preparations, i.e . the incorrect identification of some Bifidobacterium and Bacillus species and the presence of not declared microorganisms . PCR-DGGE turned out to be an appropriate culture-independent approach for a rapid detection of the predominant species in mixed probiotic cultures.

Folia Microbiol (Praha), 2002, 47(5), 477 - 80
Specific growth rate of bifidobacteria cultured on different sugars; Rada V et al.; The ability of six bifidobacterial strains (3 of human origin and 3 isolates from fermented milk products) to utilize glucose, lactose, melezitose, sucrose, raffinose, and stachyose was determined . Dairy-related bifidobacterial strains were identified as Bifidobacterium animalis (2 strains) or as B . pseudolongum (1 strain) . Human strains included B . longum (2 strains) and B . breve (1 strain) . All strains fermented lactose, sucrose, raffinose, and stachyose . Melezitose was utilized only by B . longum . B . pseudolongum did not ferment either glucose or melezitose . All isolates had a higher specific growth rate on raffinose and stachyose than on glucose . Dairy strain grew slowly on glucose compared to human strains.

J Gen Appl Microbiol, 1998 Dec, 44(6), 375 - 380
A comparison of features and the microbial constitution of the fresh feces of pigs fed diets supplemented with or without dietary microbes; Nagamine I et al.; The features and the constitution of the microbial population of fresh feces were compared between pigs fed a diet supplemented with dietary microbes and pigs given nonsupplemented diets . The former were reared on farm C and the latter on farms A and B . The concentrations of ammonia-N, indole, and skatole of fresh feces were not significantly different between pigs reared on farm C and those raised on farms A and B, but the concentrations of ammonia-N and the skatole of fresh feces were significantly different between pigs reared on farms A and B . The total VFA (volatile fatty acids) concentration of fresh feces in pigs on farm C was slightly lower than in those on farms A and B . Moreover, the molar proportion of the acetic acid in feces in pigs on farm C was lower; inversely, that of propionic and butyric acids was higher in comparison with those on farms A and B . No differences were evident in the total viable counts of feces among pigs reared on the three different farms . Clostridium perfringens was abundant in feces of pigs raised on farms A and B, but it was not detected in pigs reared on farm C . Megasphaerae, bifidobacteria, and clostridia except for C . perfringens were more abundant in the feces of pigs fed a diet supplemented with dietary microbes on farm C, compared with pigs given the nonsupplemented diets on farms A and B.

Curr Opin Clin Nutr Metab Care, 2003 Jan, 6(1), 49 - 54
Gut changes attributed to ageing: effects on intestinal microflora; Hebuterne X; PURPOSE OF REVIEW: There is increased evidence of several impaired gastrointestinal functions with ageing . In the elderly, however, most gastrointestinal functions remain relatively intact because of the large reserve capacity of the intestine and the great secretion capacity of the pancreas . This review will focus on changes in gut microflora observed in the elderly and on the potential benefit of probiotics in this population . RECENT FINDINGS: Recent studies suggest that age affects the intestinal microflora with a decrease in anaerobes and bifidobacteria population and an increase in enterobacteria . These changes and the reduced intestinal immunity of the aged may favour gastrointestinal infections that are frequent in the elderly . Clostridium difficile-associated diarrhoea, one of the most common nosocomial infections in the elderly, has a profound effect on morbidity, mortality and health costs . Probiotics may have interesting positive effects on intestinal function, and the efficacy of treatment with Lactobacilli and Saccharomyces boulardii in Clostridium difficile-associated diarrhoea has been well established in a recent meta-analysis . Studies performed in healthy elderly patients suggest that diet supplementation with probiotics may reduce the impaired immunity associated with ageing . SUMMARY: Important changes in intestinal microflora of the elderly have recently been demonstrated and may have important clinical consequences . Further studies should be conducted to determine if the consumption of probiotics is associated with a lower infection rate and a higher effectiveness of vaccines.

J Appl Microbiol, 2003, 94(1), 138 - 45
Conjugated linoleic acid biosynthesis by human-derived Bifidobacterium species; Coakley M et al.; AIMS: To assess strains of Lactobacillus, Lactococcus, Pediococcus and Bifidobacterium for their ability to produce the health-promoting fatty acid conjugated linoleic acid (CLA) from free linoleic acid . METHODS AND RESULTS: In this study, strains of Lactobacillus, Lactococcus, Pediococcus and Bifidobacterium were grown in medium containing free linoleic acid . Growth of the bacteria in linoleic acid and conversion of the linoleic acid to CLA was assessed . Of the bacteria assessed, nine strains of Bifidobacterium produced the c9, t11 CLA isomer from free linoleic acid . The t9, t11 CLA isomer was also produced by some strains, but at much lower concentrations . CONCLUSIONS: The production of CLA by bifidobacteria exhibited considerable interspecies variation . Bifidobacterium breve and B . dentium were the most efficient CLA producers among the range of strains tested, with B . breve converting up to 65% linoleic acid to c9, t11 CLA when grown in 0.55 mg ml(-1) linoleic acid . Strains also varied considerably with respect to their sensitivity to linoleic acid . SIGNIFICANCE AND IMPACT OF THE STUDY: The production of CLA by probiotic bifidobacteria offers a possible mechanism for some health-enhancing properties of bifidobacteria and provides novel opportunities for the development of functional foods.

Vet Microbiol, 2003 Mar 20, 92(1-2), 111 - 9
Interaction between probiotic lactic acid bacteria and canine enteric pathogens: a risk factor for intestinal Enterococcus faecium colonization?
Rinkinen M, Jalava K, Westermarck E, Salminen S, Ouwehand AC.
Selected probiotic lactic acid bacteria (LAB) have been shown to elicit positive health effects particularly in humans . Competitive exclusion of pathogens is one of the most important beneficial health claims of probiotic bacteria . The effect of probiotic LAB on competitive exclusion of pathogens has been demonstrated in humans, chicken and pigs . In this study we evaluated the ability of certain LAB strains (Lactobacillus rhamnosus GG, Bifidobacterium lactis Bb12, Lactobacillus pentosus UK1A, L . pentosus SK2A, Enterococcus faecium M74 and E . faecium SF273) to inhibit the adhesion of selected canine and zoonotic pathogens (Staphylococcus intermedius, Salmonella Typhimurium ATCC 14028, Clostridium perfringens and Campylobacter jejuni) to immobilised mucus isolated from canine jejunal chyme in vitro . Adhesion of C . perfringens was reduced significantly by all tested LAB strains, between 53.7 and 79.1% of the control without LAB, the LAB of canine origin yielding the best reduction . The adhesion of S . Typhimurium and S . intermedius were not significantly altered by any of the LAB included in the study . Both enterococci tested significantly enhanced the adhesion of C . jejuni, to 134.6 and 205.5% of the control without LAB . E . faecium may thus favor the adhesion and colonization of C . jejuni in the dog's intestine, making it a potential carrier and possibly a source for human infection . Enhanced C . jejuni adhesion is a new potential risk factor of enterococci . Our results further emphasize the importance of safety guidelines to be established for the probiotics intended for animal use.

Oral Microbiol Immunol, 2002 Dec, 17(6), 344 - 53
Cytokine production in response to endodontic infection in germ-free mice; Ribeiro Sobrinho AP et al.; This study evaluated the cytokine profiles (type 1 or type 2) that are triggered by and modulate endodontic periapical infections in the root canal system of germ-free mice . Microorganisms isolated from two patients with pulpal necrosis were inoculated into two groups of experimental animals: group I (Gemella morbillorum) and group II (Bifidobacterium adolescentis, Fusobacterium nucleatum and Clostridium butyricum) . In vitro, G . morbillorum induced type 1 cytokine synthesis, while the modulation processed in vivo seemed to have the opposite effect, with a reduction in the basal levels of IL-12 and IFN-gamma, IL-4-independent down-modulation . In vitro, microorganisms from group II, in poly-infection, induced a reduction of type 1 cytokine levels from day 10 to day 20, which seemed to be modulated via IL-4 . In vivo, however, a predominance of the immune response to one species over the others occurred.

Cell Immunol, 2002 Sep, 219(1), 22 - 7
Encapsulated Bifidobacterium bifidum potentiates intestinal IgA production; Park JH et al.; We asked whether Bifidobacterium bifidum regulates the synthesis of IgA by mucosal lymphoid cells . B . bifidum alone, but not Clostridium perfringens, significantly induced total IgA and IgM synthesis by both mesenteric lymph nodes (MLN) and Peyer's patch (PP) cells . We, further, investigated the mucosal antibody production following peroral administration of B . bifidum to mice . Ingested B . bifidum significantly increased the number of Ig (IgM, IgG, and IgA) secreting cells in the culture of both MLN and spleen cells . Nonetheless, B . bifidum itself does not induce the own specific antibody responses, implying that B . bifidum does not provoke unnecessary immune reaction . Subsequently, it was found that encapsulation of B . bifidum further augments the total IgA production in the culture of both MLN and spleen cells . Finally, we found that the immuno-stimulating activity of B . bifidum is due to its cellular components but not due to any actively secreting component(s) from bacteria.

Clin Nutr, 2002 Dec, 21(6), 527 - 30
A novel synbiotic therapy dramatically improved the intestinal function of a pediatric patient with laryngotracheo-esophageal cleft (LTEC) in the intensive care unit; Kanamori Y et al.; To demonstrate the beneficial effects of synbiotic therapy (combined use of probiotics and prebiotics) in critically ill patients, we designed a new protocol for such therapy.Three agents were used as synbiotics: Bifidobacterium breve, Lactobacillus casei, and galactooligosaccharides . More than 1 x 10(9) of probiotic bacteria were contained in each 1.0 g pack . We administered 3.0 g per day of each agent to the patient.A critically ill 9-month-old girl with laryngotracho-esophageal cleft (type IV) was treated by our new synbiotic therapy . Abundant amounts of synbiotic bacteria were detected in the feces which suggests that these administered bacteria affected intestinal function in situ . Bowel movements resumed soon after the commencement of synbiotic therapy and considerable amounts of short chain fatty acids were detected in the feces . Growth of the patient was satisfactory under this treatment.Our new synbiotic therapy had a beneficial effect to improve intestinal function . We recommend synbiotic therapy for critically ill patients in intensive care units as an important immunonutritional therapy.

Zhonghua Shao Shang Za Zhi, 2002 Aug, 18(4), 216 - 9
{A preliminary study of the postburn intestinal biological barrier injury in severely burned rats}; Chen J et al.; OBJECTIVE: To investigate the postburn change in the intestinal biological barrier in severely burned rats . METHODS: Wistar rats inflicted by 30% TBSA III degree scalding on the back were employed as the model . The samples were harvested at 24, 48, 72 and 96 postburn hours (PBHs), respectively with the employment of microorganism analysis, biochemical and radio-immune methods for the study . The membranous flora in cecum, the mucin and sIgA in intestinal content, the intestinal endotoxin and bacterial translocation rate and quantification analysis and the endotoxin content in cava vein were observed . RESULTS: The total intestinal membranous flora amount decreased, especially and obviously did the anaerobic bacteria such as bifidobacteria . But aerobic ones increased . In addition, The fungus and enterobacteria exhibited rapid overgrowth . This lead to evident imbalance between anaerobic and aerobic bacteria and to the destruction of intestinal biological barrier and the decrease of colonization resistance . As a result, the intestinal bacterial translocation rate increased markedly . The endotoxin content in the cava and intestinal containing increased, while the mucin and sIgA contents decreased . CONCLUSION: Intestinal biological barrier could be severely damaged after major burn, which might be one of the causes of postburn intestinal infection.

Lett Appl Microbiol, 2002, 35(6), 462 - 7
Characterization of a purified beta-fructofuranosidase from Bifidobacterium infantis ATCC 15697; Warchol M et al.; AIMS: To characterize the beta-fructofuranosidase of Bifidobacterium infantis ATCC 15697 and to compare it with other bacterial beta-fructofuranosidases . METHODS AND RESULTS: The beta-fructofuranosidase of B . infantis ATCC 15697 was purified 46.8 times over the crude extract by anion exchange chromatography, ultrafiltration and gel filtration . The sequence of 15 amino acid residues of the NH2 terminal was determined . This enzyme was a monomeric protein (Mr 70 kDa) with beta-fructofuranosidase and invertase activities . The isoelectric point was pH 4.3, the optimum pH 6.0 and pKas (4.5 and 7.2) of two active groups were obtained . The activities were inhibited by Hg2+ and p-chloromercuribenzoic acid (pCMB) . The optimal temperature was 37 degrees C and activities were unstable at 55 degrees C . beta-fructofuranosidase activity was more efficient than that of invertase with Vm/Km ratios of 0.65 and 0.025 x 10-3 l min(-1) mg(-1), respectively . The enzyme catalyses the hydrolysis of fructo-oligosaccharides, sucrose and inulin at relative velocities of 100, 10 and 6, respectively . CONCLUSIONS: The enzyme of B . infantis ATCC 15697 is an exo-inulinase which has beta-fructofuranosidase and invertase activities . This protein was different from the beta-fructofuranosidase of another strain of B . infantis (B . infantis JCM no . 7007) . SIGNIFICANCE AND IMPACT OF THE STUDY: A better knowledge of bacterial beta-fructofuranosidases, especially from bifidobacteria, has been gained.

Int J Food Microbiol, 2003 Mar 15, 81(2), 131 - 6
Evaluation of Bifidobacterium breve strain Yakult-fermented soymilk as a probiotic food; Shimakawa Y et al.; We evaluated the effects of Bifidobacterium breve-fermented soymilk on probiotic function . An administered strain of B . breve strain Yakult was capable of growing in soymilk with no additives as high as 10(9) CFU/ml . During storage of the fermented soymilk at 10 degrees C for 20 days, viable counts of the strain did not change . The growth inhibition of the strain in a bile-containing medium was lessened by the addition of soy protein . In human feeding experiments, the administered B . breve was recovered at a level of over 10(9) CFU/g faeces, accompanied by an increase in the total number of bifidobacteria . These results indicate that fermented soymilk with B . breve strain Yakult could be a novel type of probiotic food.

Appl Environ Microbiol, 2002 Dec, 68(12), 6429 - 34
Rapid identification, differentiation, and proposed new taxonomic classification of Bifidobacterium lactis; Ventura M et al.; Identification of Bifidobacterium lactis and Bifidobacterium animalis is problematic because of phenotypic and genetic homogeneities and has raised the question of whether they belong to one unique taxon . Analysis of the 16S-23S internally transcribed spacer region of B . lactis DSM10140(T), B . animalis ATCC 25527(T), and six potential B . lactis strains suggested two distinct clusters . Two specific 16S-23S spacer rRNA gene-targeted primers have been developed for specific detection of B . animalis . All of the molecular techniques used (B . lactis or B . animalis PCR primers, enterobacterial repetitive intergenic consensus PCR) demonstrated that B . lactis and B . animalis form two main groups and suggest a revision of the strains assigned to B . animalis . We propose that B . lactis should be separated from B . animalis at the subspecies level.

Zh Mikrobiol Epidemiol Immunobiol, 2002 Jul-Aug, (4), 72 - 8
{Modern methods for the evaluation of qualitative and quantitative changes in the characteristics of intestinal and vaginal microflora}; Efimov BA et al.; Disturbances in normal intestinal and vaginal microflora in women have recently become quite frequent . This accounts for the need of bacteriological laboratories for introduction of reliable methods for the diagnosis of such disturbances . Correct methodological approaches to objective evaluation of the state of intestinal and vaginal microflora are described . The methods used in the study of anaerobic microflora (lacto- and bifidobacteria, eubacteria, peptostreptococci, clostridia, bacteriods, fusobacteria) and facultative anaerobic microorganisms (enterobacteria, staphylococci, streptococci, Gardnerella, fungi of the genus Candida) have been analyzed . All stages of the study are described in consecutive order: the transportation of the material under study, its treatment in a laboratory, the spectrum of selective nutrient media for the isolation of microorganisms, methods of their identification.

Arch Pharm Res, 2002 Oct, 25(5), 681 - 4
Hypocholesterolemic activity of Bifidobacteria isolated from a healthy Korean; Rhee YK et al.; This study was undertaken to investigate the hypocholesterolemic activity of Bifidobacteria (B . breve K-110, B . breve K-111, and B . infantis K-525) isolated from a healthy Korean . The administration of B . breve K-110 and K-111 with a high cholesterol diet significantly protected the increase of serum total cholesterol and LDL cholesterol relative to that of a high cholesterol diet alone . Such a diet supplemented with 0.5% B . breve K-111 decreased serum total cholesterol and LDL cholesterol to 57 and 55%, respectively . The administration of Bifidobacteria also significantly inhibited the lipid-deposited surface in the aorta . The normalizing activity of serum cholesterol level in cholesterolemic rats was accelerated by Bifidobacteria . The normalizing activity of B . breve K-111 on serum cholesterol level was superior to that of B . breve K-110 . These results suggest that Bifidobacteria in the human intestine play a role in the prophylactics of arteriosclerosis.

Int J Antimicrob Agents, 2002 Nov, 20(5), 313 - 9
The place of probiotics in human intestinal infections; Sullivan A et al.; A number of studies have been carried out on the effect of several probiotic species on treatment and prevention of intestinal infections . The most commonly used microorganisms are lactic-acid producing bacteria such as lactobacilli and bifidobacteria belonging to the human normal microflora . In vitro and animal studies have shown that probiotic microorganisms interfere with the colonisation of Helicobacter pylori and of enteropathogenic microorganisms . In humans the significance is more uncertain . Clinically significant benefits of probiotics have been demonstrated in the treatment of rotavirus induced diarrhoea and of Saccharomyces boulardii in the prevention of antibiotic-associated diarrhoea (AAD) . In patients suffering from inflammatory bowel disease, several probiotic strains have been shown to be as effective as traditional medication in preventing relapses . Standardised and well performed studies are needed to elucidate further the mechanisms of action and the clinical significance of probiotics.

Avian Pathol, 2002 Feb, 31(1), 49 - 58
Dietary mannan-oligosaccharides and their effect on chicken caecal microflora in relation to Salmonella Enteritidis colonization; Fernandez F et al.; This study first investigates the effects of mash diet, or mash supplemented with either 2.5% mannose-oligosaccharide (MOS) or palm kernel meal (PKM), on the microflora of the hen caecal contents . Second, it investigates the effect of caecal contents of hens (HCC) fed mash or mash supplemented with MOS or PKM on the major microflora groups of chicks, and their inhibitory effect on Salmonella enterica serovar Enteritidis (PT4) colonization . Finally, this study investigates the effect over time of diets supplemented with MOS or PKM on S . Enteritidis colonization and the microflora of chicks . In hens, supplemented diets increased Bifidobacterium spp., while decreasing members of Enterobacteriaceae and Enterococcus spp., compared with the mash diet . Chicks dosed with the HCC showed, on average, increased numbers of anaerobes, while the numbers of aerobes decreased including coliforms and S . Enteritidis compared with controls without HCC . In chicks fed the MOS-supplemented or PKM-supplemented diets, S . Enteritidis colonization decreased over time, compared with mash alone . Four-week-old PKM birds showed an increase in Bifidobacterium spp . and Lactobacillus spp., with a decrease in S . Enteritidis compared with week 2 . Generally, the HCC and diets supplemented with MOS or PKM affected the birds intestinal microflora by increasing the Bifidobacterium spp . and Lactobacillus spp., while decreasing the Enterobacteriaceae groups . They also reduced susceptibility in young chickens to colonization by S . Enteritidis.

Pediatr Dev Pathol, 2003 Jan-Feb, 6(1), 6 - 23 Epub 2002 Nov 11.
Neonatal necrotizing enterocolitis: clinical considerations and pathogenetic concepts; Hsueh W et al.; Necrotizing enterocolitis (NEC), a disease affecting predominantly premature infants, is a leading cause of morbidity and mortality in neonatal intensive care units . Although several predisposing factors have been identified, such as prematurity, enteral feeding, and infection, its pathogenesis remains elusive . In the past 20 years, we have established several animal models of NEC in rats and found several endogenous mediators, especially platelet-activating factor (PAF), which may play a pivotal role in NEC . Injection of PAF induces intestinal necrosis, and PAF antagonists prevent the bowel injury induced by bacterial endotoxin, hypoxia, or challenge with tumor necrosis factor-a (TNF) plus endotoxin in adult rats . The same is true for lesions induced by hypoxia and enteral feeding in neonatal animals . Human patients with NEC show high levels of PAF and decreased plasma PAF-acetylhydrolase, the enzyme degrading PAF . The initial event in our experimental models of NEC is probably polymorphonuclear leukocyte (PMN) activation and adhesion to venules in the intestine, which initiates a local inflammatory reaction involving proinflammatory mediators including TNF, complement, prostaglandins, and leukotriene C4 . Subsequent norepinephrine release and mesenteric vasoconstriction result in splanchnic ischemia and reperfusion . Bacterial products (e.g., endotoxin) enter the intestinal tissue during local mucosal barrier breakdown, and endotoxin synergizes with PAF to amplify the inflammation . Reactive oxygen species produced by the activated leukocytes and by intestinal epithelial xanthine oxidase may be the final pathway for tissue injury . Protective mechanisms include nitric oxide produced by the constitutive (mainly neuronal) nitric oxide synthase, and indigenous probiotics such as Bifidobacteria infantis . The former maintains intestinal perfusion and the integrity of the mucosal barrier, and the latter keep virulent bacteria in check . The development of tissue injury depends on the balance between injurious and protective mechanisms.

Eur J Nutr, 2002 Nov, 41 Suppl 1, I32 - 7
The role of the intestinal microflora for the development of the immune system in early childhood; Ouwehand A et al.; The intestinal tract performs many different functions; in addition to absorption and digestion it is also the body's largest organ of host defence . Part of the intestinal mucosal barrier function is formed by a common mucosal immune system which provides communication between the different mucosal surfaces of the body . The intestine also contains a microbial ecosystem with a large body of microbes, 1-11/2 kg in an adult . The microbes and their activity have a major impact on the development and functioning of the intestinal immune system and vice versa . This mutual influence also affects the host beyond the intestine.The intestinal colonisation with a balanced microflora is of main importance for the correct development of the immune system . The importance of the intestinal microflora is most clearly seen in germfree animals, but also diseases like atopy are associated with disturbances in the intestinal microflora . This often manifests itself in a low number of bifidobacteria . The use of probiotics or prebiotics to correct this imbalance and modulate the immune activity has received increasing scientific documentation . The precise mechanisms behind these immune modulatory activities are not well understood and require further investigation.

Eur J Nutr, 2002 Nov, 41 Suppl 1, I11 - 6
Relationship of prebiotics and food to intestinal microflora; Blaut M; Dietary carbohydrates that escape digestion in the small intestine, undergo bacterial fermentation in the colon . This process affects the microbial ecology of the gastrointestinal tract and influences gut metabolism and function . Prebiotics are non-digestible but fermentable oligosaccharides that are specifically designed to change the composition and activity of the intestinal microbiota with the prospect to promote the health of the host . Dietary fiber and non-digestible oligosaccharides are the main growth substrates of gut microorganisms . Their fermentation results in the acidification of the colonic contents and the formation of short chain fatty acids which serve as fuels in different tissues and may play a role in the regulation of cellular processes . Prebiotics specifically stimulate the growth of endogenous microbial population groups such as bifidobacteria and lactobacilli which are perceived as being beneficial to human health . In spite of the interesting nutritional properties of prebiotics it is questionable whether a wholesome diet rich in fruit and vegetables needs to be supplemented with prebiotics for optimal health effects.

Carcinogenesis, 2002 Nov, 23(11), 1953 - 60
Antitumorigenic activity of the prebiotic inulin enriched with oligofructose in combination with the probiotics Lactobacillus rhamnosus and Bifidobacterium lactis on azoxymethane-induced colon carcinogenesis in rats; Femia AP et al.; Prebiotics such as fructans, and probiotics such as Lactobacilli or Bifidobacteria, or a combination of prebiotics and probiotics (synbiotics) are thought to be protective against colon cancer . Therefore, we studied whether the prebiotic inulin enriched with oligofructose (Raftilose-Synergy1, briefly, Synergy1, 10% of the diet), probiotics {Bifidobacterium lactis (Bb12) and Lactobacillus rhamnosus (LGG), each at 5x10(8) c.f.u./g diet} or synbiotics (a combination of the two) protect rats against azoxymethane (AOM)-induced colon cancer . Male F344 rats were divided into: Controls; PRE, which were fed a diet containing Synergy1; PRO, fed a diet containing LGG and Bb12; PREPRO, fed a diet containing Synergy1, LGG and BB12 . Ten days after beginning the diets, rats were treated with AOM (15 mg/kg s.c . two times); dietary treatments were continued for the entire experiment . Thirty-one weeks after AOM, rats treated with Synergy1 (PRE and PREPRO groups) had a significantly lower (P < 0.001) number of tumours (adenomas and cancers) than rats without Synergy1 (colorectal tumours/rat were 1.9 +/- 1.7, 1.1 +/- 1.1, 2.2 +/- 1.4 and 0.9 +/- 1.2 in Controls, PRE, PRO and PREPRO groups, respectively, means +/- SD) . A slight, not significant effect of probiotics in reducing malignant tumours was also observed (P = 0.079) . Caecal short-chain fatty acids (SCFA) were higher (P < 0.001) in the groups treated with Synergy1 . Apoptosis was increased in the normal mucosa of the PRO group, while no variation was observed in the tumours . Colonic proliferation was lower in the PRE group as compared with Controls . Glutathione S-transferase placental enzyme pi type expression, and to a lesser extent, inducible NO synthase were depressed in the tumours from rats in the PRE and PREPRO groups . Cycloxygenase-2 expression was increased in the tumours of control rats but not in those from PRE, PRO or PREPRO rats . In conclusion, prebiotic administration in the diet decreases AOM-induced carcinogenesis in rats.

Dig Liver Dis, 2002 Sep, 34 Suppl 2, S124 - 8
Prebiotics in infant formulas: biochemical characterisation by thin layer chromatography and high performance anion exchange chromatography; Coppa GV et al.; BACKGROUND: Breast-fed infants, unlike bottle-fed babies, have a microbic intestinal flora characterised by a marked predominance of bifidobacteria and lactic acid bacteria . This is essentially due to the prebiotic effect of oligosaccharides in human milk . Recently, oligosaccharides with a prebiotic effect have been added to formulas . Aim . To characterise the mixture of oligosaccharides contained in these new formulas . MATERIALS AND METHODS: The characterisation of oligosaccharides was performed using thin layer chromatography as well as high performance anion exchange chromatography . RESULTS: The mixture of oligosaccharides used in the formulas analysed was made up of oligosaccharides with low molecular weight (transgalactosylated oligosaccharides) and polysaccharides with high molecular weight (inulin) . CONCLUSION: With the methods employed, it was possible to characterise the mixture of oligosaccharides used as prebiotics in the formulas now available on the market.

Dig Liver Dis, 2002 Sep, 34 Suppl 2, S105 - 10
Functional food concept and its application to prebiotics; Roberfroid M; A food can be regarded as functional if it is satisfactorily demonstrated to affect beneficially one or more target functions in the body, beyond adequate nutritional effects, in a way which is relevant to either the state of wellbeing and health or the reduction of the risk of a disease . A food can be made functional by increasing the concentration, adding or improving the bioavailability of a particular component . Functional food science will serve to establish claims based either on enhanced function or disease risk reduction . Inulin and oligofructose are functional food ingredients present in miscellaneous edible plants . They are non-digestible oligosaccharides classified as dietary fibres . The target for their functional effects is the colonic microflora that ferment them and for which they serve as selective "fertilizers"; the gastrointestinal physiology; the immune functions; the bioavailability of minerals; the metabolism of lipids; and colonic carcinogenesis . The scientific data available on the nutritional effects of inulin and oligofructose provide strong evidence for a prebiotic effect (i.e., selective stimulation of growth of bifidobacteria in colonic microbiota), improvement of bowel habit (both stool frequency and stool weight) and improved calcium bioavailability.

Dig Liver Dis, 2002 Sep, 34 Suppl 2, S78 - 80
Probiotics in antibiotic-associated diarrhoea; Cremonini F et al.; Antibiotic-associated diarrhoea is a common event . In some cases, it could represent a life-threatening event . Clostridium difficile colitis is a further distinct complication of antibiotic administration . Treatment options for antibiotic-associated diarrhoea and Clostridium difficile colitis include supplementation with several types of probiotics, as overviewed in this paper . Three randomised, double-blind, controlled clinical trials show a therapeutic effect of Saccharomyces boulardii in antibiotic-associated diarrhoea . The efficacy of Lactobacillus acidophilus and bulgaricus has also been ascertained in two double-blind controlled studies . Other studies focusing on Lactobacillus as a new preventive agent for antibiotic-associated diarrhoea are not double-blind . Among these, a positive effect of Lactobacillus rhamnosus GG, Bifidobacterium longum and Enterococcus faecium SF68 has been reported . Effectiveness of probiotics in antibiotic-associated diarrhoea has, therefore, a consistent scientific rationale, however few studies have performed an assessment of bacterial recovery in stools, and this approach may be helpful in deciding a more rigorous dose standardisation.

Dig Liver Dis, 2002 Sep, 34 Suppl 2, S25 - 8
Novel probiotics for the management of allergic inflammation; von der Weid T et al.; Several pathologies of the gastrointestinal tract, particularly food allergy, are due to an exaggerated and imbalanced response of the gut mucosal immune system . The intestinal microflora is an important constituent of the gut mucosal barrier against food allergens and there is increasing evidence that one important acquired factor predisposing to food allergy in infants is the gut microflora . Indeed, the balance of bifidobacteria versus Clostridia in the neonatal flora appears to determine the allergic status in infants . In earlier studies, it was shown that the higher prevalence of allergies in infants fed standard formulas, compared to breast-fed infants, correlated with lower frequencies of bifidobacteria in their faeces . Certain Lactobacillus probiotic strains can have an inhibitory impact on allergic inflammation . The mechanisms implicated are still unclear, but it seems that they can involve both proteolytic and/or immunomodulatory functions . One challenge will be to find a probiotic strain that elicits all these functions and that fulfills all safety criteria.

J Ind Microbiol Biotechnol, 2002 Nov, 29(5), 281 - 5
Optimization of galacto-oligosaccharide production by Bifidobacterium infantis RW-8120 using response surface methodology; Roy D et al.; Oligosaccharide (OS) production, cell concentration (2 x 10(9) colony-forming unit/ml), lactose concentration (25% wt/vol), reaction time (6 h), and temperature (50 degrees C) were chosen as the central condition of the central composite design (CCD) for optimizing the production process using Bifidobacterium infantis RW-8120 in skim milk . Statistical analysis (P<0.01) revealed that the most relevant variable concerning OS production and yield was the lactose concentration . The coefficient of determination (R(2)) is good for the second-order OS production model (0.92) and fairly good for the second-order nonlinear OS yield model (0.816) . An increase of lactose concentration and temperature resulted in a higher OS production . The optimal values for OS production appear to be near the area associated with the central points of the modeling design except for the lactose concentration, which was 40% (wt/vol) of the final volume.

Appl Environ Microbiol, 2002 Nov, 68(11), 5209 - 16
Multiparametric flow cytometry and cell sorting for the assessment of viable, injured, and dead bifidobacterium cells during bile salt stress; Amor KB et al.; Using a flow cytometry-based approach, we assessed the viability of Bifidobacterium lactis DSM 10140 and Bifidobacterium adolescentis DSM 20083 during exposure to bile salt stress . Carboxyfluorescein diacetate (cFDA), propidium iodide (PI), and oxonol {DiBAC4(3)} were used to monitor esterase activity, membrane integrity, and membrane potential, respectively, as indicators of bacterial viability . Single staining with these probes rapidly and noticeably reflected the behavior of the two strains during stress exposure . However, the flow cytometry results tended to overestimate the viability of the two strains compared to plate counts, which appeared to be related to the nonculturability of a fraction of the population as a result of sublethal injury caused by bile salts . When the cells were simultaneously stained with cFDA and PI, flow cytometry and cell sorting revealed a striking physiological heterogeneity within the stressed bifidobacterium population . Three subpopulations could be identified based on their differential uptake of the probes: cF-stained, cF and PI double-stained, and PI-stained subpopulations, representing viable, injured, and dead cells, respectively . Following sorting and recovery, a significant fraction of the double-stained subpopulation (40%) could resume growth on agar plates . Our results show that in situ assessment of the physiological activity of stressed bifidobacteria using multiparameter flow cytometry and cell sorting may provide a powerful and sensitive tool for assessment of the viability and stability of probiotics.

J Microencapsul, 2002 Jul-Aug, 19(4), 485 - 94
Microencapsulation of L . acidophilus (La-05) and B . lactis (Bb-12) and evaluation of their survival at the pH values of the stomach and in bile; Favaro-Trindade CS et al.; Microcapsules were prepared using the probiotic microorganisms Lactobacillus acidophilus (La-05) and Bifidobacterium lactis (Bb-12) and the spray drying technique and cellulose acetate phthalate as the wall material . This study evaluated the resistance of these microorganisms to drying at three temperatures and also the in vitro tolerance of the free and microencapsulated form to pH values and bile concentrations similar to those found in the human stomach and intestine . With an air entry temperature of 130 degrees C and exit temperature of 75 degrees C, the number of viable cells of B . lactis was practically unaltered, whereas the population of L . acidophilus was reduced by two logarithmic cycles . B . lactis was more resistant to the drying process than L . acidophilus under all conditions tested . The morphology of the microcapsules was determined by scanning electron microscopy and the microcapsules presented a rounded external surface containing concavities, a continuous wall with no apparent porosity, average size of 22 microm, moisture content varying from 5.3 to 3.2% and water activity between 0.230 and 0.204 . After inoculation into HCl solutions with pH values adjusted to 1 and 2, incubated anaerobically at 37 degrees C, and plated after 0, 1 and 2 h of incubation, microcapsules were effective in protecting the microorganisms, while the populations of both free microorganisms were eliminated after only 1 h at the acidic conditions . Microencapsulated B . lactis and L . acidophilus, both free and microencapsulated, were also resistant after 12h to bile solutions.

J Appl Microbiol, 2002, 93(5), 794 - 9
Effect of bile on the lipid composition and surface properties of bifidobacteria; Gomez Zavaglia A et al.; AIM: The changes produced on the bacterial surface of Bifidobacteria cells when they are grown in bile were compared with those provoked by bile added to bacteria grown in the absence of bile . METHODS AND RESULTS: The adhesive properties, the zeta potential and the lipid composition of Bifidobacterial strains, isolated from human faeces and grown in MRS medium, were determined . Bacteria grown in MRS with bile showed a loss of adherence and autoaggregation in correlation with a decrease in the surface hydrophobicity in comparison to those grown in MRS without bile, concomitant with the absence of two glycolipids, the increase of sugar content and minor changes in fatty acid composition . The surface changes caused by bile shock on bacteria grown in bile-free medium were much less pronounced and, in addition, no effect on the lipid composition was apparent . CONCLUSIONS: The comparison of the results indicates that bile action on surface properties is related to metabolic changes . SIGNIFICANCE AND IMPACT OF THE STUDY: Long-term exposure of bacteria to bile may cause metabolic changes affecting their adhesive properties irreversibly . This may be taken as a criterion to define the probiotic properties of different strains.

Nutr Rev, 2002 Oct, 60(10 Pt 1), 326 - 34
Immune-stimulating and gut health-promoting properties of short-chain fructo-oligosaccharides; Bornet FR et al.; Short-chain fructo-oligosaccharides are a group of linear fructose oligomers with a degree of polymerization ranging from one up to five (oligosaccharides) . Recent observations in animal models demonstrate that prebiotics and probiotics may exert beneficial effects on gut health by enhancing gut-associated lymphoid tissue responses either directly or indirectly through the production of short-chain fatty acids and the enhanced growth of lactic bacteria such as bifidobacteria and lactobacilli . Demonstration of the potential health benefits of short-chain fructooligosaccharides on colon cancer risk is an active field of research in animal and human nutrition.

Prim Care, 2002 Jun, 29(2), 231 - 61
Respiratory and allergic diseases: from upper respiratory tract infections to asthma; Jaber R; Patients with asthma and allergic rhinitis may benefit from hydration and a diet low in sodium, omega-6 fatty acids, and transfatty acids, but high in omega-3 fatty acids (i.e., fish, almonds, walnuts, pumpkin, and flax seeds), onions, and fruits and vegetables (at least five servings a day) . Physicians may need to be more cautious when prescribing antibiotics to children in their first year of life when they are born to families with a history of atopy . More research is needed to establish whether supplementation with probiotics (lactobacillus and bifidobacterium) during the first year of life or after antibiotic use decreases the risk of developing asthma and allergic rhinitis . Despite a theoretic basis for the use of vitamin C supplements in asthmatic patients, the evidence is still equivocal, and long-term studies are needed . The evidence is stronger for exercise-induced asthma, in which the use of vitamin C supplementation at a dosage of 1 to 2 g per day may be helpful . It is also possible that fish oil supplements, administered in a dosage of 1 to 1.2 g of EPA and DHA per day, also may be helpful to some patients with asthma . Long-term studies of fish oil and vitamin C are needed for more definite answers . For the patient interested in incorporating nutritional approaches, vitamin C and fish oils have a safe profile . However, aspirin-sensitive individuals should avoid fish oils, and red blood cell magnesium levels may help in making the decision whether to use additional magnesium supplements . Combination herbal formulas should be used in the treatment of asthma with medical supervision and in collaboration with an experienced herbalist or practitioner of TCM . Safe herbs, such as Boswellia and gingko, may be used singly as adjuncts to a comprehensive plan of care if the patient and practitioner have an interest in trying them while staying alert for drug-herb interactions . No data on the long-term use of these single herbs in asthma exist . For the motivated patient, mind-body interventions such as yoga, hypnosis, and biofeedback-assisted relaxation and breathing exercises are beneficial for stress reduction in general and may be helpful in further controlling asthma . Encouraging parents to learn how to massage their asthmatic children may appeal to some parents and provide benefits for parents and children alike . Acupuncture and chiropractic treatment cannot be recommended at this time, although some patients may derive benefit because of the placebo effect . For patients with allergic rhinitis, there are no good clinical research data on the use of quercetin and vitamin C . Similarly, freeze-dried stinging nettle leaves may be tried, but the applicable research evidence also is poor . Further studies are needed to assess the efficacy of these supplements and herbs . Homeopathic remedies based on extreme dilutions of the allergen may be beneficial in allergic rhinitis but require collaboration with an experienced homeopath . There are no research data on constitutional homeopathic approaches to asthma and allergic rhinitis . Patients with COPD are helped by exercise, pulmonary rehabilitation, and increased caloric protein and fat intake . Vitamin C and n-3 supplements are safe and reasonable; however, studies are needed to establish their efficacy in COPD . On the other hand, there are convincing data in favor of N-acetyl-cysteine supplementation for the patient with COPD at doses ranging between 400 and 1200 mg daily . Red blood cell magnesium levels may guide the use of magnesium replacement . The use of L-carnitine and coenzyme Q10 in patients with COPD needs further study . The addition of essential oils to the dietary regimen of patients with chronic bronchitis is worth exploring . Patients with upper respiratory tract infections can expect a shorter duration of symptoms by taking high doses of vitamin C (2 g) with zinc supplements, preferably the nasal zinc gel, at the onset of their symptoms . Adding an herb such as echinacea or Andrographis shortens the duration of the common cold . The one study on Elderberry's use for the flu was encouraging, and the data on the homeopathic remedy Oscillococcinum interesting, but more studies should be performed . Saline washes may be helpful to patients with allergic rhinitis and chronic sinusitis . Patients also may try the German combination (available in the United States) of elderberry, vervain, gentian, primrose, and sorrel that has been tested in randomized clinical trials . Bromelain is safe to try; the trials of bromelain supplementation were promising but were never repeated . The preceding suggestions need to be grounded in a program based on optimal medical management . Patients need to be well educated in the proper medical management of their disease and skilled at monitoring disease stability and progress . Asthmatic patients need to monitor their bronchodilator usage and peak flow meter measurements to step up their medical treatment in a timely manner, if needed . Patients welcome physician guidance when exploring the breadth of treatments available today . A true patient-physician partnership is always empowering to patients who are serious about regaining their function and health.

Int J Food Microbiol, 2002 Nov 15, 79(1-2), 131 - 41
Application of cereals and cereal components in functional foods: a review; Charalampopoulos D et al.; The food industry is directing new product development towards the area of functional foods and functional food ingredients due to consumers' demand for healthier foods . In this respect, probiotic dairy foods containing human-derived Lactobacillus and Bifidobacterium species and prebiotic food formulations containing ingredients that cannot be digested by the human host in the upper gastrointestinal tract and can selectively stimulate the growth of one or a limited number of colonic bacteria have been recently introduced into the market . The aim of these products is to affect beneficially the gut microbial composition and activities . Cereals offer another alternative for the production of functional foods . The multiple beneficial effects of cereals can be exploited in different ways leading to the design of novel cereal foods or cereal ingredients that can target specific populations . Cereals can be used as fermentable substrates for the growth of probiotic microorganisms . The main parameters that have to be considered are the composition and processing of the cereal grains, the substrate formulation, the growth capability and productivity of the starter culture, the stability of the probiotic strain during storage, the organoleptic properties and the nutritional value of the final product . Additionally, cereals can be used as sources of nondigestible carbohydrates that besides promoting several beneficial physiological effects can also selectively stimulate the growth of lactobacilli and bifidobacteria present in the colon and act as prebiotics . Cereals contain water-soluble fibre, such as beta-glucan and arabinoxylan, oilgosaccharides, such as galacto- and fructo-oligosaccharides and resistant starch, which have been suggested to fulfil the prebiotic concept . Separation of specific fractions of fibre from different cereal varieties or cereal by-products, according to the knowledge of fibre distribution in cereal grains, could be achieved through processing technologies, such as milling, sieving, and debranning or pearling . Finally, cereal constituents, such as starch, can be used as encapsulation materials for probiotics in order to improve their stability during storage and enhance their viability during their passage through the adverse conditions of the gastrointestinal tract . It could be concluded that functional foods based on cereals is a challenging perspective, however, the development of new technologies of cereal processing that enhance their health potential and the acceptability of the food product are of primary importance.

Appl Microbiol Biotechnol, 2002 Oct, 60(1-2), 168 - 73 Epub 2002 Aug 21.
Bifidobacterium longum ATCC 15707 cell production during free- and immobilized-cell cultures in MRS-whey permeate medium; Doleyres Y et al.; Bifidobacterium longum ATCC 15707 cell production was studied in MRS medium supplemented with whey permeate (MRS-WP) during free-cell batch fermentations and continuous immobilized-cell cultures . Very high populations were measured after 12 h batch cultures in MRS-WP medium controlled at pH 5.5 (1.7+/-0.5x10(10) cfu/ml), approximately 2-fold higher than in non-supplemented MRS . Our study showed that WP is a low-cost source of lactose and other components that can be used to increase bifidobacteria cell production in MRS medium . Continuous fermentation in MRS-WP of B . longum immobilized in gellan gum gel beads produced the highest cell concentrations in the effluent (4.9+/-0.9x10(9) cfu/ml) at a dilution rate (D) of 0.5 h(-1) . However, maximal volumetric productivity (6.9+/-0.4x10(9) cfu ml(-1)h(-1)) during continuous cultures was obtained at D =2.0 h(-1), and was approximately 9.5-fold higher than during free-cell batch cultures at an optimal pH of 5.5 (7.2x10(8) cfu ml(-1)h(-1)).

Proc Natl Acad Sci U S A, 2002 Oct 29, 99(22), 14422 - 7 Epub 2002 Oct 15.
The genome sequence of Bifidobacterium longum reflects its adaptation to the human gastrointestinal tract; Schell MA et al.; Bifidobacteria are Gram-positive prokaryotes that naturally colonize the human gastrointestinal tract (GIT) and vagina . Although not numerically dominant in the complex intestinal microflora, they are considered as key commensals that promote a healthy GIT . We determined the 2.26-Mb genome sequence of an infant-derived strain of Bifidobacterium longum, and identified 1,730 possible coding sequences organized in a 60%-GC circular chromosome . Bioinformatic analysis revealed several physiological traits that could partially explain the successful adaptation of this bacteria to the colon . An unexpectedly large number of the predicted proteins appeared to be specialized for catabolism of a variety of oligosaccharides, some possibly released by rare or novel glycosyl hydrolases acting on "nondigestible" plant polymers or host-derived glycoproteins and glycoconjugates . This ability to scavenge from a large variety of nutrients likely contributes to the competitiveness and persistence of bifidobacteria in the colon . Many genes for oligosaccharide metabolism were found in self-regulated modules that appear to have arisen in part from gene duplication or horizontal acquisition . Complete pathways for all amino acids, nucleotides, and some key vitamins were identified; however, routes for Asp and Cys were atypical . More importantly, genome analysis provided insights into the reciprocal interactions of bifidobacteria with their hosts . We identified polypeptides that showed homology to most major proteins needed for production of glycoprotein-binding fimbriae, structures that could possibly be important for adhesion and persistence in the GIT . We also found a eukaryotic-type serine protease inhibitor (serpin) possibly involved in the reported immunomodulatory activity of bifidobacteria.

Can J Microbiol, 2002 Aug, 48(8), 759 - 63
Fermentation of chicory fructo-oligosaccharides in mixtures of different degrees of polymerization by three strains of bifidobacteria; Perrin S et al.; We estimated and compared the action of three selected strains of bifidobacteria in a semi-synthetic medium for different degrees of polymerization of fructo-oligosaccharides contained in three commercial products derived from chicory inulin: Fibrulose F97 (shorter chains), Fibruline Instant (native chains), Fibruline LC (longer chains) . Biomass and production of lactate and acetate were greater when the substrate contained mostly shorter chain fructo-oligosaccharides . Shorter chains were first to be consumed, and one strain could use longer chains . As the degree of polymerization increased, residual fructo-oligosaccharides increased after growth of the strains, and the rate of consumption of fructo-oligosaccharides decreased.

Antonie Van Leeuwenhoek, 2002 Aug, 82(1-4), 341 - 52
The intestinal LABs; Vaughan EE et al.; The complete gastrointestinal (GI) tract of humans is colonised soon after birth by a myriad of microbial species with a characteristic distribution depending on the location . GI-tract ecology has been experiencing a revival due to the development of molecular techniques, especially those based on 16S RNA (zRNA) genes . A richer ecosystem than previously imagined of novel species is being discovered that is significantly influenced by our host genotype . Special attention has been focused on the bifidobacteria and the lactic acid bacterial (LAB) populations, both those that are naturally present within this complex ecosystem and those that are ingested as probiotics in functional foods . Overall this interest stems from a increasing awareness of interplay between microflora, diet and the health of the host, and is further stimulated by an increasing incidence of gastrointestinal illnesses, and atopy . Substantial documentation of benefits to host health has especially distinguished the LAB for multidisciplinary research aimed to determine the molecular mechanisms involved . Recent advances in molecular technologies, including high-throughput genomics-based approaches, can significantly advance our understanding of the microbe--diet--host interactions and offer valuable information for design and application of health-targeted microbes.

Antibiot Khimioter, 2002, 47(4), 16 - 9
{Oral administration of pectins for prophylaxis and treatment of purulent septic complications in patients with burns}; Lazareva EB et al.; Clinical and laboratory evaluation of pectins application efficacy demonstrated that the patients treated with pectines has lower frequency of bacteriemia, intoxication, infectious complications and lethality . Pectins use per os resulted by acceleration of burns wounds healing and by lower microbial dissemination . Frequency of coagulase-positive staphylococci, streptococci and enterococci isolation was also lower in the treated group of patients when compared to the control group . Feces microbial profile also demonstrated positive trend--normal microflora (bifidobacteria and lactobacilli) enhanced, opportunistic bacteria diminished . Pectins application provided normalization of leucocytes, lymphocytes, B-lymphocytes, immunoglobulins A and G . The results of investigation demonstrated pectins efficacy at the complex treatment regimes of the patients with burns.

Microbiol Immunol, 2002, 46(8), 535 - 48
Phylogenetic analysis of the human gut microbiota using 16S rDNA clone libraries and strictly anaerobic culture-based methods; Hayashi H et al.; The human gut microbiota from three healthy subjects were compared by the use of a sequence analysis of 16S rDNA libraries and a culture-based method . Direct counts ranged from 1.9 X 10" to 4.0 X 10" cells/g (wet weight), and plate counts totaled 6.6 X 10(10) to 1.2 X 10(11) CFU/g (wet weight) . Sixty to seventy percent of the bacteria in the human intestinal tract cannot be cultured with currently available methods . The 16S rDNA libraries from three subjects were generated from total community DNA in the intestinal tract with universal primer sets . Randomly selected clones were partially sequenced . All purified colonies detected from the surface of the agar plate were used for a partial sequencing of 16S rDNA . On the basis of sequence similarities, the clones and colonies were classified into several clusters corresponding to the major phylum of the domain Bacteria . Among a total of 744 clones obtained, approximately 25% of them belonged to 31 known species . About 75% of the remaining clones were novel "phylotypes" (at least 98% similarity of clone sequence) . The predominant intestinal microbial community consisted of 130 species or phylotypes according to the sequence data in this study . The 16S rDNA libraries and colonies included the Bacteroides group, Streptococcus group, Bifidobacterium group, and Clostridium rRNA clusters IV, IX, XIVa, and XVIII . Moreover, several previously uncharacterized and uncultured microorganisms were recognized in clone libraries and colonies . Our results also showed marked individual differences in the composition of intestinal microbiota.

J Ind Microbiol Biotechnol, 2002 Oct, 29(4), 196 - 9
Glucooligosaccharides from Leuconostoc mesenteroides B-742 (ATCC 13146): a potential prebiotic; Chung CH et al.; There is an emerging market for functional oligosaccharides for use in foods . Currently, technology for the production of oligosaccharides is limited to extraction from plant sources, acid or enzymatic hydrolysis of polysaccharides or synthesis by transglycosylation reactions . Oligosaccharides can also be produced using a Leuconostoc fermentation and restricting the polymer size by addition of maltose . Maltose limits the dextransucrase reaction, yielding high concentrations of alpha-glucooligosaccharides . Branched oligomers produced by this process were readily catabolized by bifidobacteria and lactobacilli but were not readily utilized by either Salmonella sp . or Escherichia coli, pointing toward their use in intestinal microflora modification.

Syst Appl Microbiol, 2002 Aug, 25(2), 241 - 8
PCR-ELISA I: Application to simultaneous analysis of mixed bacterial samples composed of intestinal species; Laitinen R et al.; Sixteen oligonucleotide identification probes, designed in this study or adapted from literature, were tested for a PCR-ELISA application to simultaneously detect under standardised conditions selected intestinal bacteria, lactobacilli and bifidobacteria . The level of specificity obtained with most of the probes fulfilled the set criteria . The lack of efficiency of PCR performed with the primers, proposed to be specific for the entire eubacteria domain, and compromises made in hybridisation conditions due to simultaneous usage of multiple probes reduced the sensitivity of the PCR-ELISA test . The method was, however, found to be suitable for detecting predominant members of the intestinal flora . Applicability of the PCR-ELISA test could be further widened using primers with a more restricted specificity in the PCR step, as was demonstrated for the detection of Bifidobacterium with genus-specific primers . Advantages of the PCR-ELISA method include convenient performance and the possibility to test rapidly large amounts of samples with a multitude of probes.

J Urol, 2002 Oct, 168(4 Pt 1), 1512 - 7
The potential role of probiotics in pediatric urology; Reid G; PURPOSE: The application of select microbial strains to increase the host population of good bacteria is called probiotics, a term defined as "live microorganisms which when administered in adequate amounts confer a health benefit on the host." This review was done to evaluate the potential role that probiotic therapy may have in pediatric urology . MATERIALS AND METHODS: Many children around the world die of diseases, such as gastrointestinal infection and HIV, while many have urinary tract infections that subsequently recur frequently in adulthood . Until recently the role of intestinal and urogenital (vaginal, urethral and perineal) microflora in health and disease has received scant attention . The data available in the literature on this topic were examined and a personal viewpoint is presented on how they may relate to urology . RESULTS: There is mounting evidence that certain strains of lactobacilli and bifidobacteria have a major part in the maintenance and restoration of health in children and adults . CONCLUSIONS: Implications for pediatric urology include a decreased risk of infection and stone disease as well as possible positive effects on preventing and managing inflammatory and some carcinogenic diseases.

Appl Environ Microbiol, 2002 Oct, 68(10), 4986 - 95
Culture-independent microbial community analysis reveals that inulin in the diet primarily affects previously unknown bacteria in the mouse cecum; Apajalahti JH et al.; Inulin is a well-known fructose-based prebiotic which has been shown to stimulate the growth of bifidobacteria, a bacterial group generally considered beneficial for intestinal health . In the present study, we analyzed inulin-associated shifts in the total bacterial community of wild-type mice and mice carrying a genetically inactivated adenomatous polyposis coli tumor suppressor gene by using DNA-based approaches independent of bacterial culturability . Mice were fed a high-fat, nonfiber diet with or without inulin inclusion at a 10% (wt/wt) concentration . Cecal contents were analyzed after 0, 3, and 9 weeks on the experimental diets . Inulin inclusion significantly affected the total bacterial community structure of the cecum as determined by both a nonselective percent-guanine-plus-cytosine-based profiling analysis and a more specific 16S ribosomal DNA sequence analysis . The shifts included stimulation of bifidobacteria and suppression of clostridia, but sequence comparison revealed that the major shifts were within previously unknown bacterial taxa . Concomitantly, significantly higher bacterial densities, determined by flow cytometry, were observed with the inulin-amended diet, and the metabolism of the cecal bacterial community was altered, as indicated by higher levels of residual short-chain fatty acids, particularly lactic acid . With regard to all of the microbiological parameters measured, the wild-type mice and mice carrying a genetically inactivated adenomatous polyposis coli tumor suppressor gene were essentially identical . Studies of the implications of pre- and probiotics may need to be expanded to include careful analysis of their effects on the entire microbial community, rather than just a few well-known species . Further studies are needed to increase our understanding of the possible roles of currently unknown gastrointestinal bacteria in health and disease.

Zhongguo Zhong Yao Za Zhi, 1998 Dec, 23(12), 743 - 5, back cover
{Influence of alum on intestinal flora in mice}; Yan M et al.; OBJECTIVE: To observe the influence of alum on the intestinal microecological balance in normal microorganisms . METHOD: The mice were administered orally with alum of a small dosage(0.25/kg) and a large dosage(1 g/kg) for half a month, two months and three months, and a micro flora analysis of the mice was carried out at intervals of the above mentioned administrations . RESULT: The intestinal flora in the animals administered with alum was imbalanced . The counts of bifidobacteria and lactobacilli closely related to human physiological activities were decreased . The counts of pathogenic E . Coli significantly increased; and the longer the animals were treated with alum, the stronger the microecological balance was influenced . CONCLUSION: Alum could induce imbalance of the normal intestinal flora in mice.

Crit Rev Biochem Mol Biol, 2002, 37(4), 259 - 81
Effects of consumption of probiotics and prebiotics on serum lipid levels in humans; Pereira DI et al.; The objective of this article is to review existing studies concerning the effects of probiotics and prebiotics on serum cholesterol concentrations, with particular attention on the possible mechanisms of their action . Although not without exception, results from animal and human studies suggest a moderate cholesterol-lowering action of dairy products fermented with appropriate strain(s) of lactic acid bacteria and bifidobacteria . Mechanistically, probiotic bacteria ferment food-derived indigestible carbohydrates to produce short-chain fatty acids in the gut, which can then cause a decrease in the systemic levels of blood lipids by inhibiting hepatic cholesterol synthesis and/or redistributing cholesterol from plasma to the liver . Furthermore, some bacteria may interfere with cholesterol absorption from the gut by deconjugating bile salts and therefore affecting the metabolism of cholesterol, or by directly assimilating cholesterol . For prebiotic substances, the majority of studies have been done with the fructooligosaccharides inulin and oligofructose, and although convincing lipid-lowering effects have been observed in animals, high dose levels had to be used . Reports in humans are few in number . In studies conducted in normal-lipidemic subjects, two reported no effect of inulin or oligofructose on serum lipids, whereas two others reported a significant reduction in serum triglycerides (19 and 27%, respectively) with more modest changes in serum total and LDL cholesterol . At present, data suggest that in hyperlipidemic subjects, any effects that do occur result primarily in reductions in cholesterol, whereas in normal lipidemic subjects, effects on serum triglycerides are the dominant feature.

J Dairy Res, 2002 May, 69(2), 293 - 302
Effect of bile components on the surface properties of bifidobacteria; Kociubinski G et al.; The action of bile on bifidobacteria was studied by measuring changes in zeta potential . hydrophobicity and adherence to enterocyte-like cells in vitro . Highly hydrophobic strains shocked with bile displayed more negative zeta potential values and a decrease in adherence . When a non-hydrophobic non-adherent strain (CIDCA 5324) was shocked with bile, an increase in hydrophobicity was observed . However, no changes of zeta potential or adherence properties were apparent . The action of the bile components was different from the action of whole bile . Cholate and deoxycholate produced a decrease in the negativity of zeta potential values of all strains studied whereas taurocholate displayed a shift in zeta potential of hydrophobic strains to more negative values, thus explaining the decrease in the autoaggregation by charge repulsion . However, the decrease in zeta potential caused by cholate and deoxycholate did not increase autoaggregation in a hydrophobic non-adherent strain (CIDCA 531) . This suggests that other forces are contributing to autoaggregation.

Space Med Med Eng (Beijing), 2002 Jun, 15(3), 186 - 8
{Effects of head down bed rest or drinking water containing electrolytic silver-ion on intestinal bifidobacteria}; Li ZX et al.; Objective . To investigate the effect of head down bed rest (HDBR) or electrolytic silver-ion on intestinal bifidobacteria in men and mice . Method . Three tests were conducted . Counts of bifidobacteria in feces were made in: 1) 5 male healthy volunteers after 21 d -6 degrees HDBR . 2) 4 groups of mice drank water containing no Ag+, 0.20, 0.40 and 0.60 mg/ L of Ag+ respectively for 1 month . 3) Seven male healthy volunteers drank water containing 0.20 mg/L of silver-ion for 16 days . Result . 1) The count of bifidobacteria after HDBR {8.08 +/- 0.82 (lgn/g)} was significantly decreased as compared with the control group {8.72 +/- 0.82 (lgn/g)} (P<0.01) . It suggests that HDT -6 degrees bed-rest might reduce the number of bifidobacteria in the human colon . 2) The count of bifidobacteria in feces was negatively correlated with the silver-ion concentration in the water (r= -0.992, P<0.01) . 3) The count of bifidobacteria in feces after test {7.83 +/- 0.71(lgn/g)} was significantly decreased as compared with that before the test (8.91 +/- 0.84) (P<0.05) . Conclusion . It demonstrates that HDT -6 degrees bed-rest and electrolytic silver-ion all might reduce the count of bifidobacteria in feces.

Br J Nutr, 2002 Sep, 88 Suppl 1, S95 - 9
Improvement of the probiotic effect of micro-organisms by their combination with maltodextrins, fructo-oligosaccharides and polyunsaturated fatty acids; Bomba A et al.; Probiotics could represent an effective alternative to the use of synthetic substances in nutrition and medicine . The data concerning the efficacy of probiotics are often contradictory . This paper focuses on the enhancement of the efficacy of probiotics by their combination with synergistically acting components of natural origin . Maltodextrins can be obtained by enzymatic hydrolysis of starch and are suitable for consumption . Administration of Lactobacillus paracasei together with maltodextrin decreased the number of Escherichia coli colonising the jejunal mucosa of gnotobiotic piglets by 1 logarithm compared to the control group . Fructo-oligosaccharides (FOS) are naturally occurring oligosaccharides, mainly of plant origin . L . paracasei administered in combination with FOS significantly increased counts of Lactobacillus spp., Bifidobacterium spp., total anaerobes and total aerobes compared to the control group as well as the L . paracasei group . It also significantly decreased Clostridium and Enterobacterium counts in the faeces of the weanling piglets compared with the control group . Dietary lipids influence the gastrointestinal microbiota and specifically the population of lactic acid bacteria . In gnotobiotic piglets the oral administration of an oil containing polyunsaturated fatty acids (PUFA) significantly increased the number of L . paracasei adhering to jejunal mucosa compared to the control group . Our results showed that maltodextrin KMS X-70 and PUFA can be used to enhance the effect of probiotic micro-organisms in the small intestine, and similarly FOS enhance the effect of probiotic micro-organisms in the large intestine.

Aliment Pharmacol Ther, 2002 Sep, 16(9), 1669 - 75
Impact of supplement with Lactobacillus- and Bifidobacterium-containing yogurt on triple therapy for Helicobacter pylori eradication; Sheu BS et al.; AIM: To test whether supplements of Lactobacillus- and Bifidobacterium-containing yogurt (AB-Yogurt) affect the success of Helicobacter pylori eradication . METHODS: One hundred and sixty H . pylori-infected patients were randomized into a triple-plus-yogurt group or a triple-only group, receiving 1 week of triple therapy with and without supplements of AB-Yogurt, respectively . In the triple-plus-yogurt group, AB-Yogurt was continued for 4 weeks after triple therapy . Eight weeks later, patients were assessed for the success of H . pylori eradication . The stool samples of 22 randomly selected patients, 11 from each group, were provided on enrolment, at the first week and at the fifth week for evaluation of the percentage of Bifidobacterium in anaerobes . RESULTS: By intention-to-treat analysis, the triple-plus-yogurt group had a higher H . pylori eradication rate than the triple-only group (91% vs . 78%, P < 0.05) . The per protocol H . pylori eradication rates were similar for both groups (93.5% vs . 89%, P = N.S.) . Only patients supplemented with AB-Yogurt showed restoration of the percentage of Bifidobacterium in the anaerobes of stools at the fifth week to the level in the stools on enrolment . CONCLUSIONS: Supplement with AB-Yogurt can improve the intention-to-treat eradication rates of H . pylori, and can restore the depletion of Bifidobacterium in stools after triple therapy.

Int Endod J, 2002 Aug, 35(8), 698 - 702
Combinations of bacterial species in endodontic infections; Peters LB et al.; AIM: This study was undertaken to investigate combinations of bacteria found in root-canal infections of teeth with periapical bone destruction without clinical signs and symptoms . METHODOLOGY: Endodontic samples from 58 root canals were cultured anaerobically and microorganisms were counted and identified . Eighty-one combinations of microorganisms were found and tested for a symbiotic relationship using the Fisher's exact test and Odds ratio calculation . RESULTS: All samples contained microorganisms with a median CFU mL(-1) of 8x10(4) per sample . Strict anaerobic species accounted for 87% of the microflora . The most prevalent bacteria were Prevotella intermedia, Peptostreptococcus micros and Actinomyces odontolyticus, present in 33, 29 and 19%, respectively, of the cultured canals . A significant relationship (P<0.05) and an Odds ratio >2 were found between P . intermedia and P . micros, P . intermedia and P . oralis, A . odontolyticus and P . micros, Bifidobacterium spp . and Veillonella spp . Conclusions: These results indicate that endodontic pathogens do not occur at random but are found in specific combinations.

J Appl Microbiol, 2002, 93(3), 390 - 7
Manipulation of colonic bacteria and volatile fatty acid production by dietary high amylose maize (amylomaize) starch granules; Wang X et al.; AIMS: To study the effects of amylomaize starch and modified (carboxymethylated and acetylated) amylomaize starches on the composition of colonic bacteria and the production of volatile fatty acids, in mice . METHODS AND RESULTS: Balb/c mice were fed with experimental diets containing various amount of amylomaize and modified amylomaize starches . Colonic bacterial populations and short-chain fatty acids were monitored . Results showed that the increases in indigenous bifidobacteria were detected in mice fed all starches tested; however, the highest numbers were observed in the group fed with 40% unmodified amylomaize starch . The starch type influenced the populations of indigenous Lactobacillus, Bacteroides and coliforms . High Lactobacillus numbers were achieved in the colon of mice fed with high concentration of amylomaize starch . Acetylated amylomaize starch significantly reduced the population of coliforms . In addition, orally dosed amylomaize utilizing bifidobacteria reached their highest levels when fed together with amylomaize or carboxymethylated amylomaize starch and in both cases butyrate levels were markedly increased . CONCLUSIONS: These results indicate that different amylomaize starches could generate desirable variation in gut microflora and that particular starches may be used to selectively modify gut function . SIGNIFICANCE AND IMPACT OF STUDY: Amylomaize starch appeared to enhance the desirable composition of colonic bacteria in mice, and suggested it possessed the potential prebiotic properties . Therefore, resistant starch and its chemical derivatives may exert beneficial impacts to the human colon.

J Gastroenterol Hepatol, 2002 Aug, 17(8), 818 - 24
A new prebiotic from germinated barley for nutraceutical treatment of ulcerative colitis; Bamba T et al.; A germinated barley foodstuff (GBF) containing glutamine-rich protein and hemicellulose-rich fiber was made from brewer's spent grain, by physical isolation . Our previous studies demonstrated that GBF supported maintenance of epithelial cell populations, facilitated epithelial repair, and suppressed epithelial nuclear factor kappaB-DNA-binding activity through generating increased short-chain fatty acid (especially butyrate) production by luminal microflora, which includes Bifidobacterium and Eubacterium, thereby preventing experimental colonic injury . The fiber fraction also modulates stool water content because of its high water-holding capacity . The patients with mild to moderate active ulcerative colitis who had been unresponsive to or intolerant of standard treatment received 20-30 g GBF, feeding daily in a non-randomized, open-label fashion . At 4 weeks, this treatment resulted in a significant clinical and endoscopic improvement . The improvement was associated with an increase in stool butyrate concentrations . These results indicate that GBF feeding is a potentially new, attractive prebiotic treatment in patients with ulcerative colitis . The potency of GBF on modulating microflora, as well as the high water-holding capacity, may play an important role in treatment and prolongation of remission in ulcerative colitis.

Biosci Biotechnol Biochem, 2002 Jun, 66(6), 1419 - 22
Two novel oligosaccharides formed by 1F-fructosyltransferase purified from roots of asparagus (Asparagus officinalis L.); Yamamori A et al.; Two novel oligosaccharides, tetra-and penta-saccharides were synthesized by fructosyl transfer from 1-kestose to 4G-beta-D-galactopyranosylsucrose with a purified 1F-fructosyltransferase of asparagus roots and identified as 1F-beta-D-fructofuranosyl-4G-beta-D-galactopyranosylsucrose, O-beta-D-fructofuranosyl-(2-->1)-beta-D-fructofuranosyl-O-{beta-D-galactopyranosyl-(1-->4)}-alpha-D-glucopyranoside and 1F(1-beta-D-fructofuranosyl)2-4G-beta-D-galactopyranosylsucrose, {O-beta-D-fructofuranosyl-(2-->1)}2-beta-D-fructofuranosyl-O-{beta-D-galactopyranosyl-(1-->4)}-alpha-D-glucopyranoside, respectively . Both oligosaccharides were scarcely hydrolyzed by carbohydrase from rat small intestine . Human intestinal bacterial growth by 1F-beta-D-fructofuranosyl-4G-beta-D-galactopyranosylsucrose was compared with that by the tetrasaccharides, stachyose and nystose . Bifidobacteria utilized 1F-beta-D-fructofuranosyl-4G-beta-D-galactopyranosylsucrose to the same extent as stachyose or nystose . On the other hand, the unfavorable bacteria, Clostridium perfringens, Escherichia coli and Enterococcusfaecalis, that produce mutagenic substances did not use the synthetic oligosaccharide.

J Dairy Sci, 2002 Jun, 85(6), 1383 - 9
Partial characterization of bifidobacterium breve C50 cell-free whey compounds inducing modifications to the intestinal microflora; Mullie C et al.; Cell-free whey obtained from milk fermented with Bifidobacterium breve C50 (Bb C50) has been shown to modify the intestinal flora in humans and mice . Previous work showed that no antibiotic-like or barrier effect due to overgrowing bifidobacteria was implied in the microbial modifications . The present study was focused on characterizing the compounds and mechanisms involved . Protein, sugar, and enzymatic profiles of Bb C50 whey were therefore determined and compared to those of a whey unable to modify the intestinal flora of humans and mice . No remarkable difference was noted except for a higher lactosidase activity in Bb C50 whey . Various physical treatments were then applied to fractions of Bb C50 whey . Activity was assessed in C3H mice by analyzing changes in the intestinal flora balance throughout a 15-d administration of each treated whey . Heating at 80 degrees C and aerobic storage for 2 wk completely abolished Bb C50 whey activities . In contrast, the addition of a reducing agent (cysteine hydrochloride), either at the beginning of a 15-d aerobic storage or prior to administration, as well as preserved these activities . Susceptibility to heating and oxidation suggested that an enzyme might play a role in the induced microbial changes . Since the Bb C50 lactosidase was partly inactivated by the oxidative treatment, it could support the in vivo activity . The enzyme might reach the intestinal lumen and partly degrade substrates, such as mucins, usually used by the intestinal flora . The released molecules might then favor the development of a new microbial balance.

Eur J Clin Nutr, 2002 Aug, 56 Suppl 3, S60 - 4
Interactions between the microbiota and the intestinal mucosa; Schiffrin EJ et al.; The intestinal microflora can be considered as a postnatally aquired organ composed of a large diversity of bacterial cells that can perform different functions for the host . This organ is highly exposed to environmental influences and thus modulated in its composition and functions by external 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 . In addition, the microflora seem to play a significant role in the maintenance of intestinal immune homeostasis and prevention of inflammation . At the present time, the contribution of intestinal epithlial cell in the first line of defence against pathogenic bacteria and microbial antigens has been recognized, in contrast, the interactions of intestinal epithelial cells with commensal bacteria are less understood . The present work summarizes the increasing scientific attention for mechanisms of the innate immune response of the host to different components of the autochthonous microflora and suggests a potential role for selected probiotic bacteria in the regulation of intestinal inflammation.

Surg Today, 2002, 32(7), 581 - 6
Changes in the intestinal flora after the administration of prophylactic antibiotics to patients undergoing a gastrectomy; Takesue Y et al.; PURPOSE: Changes in the intestinal flora were investigated in patients administered antibiotics for 4 days after a gastrectomy . METHODS: Twenty-four patients were divided into each of the following groups: cefazolin, 1 g every 8 h; cefozopran, 0.5 g every 8 h; flomoxef, 1 g every 8 h . All drugs were administered intravenously for a 4-day period . RESULTS: Antibiotic prophylaxis caused no major change in the total number of anaerobes because of the preservation of the Bacteroides fragilis group . However, the number of Bifidobacterium, Lactobacillus, Eubacterium, and Veillonella spp . decreased by an average of 100-fold . These changes were accompanied by an overgrowth of Enterococcus spp . and Pseudomonas aeruginosa . CONCLUSIONS: Four days of antibiotic prophylaxis after surgery caused a suppression of colonization resistance in the intestinal flora . Certain genera of anaerobes may thus play an important role in preventing overgrowth of antibiotic-resistant organisms after surgery . Surgeons should decide the duration of antibiotic use after carefully considering its influence on the intestinal flora.

Appl Microbiol Biotechnol, 2002 Jul, 59(2-3), 297 - 302 Epub 2002 Apr 20.
Quantitative determination of the spatial distribution of pure- and mixed-strain immobilized cells in gel beads by immunofluorescence; Doleyres Y et al.; A new method was developed to detect and quantify two strains, Lactococcus lactis subsp . lactis biovar . diacetylactis MD and Bifidobacterium longum ATCC 15707, immobilized separately and co-immobilized in gel beads, using specific polyclonal antibodies and confocal laser-scanning microscopy . The establishment of biomass concentration profiles for each strain was measured during colonization of beads using successive pH-controlled batch fermentations . Growth occurred preferentially in 200- and 300-microm peripheral layers of the beads for L . diacetylactis and B . longum, respectively . Repeated-batch cultures with immobilized cells permitted the production of a mixed culture containing a non-competitive strain of bifidobacteria, as a result of immobilized-cell growth and high cell-release activity from the beads . During co-immobilized fermentations, there were no apparent interactions between the strains.

Lett Appl Microbiol, 2002, 35(2), 126 - 30
Fermentation of pectin and glucose, and activity of pectin-degrading enzymes in the rabbit caecal bacterium Bifidobacterium pseudolongum; Slovakova L et al.; AIMS: In a rabbit caecal bacterium Bifidobacterium pseudolongum, metabolites of pectin and glucose, and activities of enzymes involved in the degradation of pectin were assayed . Simultaneously, activities of these enzymes were assayed in a rumen pectinolytic strain of Streptococcus bovis . METHODS AND RESULTS: A strain B . pseudolongum P6 which grew best on pectin was selected among bifidobacteria isolated from the rabbit caecum . Cultures of B . pseudolongum P6 grown on pectin produced significantly less formate, lactate and ethanol, and more acetate and succinate than cultures grown on glucose . No CO2 production on pectin was observed . Pectin macromolecule was degraded by extracellular pectinase (EC 3.2.1.15) . Cell extracts possessed the activity of 2-keto-3-deoxy-6-phosphogluconate (KDPG) aldolase (EC 4.1.2.14) . Streptococcus bovis X4, possessed activity of exopectate lyase and pectinase, but not that of KDPG aldolase . CONCLUSIONS: Our results are consistent with the assumption that in B . pseudolongum P6 acidic products of pectin degradation are catabolized via a modified Entner-Doudoroff pathway, as shown previously in rumen pectin-utilizing bacteria . The missing KDPG aldolase activity in Strep . bovis X4 seems to be the reason for the absence of growth of this bacterium on pectin . SIGNIFICANCE AND IMPACT OF THE STUDY: Information on polysaccharide metabolism in bifidobacteria is fragmentary . This study extends the knowledge on pectin metabolism in intestinal bacteria.

J Nutr, 2002 Jul, 132(7), 2048 - 54
A combination of dietary fructooligosaccharides and isoflavone conjugates increases femoral bone mineral density and equol production in ovariectomized mice; Ohta A et al.; Fructooligosaccharides (FOS) stimulate the growth of bifidobacteria, which cleave isoflavone conjugates to yield the corresponding aglycones and metabolites . In a previous study, FOS modified the absorption and enterohepatic recirculation of isoflavones in rats . In the present study, we determined the effect of the combination of dietary FOS and isoflavone conjugates on bone mass in ovariectomized (OVX) and surgical control mice . After undergoing OVX or sham operation, female ddY mice (8 wk old, n = 64) were randomly assigned to four groups: a purified control diet (AIN-93G) group, a FOS diet (AIN-93G + 5% FOS) group, an isoflavone diet (AIN-93G + 0.2% isoflavone conjugates) group, or a FOS and isoflavone diet (AIN-93G + 5% FOS + 0.2% isoflavone conjugates) group . After 6 wk, the mice were killed and the blood and femora were sampled immediately . In OVX mice, both isoflavone conjugates and FOS prevented femoral bone loss . An additive effect of dietary isoflavone conjugates and FOS was observed by dual-energy X-ray absorptiometry in the distal part of the femur and in trabecular bone, by peripheral quantitative computed tomography . Moreover, FOS increased cecal beta-glucosidase activity and equol production from daidzein in both OVX and surgical control mice fed isoflavone conjugates . These results suggest that FOS increase the bioavailability of isoflavones, leading to cooperative effects in the prevention of osteopenia in OVX mice.

Crit Rev Oral Biol Med, 2002, 13(2), 171 - 83
Invasion of dentinal tubules by oral bacteria; Love RM et al.; Bacterial invasion of dentinal tubules commonly occurs when dentin is exposed following a breach in the integrity of the overlying enamel or cementum . Bacterial products diffuse through the dentinal tubule toward the pulp and evoke inflammatory changes in the pulpo-dentin complex . These may eliminate the bacterial insult and block the route of infection . Unchecked, invasion results in pulpitis and pulp necrosis, infection of the root canal system, and periapical disease . While several hundred bacterial species are known to inhabit the oral cavity, a relatively small and select group of bacteria is involved in the invasion of dentinal tubules and subsequent infection of the root canal space . Gram-positive organisms dominate the tubule microflora in both carious and non-carious dentin . The relatively high numbers of obligate anaerobes present-such as Eubacterium spp., Propionibacterium spp., Bifidobacterium spp., Peptostreptococcus micros, and Veillonella spp.-suggest that the environment favors growth of these bacteria . Gram-negative obligate anaerobic rods, e.g., Porphyromonas spp., are less frequently recovered . Streptococci are among the most commonly identified bacteria that invade dentin . Recent evidence suggests that streptococci may recognize components present within dentinal tubules, such as collagen type I, which stimulate bacterial adhesion and intra-tubular growth . Specific interactions of other oral bacteria with invading streptococci may then facilitate the invasion of dentin by select bacterial groupings . An understanding the mechanisms involved in dentinal tubule invasion by bacteria should allow for the development of new control strategies, such as inhibitory compounds incorporated into oral health care products or dental materials, which would assist in the practice of endodontics.

J Nutr Biochem, 2002 Jun, 13(6), 364 - 369
Influence of probiotic supplemented infant formula on composition of plasma lipids in atopic infants; Kankaanpaa PE et al.; Probiotic therapy is a new, successful approach to alleviating allergic symptoms . In this study, our aim was to investigate whether the positive results obtained with probiotic therapy would be associated with the differential absorption and utilization of dietary PUFA . 15 infants referred to a pediatric clinic on the basis of atopic eczema were weaned to Bifidobacterium Bb-12 or Lactobacillus GG supplemented infant formula, or to the same formula without probiotics (randomized, placebo-controlled, double blind study design) . In plasma neutral lipids, alpha-linolenic acid (18:3 n-3) proportions were reduced by the probiotic supplementation . In phospholipids, Lactobacillus GG supplemented formula did not influence alpha-linolenic acid proportions, while Bifidobacterium Bb-12 supplemented formula increased the proportion of alpha-linolenic acid; from 0.13 +/- 0.03 to 0.24 +/- 0.03 (mean +/- SEM) (P = 0.002) . These results show that some physiological effects of probiotics may be associated with physiological interactions between probiotics and dietary PUFA.

Br J Nutr, 2002 May, 87 Suppl 2, S293 - 6
Oligosaccharides in infant formula; Vandenplas Y; Breast-feeding is the golden standard for infant feeding . However, the majority of a few week old infants are fed with a second choice infant feeding, cow's milk based formula . Amongst the multiple differences between human and cow's milk regards the development of the gastro-intestinal flora: the flora of the breast-fed infant being richer in bifidobacteria and lactobacilli . Both species are known to be potentially beneficial for the health of the host . The absence of oligosaccharides, the third largest component in human milk, in cow's milk is likely to account for the differences in colonic flora . The oligosaccharide content and concentration in breast milk is - just as for the other macronutrients - a dynamic process, making it impossible for industry to mimic nature . However, if the composition cannot be mimicked, the effect and function can be imitated . The addition of two oligosaccharides, galacto-oligosaccharides and inulin, to cow's milk based infant formula has been shown to have a bifidogenic effect, and to stimulate the growth of bifidi and lactobacilli . In conclusion, the addition of oligosaccharides to cow's milk based infant formula brings this alternative, second choice infant feeding one step closer to the golden standard of human milk . But, prolonged breast-feeding should still be promoted with maximum effort.

Br J Nutr, 2002 May, 87 Suppl 2, S273 - 81
Experimental evidences on the potential of prebiotic fructans to reduce the risk of colon cancer; Pool-Zobel B et al.; Inulin is extracted from the chicory root . It is a set of fructans with its monomers linked by means of beta(2-1) bonds . This linkage cannot be hydrolysed by either pancreatic or by brush border digestive enzymes in the upper intestinal tract of humans . As such the carbohydrates arrive in the colon, where they are fermented by bifidobacteria and other lactic acid producing bacteria, thus enhancing their relative populations in the gut . Recent research in experimental animal models revealed that inulin has significant anticarcinogenic properties . It acts chemopreventively by reducing the incidence of azoxymethane (AOM) - induced aberrant crypt foci and tumours in the colon . These effects may be due to the stimulation of bifidobacteria, which themselves have been shown to act as antigenotoxic in the colon and to reduce AOM-induced tumours . Also fermentation products, including the short-chain fatty acid butyrate, could contribute to the protective effects . In this case a mechanism may be the induction of apoptosis of already transformed cells . The experimental evidence from animal studies and from studies elucidating potential mechanisms strongly supports the possibility that inulin will contribute to reducing risks for colon cancer in humans . In order to obtain more insight into this possibility, human dietary intervention studies relating biomarkers of reduced risk to inulin consumption are needed.

Br J Nutr, 2002 May, 87 Suppl 2, S213 - 9
Oligofructose and experimental model of neonatal necrotising enterocolitis; Butel MJ et al.; The gut of preterm neonates is colonised with a paucity of bacterial species originating more from the environment than from the mother . Furthermore, a delayed colonisation by bifidobacteria promotes colonisation by potentially pathogenic bacteria . This may contribute towards the development of neonatal necrotising enterocolitis (NEC) . The physiopathology of NEC is still unclear but immaturity of the gut, enteral feeding and bacterial colonisation are all thought to be involved . None of the current preventive treatments are considered satisfactory . Modulating the autochthonous microflora by probiotics or prebiotics could be a more reliable approach to prevention . Using gnotobiotic quails as an experimental model of NEC we have shown that onset of intestinal lesions requires a combination of low endogenous lactase activity, lactose in diet, and colonisation by lactose-fermenting bacteria such as the clostridia . The protective role of bifidobacteria was demonstrated in this model through a decrease in clostridial populations and in butyric acid . Oligofructose dietary supplementation was shown to enhance this effect with an increase in the bifidobacterial level and consequently a greater decrease in clostridia . However, oligofructose was unable to promote a bifidobacterial acquisition when the microflora was initially deprived of this group . Nevertheless, oligofructose can act as an anti-infective agent and decrease the occurrence or severity of the lesions depending on the bacteria involved . According to these results and to the fact that oligosaccharides are a major component of breast milk, the addition of oligofructose in formula milks may be a nutritional approach to favouring colonisation by a beneficial flora.

J Dairy Sci, 2002 May, 85(5), 1058 - 69
Production of oligosaccharides in yogurt containing bifidobacteria and yogurt cultures; Lamoureux L et al.; Yogurts were prepared by using yogurt cultures combined to mixed cultures of bifidobacteria (Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium longum) and by adding a preincubation step (1.5 h at 50 degrees C) with bifidobacteria to the conventional method of manufacture in order to produce oligosaccharides . The survival of bifidobacteria was drastically affected during storage of yogurts, except for products containing B . animalis, in which viable counts remained at >10(6) cfu/g after 28 d of storage at 4 degrees C . Oligosaccharides with a degree of polymerization of 3 were produced during the preincubation step (0.31 to 0.68%), and the amount in the final products varied according to the species of bifidobacteria inoculated during the preincubation step or the concentration of bifidobacteria used as second inoculum during the fermentation process . In fact, the higher concentration of oligosaccharides measured at the end of the fermentation process (0.72%) and the 28 d-storage period (0.67%) was obtained for yogurts containing B . infantis . However, yogurts containing B . breve showed higher beta-galactosidase activities and had lower lactose concentrations after the fermentation process and the storage period than the other yogurts . The use of a mixed cultures of bifidobacteria (B . animalis, B . infantis, or B . breve) thus allows the production of yogurts in which bifidobacteria can survive in relatively high cell numbers and contain appreciable amount of oligosaccharides.

Appl Microbiol Biotechnol, 2002 Jun, 59(1), 72 - 8 Epub 2002 Apr 12.
2-Amino-3-carboxy-1,4-naphthoquinone affects the end-product profile of bifidobacteria through the mediated oxidation of NAD(P)H; Yamazaki S et al.; Glucose metabolism of bifidobacteria in the presence of 2-amino-3-carboxy-1,4-naphthoquinone (ACNQ), a specific growth stimulator for bifidobacteria, and ferricyanide (Fe(CN)(6)(3-)) as an extracellular electron acceptor was examined using resting cells of Bifidobacterium longum and Bifidobacterium breve . NAD(P)H in the cells is oxidized by ACNQ with the aid of diaphorase activity, and reduced ACNQ donates the electron to Fe(CN)(6)(3-) . Exogenous oxidation of NADH by the ACNQ/Fe(CN)(6)(3-) system suppresses the endogenous lactate dehydrogenase reaction competitively, which results in the remarkable generation of pyruvate and a decrease in lactate production . In addition, a decrease in acetate generation is also observed in the presence of ACNQ and Fe(CN)(6)(3-) . This phenomenon could not be explained in terms of the fructose-6-phosphate phosphoketolase pathway, but suggests rather that glucose is partially metabolized via the hexose monophosphate pathway . This was verified by NADP(+)-induced reduction of Fe(CN)(6)(3-) in cell-free extracts in the presence of ACNQ . Effects of the ACNQ/Fe(CN)(6)(3-) system on anaerobically harvested cells were also examined . Stoichiometric analysis of the metabolites from the pyruvate-formate lyase pathway suggests that exogenous oxidation of NADH is an efficient method to produce ATP in this pathway.

J Microbiol Methods, 2002 Sep, 51(1), 57 - 62
Detection of aminotransferase activity of Propionibacterium freudenreichii after SDS-PAGE; Thierry A et al.; Aminotransferases (ATs) had previously been detected after native electrophoresis . We show now that aminotransferase(s) of Propionibacterium freudenreichii can be detected after sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) . Moreover, it retained a high activity (84%) in the presence of 0.23% SDS, contrary to what was observed for aminotransferase(s) of Bifidobacterium bifidum (54%) and of six other cheese-related species (0-20%).

Dis Colon Rectum, 2002 Mar, 45(3), 384 - 8
Sulfate-reducing bacteria colonize pouches formed for ulcerative colitis but not for familial adenomatous polyposis; Duffy M et al.; PURPOSE: Ileal pouch-anal anastomosis remains the "gold standard" in surgical treatment of ulcerative colitis and familial adenomatous polyposis . Pouchitis occurs mainly in patients with a background of ulcerative colitis, although the reasons for this are unknown . The aim of this study was to characterize differences in pouch bacterial populations between ulcerative colitis and familial adenomatous pouches . METHODS: After ethical approval was obtained, fresh stool samples were collected from patients with ulcerative colitis pouches (n = 10), familial adenomatous polyposis (n = 7) pouches, and ulcerative colitis ileostomies (n = 8) . Quantitative measurements of aerobic and anaerobic bacteria were performed . RESULTS: Sulfate-reducing bacteria were isolated from 80 percent (n = 8) of ulcerative colitis pouches . Sulfate-reducing bacteria were absent from familial adenomatous polyposis pouches and also from ulcerative colitis ileostomy effluent . Pouch Lactobacilli, Bifidobacterium, Bacteroides sp, and Clostridium perfringens counts were increased relative to ileostomy counts in patients with ulcerative colitis . Total pouch enterococci and coliform counts were also increased relative to ileostomy levels . There were no significant quantitative or qualitative differences between pouch types when these bacteria were evaluated . CONCLUSIONS: Sulfate-reducing bacteria are exclusive to patients with a background of ulcerative colitis . Not all ulcerative colitis pouches harbor sulfate-reducing bacteria because two ulcerative colitis pouches in this study were free of the latter . They are not present in familial adenomatous polyposis pouches or in ileostomy effluent collected from patients with ulcerative colitis . Total bacterial counts increase in ulcerative colitis pouches after stoma closure . Levels of Lactobacilli, Bifidobacterium, Bacteroides sp, Clostridium perfringens, enterococci, and coliforms were similar in both pouch groups . Because sulfate-reducing bacteria are specific to ulcerative colitis pouches, they may play a role in the pathogenesis of pouchitis.

Microbiol Immunol, 2002, 46(4), 293 - 7
Adhesion of lactic acid bacteria to caco-2 cells and their effect on cytokine secretion; Morita H et al.; Cytokines secreted by human enterocytes play a critical role in mucosal and systemic immunity . Intestinal microorganisms can influence this secretion . In the present study, 30 strains of lactic acid bacteria were characterized for their adhesion to Caco-2 cells and their potential to stimulate proinflammatory cytokine secretion by this cell line . The bacteria adhered in a strain-dependent manner to Caco-2 cells . Contact with lactobacilli did not result in the production of IL-6 or IL-8 . A slight IL-6 and IL-8 production by a Caco-2 cell was detected after exposure to 8 of the tested Bifidobacterium strains . No correlation was found between adhesion and cytokine induction among the bacteria tested . This indicates that lactic acid bacteria, even those with strong adhesive properties, are not very likely to trigger an inflammatory response in human enterocytes.

Berl Munch Tierarztl Wochenschr, 2002 May-Jun, 115(5-6), 173 - 8
{Bifidobacterium adolescentis suppresses the humoral immune response to an autochthonous intestinal bacterium--experiments with gnotobiotic rats}; Scharek L et al.; On the basis of association-experiments with gnotobiotic rats, we described the immunogenicity of two selected bacterial species (Bifidobacterium adolescentis and Bacteroides thetaiotaomicron) . B . adolescentis is a gram-positive lactic acid producing bacterium, strains of which are claimed to have probiotic properties . B . thetaiotaomicron is a gram-negative rod, autochthonous to the human as well as to the rats' intestinal tract . Colonization of the gut was monitored by determination of bacterial cell counts in the animals' feces . In order to investigate the systemic immune reaction, the amounts of specific serum-IgG and -IgA against both bacterial species were measured in the serum . The intestinal immune reaction was examined by measuring the specific IgA in the rats' feces . Knowing about the antibody levels in gnotobiotic rats induced by monoassociation we subsequently disassociated two groups of rats in order to investigate the impact of B . adolescentis on the immune reaction against B . thetaiotaomicron . One group was disassociated simultaneously with B . adolescentis and B . thetaiotaomicron, the second group was disassociated with these bacteria in sequence . B . adolescentis was merely able to induce a mucosal immune reaction, while B . thetaiotaomicron challenged the mucosal as well as the systemic immune system . Furthermore B . adolescentis obviously suppressed the systemic and mucosal immune reaction against the autochthonous B . thetaiotaomicron.

Int J Syst Evol Microbiol, 2002 May, 52(Pt 3), 995 - 9
Bifidobacterium scardovii sp . nov., from human sources; Hoyles L et al.; Five strains of an unusual catalase-negative Gram-positive asporogenous rod-shaped bacterium from human sources were subjected to a polyphasic taxonomic study . The presence of fructose-6-phosphate phosphoketolase, a key enzyme of bifidobacterial hexose metabolism, indicated the strains were members of the genus Bifidobacterium but they did not correspond to any of the recognized species of this genus on the basis of biochemical profiles and whole-cell protein analyses . Comparative 16S rRNA gene sequencing confirmed the placement of the isolates in the genus Bifidobacterium, and demonstrated they represent a hitherto unknown subline within the genus displaying > 5% sequence divergence with recognized species . Based on both phenotypic and phylogenetic criteria, it is proposed that the isolates recovered from human sources be classified as a new species, Bifidobacterium scardovii sp . nov.; the type strain is CCUG 13008T (= DSM 13734T).

Int J Syst Evol Microbiol, 2002 May, 52(Pt 3), 809 - 12
Transfer of Bifidobacterium inopinatum and Bifidobacterium denticolens to Scardovia inopinata gen . nov., comb . nov., and Parascardovia denticolens gen . nov., comb . nov., respectively; Jian W et al.; Bifidobacterium inopinatum Crociani et al . 1996 and Bifidobacterium denticolens Crociani et al . 1996 have distinct phenotypic characteristics and low G+C contents compared with other bifidobacteria . In the 16S rRNA phylogenetic tree, these two species grouped in an independent subcluster . In our previous work, partial heat-shock protein 60 (HSP60) gene-sequence analysis also indicated that these two species had distinct taxonomic positions . In this work, the complete HSP60 genes of five representative bacterial strains were sequenced by using an inverse PCR method . The complete sequence similarities turned out to be at the same level as those of the partial genes, thus confirming the result based on partial sequence analysis . On the basis of all the evidence mentioned above, it is proposed that these two species should be transferred to two new genera as Scardovia inopinata gen . nov., comb . nov., and Parascardovia denticolens gen . nov., comb . nov.

Arch Pediatr, 2002 May, 9(5), 543 - 8
{Human milk: a perpetual (re)appreciation}; Langhendries JP; Based upon recent studies, it appears more and more evident that exclusive breast-feeding can be said as the best functional food, the last being defined as: "a food that has components or ingredients bringing a specific medical or physiological benefit to the host, other than purely nutritional effects" . In particular, human breast milk does favour optimal bacterial intestinal colonization with bifidobacteria and lactic acid bacteria in the neonates and contains a large number of components with antimicrobial activity.

Int J Food Microbiol, 2002 Jun 5, 76(1-2), 75 - 81
Application of nontraditional meat starter cultures in production of Hungarian salami; Pidcock K et al.; Listeria monocytogenes and Escherichia coli O111 have been implicated in several outbreaks of food-borne disease linked to smallgoods products . Traditional meat starter cultures, containing a mixture of lactic acid bacteria (LAB) and staphylococci, are used to maintain safety and sensory properties of Hungarian salami . The present study investigated if nontraditional meat starter (NTMS) cultures can be used for improving the safety of Hungarian salami . Salami batter was inoculated with List . monocytogenes and E . coli and subsequently fermented with NTMS cultures and a commercially available meat starter . A total of 15 NTMS cultures were tested . The salami was monitored for levels of pathogen, LAB and pH . When used in conjunction with the commercial meat starter, 9 NTMS cultures reduced the E . coli O111 count by more than 2.5 log units, whereas 10 of the NTMS cultures reduced List . monocytogenes by more than 2.5 log units . The commercial meat starter alone reduced E . coli and List . monocytogenes by 1.2 and 1.3 log units, respectively . Some NTMS cultures reduced the pathogen count without affecting pH of the salami batter . All NTMS cultures survived in salami throughout fermentation and maturation . It was concluded that NTMS cultures, including Lactobacillus acidophilus LAFTI L10, L . paracasei LAFTI L26, L . paracasei 5119, Lactobacillus sp . L24 and Bifidobacterium lactis LAFTI B94, may be used to increase the safety of Hungarian salami because these cultures gave strong inhibition of both E . coli O111 and List . monocytogenes.

Pediatr Res, 2002 Jun, 51(6), 750 - 5
Immune responses in rhesus rotavirus-challenged BALB/c mice treated with bifidobacteria and prebiotic supplements; Qiao H et al.; Bifidobacterium species (B . bifidum and B . infantis), with or without prebiotic compounds (arabino-galactan, short-chain fructo-oligosaccharide, iso-malto-dextrins), were orally fed to Balb/c pups (n = 192) to evaluate their potential synergistic effects on modulating the course of rhesus rotavirus (RRV) infection, as well as their ability to mediate the associated mucosal and humoral immune responses . Rotavirus-specific IgA and IgG in serum, rotavirus antigen, and specific IgA in feces were measured by ELISA . Mucosal total IgA and IgG levels were determined in Peyer's patches by flow cytometry . Significantly delayed onset (p = 0.001) and early resolution (p < 0.001) of diarrhea were observed in bifidobacteria-treated, RRV-infected mice compared with RRV-infected control mice . Supplementation with prebiotic compounds did not shorten the clinical diarrhea course more than that observed with bifidobacteria treatment alone . Rotavirus-specific IgA in feces was 16-fold elevated on d 5 postinfection in bifidobacteria-treated, RRV-infected mice compared with the RRV-infected alone group . In addition, the level of rotavirus-specific IgA in serum was four-fold higher in bifidobacteria-treated, RRV-infected litters versus mice challenged with RRV alone on 28 and 42 d postinfection . No enhancement of the immune response was found in RRV-infected mice that were treated with both bifidobacteria and prebiotic compounds over those treated with bifidobacteria only . The findings suggest that bifidobacteria may act as an adjuvant by modulating early mucosal and strong humoral rotavirus-specific immune responses, and mitigate severity of rotavirus-induced diarrhea.

Mikrobiologiia, 2002 Mar-Apr, 71(2), 205 - 10
{Characterization of the cell-bound polysaccharides of Bifidobacterium adolescentis 94 BIM}; Novik GI et al.; The cell-bound polysaccharides (PSs) of Bifidobacterium adolescentis 94 BIM were stripped from the cell surface with 2% sodium dodecyl sulfate (SDS), 1.5% Cetavlon, and 1% Triton X-100 and purified by precipitation with 5 volumes of ethanol . According to the extraction conditions used, the polysaccharide preparations were designated as PS-SDS-6 degrees C, PS-SDS-100 degrees C, PS-Cet, and PS-Trit . The gel-permeation chromatography of the first two preparations with the use of a Bio-Gel P-10 column and 1% acetic acid as the eluant yielded two peaks, F1 and F2, which contained carbohydrates and no phosphorus . All polysaccharides were primarily composed of glucose and galactose . The polysaccharides PS-Cet and PS-Trit were found to be branched and contain glucose residues at the terminal position, position 4, and position 6, and galactose residue at position 3 . PS-SDS-6 degrees C has a glucose residue at position 4.

Am J Gastroenterol, 2002 May, 97(5), 1182 - 6
Effect of probiotic strains on interleukin 8 production by HT29/19A cells; Lammers KM et al.; OBJECTIVES: Promising results from clinical studies on the effect of probiotics as maintenance therapy in inflammatory bowel disease and in the prevention of onset of pouchitis ask for studies to unravel the still poorly understood mechanism of action of probiotics . METHODS: To evaluate whether the probiotic bacteria that were used in the clinical studies (VSL#3, Escherichia coli Nissle 1917, and Lactobacillus GG) are able to induce chemokine production in epithelial cells, HT29/19A monolayers were incubated with cell debris and cell extract fractions of single strains of the probiotic bacteria in doses ranging from 10(3) to 10(9) colony-forming units/ml for 32 h . Supernatants were measured for interleukin 8 by ELISA . RESULTS: Lactobacilli and bifidobacteria strains from VSL#3 and Lactobacillus GG did not induce interleukin 8, whereas both cell debris and cell extracts from E . coli Nissle 1917 induced interleukin 8 production in a dose-dependent way . Cell extracts from streptococcal strains induced interleukin 8 when applied at high concentrations . CONCLUSIONS: Probiotic Gram-positive bacteria did not induce interleukin 8, whereas the nonpathogenic, Gram-negative E . coli Nissle 1917 strain induced interleukin 8 in a dose-dependent way in this culture model . These results suggest that probiotic Gram-positive bacteria and E . coli Nissle 1917 may exert their beneficial effects on the host by a different mechanism of action.

Biosci Biotechnol Biochem, 2002 Mar, 66(3), 679 - 81
Isolation and identification of a new bifidogenic growth stimulator produced by Propionibacterium freudenreichii ET-3; Isawa K et al.; We have found a new growth stimulator for bifidobacteria in the culture broth of Propionibacterium freudenreichii ET-3 . The bifidogenic growth stimulator (BGS) was purified by Diaion HP-20 column chromatography and preparative HPLC . Spectroscopic methods including 1H-NMR, UV, and LC-ESI-MS experiments indicated that the chemical structure of the bifidogenic growth stimulator was 1,4-dihydroxy-2-naphthoic acid (DHNA) . Approximately 10 mg/L of DHNA was found to be produced in the culture broth of P . freudenreichii ET-3.

Biosci Biotechnol Biochem, 2002 Mar, 66(3), 598 - 603
Cloning and expression in Escherichia coli of a gene, hup, encoding the histone-like protein HU of Bifidobacterium longum; Takeuchi A et al.; A genomic DNA library of Bifidobacterium longum ATCC15707 was transfected into an Escherichia coli strain deficient in both HU and IHF, the growth of which is cold-sensitive because of the deficiency in these proteins . Cold-resistant colonies were selected and the DNA was cloned and sequenced . A polypeptide consisted of 93 amino acids, a predicted molecular mass of 9983 Da with an isoelectric point of 10.35, was deduced from an orf in the middle of the DNA fragment . The amino acid sequence was highly similar to HU family proteins, and 26 aas of N terminal was identical to a histone-like protein, HBI, a HU family protein of B . longum . Incapabilities of Mu phage propagation in an E . coli mutant deficient in HU or IHF could be suppressed by DNA bearing this orf . These results showed that the orf is a gene hup encoding HBI, a histone-like protein HU of B . longum.

Am J Gastroenterol, 2002 Apr, 97(4), 939 - 46
Changes in the bacterial flora of the neoterminal ileum after ileocolonic resection for Crohn's disease; Neut C et al.; OBJECTIVE: Bacterial agents have been implicated in the early recurrence of Crohn's disease after ileocolectomy . The aim of our study was to identify and quantify bacteria associated with the ileal mucosa in patients and controls and to correlate specific bacteria with recurrence . METHODS: The predominant bacterial microflora of the ileum were enumerated and identified, aerobically and anaerobically, in biopsies obtained at the time of surgery or by endoscopy from 61 patients with Crohn's disease and 10 ileocolectomy controls . The 61 specimens were comprised of 13 ileal biopsies taken from resection specimens, seven taken after ileostomy, and 41 taken after ileocolectomy . RESULTS: Ileocolectomy induced a significant increase in bacterial counts and variety in the neoterminal ileum in both patients and controls that was not observed in ileostomy biopsies . Comparison between patients and controls revealed greater numbers of Escherichia coli and enterococci in Crohn's disease and of bifidobacteria and ruminococci in controls . Early recurrence was associated with high counts of E . coli and bacteroides and the frequent isolation of fusobacteria . CONCLUSION: After ileocolectomy, colonization of the neoterminal ileum is increased . Our data suggest that increases in the populations of specific bacteria such as E . coli, enterococci, bacteroides, and fusobacteria may be important in postoperative recurrence of Crohn's disease.

J Pediatr Surg, 2002 May, 37(5), 699 - 702
Probiotic supplementation reduces the risk of bacterial translocation in experimental short bowel syndrome; Eizaguirre I et al.; BACKGROUND/PURPOSE: Probiotics are live organisms that survive passage through the gastrointestinal tract and have beneficial effects on the host . Lactobacillus and Bifidobacterium have been recommended for cholesterol lowering, acute diarrhea, prevention of cancer, or inflammatory bowel disease . On the other hand, after massive bowel resection, bacterial overgrowth is frequent and favors bacterial translocation (BT) . The possible beneficial effects of Bifidobacterium lactis (BL) administration on BT in experimental short bowel syndrome (SBS), have not been investigated . The aim of this study was to test the hypothesis that BL administration decreases BT in SBS in animals fed orally . METHODS: One hundred twenty-eight adult Wistar rats fed orally with standard rat chow and tap water "ad libitum" were maintained in individual metabolic cages for 10 days and divided into 3 groups: control group (n = 71): nonmanipulated animals; RES group (n = 39): 80% gut resection from 10 cm beyond the angle of Treitz to 10 cm above the cecum; RES-PRO group (n = 18): same resection and daily 7.8 x 10(8) CFU B Lactis administration, after orogastric intubation . At the end of the experiment they were killed, 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 6% of control group rats . The incidence of BT in the RES group was 87% (34 of 39), whereas only 50% (9 of 18) of RES-PRO animals had BT (P <.05) . The relative risk reduction (RRR) was 0.43 (95% Cl 0.14 to 0.72), and the number needed to treat (NNT) was 3 (95% Cl 2 to 8) . In other words, animals that received BL had the risk of BT reduced by 43% (RRR of 0.43), and of every 3 animals treated, 1 is expected to be free of BT (NNT of 3) . CONCLUSION: Administration of B Lactis reduces the incidence of BT in adult Wistar rats after 80% gut resection .

Cardiology, 2002, 97(2), 55 - 8
Pericarditis due to anaerobic bacteria; Brook I; This review describes the microbiology, diagnosis and management of pericarditis due to anaerobic bacteria . The predominant anaerobes recovered from patients with pericarditis were: gram-negative bacilli (mostly of the Bacteroides fragilis group), anaerobic streptococci, Clostridium spp., Fusobacterium spp., and Bifidobacterium spp . Anaerobic bacteria can be isolated in pericarditis resulting from the following mechanisms: (1) spread from a contiguous focus of infection, either de novo or after surgery or trauma (pleuropulmonary, esophageal fistula or perforation, and odontogenic); (2) spread from a focus of infection within the heart, most commonly from endocarditis; (3) hematogenous infection, and (4) direct inoculation as a result of a penetrating injury or cardiothoracic surgery . No differences were found in the clinical diagnostic features between cases of pericarditis due to anaerobic bacteria and those due to aerobic and facultative bacteria . Anaerobic gram-negative bacilli have increased their resistance to penicillins and other antimicrobials in the last decade . Complete identification and testing for antimicrobial susceptibility and lactamase production are therefore essential for the management of infections caused by these bacteria . Treatment of pericarditis involving anaerobic bacteria includes the use of antibiotic therapy effective against these organisms .

J Appl Microbiol, 2002, 92(5), 958 - 65
Metabolism by bifidobacteria and lactic acid bacteria of polysaccharides from wheat and rye, and exopolysaccharides produced by Lactobacillus sanfranciscensis; Korakli M et al.; AIMS: The metabolism by bifidobacteria of exopolysaccharide (EPS) produced by Lactobacillus sanfranciscensis was investigated . To evaluate the significance of the EPS produced by Lact . sanfranciscensis during dough fermentation on the overall prebiotic properties of bread, metabolism by bifidobacteria of water-soluble polysaccharides (WSP) from wheat and rye was investigated . METHODS AND RESULTS: Polyglucose and polyfructan contained in WSP from wheat and rye were metabolized by bifidobacteria . In contrast, WSP isolated from fermented doughs were not metabolized by bifidobacteria . The arabioxylan fraction of WSP was metabolized neither by bifidobacteria nor by lactobacilli . All the bifidobacteria tested were able to metabolize fructan from Lact . sanfranciscensis . The kinetics of EPS metabolism by various bifidobacteria were characterized by diauxic utilization of fructose and EPS . CONCLUSIONS: Bifidobacteria metabolize fructan from Lact . sanfranciscensis . Polyfructan and the starch fractions from wheat and rye, which possess a bifidogenic effect, were degraded by cereal enzymes during dough fermentation, while the EPS were retained . SIGNIFICANCE AND IMPACT OF THE STUDY: EPS produced by sourdough lactic acid bacteria will improve the nutritional properties of sourdough fermented products.

Appl Microbiol Biotechnol, 2002 Mar, 58(4), 439 - 45 Epub 2002 Feb 01.
Purification and characterization of a recombinant beta-galactosidase with transgalactosylation activity from Bifidobacterium infantis HL96; Hung MN et al.; A beta-galactosidase isoenzyme, beta-Gall, from Bifidobacterium infantis HL96, was expressed in Escherichia coli and purified to homogeneity . The molecular mass of the beta-Gall subunit was estimated to be 115 kDa by SDS-PAGE . The enzyme appeared to be a tetramer, with a molecular weight of about 470 kDa by native PAGE . The optimum temperature and pH for o-nitrophenyl-beta-D-galactopyranoside (ONPG) and lactose were 60 degrees C, pH 7.5, and 50 degrees C, pH 7.5, respectively . The enzyme was stable over a pH range of 5.0-8.5, and remained active for more than 80 min at pH 7.0, 50 degrees C . The enzyme activity was significantly increased by reducing agents . Maximum activity required the presence of both Na+ and K+, at a concentration of 10 mM . The enzyme was strongly inhibited by p-chloromercuribenzoic acid, divalent metal cations, and Cr3+, and to a lesser extent by EDTA and urea . The hydrolytic activity using lactose as a substrate was significantly inhibited by galactose . The Km, and Vmax values for ONPG and lactose were 2.6 mM, 262 U/mg, and 73.8 mM, 1.28 U/mg, respectively . beta-Gall possesses strong transgalactosylation activity . The production rate of galactooligosaccharides from 20% lactose at 30 and 60 degrees C was 120 mg/ml, and this rate increased to 190 mg/ml when 30% lactose was used.

Zh Mikrobiol Epidemiol Immunobiol, 2002 Jan-Feb, (1), 68 - 9
{Use of lactusan for the correction of microecological intestinal disturbances in children}; Rodoman VE et al.; The study demonstrated that lactusan (lactulose) could be used as an effective remedy in dysbacteriosis . Twenty four sick children aged 1 month to 15 years were examined . Administration of lactusan to 21 patients resulted in perceptible improvement of clinical symptoms, mainly to the cessation of diarrhea . In 14 children an increase in the population level of bifidobacteria and lactic acid bacteria and a decrease in the number of opportunistic microorganisms were observed . Lactulose should be referred to prebiotics.

Zh Mikrobiol Epidemiol Immunobiol, 2002 Jan-Feb, (1), 64 - 7
{State of normal intestinal microflora in preschool children living in the ecologically adverse region}; Levanova LA et al.; In children living in an ecologically unfavorable area the quantitative content of bifidobacteria and enterococci appeared to be considerably decreased while the level of sulfate-reducing clostridia, on the contrary, elevated . The suppression of bifidoflora leads to decreased immune responsiveness that promotes different somatic diseases . Bronchitis, pneumonia, tonsillitis, pharyngitis, otitis, allergic diseases, diseases of digestive organs, acute respiratory virus infections are more often registered in children living in ecologically unfavorable Central district than in children living in the ecologically favorable Lenin district.

Int J Vitam Nutr Res, 2002 Mar, 72(2), 101 - 8
Effects of a partially hydrolyzed curdlan on serum and hepatic cholesterol concentration, and cecal fermentation in rats; Shimizu J et al.; A significant reduction was observed for serum and hepatic cholesterol concentrations in the rats fed diet containing a 5% partially hydrolyzed curdlan (PHCD), whereas only the hepatic cholesterol concentration was decreased in the curdlan (CD)-fed rats . The cecal contents in the CD group contained a significantly larger amount of short-chain fatty acids, but not those in the PHCD group . CD, but not PHCD, significantly increased the population of cecal bifidobacteria . From the in vitro fermentation test with cecal contents from cellulose powder (CP) and CD-fed rats, PHCD proved to be easily fermented by both cecal contents; incidentally CD was more susceptible to the cecal contents from CD-fed rats than to those from CP-fed rats . These results suggest that PHCD is involved in the modulation of lipid metabolism and intestinal microflora through a different manner from the native CD in rats.

J Microbiol Methods, 2002 Jun, 50(1), 91 - 5
Purification and characterization of 3,3-dihydroxyazetidine from culture medium of Bacillus mesentericus and B . subtilis; Ozone F et al.; A method is described for the purification of 3,3-dihydroxyazetidine (DHA), which accelerates the growth of Bifidobacterium spp., from the culture medium of Bacillus mesentericus (BM) . Purification involved adsorption to an ion-exchange resin; it required less time and gave a higher yield than a previously reported method . Monitoring the inhibition of E . coli NIHJ JC-2, we searched for other strains that produced 3,3-dihydroxyazetidine . We found that not only B . mesentericus TO-A but also B . subtilis IFO13719 produced the compound of interest.

Int J Food Microbiol, 2002 Mar 25, 74(1-2), 79 - 86
Survival of bifidobacteria after spray-drying; Lian WC et al.; To investigate the survival of bifidobacteria after spray-drying, Bifidobacterium infantis CCRC 14633, B . infantis CCRC 14661, B . longum ATCC 15708, B . longum CCRC 14634 and B . longum B6 were first spray-dried with different carrier media including 10% (w/w) gelatin, gum arabic and soluble starch . B . infantis CCRC 14633 and B . longum were also determined in skim milk . It was found that survival of bifidobacteria after spray-drying varied with strains and is highly dependent on the carriers used . Among the test organisms, B . longum B6 exhibited the least sensitivity to spray-drying and showed the highest survival of ca . 82.6% after drying with skim milk . Comparisons of the effect of carrier concentrations revealed that spray-drying at 10% (w/w) gelatin, gum arabic or soluble starch resulted in the highest survival of bifidobacteria . In addition, among the various outlet-air temperatures tested, bifidobacteria showed the highest survival after drying at 50 degrees C . Elevation of outlet-air temperature caused increased inactivation of bifidobacteria . However, the inactivation caused by increased outlet-air temperature varied with the carrier used, with the greatest reduction observed using soluble starch and the least with skim milk.

Curr Opin Drug Discov Devel, 2002 Mar, 5(2), 200 - 3
The genus Bifidobacterium for cancer gene therapy; Fujimori M et al.; A fundamental obstacle in cancer gene therapy is the specific targeting of therapy to solid tumor; as yet, no systemic delivery system exists . Bifidobacterium longum, a strain of domestic bacteria that is non-pathogenic and anaerobic, selectively localized to and proliferated in solid tumors after systemic application . We propose a novel approach to cancer gene therapy in which anaerobic bacteria of the genus B longum are used to achieve tumor-specific gene delivery and enzyme-prodrug therapy.

Curr Opin Drug Discov Devel, 2002 Mar, 5(2), 194 - 9
Live bacteria as anticancer agents and tumor-selective protein delivery vectors; Bermudes D et al.; The development of novel cancer therapies that are selective for cancer cells with limited toxicity to normal tissues is a challenge for oncology researchers . Microorganisms, such as viruses with selectivity for tumor cells or tumor micro-environments, have been investigated as potential arsenals for decades . Genetically-modified, non-pathogenic bacteria have begun to emerge as potential antitumor agents, either to provide direct tumoricidal effects or to deliver tumoricidal molecules . Attenuated Salmonella, Clostridium and Bifidobacterium are capable of multiplying selectively in tumors and inhibiting their growth, representing a new approach for cancer treatment . Because of their selectivity for tumor tissues, these bacteria would also be ideal vectors for delivering therapeutic proteins to tumors . VNP20009, an attenuated strain of Salmonella typhimurium, and its derivative, TAPET-CD, which expresses an Escherichia coli cytosine deaminase (CD), are particularly promising, and are currently undergoing phase I clinical trials in cancer patients.

Biochem Cell Biol, 2002, 80(1), 91 - 4
Lactoferrin-binding proteins in Bifidobacterium bifidum; Kim WS et al.; Lactoferrin is an iron-binding glycoprotein and its bacteriostatic and bactericidal effects on gram-positive and gram-negative bacteria are well known . On the other hand, it is known that certain kinds of lactic acid bacteria are resistant to its antibacterial effects . Moreover, it is reported that lactoferrin promotes the growth of bifidobacteria in in vitro and in vivo experiments . In our experiments, lactoferrin-binding protein was found both in the membrane and cytosolic fractions of Bifidobacterium bifidum Bb-11 . The bifidobacteria were grown in anaerobic conditions with lactobacilli MRS broth containing cysteine, harvested by centrifugation, and processed by sonication . The lactoferrin-binding proteins on the PVDF-membrane transferred after SDS-PAGE were detected by far-Western (western-Western) method using biotinylated lactoferrin and streptavidin-labelled horse radish peroxidase . The molecular weights of the lactoferrin binding protein detected in the membrane fraction were estimated to be 69 kDa and those in cytosolic fractions were 20, 35, 50, and 66 kDa.

J Agric Food Chem, 2002 Mar 27, 50(7), 1840 - 4
Selective responses of three Ginkgo biloba leaf-derived constituents on human intestinal bacteria; Lee HS et al.; The selective responses of Ginkgo biloba leaf-derived materials against six intestinal bacteria was examined using an impregnated paper disk method and compared with that of bilobalide, ginkgolides A and B, kaempferol, and quercetin . The components of G . biloba leaves were characterized as kaempferol 3-O-alpha-(6' "-p-coumaroylglucosyl-beta-1,4-rhamnoside), kaempferol 3-O-(2' '-O-beta-D-glucopyranosyl)-alpha-L-rhamnopyranoside, and quercetin 3-O-alpha-(6' "-p-coumaroylglucosyl-beta-1,4-rhamnoside) by spectroscopic analysis . The growth responses varied with each bacterial strain tested . At 2 mg/disk, kaempferol 3-O-alpha-(6' "-p-coumaroylglucosyl-beta-1,4-rhamnoside) and quercetin 3-O-alpha-(6' "-p-coumaroylglucosyl-beta-1,4-rhamnoside) revealed potent inhibition against Clostridium perfringens, and kaempferol 3-O-(2' '-O-beta-D-glucopyranosyl)-alpha-L-rhamnopyranoside showed a clear inhibitory effect on Escherichia coli . At 0.5 mg/disk, quercetin 3-O-alpha-(6' "-p-coumaroylglucosyl-beta-1,4-rhamnoside) showed a strong activity against C . perfringens, but weak activity was exhibited by kaempferol 3-O-alpha-(6' "-p-coumaroylglucosyl-beta-1,4-rhamnoside) against C . perfringens and kaempferol 3-O-(2' '-O-beta-D-glucopyranosyl)-alpha-L-rhamnopyranoside against E . coli . No inhibition was observed from treatments conducted with bilobalide, ginkgolides A and B, kaempferol, or quercetin . Furthermore, these isolated compounds did not inhibit Bifidobacterium bifidum, B . longum, B . adolescentis, or Lactobacillus acidophilus.

Folia Microbiol (Praha), 2001, 46(6), 573 - 6
Bifidobacterium bifidum monoassociation of gnotobiotic mice: effect on enterocyte brush-border enzymes; Kozakova H et al.; The effect of intestinal colonization with Bifidobacterium bifidum (Gram-positive anaerobic bacterium colonizing the intestine of healthy new-born mammals, exhibiting a probiotic effect, protecting the intestinal mucosa against colonization by pathogenic microflora) on enterocyte brush-border enzymes was examined in weaned 23-d- and in 2-month-old gnotobiotic inbred mice and compared with that in corresponding germ-free (GF) and conventional (CV) controls . The two groups of GF mice were associated with human B . bifidum 11 d before the end of the experiment . Specific activity of enterocyte brush-border enzymes--lactase, alkaline phosphatase and gamma-glutamyltranspeptidase was significantly higher in both age groups of GF mice in comparison with CV ones; on the other hand, sucrase and glucoamylase activities were higher in CV mice . Monoassociation with B . bifidum accelerates biochemical maturation of enterocytes resulting in a shift of specific activities of brush-border enzymes between the values found for GF and CV mice . This effect of B . bifidum supplementation was less pronounced for alkaline phosphatase, sucrase, glucoamylase and dipeptidyl peptidase i.v . in immature gut of weaned mice than of 2-month-old ones.

Nutr Metab Cardiovasc Dis, 2001 Aug, 11(4 Suppl), 87 - 93
Evaluation in human volunteers of the potential anticarcinogenic activities of novel nutritional concepts: prebiotics, probiotics and synbiotics (the SYNCAN project QLK1-1999-00346); Van Loo J et al.; Prebiotics is a recent novel food concept that includes food ingredients that are not digested in the human upper intestinal tract and hence arrive in the colon where they are selectively fermented by a limited number of colonic bacteria . Amongst these are bifidobacteria and lactobacilli, which are considered indicators of a well-balanced intestinal flora . Probiotics are bacteria that, while passing through the intestine, may exert specific beneficial effects on the host's physiology . In general, probiotics are members of the group of the lactic acid-producing bacteria . By means of a variety of experimental models it was demonstrated that prebiotic carbohydrates and probiotics consistently reduced processes of carcinogenesis and tumorigenesis . Synergistic chemopreventive actions were observed with combinations of the two, which together are called synbiotics . One of the most important causes of death in the ageing western population is colon cancer, which is typically associated with a western-style diet . On the basis of the available experimental data, an EU-funded research project (the SYNCAN project QLK1-1999-00346) was set up to evaluate whether synbiotics and prebiotics can be added to food without detriment to (and hopefully eventually improving) organoleptic properties . They are, as such, a good vector for importing nutritionally interesting properties into our diet.

J Clin Microbiol, 2002 Mar, 40(3), 1001 - 9
Molecular analysis of bacterial species associated with childhood caries; Becker MR et al.; Although substantial epidemiologic evidence links Streptococcus mutans to caries, the pathobiology of caries may involve more complex communities of bacterial species . Molecular methods for bacterial identification and enumeration now make it possible to more precisely study the microbiota associated with dental caries . The purpose of this study was to compare the bacteria found in early childhood caries (ECC) to those found in caries-free children by using molecular identification methods . Cloning and sequencing of bacterial 16S ribosomal DNAs from a healthy subject and a subject with ECC were used for identification of novel species or uncultivated phylotypes and species not previously associated with dental caries . Ten novel phylotypes were identified . A number of species or phylotypes that may play a role in health or disease were identified and warrant further investigation . In addition, quantitative measurements for 23 previously known bacterial species or species groups were obtained by a reverse capture checkerboard assay for 30 subjects with caries and 30 healthy controls . Significant differences were observed for nine species: S . sanguinis was associated with health and, in order of decreasing cell numbers, Actinomyces gerencseriae, Bifidobacterium, S . mutans, Veillonella, S . salivarius, S . constellatus, S . parasanguinis, and Lactobacillus fermentum were associated with caries . These data suggest that A . gerencseriae and other Actinomyces species may play an important role in caries initiation and that a novel Bifidobacterium may be a major pathogen in deep caries . Further investigation could lead to the identification of targets for biological interventions in the caries process and thereby contribute to improved prevention of and treatment for this significant public health problem.

Eur J Nutr, 2001 Dec, 40(6), 293 - 300
Effect of lactobacilli, bifidobacteria and inulin on the formation of aberrant crypt foci in rats; Bolognani F et al.; BACKGROUND: Our studies were aimed at investigating the effect of lactic acid producing bacteria (LAB) or inulin, a natural source of non-digestible oligosaccharides derived from chicory, on the induction by carcinogens of aberrant crypt foci (ACF) in the colon, which are considered to be early precursor lesions of neoplasia . METHODS: Strains of Bifidobacterium longum, Lactobacillus casei and Lactobacillus acidophilus were administered to rats fed a purified high starch diet, under a variety of treatment protocols including daily gavage, via the drinking water and in the diet . The rats were treated with methyl-N-nitrosourea, 1,2-dimethylhydrazine, or azoxymethane (AOM) to induce ACF . RESULTS: In general, no consistent significant changes in ACF numbers were detected in these experiments . In one study, the basal diet of the rats was changed to one containing a higher level of fat (corn oil) . Under these conditions, a significant decrease in AOM-induced colonic ACF was seen in rats given L . acidophilus or inulin . In a concurrent group of animals fed a low fat diet, no significant decrease in ACF was observed . CONCLUSIONS: The results indicate that the type of diet fed can influence the detection of protective effects of LAB and oligosaccharides and that against the background of a diet with a level of fat typical of a Western diet, evidence for a protective effect of L . acidophilus and inulin towards colon cancer was obtained

Zh Mikrobiol Epidemiol Immunobiol, 2001 Sep-Oct, (5), 65 - 8
{Dysbacteriosis in patients with colon polyposis}; Chesnokova MG; The study revealed the most profound changes in the composition of intestinal microflora in patients with polyposis of the large intestine . In these patients anaerobic microflora (bifidobacteria, lactic acid bacteria) was more often suppressed than in other examined groups, in particular, patients with cholelithic disease . The associations of hemolytic Escherichia with Klebsiella pneumoniae, Pseudomonas aeruginosa, Citrobacter freundii were often observed as well as the increased content of enterococci and fungi of the genus Candida . The determination of frequency and degree of manifestations showed that dysbacteriosis was registered in the absolute majority of patients (97.4%) with polyposis of the large intestine . Among patients with cholelithic disease disturbances in microbiocenosis were detected in 60.0% of cases, the profundity and character of the microflora composition changes being less pronounced.

Zhonghua Yu Fang Yi Xue Za Zhi, 1999 Nov, 33(6), 337 - 9
{Roles of bifidobacterium on prevention of experimental colorectal carcinoma and induction of apoptosis}; Wang L et al.; OBJECTIVE: To explore the effect of bifidobacterium adolescence on prevention from experimental colorectal carcinoma and its antitumor mechanism . METHODS: An animal model of nude mouse tumor transplanted from colorectal carcinoma was set up . Bifidobacteria adolescence were intraperitoneally injected into nude mice in advance . The growth rate, ultrastructure, density of apoptotic cells, rate of bcl-2 and bax oncoprotein expression and its positive cell density in transplanted tumor of nude mice from colorectal carcinoma were investigated by employing transmission microscopy, in situ end labeling technique and immunohistochemical staining . RESULTS: The growth rate of transplanted tumors from colorectal carcinoma in the group with bifidobacterium prevention was slower than that in the control group . Many tumor cells at varied apoptotic stages could be observed in transplantation tumor tissue in the bifidobacterium prevention group and their density of apoptotic cells, rate of bcl-2 and bax oncoprotein expression and its positive cell density were significantly higher in the group with bifidobacterium prevention, as compared with the control group (P < 0.01) . But, expression rate of bcl-2 oncoprotein turned contrary to that of bax oncoprotein . CONCLUSION: Bifidobacteria adolescence could markedly prevent from occurrence and development of colorectal carcinoma in vivo and induce apoptosis of tumor.

Clin Microbiol Infect, 1997 Feb, 3(1), 124 - 132
Oral supplementation with lactic acid-producing bacteria during intake of clindamycin; Nord CE et al.; OBJECTIVE: To study the effect of administration of clindamycin with or without supplementation of the intestinal microflora with Bifidobacterium bifidum and Lactobacillus acidophilus . METHODS: Twenty-three healthy subjects received clindamycin by mouth for 7 days . Eleven of the subjects also received capsules containing lyophilized L . acidophilus and B . bifidum for 14 days . The other 12 subjects received placebo . RESULTS: There was a marked decrease in total numbers of anaerobic bacteria during the administration of clindamycin . In the lactic acid bacteria-supplemented group, a tendency towards delayed reduction and earlier increase in bifidobacteria was observed, and two of 11 subjects (18%) were colonized with Clostridium difficile, in comparison with five of 12 (41%) in the placebo group . The total number of microorganisms was significantly higher in the lactic acid bacteria-supplemented group than in the placebo group (p=0.02) 4 days after the end of clindamycin administration . The difference was mainly due to higher counts of Escherichia coli and enterococci . Mean levels of other enterobacteria increased less in the lactic acid bacteria-supplemented group than in the placebo group between days 0 and 14 . CONCLUSIONS: The recolonization with aerobic and anaerobic microorganisms was faster in the lactic acid bacteria-supplemented group than in the placebo group . This may be of importance in preventing colonization with C . difficile.

Eur J Biochem, 2002 Jan, 269(2), 712 - 8
Human milk provides peptides highly stimulating the growth of bifidobacteria; Liepke C et al.; The large intestine of breast-fed infants is colonized predominantly by bifidobacteria, which have a protective effect against acute diarrhea . In this study we report for the first time the identification of human milk peptides that selectively stimulate the growth of bifidobacteria . Several bifidogenic peptides were purified chromatographically from pepsin-treated human milk and identified as proteolytically generated fragments from the secretory component of the soluble polyimmunoglobulin receptor and lactoferrin; both of these proteins exhibit antimicrobial effects . Hydrolysis of the identified peptides with the gastrointestinal proteases pepsin, trypsin and chymotrypsin did not lead to the loss of bifidogenic activity, indicating their potential function in vivo . Sequential comparison revealed a similar structural motif within the identified peptides . A correspondingly designed small peptide (prebiotic lactoferrin-derived peptide-I, PRELP-I) was found to stimulate the growth of bifidobacteria as effectively as the native peptides . The combination of antimicrobial and bifidobacterial growth stimulatory activity in human milk proteins leads to highly specific compounds capable of regulating the microbial composition of infants' large intestine.

J Appl Microbiol, 2001 Dec, 91(6), 1110 - 7
The regulatory effects of whey retentate from bifidobacteria fermented milk on the microbiota of the Simulator of the Human Intestinal Microbial Ecosystem (SHIME); Meddah AT et al.; AIMS: To investigate the effects of whey retentate from Bifidobacteria fermented milk . METHODS AND RESULTS: The simulator of the human intestinal microbial ecosystem (SHIME) was used . The composition of the microbiota and its metabolic activities were analysed . Changes in the microbial composition became apparent within 15 days of the treatment in the vessels representing the ileum and the large intestine . The whey retentate favoured the growth of endogenous bifidobacteria and induced a decrease in Bacteroides fragilis and in sulphite-reducing clostridia, especially Clostridium perfringens . After the administration was stopped, these populations tended to revert to their original levels, except for the streptococci and the staphylococci populations . The treatment also led to an increase in acetic acid, CH4 and CO2 production, suggesting overgrowth of some anaerobic bacteria . Ammonium, generally considered as undesirable, declined . CONCLUSIONS: The whey retentate clearly altered the microbial community in the SHIME . SIGNIFICANCE AND IMPACT OF THE STUDY: Whey retentate appears to exert a beneficial effect on the in vitro gastrointestinal system; these findings warrant confirmation by in vivo studies.

J Appl Microbiol, 2001 Dec, 91(6), 1059 - 66
Evaluation of microencapsulation of a Bifidobacterium strain with starch as an approach to prolonging viability during storage; O'Riordan K et al.; AIMS: To optimize a spray coating process for the production of encapsulated microspheres containing viable Bifidobacterium cells and to determine whether the readily gelatinized modified starch coating used in this study improved bacterial survival in foods or under acid conditions . METHODS AND RESULTS: An air inlet temperature of 100 degrees C was demonstrated to be optimal for the spray drying process, as it afforded good drying, low outlet temperatures (45 degrees C) and resulted in less than 1 log reduction in bifidobacteria numbers during drying . Maximum recovery yields of 30% were obtained after optimizing the air aspiration conditions . The average size of the Bifidobacterium PL1-containing starch microparticles was determined by scanning electron microscopy to be of the order of 5 microm . The starch-coated cells did not display any enhanced viability compared with free PL1 cells when exposed to acid conditions for 6 h or in two dry food preparations over 20 d storage at ambient temperature (19-24 degrees C) . Determination of 1491 nucleotides of the 16S rRNA gene from PL1 indicated that it shared 97% homology with a previously sequenced Bifidobacterium ruminantium strain . CONCLUSIONS: Our data demonstrated that, although spray drying is a valuable process for encapsulating bifidobacteria, further work is required to ascertain a more appropriate coating material that will protect this strain against adverse environmental conditions . SIGNIFICANCE AND IMPACT OF THE STUDY: The production of small, uniformly coated microspheres containing viable bifidobacteria using an affordable and industrially convenient process, such as spray drying, has commercial implications for the production of probiotic products . Although popular for use as a coating polymer by the food industry, this study indicated that modified starches might not be suitable for use as an encapsulating material for probiotic strains.

J Appl Microbiol, 2001 Dec, 91(6), 985 - 96
Increased resistance of mice to Salmonella enterica serovar Typhimurium infection by synbiotic administration of Bifidobacteria and transgalactosylated oligosaccharides; Asahara T et al.; AIMS: The anti-infectious activity of Bifidobacteria in combination with transgalactosylated oligosaccharides (TOS) against Salmonella enterica serovar Typhimurium LT-2 in an opportunistic antibiotic-induced murine infection model in mice was examined . METHODS AND RESULTS: B . breve (strain Yakult) with natural resistance to streptomycin sulphate (SM, MIC: > 4 mg ml(-1)), when given daily at a dose of 108 cfu/mouse orally under SM treatment was constantly excreted at 10(10) cfu g(-1) faeces so long as SM was administered, even at 2 weeks after discontinuing administration of B . breve . Explosive intestinal growth and subsequent extra-intestinal translocation of orally infected LT-2 under SM treatment were inhibited by B . breve colonization, and this anti-infectious activity was strengthened by synbiotic administration of TOS with B . breve . Comparison of anti-Salmonella activity among several Bifidobacterium strains with natural resistance to SM revealed that strains such as B . bifidum ATCC 15696 and B . catenulatum ATCC 27539T conferred no activity, even when they reached high population levels similar those of effective strains such as strain Yakult and B . pseudocatenulatum DSM 20439 . Both the increase in the concentration of organic acids and the lowered pH in the intestine due to bifidobacterial colonization correlated with the anti-infectious activity . Moreover, the crude cecal extract of B . breve-colonized mice exerted growth-inhibitory activity against LT-2 in vitro, whereas that of the ineffective B . bifidum-colonized cecum showed much lower activity . CONCLUSIONS: Intestinal colonization by bifidobacteria given exogenously together with TOS during antibiotic treatment prevents the antibiotic-induced disruption of colonization resistance to oral infection with S . enterica serovar Typhimurium, and the metabolic activity needed to produce organic acids and lower the intestinal pH is important in the anti-infectious activity of synbiotics against enteric infection with Salmonella . SIGNIFICANCE AND IMPACT OF THE STUDY: These results indicate that certain bifidobacteria together with prebiotics may be used for the prophylaxis against opportunistic intestinal infections with antibiotic-resistant pathogens.

Scand J Gastroenterol, 2002 Feb, 37(2), 215 - 21
Effects of lactic acid bacteria on the uptake and distribution of the food mutagen Trp-P-2 in mice; Orrhage KM et al.; BACKGROUND: Lactic acid bacteria have been reported to have antimutagenic and anticarcinogenic properties in vivo and in vitro . Lactobacillus acidophilus and Bifidobacterium longum have earlier been shown to bind the food mutagen Trp-P-2 in vitro . METHODS: The influence of oral supplementation with L . acidophilus NCFB 1748 and B . longum BB 536 on the uptake and distribution of 14C-labelled Trp-P-2 in several mouse tissues was quantified by liquid scintillation measurements and examined by tape section autoradiography (gives an unbiased qualitative registration of differences in overall tissue distribution) in the present investigation . Furthermore, the effect of 13-naphthoflavone (BNF), a cytochrome P4501A (CYP1A)-inducing agent, on the distribution of 14C-labelled Trp-P-2 was examined . RESULTS: After oral (6 mg/kg; 5 microCi) or iv (1.2 mg/kg; 1 microCi) administration of 14C-labelled Trp-P-2, high levels of radioactivity were observed in the bile, urine and contents of the gastrointestinal tract . Lower levels were present in the liver, lung, kidney, intestines, brown fat, submaxillary salivary gland and thymus . In mice supplemented with lactic acid bacteria there was a significantly decreased level (29%-73%) of radioactivity in the lung, thymus, liver, kidney, submaxillary salivary gland and small intestine as compared with controls . In mice pretreated with BNF, a low but distinct localization of radioactivity in the lung was observed, whereas no similar localization occurred in controls . CONCLUSIONS: The results suggest that (i) there is a decreased bioavailability of the Trp-P-2 in the majority of the tissues examined in bacteria supplemented mice and (ii) there is a low but distinct CYP1A-dependent activation of Trp-P-2 in the lung of BNF-treated mice.

Nutr Hosp, 2001 Nov-Dec, 16(6), 239 - 56
Use of pro-, pre- and synbiotics in the ICU--future options; Bengmark S et al.; Although the word synbiotics was coined to describe the combined action of pre- and probiotics, the ability to, like antibiotics, control infection, the term is now increasingly used in a wider sense, as a name for all the substances released by microbial fermentation in the lower gut . One obvious reason is that most of the substances released seem to influence the immune defense, increase resistance to disease, and, most important, prevent complications to surgery such as infections and thrombosis . Protection layer of lactobacillus does not exist only on the GI tract mucosa, it is important at all exterior body surfaces including those of the eye, the nose, the mouth, the respiratory tract, the vagina, not to forget the skin . It is clearly reduced at all sites when the patient is in the settings of ICU . Each human being has his/her own unique microbial collection, especially of strains of Bifidobacterium and Lactobacillus, and it should be possible to identify an individual on the basis of his/her personal intestinal microflora . The flora seems always to be significantly reduced in the sick, especially in connection with severe disease, care in ICU, and in patients with little food intake or on parenteral nutrition . Supply of both pre- and probiotics can modify functions such as appetite, sleep, mood and circadian rhythm, and this most likely through metabolites produced by microbial fermentation in the gut . Supply of lactic acid bacteria (LAB) can also significantly reduce serum levels of a variety of toxins such as endotoxin . An umbrella of supplemented probiotics could provide to the patients with liver cirrhosis a tool to reduce septic manifestations and the incidence of bleeding . LAB are effective in controlling diarrhea of both bacterial and viral origin . A series of experimental studies and several uncontrolled clinical studies support the idea of using probiotics in patients with IBD . Ecoimmunonutrition with pre- pro- and synbiotics offer to be suitable tools in the new millennium.

Zhonghua Wai Ke Za Zhi, 2000 Aug, 38(8), 625 - 9
{The prevention of infection complicating acute necrotizing pancreatitis:an experimental study}; Deng Q et al.; OBJECTIVE: To observe the effects of 5 intervention measures on infection complicating acute necrotizing pancreatitis (ANP) in dogs and rats . METHODS: A lethal model of ANP was reproduced by infusion of artificial bile into the biliopancreatic duct . Animals were divided randomly into: ANP group (no treatment); Chinese medicine group ("Qing Yi Tang"); Bifidobacterium mixture group; purgation group (MgSO(4)); selective decontamination of the digestive tract (SDD) group; and somatostatin group . The pancreas and intestine were observed morphologically and tight junction on ileum epithelia was assessed on cryofracture replicas . Blood and/or tissue levels of DAO and D-lactic acid and uric contents of lactulose/mannitol (L/M), served as indicators of gut barrier function, were measured at various time points . Intestinal flora and incidence of bacterial translocation (BT) to organs were examined . RESULTS: In early stage of ANP, mucosal and epithelial tight junction damage and flora disturbance occurred in the gut . In addition, the gut barrier function indicators deteriorated . The BT rates were as high as 78.6% (canine) and 59.5% (rat) . Treatment with Chinese medicine markedly improved gut barrier function and reduced BT rate (32.1% - 37.0%) . Having similar purgative response as Chinese medicine, MgSO4 did not show any beneficial effect on gut barrier protection and the incidence of BT was not reduced . In bifidobacterium mixture and SDD group, flora balance was preserved well and similar results as Chinese medicine were obtained . The BT rates of these two groups decreased to 33.9% and 33.3%, respectively . Somatostatin markedly blunted pancreatic tissue injury and ameliorated gut barrier damage during early phase of ANP . All intervention measures except for purgation also decreased mortalities (14.3% - 35.3%, compared with 58.8% of the no treatment group) . CONCLUSIONS: Treatment with Chinese medicine, bifidobacterium mixture, SDD and somatostatin attenuated gut barrier damage and BT after ANP, and could be used to prevent secondary infection after ANP, but purgation alone is not effective.

Zhonghua Wai Ke Za Zhi, 1998 Apr, 36(4), 230 - 3
{The pathogenesis of infection complicated by acute necrotizing pancreatitis: an experimental study}; Wu C et al.; OBJECTIVE: To observe the injury of gut barrier function and intestinal bacterial translocation following acute necrotizing pancreatitis (ANP), and to investigate the pathogenesis of infection complicated by ANP . METHOD: Fifteen dogs were colonized with a strain of E . coli JM109 bearing ampicillin-resistant plasmid PUC18, then divided into two groups: control group (n = 7) and ANP group (n = 8) . Acute necrotizing pancreatitis was induced by injection of sodium taurocholate and trypsin into the pancreatic duct . RESULT: In the ANP group, as compared with the control group, serum amylase level increased significantly, the lactulose/mannitol ratio was 2-12 times higher, blood and ileum diamine oxidase activity decreased significantly . E . coli population in intestinal mucosal flora increased, while bifidobacterium and lactobacillus reduced significantly . Bifidobacterium/E . coli ratio was reversed . Morphological study revealed areas of hemorrhage and necrosis in the pancreas and shedding of ileal villi . In the control group, blood cultures were negative, and no organism was found in organs except for mesenteric lymph nodes in two dogs . In the ANP group, blood and organ cultures were positive in all dogs and JM109 could be found uniformly too . CONCLUSION: ANP impaired the gut barrier function, which led to bacterial translocation from the gut to the pancreas and other organs, serving as a primary source of infection secondary to ANP.

Biol Pharm Bull, 2002 Jan, 25(1), 68 - 71
Metabolism of kalopanaxsaponin K by human intestinal bacteria and antirheumatoid arthritis activity of their metabolites; Kim DH et al.; When kalopanaxsaponin K (KPK) from Kalopanax pictus was incubated for 24 h at 37 degrees C with human intestinal microflora, KPK was mainly metabolized to kalopanaxsaponin I (KPI) via kalopanaxsaponin H (KPH) rather than via kalopanaxsaponin J (KPJ), and then transformed to kalopanaxsaponin A (KPA) and hederagenin . Bacteroides sp., and Bifidobacterium sp . and Fusobacterium sp . transformed KPK to KPI and KPA and hederagenin via KPH or KPJ . However, Lactobacillus sp . and Streptococcus sp . transformed KPK to KPI, KPA, and hederagenin only via KPJ . The metabolite KPA of KPK showed potent antirheumatoid arthritis activity.

Appl Environ Microbiol, 2002 Feb, 68(2), 831 - 7
Expression of the xylulose 5-phosphate phosphoketolase gene, xpkA, from Lactobacillus pentosus MD363 is induced by sugars that are fermented via the phosphoketolase pathway and is repressed by glucose mediated by CcpA and the mannose phosphoenolpyruvate phosphotransferase system; Posthuma CC et al.; Purification of xylulose 5-phosphate phosphoketolase (XpkA), the central enzyme of the phosphoketolase pathway (PKP) in lactic acid bacteria, and cloning and sequence analysis of the encoding gene, xpkA, from Lactobacillus pentosus MD363 are described . xpkA encodes a 788-amino-acid protein with a calculated mass of 88,705 Da . Expression of xpkA in Escherichia coli led to an increase in XpkA activity, while an xpkA knockout mutant of L . pentosus lost XpkA activity and was not able to grow on energy sources that are fermented via the PKP, indicating that xpkA encodes an enzyme with phosphoketolase activity . A database search revealed that there are high levels of similarity between XpkA and a phosphoketolase from Bifidobacterium lactis and between XpkA and a (putative) protein present in a number of evolutionarily distantly related organisms (up to 54% identical residues) . Expression of xpkA in L . pentosus was induced by sugars that are fermented via the PKP and was repressed by glucose mediated by carbon catabolite protein A (CcpA) and by the mannose phosphoenolpyruvate phosphotransferase system . Most of the residues involved in correct binding of the cofactor thiamine pyrophosphate (TPP) that are conserved in transketolase, pyruvate decarboxylase, and pyruvate oxidase were also conserved at a similar position in XpkA, implying that there is a similar TPP-binding fold in XpkA.

Curr Microbiol, 2002 Mar, 44(3), 212 - 5
Adhesive property of Bifidobacterium lactis LKM512 and predominant bacteria of intestinal microflora to human intestinal mucin; Matsumoto M et al.; The adhesive property to the intestinal mucin of Bifidobacterium lactis LKM512, B . longum, B . breve, B . bifidum, B . adolescentis, B . infantis, Bacteroides vulgatus, Bacteroides distasonis, Eubacterium aerofaciens, Clostridium perfringens, Escherichia coli, and Lactobacillus acidophilus were examined . Adhesive rate of LKM512 to the mucin was significantly (p < 0.05, 0.01, or 0.001) stronger than the other strains from 2 to 100 time . Though the adhesive property of many strains was almost same to the mucin of 20-year-old and 50-year-old generations, in case of 4-month-old was different . Adhesive inhibitory effect of C . perfringens to the mucin by LKM512 was examined . Under the condition that LKM512 was 108/ml and that C . perfringens was 106/ml, adhesion of C . perfringens to the mucin was inhibited at 99.6%, when LKM512 adhered in advance . There was the strong inhibition of adhesion at 74.0%, when C . perfringens adhered to mucin in advance . Thus, LKM512 can inhibit the adhesion of harmful bacteria to the intestinal mucin, the possibility of using as a probiotic strain has to be verified.

World J Gastroenterol, 1998 Jun, 4(3), 242 - 245
Relationship between enteric microecologic dysbiosis and bacterial translocation in acute necrotizing pancreatitis; Wu CT et al.; AIM:To investigate the potential role of intestinal microflora barrier in the pathogenesis of pancreatic infection.METHODS:Fifteen dogs were colonized with a strain of E.coli JM109 bearing ampicillin resistance plasmid PUC18.The animals were divided into two groups . In experimental group (n = 8), acute necrotizing pancreatitis (ANP) was induced by injection of 0.5ml/kg of sodium tarocholate with 3000U/kg trypsin into the pancreatic duct . The control group (n = 7) underwent laparotomy only . All animals were sacrificed 7 days later . Mucosal and luminal microflora of intestine were analyzed quantitatively, and various organs were harvested for culturing, blood samples were obtained for determination of serum amylase activities and plasma lipopolysaccharide (LPS) concentrations.RESULTS:In the experimental group, the number of E . coli in the intestine was much higher than those of the controls, while bifidobacterium and lactobacillus were decreased significantly (Jejunum, 1.75 +/- 0.95 vs 2.35 +/- 0.79,P <0.05; 1.13 +/- 0.8 vs 1.83 +/- 0.64,P <0.05; ileum, 2.89 +/- 0.86 vs 3.87 +/- 1.05,P <0.05; 1.78 +/- 0.79 vs 3.79 +/- 1.11,P <0.01;cecum, 2.70 +/- 0.88 vs 4.89 +/- 0.87,P <001; 2.81 +/- 0.73 vs 3.24 +/- 0.84,P <0.05 . Content of Cecum, 3.06 +/- 0.87 vs 5.15 +/- 1.44,P <0.01; 2.67 +/- 0.61 vs 4.25 +/- 0.81,P <0.01), resulting in reversal of bifido-bacterium/E . coli ratio as compared with the control group (jejunum,0.51 +/- 0.76 vs 1.23 +/- 0.53, P <0.05; ileum, 0.62 +/- 0.68 vs 1.16 +/- 0.32,P <0.05; cecum,0.46 +/- 0.44 vs 1.03 +/- 0.64, P < 0.05) . In addition, intestinal bacteria were isolated from organs of all animals in the experimental group, and JM109 was also detected in most cases . Positive blood culture was 75.0% and 62.5% on day 1 and 2 after induction of ANP, respectively, but no bacterium was found in the controls . As compared with the control group, blood LPS levels and serum amylase activities increased 1-3 times and 3-8 times respectively.CONCLUSION: Microecological disturbance could occur in ANP, and overgrowth of intestinal gram-negative bacteria may lead to translocation to the pancreas and other organs, becoming the source of pancreatic and peripancreatic infection.

J Infect Chemother, 1999 Mar, 5(1), 52 - 57
Changes in intestinal flora after administration of panipenem/betamipron or sulbactam/cefoperazone for treatment of postoperative infections in gastrectomy patients; Takesue Y et al.; To determine the effect of antibiotics used for the treatment of postoperative infections on the normal intestinal flora in patients who have undergone gastroenterological surgery, we performed a prospective randomized study in patients who had undergone distal gastrectomy . Cefazoline was administered up to postoperative day 3 for prophylaxis . For the treatment of postoperative infections, either panipenem/betamipron (PAPM/BP; n = 5) or sulbactam/cefoperazone (SBT/CPZ; n = 5) was adminstered to ten patients with signs of postoperative infections . Feces were examined for bacteria before and 4 and 7 days after the commencement of administration of these therapeutic antibiotics . The anaerobic bacterial count decreased significantly in four of five patients treated with SBT/CPZ, from 1010.2+/-0.36/g before treatment to 105.22+/-3.51/g . In the PAPM/BP-treated group, the anaerobic bacterial count did not change after antibiotic therapy . We also investigated the major genera of colonic anaerobes, and found that the numbers of bifidobacteria were maintained during PAPM/BP administration . However, in the SBT/CPZ group, the number of bifidobacteria decreased to below the detection limit in four of five patients . Little change was observed in the aerobic bacterial count in either group . However, Enterobacteriaceae were replaced by enterococcus or candida in all patients from the SBT/CPZ group and in one of five patients from the PAPM/BP group . Physicians should be alert for the possible clinical consequences of changes in intestinal microflora after antibiotic treatment of postoperative infections.

J Food Prot, 2002 Jan, 65(1), 214 - 8
Effect of honey on the growth of and acid production by human intestinal Bifidobacterium spp.: an in vitro comparison with commercial oligosaccharides and inulin; Kajiwara S et al.; Five human intestinal Bifidobacterium spp., B . longum, B . adolescentis, B . breve, B . bifidum, and B . infantis, were cultured in reinforced clostridial medium (control) and in reinforced clostridial medium supplemented with 5% (wt/vol) honey, fructooligosaccharide (FOS), galactooligosaccharide (GOS), and inulin . Inoculated samples were incubated anaerobically at 37degrees C for 48 h . Samples were collected at 12-h intervals and examined for specific growth rate . Levels of fermentation end products (lactic and acetic acids) were measured by high-pressure liquid chromatography . Honey enhanced the growth of the five cultures much like FOS, GOS, and inulin did . Honey, FOS, GOS, and inulin were especially effective (P < 0.05) in sustaining the growth of these cultures after 24 h of incubation as compared with the control treatment . Overall, the effects of honey on lactic and acetic acid production by intestinal Bifidobacterium spp . were similar to those of FOS, GOS, and inulin.

Eur J Pharm Sci, 2002 Feb, 15(1), 1 - 9
Probiotics: potential pharmaceutical applications; Kaur IP et al.; Realisation of the importance of human gut microbiota in health restoration and maintenance has kindled an interest in probiotics . Probiotics are defined as the microbial food supplements, which beneficially affect the host by improving its intestinal microbial balance . Probiotics are the health enhancing functional food ingredients used therapeutically to prevent diarrhea, improve lactose tolerance and modulate immunity . They may also have potential to prevent cancer and lower serum cholesterol levels . Lactobacillus, Bifidobacterium and several other microbial species are perceived to exert such effects by changing the composition of the gut microbiota . However, it is important that exogenously administered bacteria reach and establish themselves in the large intestine in an intact form . The use of non-digestible oligosaccharides ('prebiotics') can fortify intestinal microflora and stimulate their growth . The present review encompasses information regarding the probiotics and their proposed uses . It addresses the concepts of prebiotics and synbiotics, the application of genetic engineering to produce newer probiotics . Finally, the list of commercially available products are reviewed with discussion of questions regarding the reliability, utility and the safety of these products.

Curr Opin Lipidol, 2002 Feb, 13(1), 61 - 7
Prebiotics and lipid metabolism; Delzenne NM et al.; Prebiotics are defined as nondigestible food ingredients that beneficially affect the host by selectively stimulating the growth or the activity of one or a limited number of bacteria (bifidobacteria, lactobacilli) in the colon . Dietary fructans are nutritionally interesting oligosaccharides that strictly conform to the definition of prebiotics and (in view of experimental studies in animals and of less numerous studies in humans) exhibit interesting serum or hepatic lipid lowering properties . Other nondigestible/fermentable nutrients, which also modulate intestinal flora activity, exhibit cholesterol or triglyceride lowering effects . Are changes in intestinal bacterial flora composition or fermentation activity responsible for those effects? What is the future of prebiotics in the nutritional control of lipidaemia and cardiovascular disease risk in humans? Those questions only receive partial response in the present review because studies of the systemic effects of prebiotics are still in their infancy, and require fundamental research devoted to elucidating the biochemical and physiological events that allow prebiotics to exert systemic effects on lipid metabolism.

Zhongguo Zhong Xi Yi Jie He Za Zhi, 1999 Sep, 19(9), 541 - 3
{Effect of alum on intestinal microecological balance in mice}; Yan M et al.; OBJECTIVE: To observe the degree of microecological imbalance induced by alum in normal intestine of mice, and the bacterial adherence activity in the intestine of the mice administered orally with alum . METHODS: The mice were medicated orally with alum of small and large doses (0.25 g/kg and 1 g/kg) for 8 weeks, then 5 weeks after stopping administration of alum, microflora analysis and bacterial adherence to intestinal mucosal epithelial cells were carried out respectively . RESULTS: Eight weeks after administration with alum, the counts of Bifidobacteria and Lactobacilli decreased significantly, but the numbers of pathogenetic E . Coli increased significantly . The adherence rate of Bifidobacteria to the enterocytes of mice reduced markedly, but E . Coli was on the contrary of Bifidobacteria . Five weeks after the ceasing of medication, the intestinal flora were balanced, the adherence rate of both strains as mentioned above recovered to normal level . CONCLUSION: The imbalance of intestinal flora of mice administered with alum for a long time were only the transient change of bacterial counts . The imbalance of intestinal flora and the adherence rate of bacteria regained normal status five weeks after cancelling action factor.

J Agric Food Chem, 2002 Jan 16, 50(2), 384 - 9
Biological activity of alpha-galactoside preparations from Lupinus angustifolius L . and Pisum sativum L . seeds; Gulewicz P et al.; Biological activity tests were performed on alpha-galactoside preparations obtained from Lupinus angustifolius L . cv . Mirela (alkaloid-rich) and Pisum sativum L . cv . Opal seeds . The studies included the following tests: acute toxicity, cytotoxic test, delayed type hypersensitivity (DTH), plaque-forming cell number (IgM-PFC), and influence on the growth of bifidobacteria and coliform presence in rat colon . Results of these studies showed that alpha-galactosides from lupin and pea seeds were essentially nontoxic . Their acute toxicity (LD(50)) in mice was >4000 mg kg(-1) of body weight . alpha-galactoside preparations were not cytotoxic for mouse thymocytes in vitro . The in vitro test shows that oligosaccharides from lupin and pea are utilized by selected beneficial colon bacterium strains . The in vivo experiment demonstrated that alpha-galactosides from legume significantly influenced the growth of bifidobacteria in rats colon . Simultaneously, the decrease of the coliform presence was observed . The chemical composition of the tested preparations had no significant effect on their biological activity.

Appl Microbiol Biotechnol, 2001 Dec, 57(5-6), 647 - 52
High-efficiency synthesis of oligosaccharides with a truncated beta-galactosidase from Bifidobacterium bifidum; Jorgensen F et al.; An exceptionally large beta-galactosidase, BIF3, with a subunit molecular mass of 188 kDa (1,752 amino acid residues) was recently isolated from Bifidobacterium bifidum DSM20215 {Moller et al . (2001) Appl Environ Microbiol 67:2276-2283} . The BIF3 polypeptide comprises a signal peptide followed by an N-terminal beta-galactosidase region and a C-terminal galactose-binding motif . We have investigated the functional importance of the C-terminal part of the BIF3 sequence by deletion mutagenesis and expression of truncated enzyme variants in Escherichia coli . Deletion of approximately 580 amino acid residues from the C-terminal end converted the enzyme from a normal, hydrolytic beta-galactosidase into a highly efficient, transgalactosylating enzyme . Quantitative analysis showed that the truncated beta-galactosidase utilised approximately 90% of the reacted lactose for the production of galacto-oligosaccharides, while hydrolysis constituted a 10% side reaction . This 9:1 ratio of transgalactosylation to hydrolysis was maintained at lactose concentrations ranging from 10% to 40%, implying that the truncated beta-galactosidase behaved as a "true" transgalactosylase even at low lactose concentrations.

Immunobiology, 2001 Dec, 204(4), 442 - 57
The failure of oral tolerance induction is functionally coupled to the absence of T cells in Peyer's patches under germfree conditions; Maeda Y et al.; Although intestinal bacterial flora has been thought to play a role in the induction of oral tolerance, the mechanism has yet to be elucidated . We therefore examined the bacterial flora-dependent acquisition of susceptibility to oral tolerance induction using a gnotobiotic murine model . Germ-free (GF) mice exhibited a significant shortage of T cells in the PPs in comparison to SPF mice . A recovery in the number of such T cells was accomplished in the gnotobiotic mice associated with Bifidobacterium infantis or Escherichia coli but not in the gnotobiotic mice with Clostridium perfringens or Staphylococcus aureus . To examine the susceptibility to oral tolerance induction, these mice were orally given ovalbumin (OVA) as a tolerogen and then injected i.p . with the Ag . The Ag-specific IgG1 in the serum remained at a low level in both SPF and those gnotobiotic mice groups containing a sufficient number of T cells in the PPs . However, no such unresponsiveness in the Ab response was observed in GF or the other gnotobiotic mice groups containing only a few T cells in the tissues . Adoptive cell transfer analysis clearly showed that a sufficient number of T cells in the PPs is required for the induction of oral tolerance . Furthermore, the reduced expression of SLC (secondary lymphoid-tissue chemokine), which is responsible for T-cell migration to lymphoid organs, was observed in the PPs of GF mice, resulting in a shortage of T cells in the tissues . However, the reduced expression of SLC was restored even in the GF mice after conventionalization, thus suggesting that the failure of oral tolerance induction is functionally coupled to the innate absence of T cells under the GF condition.

J Clin Microbiol, 2002 Jan, 40(1), 265 - 7
Identification by 16S rRNA gene sequencing of Lactobacillus salivarius bacteremic cholecystitis; Woo PC et al.; An anaerobic, nonsporulating, gram-positive bacterium was isolated from blood and bile pus cultures of a 70-year-old man with bacteremic acute cholecystitis . The API 20A system showed that it was 70% Actinomyces naeslundii and 30% Bifidobacterium species, whereas the Vitek ANI system and the ATB ID32A Expression system showed that it was "unidentified." The 16S rRNA gene of the strain was amplified and sequenced . There were 3 base differences between the nucleotide sequence of the isolate and that of Lactobacillus salivarius subsp . salivarius or L . salivarius subsp . salicinius, indicating that the isolate was a strain of L . salivarius . The patient responded to cholecystectomy and a 2-week course of antibiotic treatment . Identification of the organism in the present study was important because the duration of antibiotic therapy would have been entirely different depending on the organism . If the bacterium had been identified as Actinomyces, penicillin for 6 months would have been the regimen of choice . However, it was Lactobacillus, and a 2-week course of antibiotic was sufficient.

Appl Environ Microbiol, 2002 Jan, 68(1), 124 - 37
16S rRNA-based analysis of microbiota from the cecum of broiler chickens; Zhu XY et al.; The microbiota of the intestinal tract of chickens plays an important role in inhibiting the establishment of intestinal pathogens . Earlier culturing and microscopic examinations indicated that only a fraction of the bacteria in the cecum of chickens could be grown in the laboratory . Therefore, a survey of cecal bacteria was done by retrieval of 16S rRNA gene sequences from DNA isolated from the cecal content and the cecal mucosa . The ribosomal gene sequences were amplified with universal primers and cloned or subjected to temporal temperature gradient gel electrophoresis (TTGE) . Partial 16S rRNA gene sequences were determined from the clones and from the major bands in TTGE gels . A total of 1,656 partial 16S rRNA gene sequences were obtained and compared to sequences in the GenBank . The comparison indicated that 243 different sequences were present in the samples . Overall, sequences representing 50 phylogenetic groups or subgroups of bacteria were found, but approximately 89% of the sequences represented just four phylogenetic groups (Clostridium leptum, Sporomusa sp., Clostridium coccoides, and enterics) . Sequences of members of the Bacteroides group, the Bifidobacterium infantis subgroup, and of Pseudomonas sp . each accounted for less than 2% of the total . Sequences related to those from the Escherichia sp . subgroup and from Lactobacillus, Pseudomonas, and Bifidobacterium spp . were generally between 98 and 100% identical to sequences already deposited in the GenBank . Sequences most closely related to those of the other bacteria were generally 97% or less identical to those in the databases and therefore might be from currently unknown species . TTGE and random cloning indicated that certain phylogenetic subgroups were common to all birds analyzed, but sequence data from random cloning also provided evidence for qualitative and quantitative differences among the cecal microbiota of individual birds reared under very similar conditions.

J Food Prot, 2001 Dec, 64(12), 2001 - 6
Enzyme-based most probable number method for the enumeration of Bifidobacterium in dairy products; Bibiloni R et al.; An enzyme-based assay in combination with the most probable number (MPN) technique was developed for the enumeration of bifidobacteria . The assay employs the detection of fructose-6-phosphate phosphoketolase (F6PPK) activity as an indicator of the presence of bifidobacteria . The method was validated against viable counts and optimized with respect to selective media in order to quantitatively assess bifidobacteria in dairy products and other probiotic preparations . Several commercial products and homemade fermented milks were analyzed . Counts of bifidobacteria ranged from 10(7) to 10(8) MPN/ml in commercial products and homemade fermented milks . Commercial starters provided by Argentinean industries had between 10(7) and 10(11) MPN/ml . The results obtained in this study suggest that the combination of F6PPK activity determination and the MPN methodology allows an accurate determination of Bifidobacterium in pure cultures, dairy products, and other probiotic preparations.

Berl Munch Tierarztl Wochenschr, 2001 Nov-Dec, 114(11-12), 410 - 9
{Probiotics--possibilities and limitations of their application in food, animal feed, and in pharmaceutical preparations for men and animals}; Reuter G; Probiotics are cultures of special microorganisms, which have been used as feed additives since the seventies of the past century and already since the twenties in food specimen and in pharmaceuticals . The definition of "Probioticum" was formulated in 1974 simultaneously with the use of living cultures in feed for various animals in order to substitute the application of nutritive antibiotics or chemotherapeutics . In the meantime probiotics are applied not only as feed supplements or pharmaceuticals but increasingly in suitable food specimens such as dairy products, fruit juices, chocolates, and even meat products . Of course, heating of such products prior to consumption or application must be ruled out . The selection of a suitable strain of a microorganism can be regarded as the primary requirement for the use as a probiotic . These cultures must be able to pass the stomach-duodenum barrier in a viable state and to multiply at the site of destination in the intestine . Additionally, they must be capable of producing antagonistic metabolites against a dominating saprophytic microflora resulting in a competitive growth . These abilities are common among lactic acid bacteria, e.g . lactobacilli and bifidobacteria . Yet, intended autochthonous species do not possess very good technological features for the enrichment in food specimens, because their viability will decrease rapidly under unfavourable conditions as it is the case with mineral supplements in feed or with low acidity in fermented dairy products . Therefore, some other microorganisms like spore-forming species or yeast cultures were introduced as probiotic components . These possess some similar features which render them suitable for probiotic use . But their physiological and ecological traits do not qualify them as probiotics of first choice . The special efficacy of probiotics must be strictly verified in animal nutrition due to restrictive EC-regulations, in pharmacy due to legal restraints, and in food applications in accordance with food law regulations . Safety aspects are considered very restrictively in feed applications, in the food and pharmaceutical sector they should be in accordance with the intended purpose of "fulfilling health claims" . In the presented review the different requirements for the application of probiotics in animal nutrition, in food, and in pharmaceuticals will be provided . The special effect of competitive exclusion of pathogenetic and toxinogenic microorganisms in fowl performance are mentioned and even the application of probiotics as marine aquacultures of fish and Crustaceae will be included . Furthermore, the safety aspects resulting from the tremendous amount of industrially produced cultures which are distributed into the environment will be discussed . In conclusion it may be pointed out that probiotics may serve to partially replace the presently reduced or even prohibited application of nutritive antibiotics or chemotherapeutics in animal nutrition and in fulfillment of health claims in man and animals . Economic and environmental aspects will reduce the overall application of probiotics world-wide . Restrictions of use and controls of efficacy and safety are essential and must be implemented periodically.

Can J Microbiol, 2001 Nov, 47(11), 1048 - 52
Enteric bifidobacteria: isolation from human infants and challenge studies in mice; Locascio M et al.; Bifidobacteria from breast-fed infants, formula-fed infants, or premature babies fed by parenteral methods were isolated and identified . The persistence of these microorganisms in the gastrointestinal tract of mice, after oral administration, was studied to determine the optimal dose and frequency of translocation to the liver and spleen . The rate of isolation among infants varied between 19 and 82% depending on the origin of the samples, with the highest values seen in breast-fed babies . The predominant species found in all cases was Bifidobacterium adolescentis . The optimal dose for oral administration of bifidobacteria to mice was 10(7) cells per day per animal for up to 2, 5, or 7 days . These bacteria remained up to 5 days postfeeding, even if feeding was interrupted . The results of bacterial translocation assays showed differences for the different strains and doses tested.

Int J Food Microbiol, 2001 Nov 8, 70(3), 213 - 20
Effects of different probiotic strains of Lactobacillus and Bifidobacterium on bacterial translocation and liver injury in an acute liver injury model; Adawi D et al.; Septic complications represent frequent causes of morbidity in liver diseases and following hepatic operations . Most infections are caused by the individual own intestinal microflora . The intestinal microflora composition is important in physiological and pathophysiological processes in the human gastrointestinal tract, but their influence on liver in different situations is unclear . We therefore studied the effect of different Lactobacillus strains and a Bifidobacterium strain on the extent of liver injury, bacterial translocation and intestinal microflora in an acute liver injury model . Sprague-Dawley rats were divided into five groups: acute liver injury control, acute liver injury + B . animalis NM2, acute liver injury + L . acidophilus NMI, acute liver injury + L . rhamnosus ATCC 53103, and acute liver injury + L . rhamnosus DSM 6594 and L . plantarum DSM 9843 . The bacteria were administered rectally daily for 8 days . Liver injury was induced on the 8th day by intraperitoneal injection of D-galactosamine (1.1 g/kg BW) . Samples were collected 24 h after the liver injury . Liver enzymes and bilirubin serum levels, bacterial translocation (to arterial and portal blood, liver and mesenteric lymph nodes (MLNs)), and intestinal microflora were evaluated . L . acidophilus NM1; L . rhamnosus ATCC 53103, and L . rhamnosus DSM 6594 + L . plantarum DSM 9843 decreased bacterial translocation compared to the liver injury control group . B . animalis NM2 increased bacterial translocation to the mesenteric lymph nodes . The levels of alanine aminotransferase (ALAT) were significantly lower in the L . acidophilus, L . rhamnosus ATCC 53103, L . rhamnosus DSM 6594 + L . plantarum DSM 9843 groups compared to the liver injury group . The L . rhamnosus and L . rhamnosus + L . plantarum groups significantly reduced ALAT levels compared to the B . animalis group . All administered bacteria decreased the Enterobacteriaceae count in the cecum and colon . Administration of different lactobacilli and a Bifidobacterium strain in an acute liver injury rat model, has shown different effects on bacterial translocation and hepatocellular damage . L . acidophilus, L . rhamnosus, and L . rhamnosus + L . plantarum reduced bacterial translocation and hepatocellular damage . B . animalis NM2 increased bacterial translocation to the mesenteric lymph nodes and did not affect hepatocellular damage.

Early Hum Dev, 2001 Nov, 65 Suppl, S43 - 52
Oligosaccharides: application in infant food; Rivero-Urgell M et al.; Oligosaccharides are a complex mixture of approximately 130 compounds present in human milk . It has been shown that human milk oligosaccharides induce an increase in the number of bifidobacteria of colonic flora in breast-fed infants, accompanied with a significant reduction in the number of pathogenic potential bacteria, due to their bifidogenic activity.Complex oligosaccharides have the capacity of inhibiting the binding of pathogens to cell surface because they act as competitive receptors . They are associated with a lower risk of infections and diarrhoea and an improvement of the immune system response.Due to the decrease of the pH intestinal caused by their fermentation, oligosaccharides provoke a reduction of the flora pathogens, an increase of bifidobacteria and an increase of the availability of minerals.In the food industry, simple oligosaccharides such as fructooligosaccharides and galactooligosaccharides are used as bifidogenic oligosaccharides and some infant products contain them in the hope that this might provide some of the benefits attributed to oligosaccharides in human milk.This paper reviews characteristics of oligosaccharides, their beneficial effects and use of oligosaccharides in the food industry.In adults, the benefits of some of the oligosaccharides have been established in several clinical trials, but in infants more fundamental research is needed to establish the metabolic role of these components and the daily intake with bifidogenic activity.

Int J Mol Med, 2002 Jan, 9(1), 65 - 70
Prebiotic treatment of experimental colitis with germinated barley foodstuff: a comparison with probiotic or antibiotic treatment; Fukuda M et al.; There is increasing evidence that intestinal microflora play an important role in the pathogenesis of ulcerative colitis . Therefore, modification of the microflora by prebiotics, probiotics, and antibiotics may be a rational approach for controlling intestinal inflammation . Germinated barley food-stuff (GBF) is an insoluble mixture of glutamine-rich protein and hemicellulose-rich dietary fiber . GBF is utilized efficiently by Bifidobacterium, Lactobacillus, and Eubacterium and converted by them into lactate, acetate, and butyrate . These bacterial organic acids preserve a favorable intestinal condition . We have previously shown that GBF has attenuated intestinal inflammation in patients with ulcerative colitis and experimental colitis models through prebiotic actions . The aim of this study was to compare the effect of GBF with that of probiotics and antibiotics in an experimental colitis model . Colitis was induced by feeding male SD rats with a diet containing 3.0-3.5% dextran sodium sulfate (DSS) . The therapeutic effect of oral administration of a prebiotic (GBF), probiotics (mixture of Lactobacillus and Clostridium butyricum), antibiotics (vancomycin, metronidazole), and the vehicle was determined by assessing clinical and pathological scores on day 6 after initiation of colitis . Butyrate concentrations in the cecal content were also determined . GBF treatment significantly reduced colonic inflammation as assessed by clinical scores with an increase in cecal butyrate levels . Probiotic treatment with a mixture of Lactobacillus and Clostridium butyricum did not show such an effect . Both antibiotic treatments significantly attenuated clinical and pathological scores . However, in contrast to GBF, this treatment led to a significant decrease in cecal butyrate levels . These data suggest that modification of the intestinal microflora by prebiotics, including GBF, may serve as a useful adjunct in the treatment of ulcerative colitis as well as antibiotic treatment.

Acta Paediatr Taiwan, 2001 Sep-Oct, 42(5), 301 - 5
Oral bacterial therapy promotes recovery from acute diarrhea in children; Lee MC et al.; Lactobacilli in the intestines play an important role in developing natural defenses against both intestinal bacterial and viral infections . So a prospective clinical study was carried out at Cathay General Hospital to determine the effect of Lactobacillus acidophilus and Bifidobacterium infantis on the course of acute diarrhea in hospitalized children . Altogether 100 children between 6 and 60 months of age were collected and randomly allocated into 2 groups . Study group (n = 50) was given Infloran Berna, which contains 10(9) viable Lactobacillus acidophilus and 10(9) Bifidobacterium infantis, one capsule tid for 4 days and control group (n = 50) received parenteral rehydration only without any medication . Only 20 stool cultures in study group had positive culture results for Lactobacillus or Bifidobacterium . All children were evaluated for the degree of dehydration before rehydration . The clinical course of diarrhea was followed during the treatment period . Features on admission were similar between the study group and control group in age, duration of diarrhea at home, serum sodium & potassium and dehydration degree . The duration of diarrhea was defined as the time until the last appearance of watery stool . There was no difference between the study group and control group in the frequency of diarrhea stools on the day before admission (p > 0.05) . However, the frequency of diarrhea for study group improved on the first and second day of hospitalization with statistical difference (p < 0.01) . The duration of diarrhea during hospitalization in study group also decreased (3.1 vs . 3.6 days, p < 0.01) . Oral bacterial therapy is an effective adjuvant therapy in rotavirus positive and negative children with diarrhea and can safely be administered during an episode of acute diarrhea.

J Food Prot, 2001 Nov, 64(11), 1775 - 9
Growth and viability of commercial Bifidobacterium spp . in honey-sweetened skim milk; Ustunol Z et al.; Two commercial Bifidobacterium bifidum (Bf-1 and Bf-6) were cultured in 12% (wt/vol) reconstituted nonfat dry milk (NDM) containing 5% (wt/wt) honey, sucrose, fructose, or glucose . Inoculated samples were incubated anaerobically at 37 degrees C for 48 h . Samples were collected at 12-h intervals and examined for (i) specific growth rate, (ii) pH, and (iii) levels of fermentation end products (lactic and acetic acids) as measured by high-performance liquid chromatography (HPLC) . Viability of the organisms during 28 days of refrigerated storage at 4 degrees C was also assessed at 7-day intervals . Growth promotion and acid production were greatest when Bf-1 and Bf-6 were grown in the presence of honey . For both Bf-1 and Bf-6, retention of viability was greatest up to 14 days of refrigerated storageat 4 degrees C when they were grown and stored in the presence of honey compared to other sweeteners.

J Food Prot, 2001 Nov, 64(11), 1667 - 73
Inhibitory activity of Bifidobacterium longum HY8001 against Vero cytotoxin of Escherichia coli O157:H7; Kim SH et al.; Vero cytotoxin (VT)-producing Escherichia coli (VTEC), such as E . coli O157:H7, are emerging foodborne pathogens worldwide . VTs are associated with hemorrhagic colitis and hemolytic uremic syndrome in humans . Attachment of the B subunit of VTs to its receptor, globotriaosylceramide (Gb3), at gut epithelium is the primary step and, consequently, the A subunit of VTs inhibits protein synthesis in the target cell . Proinflammatory cytokines, such as tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta, up-regulate Gb3 expression, increase sensitivity to VTs, and enhance VT action in developing disease . Currently, there is a growing interest in probiotics, given the increasing occurrence of antibiotic-resistant bacteria . In particular, much work on bifidobacteria among probiotics, regarded as microorganisms targeted for technological and therapeutic applications, has been performed . In Korea, the neutralizing effect of the culture supernatant of Bifidobacterium longum HY8001, Korean isolate, against the VTs from E . coli O157:H7 was found . Therefore, this study focused on the raveling of the inhibitory effect of B . longum HY8001 against VTs, through the interference B subunit of VTs and Gb3 interaction . Mice were inoculated intragastrically with B . longum HY8001 culture supernatant before and after challenge with E . coli O157:H7 . Control mice were inoculated intragastrically only with E . coli O157:H7 . Cytokine, TNF-alpha, and IL-1beta levels in sera and expression of their mRNA were decreased, and expression of Gb3 in renal tubular epithelial cells was reduced in mice treated with B . longum HY8001 culture supernatant . In competitive enzyme-linked immunosorbent assays (ELISAs), the culture supernatant of B . longum HY8001 primarily binds VTs to interfere the VTs with Gb3 interaction . These results suggest that soluble substance(s) in B . longum HY8001 culture supernatant may have inhibitory activity on the expression of Gb3, VT-Gb3 interaction, or both . Further study should be done to elucidate the property of soluble substances in B . longum HY8001 culture supernatant.

Am J Clin Nutr, 2001 Dec, 74(6), 833 - 9
Enhancement of immunity in the elderly by dietary supplementation with the probiotic Bifidobacterium lactis HN019; Gill HS et al.; BACKGROUND: The aging process can lead to a decline in cellular immunity . Therefore, the elderly could benefit from safe and effective interventions that restore cellular immune functions . OBJECTIVE: We determined whether dietary supplementation with the known immunostimulating probiotic Bifidobacterium lactis HN019 could enhance aspects of cellular immunity in elderly subjects . DESIGN: Thirty healthy elderly volunteers (age range: 63-84 y; median: 69 y) participated in a 3-stage dietary supplementation trial lasting 9 wk . During stage 1 (run-in), subjects consumed low-fat milk (200 mL twice daily for 3 wk) as a base-diet control . During stage 2 (intervention), they consumed milk supplemented with B . lactis HN019 in a typical dose (5 x 10(10) organisms/d) or a low dose (5 x 10(9) organisms/d) for 3 wk . During stage 3 (washout), they consumed low-fat milk for 3 wk . Changes in the relative proportions of leukocyte subsets and ex vivo leukocyte phagocytic and tumor-cell-killing activity were determined longitudinally by assaying peripheral blood samples . RESULTS: Increases in the proportions of total, helper (CD4(+)), and activated (CD25(+)) T lymphocytes and natural killer cells were measured in the subjects' blood after consumption of B . lactis HN019 . The ex vivo phagocytic capacity of mononuclear and polymorphonuclear phagocytes and the tumoricidal activity of natural killer cells were also elevated after B . lactis HN019 consumption . The greatest changes in immunity were found in subjects who had poor pretreatment immune responses . In general, the 2 doses of B . lactis HN019 had similar effectiveness . CONCLUSION: B . lactis HN019 could be an effective probiotic dietary supplement for enhancing some aspects of cellular immunity in the elderly.

J Appl Microbiol, 2001 Nov, 91(5), 878 - 87
A comparative in vitro evaluation of the fermentation properties of prebiotic oligosaccharides; Rycroft CE et al.; AIMS: Comparison of in vitro fermentation properties of commercial prebiotic oligosaccharides . METHODS AND RESULTS: Populations of predominant gut bacterial groups were monitored over 24 h of batch culture through fluorescent in-situ hybridization . Short-chain fatty acid and gas production were also measured . All prebiotics increased the numbers of bifidobacteria and most decreased clostridia . Xylo-oligosaccharides and lactulose produced the highest increases in numbers of bifidobacteria whilst fructo-oligosaccharides produced the highest populations of lactobacilli . Galacto-oligosaccharides (GOS) resulted in the largest decreases in numbers of clostridia . Short-chain fatty acid generation was highest on lactulose and GOS . Gas production was lowest on isomalto-oligosaccharides and highest on inulin . CONCLUSIONS: The oligosaccharides differed in their fermentation characteristics . Isomalto-oligosaccharides and GOS were effective at increasing numbers of bifidobacteria and lactate whilst generating the least gas . SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides comparative data on the properties of commercial prebiotics, allowing targeting of dietary intervention for particular applications and blending of oligosaccharides to enhance overall functionality.

Curr Issues Intest Microbiol, 2001 Sep, 2(2), 43 - 53
The Lactobacillus and Bifidobacterium microflora of the human intestine: composition and succession; Reuter G; Lactobacillus and bifidobacterial cultures are increasingly used as probiotics in pharmaceuticals and in foods . The selection of strains is performed often for technological rather than for microecological reasons . Detailed reports about species and strains composition of these microorganisms in the intestinal microflora of man are rare . Our investigations were performed with samples originating from infants and adults, taken from faeces and from upper sections of the intestinal tract including mouth and stomach, and from caecum and colon . Post mortem cases as well as test subjects under physiological conditions were analyzed using an automatic capsule system sampling at defined times in different parts of the intestinal tract . The fate of selected strains after oral intake was studied, too . Furthermore, influences of the microflora originating from food were considered . The identification of autochthonous (indigenous) and allochthonous (transient) species could be achieved with descriptions of new species in the genera Lactobacillus and Bifidobacterium . L . gasseri and L . reuteri proved to be predominant autochthonous Lactobacillus species in infants as well as in adults . Both species were occasionally present even in the stomach . This was also the case with an anaerobic lactic acid bacterium, previously named Catenabacterium catenaforme, later classified as L . ruminis, a non-motile variant of this species . The bifidobacterial microflora differed in composition between infants and adults and in different stages of the host's life . Up to 5 species or special strains of bifidobacteria could be present in different, individually fixed, combinations . Species typical for infants were B . bifidum, B . infantis, B . breve, and B . parvulorum . Typical for adults were 4 different variants of B . adolescentis . B . bifidum and B . longum could often be found in both groups, but in lower numbers . B . longum showed some oxygen tolerance whereas B . bifidum and B . adolescentis required strict anaerobic and fastidious conditions for cultivation . The autochthonous Lactobacillus and Bifidobacterium microflora in man will remain stable life-long . With lactobacilli, however, some successions may be caused by transient species derived from food or from the oral cavity, thus giving the impression of an altered microflora . Nevertheless L . gasseri, L . reuteri, L . ruminis, and to some degree, L . salivarius, may be present as autochthonous species all of the time . With bifidobacteria, a decreasing tendency in counts and in multiple composition in elderly people exists . Furthermore, this microflora is also influenced by consumption habits, which are probably caused by geographical circumstances.

J Dairy Sci, 2001 Oct, 84(10), 2152 - 6
Aflatoxin B1 binding by dairy strains of lactic acid bacteria and bifidobacteria; Peltonen K et al.; Various food commodities including dairy products may be contaminated with aflatoxins, which, even in small quantities, have detrimental effects on human and animal health . Several microorganisms have been reported to bind or degrade aflatoxins in foods and feeds . This study assessed the binding of aflatoxin B1 (AFB1) from contaminated solution by 20 strains of lactic acid bacteria and bifidobacteria . The selected strains are used in the food industry and comprised 12 Lactobacillus, five Bifidobacterium, and three Lactococcus strains . Bacteria and AFB1 were incubated (24 h, +37 degrees C) and the amount of unbound AFB1 was quantitated by HPLC . Between 5.6 and 59.7% AFB1 was bound from solution by these strains . Two Lactobacillus amylovorus strains and one Lactobacillus rhamnosus strain removed more than 50% AFB1 and were selected for further study . Bacterial binding of AFB1 by these strains was rapid, and more than 50% AFB1 was bound throughout a 72-h incubation period . Binding was reversible, and AFB1 was released by repeated aqueous washes . These findings further support the ability of specific strains of lactic acid bacteria to bind selected dietary contaminants.

Curr Microbiol, 2001 Nov, 43(5), 351 - 4
Differences in composition and mucosal adhesion of bifidobacteria isolated from healthy adults and healthy seniors; He F et al.; Fifty-one Bifidobacterium strains were isolated from the feces of healthy adults (30-40 years old) and seniors (older than 70 years of age) . B . adolescentis, B . breve, B . infantis, and B . longum were isolated from the healthy adults and B . adolescentis and B . longum from elderly subjects . The tested bacteria bound, in vitro, to intestinal mucus in a strain dependent manner . The strains isolated from healthy adults, and especially B . adolescentis, bound better to intestinal mucus than those isolated from seniors . These results indicate that the mucosal adhesive properties of the human Bifidobacterium flora were reduced with the aging of the host . This shift to a Bifidobacterium flora with reduced adhesive abilities may explain the decrease in bifidobacteria levels in the intestinal microflora of aging people.

Scand J Gastroenterol, 2001 Nov, 36(11), 1185 - 92
Sequential changes in luminal microflora and mucosal cytokine expression during developing of colitis in HLA-B27/beta2-microglobulin transgenic rats; Hata K et al.; BACKGROUND: Transgenic rats expressing HLA-B27 and human beta2-microglobulin (HLA-B27 rats) spontaneously develop chronic colitis resembling human inflammatory bowel disease . We investigated the sequential changes in the luminal bacterial flora and mucosal cytokine mRNA expression in this model . METHODS: HLA-B27 rats were maintained in a specific pathogen-free environment, and luminal microflora was evaluated by standard bacterial culture technique . The expression of mucosal cytokine mRNA was analysed by RT-PCR methods . RESULTS: Clinical symptoms of colitis appeared at 8 weeks of age . The total number of obligate anaerobes was higher than those of facultative anaerobes during the experimental period . At 6 weeks of age, the colonization of Bacteroides spp., Bifidobacterium spp . and Lactobacillus spp . was already detectable at high concentrations, whereas Clostridium spp . and Eubacterium spp . were not detected . The expression of proinflammatory cytokines (IL-Ibeta, IL-8 and TNF-alpha) appeared at 8 weeks of age, and these were detectable until 17 weeks . A similar pattern was observed in the expression of Th1 cytokines (IL-2, IL-12 and IFN-gamma) . On the other hand, the expression of Th2 cytokines (IL-4, IL-10 and TGF-beta) was weak . IL-4 mRNA expression was weakly detectable only at 6 and 8 weeks of age . The expression of IL-10 and TGF-beta mRNA was scarcely detectable throughout the experimental period . CONCLUSION: The development of colitis may be mediated by both the predominant expression of Th1 cytokines and the weakness of Th2 cytokine expression in the mucosa . The colonization of anaerobic bacteria, especially Bacteroides spp., may be initiating and promoting these cytokine responses.

Digestion, 2001, 64(2), 92 - 9
Preventive effects of Bifidobacterium- and Lactobacillus-fermented milk on the development of inflammatory bowel disease in senescence-accelerated mouse P1/Yit strain mice; Matsumoto S et al.; BACKGROUND/AIMS: We recently discovered inflammatory bowel disease (IBD) in the ileum and cecum of SAMP1/Yit strain mice under specific pathogen-free (SPF) conditions . To determine the effect of lactic acid bacteria (LAB - Bifidobacterium breve, Bifidobacterium bifidum and Lactobacillus acidophilus)-fermented milk on the prevention of IBD in SAMP1/Yit strain mice, we compared the disease severity on the intestinal inflammation among the group of mice fed saline, unfermented milk or LAB-fermented milk . METHODS: Three-week-old SAMP1/Yit strain mice (n = 72) were subdivided into three groups, that were fed saline, unfermented milk and LAB-fermented milk, respectively . The diets were orally administered daily via a gastric tube . When the mice reached 20 weeks of age, they were sacrificed and the IBD scores were compared among the three groups . RESULTS: Administration of the LAB-fermented milk to SAMP1/Yit strain mice reduced histological injury score, compared with those in saline-treated or unfermented milk-treated SAMP1/Yit strain mice . Ileal tissue weight and myeloperoxidase activity also reduced by treatment of LAB-fermented milk . Moreover, the tissue contents of immunoglobulins such as IgG1 and IgG2a were lower in the inflammatory regions in the SAMP1/Yit strain group fed LAB-fermented milk than that fed saline and unfermented milk . Cytokine-specific ELISA assays indicated the production of T-helper 1 cytokines such as interferon-gamma and tumor necrosis factor-alpha in the culture supernatants of MLN cells in the SAMP1/Yit strain group fed LAB-fermented milk was lower than those fed saline and unfermented milk . On the contrary, the production of interleukin-10 in MLN cells increased by prevention with LAB-fermented milk . CONCLUSION: LAB-fermented milk is beneficial for the treatment of murine IBD and this effect may be modulated through stabilization of the mucosal immunity by LAB .

Int J Food Microbiol, 2001 Sep 28, 69(3), 167 - 82
Media for the isolation and enumeration of bifidobacteria in dairy products; Roy D; Bifidobacteria are commonly used for the production of fermented milks, alone or in combination with other lactic acid bacteria . Bifidobacteria populations in fermented milks should be over 10(6) bifidobacteria/g at the time of consumption of strain added to the product . Hence, rapid and reliable methods are needed to routinely determine the initial inoculum and to estimate the storage time period bifidobacteria remain viable . Plate count methods are still preferable for quality control measurements in dairy products . It is, therefore, necessary to have a medium that selectively promotes the growth of bifidobacteria, whereas other bacteria are suppressed . The present paper is an overview of media and methods including summaries of published comparisons between different selective media . Culture media for bifidobacteria may be divided into basal, elective, differential and selective culture medium . Non-selective media are useful for routine enumeration of bifidobacteria when present in non-fermented milks . Reinforced Clostridial Agar and De Man Rogosa Sharpe (MRS) supplemented with cysteine and agar available commercially are the media of choice for industrial quality control laboratories . Several media for selective or differential isolation have been described for enumeration of bifidobacteria from other lactic acid bacteria . From the large number of selective media available, it can be concluded that there is no standard medium for the detection of bifidobacteria . However, Columbia agar base media supplemented with lithium chloride and sodium propionate and MRS medium supplemented with neomycin, paromomycin, nalidixic acid and lithium chloride can be recommended for selective enumeration of bifidobacteria in dairy products.

J Agric Food Chem, 2001 Oct, 49(10), 4656 - 61
Selective growth inhibitor toward human intestinal bacteria derived from Pulsatilla cernua root; Lee HS et al.; Among 21 medicinal plants, the growth-inhibiting activity of Pulsatilla cernua root-derived materials toward human intestinal bacteria was examined by using an impregnated paper disk method . The biologically active components of P . cernua roots were characterized as 4-hydroxy-3-methoxycinnamic acid and 3,4-dihydroxycinnamic acid by spectroscopic analysis . The activity was compared with that of six commercially available cinnamic acid derivatives trans-cinnamaldehyde, trans-cinnamic acid, cinnamyl alcohol, 2-methoxycinnamic acid, 3-methoxycinnamic acid, and 4-methoxycinnamic acid . The growth responses varied with each bacterial strain tested . Two isolated compounds revealed a potent inhibition against Clostridium perfringens, and moderate to weak activity against Escherichia coli was exhibited by 4-hydroxy-3-methoxycinnamic acid . Weak or no inhibitory activity was obtained against the bifidobacteria or Lactobacillus acidophilus . The inhibitory effect was much more pronounced in C . perfringens and E . coli as compared to B . adolescentis, B . bifidum, B . fragilis, B . longum, or L . acidophilus . Cinnamaldehyde exhibited a strong growth-inhibiting activity, but no inhibition was observed from treatments with trans-cinnamic acid, cinnamyl alcohol, 2-methoxycinnamic acid, 3-methoxycinnamic acid, and 4-methoxycinnamic acid . These results may be an indication of at least one of the pharmacological actions of P . cernua root.

Int J Syst Evol Microbiol, 2001 Sep, 51(Pt 5), 1633 - 8
New approach to phylogenetic analysis of the genus Bifidobacterium based on partial HSP60 gene sequences; Jian W et al.; The partial 60 kDa heat-shock protein (HSP60) genes of 36 Bifidobacterium strains representing 30 different Bifidobacterium species and subspecies and of the type strain of Gardnerella vaginalis were cloned and sequenced using a pair of universal degenerate HSP60 PCR primers . The HSP60 DNA sequence similarities were determined for the taxa at various ranks as follows: 99.4-100% within the same species, 96% at the subspecies level, and 73-96% (mean 85%) at the interspecies level (and 98% in the case of two groups of closely related species, Bifidobacterium animalis and Bifidobacterium lactis, Bifidobacterium infantis, Bifidobacterium longum and Bifidobacterium suis, whose 165 rRNA sequence similarities are all above 99%) . The HSP60 DNA sequence similarities between different Bifidobacterium species and G . vaginalis, a closely related bacterium according to 16S rRNA analysis, ranged from 71 to 79% (mean 75%) . Although the topology of the phylogenetic tree constructed using the HSP60 sequences determined was basically similar to that for 16S rRNA, it seemed to be more clear-cut for species delineation, and the clustering was better correlated with the DNA base composition (mol% G+C) than that of the 16S rRNA tree . In the HSP60 phylogenetic tree, all of the high-G+C (55-67 mol%) bifidobacteria were grouped into one cluster, whereas the low-G+C species Bifidobacterium inopinatum (45 mol %) formed a separate cluster with G . vaginalis (42 mol%) and Bifidobacterium denticolens (55 mol%); a Bifidobacterium species of intermediate G+C content formed another cluster between the two . This study demonstrates that the highly conserved and ubiquitous HSP60 gene is an accurate and convenient tool for phylogenetic analysis of the genus Bifidobacterium.

J Endod, 2001 Oct, 27(10), 605 - 9
Implantation of bacteria from human pulpal necrosis and translocation from root canals in gnotobiotic mice; Ribeiro Sobrinho AP et al.; The aim of this study was to determine whether microorganisms recovered from infected human root canals were able to survive and translocate to a local lymph node when experimentally inoculated into the root canal system of germ-free mice . The microorganisms isolated from two patients with pulpal necrosis were inoculated in two groups of experimental animals; group I (Gemella morbillorum) and group II (Bifidobacterium adolescentis, Fusobacterium nucleatum, and Clostridium butyricum) . G . morbillorum showed the highest frequency of colonization and translocation to the draining lymph node . In group II only F . nucleatum and C . butyricum colonized and translocated when inoculated in tri-association . When the bacteria from group II were inoculated in monoinfection all three species colonized the root canal of germ-free mice and translocated to the draining lymph node, but with different frequencies . We conclude that selective mechanisms occur in which some bacterial species are fit to survive, multiply, and translocate in the germ-free mouse model.

J Allergy Clin Immunol, 2001 Oct, 108(4), 516 - 20
Allergy development and the intestinal microflora during the first year of life; Bjorksten B et al.; BACKGROUND: The intestinal microflora is a likely source for the induction of immune deviation in infancy . OBJECTIVE: The purpose of this study was to prospectively relate the intestinal microflora to allergy development in 2 countries differing with respect to the prevalence of atopic diseases . METHODS: Newborn infants were followed prospectively through the first 2 years of life in Estonia (n = 24) and Sweden (n = 20) . By that age, 9 Estonian and 9 Swedish infants had developed atopic dermatitis and/or positive skin prick test results . Stool samples were obtained at 5 to 6 days and at 1, 3, 6, and 12 months, and 13 groups of aerobic and anaerobic microorganisms were cultivated through use of standard methods . RESULTS: In comparison with healthy infants, babies who developed allergy were less often colonized with enterococci during the first month of life (72% vs 96%; P <.05) and with bifidobacteria during the first year of life (17% to 39% vs 42% to 69%; P <.05) . Furthermore, allergic infants had higher counts of clostridia at 3 months (median value, 10.3 vs 7.2 log(10); P <.05) . The prevalence of colonization with Staphylococcus aureus was also higher at 6 months (61% vs 23%; P <.05), whereas the counts of Bacteroides were lower at 12 months (9.9 vs 10.6 log(10); P <.05) . CONCLUSION: Differences in the composition of the gut flora between infants who will and infants who will not develop allergy are demonstrable before the development of any clinical manifestations of atopy . Because the observations were made in 2 countries with different standards of living, we believe that our findings could indicate a role for the intestinal microflora in the development of and protection from allergy.

Zh Mikrobiol Epidemiol Immunobiol, 2001 Jul-Aug, (4), 65 - 6
{Study of the intestinal microflora in residents of the city of Vladikavkaz}; Ryndina EI; Microbiological study revealed high occurrence of dysbacteriosis with the decreased amount of Escherichia coli (80.9%) and the presence of E . coli hemolytic forms (76.6%) . Rather often the elevated content of E . coli with decreased fermentative properties was noted (51.1%) . In all cases E . coli with decreased biochemical activity had hemolytic activity, which was accompanied by the deficiency of bifidobacteria (79.2% of cases) and the combined deficiency of lactic acid bacteria and bifidobacteria (75% of cases).

Zh Mikrobiol Epidemiol Immunobiol, 2001 Jul-Aug, (4), 47 - 50
{Formation of intestinal microflora in children during the first year of their life}; Levanova LA et al.; In 525 young children the state of intestinal microbiocenosis was studied every month of the first year their life . The study revealed that the process of the microflora formation lasted throughout the first year of their life and was characterized by dysbiotic disturbances . During this period the aggravation of dysbiotic changes in the intestine of these children on months 3, 6-7 and 11-12 was of particular importance . The formation of stable dysbacteriosis led to a decrease in the immunological status of the child, which was manifested by the increased content of such microorganisms as hemolytic cocci, Proteus and a decrease in the quantitative level of bifidobacteria in the total intestinal microbiocenosis by the end of the first year of child's life.

Zh Mikrobiol Epidemiol Immunobiol, 2001 Jul-Aug, (4), 100 - 4
{Hormonal correction of the resident microflora of the vagina and uterus cervix in women with chronic cervicitis}; Dolgushin II et al.; Clinical, microbiological and hormonal examination of women with chronic cervicitis revealed lesions in the upper section of the reproductive tract in a high proportion of those examined, hormonal disturbances being registered in 96.7% of women . Dysbiotic manifestations (suppression of lacto- and bifidoflora and the excessive growth of opportunistic microorganisms) in the uterus cervix and vagina observed in patients with chronic cervititis were not associated with the etiology of the inflammatory process . The degree of dysmicrobiocenosis in the lower section of the genital tract in women with chronic cervicitis depends on the character of hormonal disturbances . The most significant inhibition of the resident flora was observed when ovarian dysfunction occurred and less significant--in cases of hyperprolactinemia and changes in the level of hypophysial hormones . Hormonal disturbances led to contamination of vagina and cervical canal with opportunistic microorganisms that was inversely proportional to the presence of lactic acid bacteria and bifidobacteria in these organs . Complex therapy of women with chronic cervicitis with the use of preparations for the correction of hormonal disturbances made it possible to restore the normal microflora of the genital tract and to improve the results of treatment.

Mikrobiologiia, 2001 Jul-Aug, 70(4), 495 - 502
{Study of the physiological and biochemical characteristics of bifidobacteria at the late stages of population development}; Novik GI et al.; An investigation of the physiological and biochemical characteristics of the Bifidobacterium bifidum no . 1, B . adolescentis MC-42, and B . adolescentis 94-BIM strains showed that bifidobacteria with a higher growth rate produced greater amounts of the end fermentation products, acetate and lactate . The growth of the strains in batch cultures was found to be inhibited by acidic fermentation products . The growth of B . bifidum no . 1 in a batch mode lasted 100 h at a population density of 10(6) CFU/ml and the growth of B . adolescentis MC-42 and 94-BIM lasted 96-120 h at population densities from 10(4) to 10(7) CFU/ml . Analysis of the bifidobacterial populations by light and electron microscopy showed that they represent conglomerates of cells with lysed cytoplasm in the cell center and intact cytoplasm in the apical parts of the cells . The maximum production of extra-cellular and cell-bound proteinases was observed in the logarithmic growth phase . By the 120th h of cultivation, the metabolic activity of cells, the production of proteinases, and the protein content of bifidobacterial cultures considerably decreased . In the first, second, and third subcultures of 96-h-old bifidobacterial cells on fresh nutrient media, the population density of bifidobacteria and their normal physiological and biochemical characteristics were restored after 48 to 72 h of cultivation.

Zh Mikrobiol Epidemiol Immunobiol, 2001 May-Jun, (3), 97 - 9
{Intestinal microbiocenosis in children with intestinal enzymopathy}; Kamilova AT et al.; 141 children with different kinds of intestinal enzymopathy were examined; of these, 33 had celiac disease, 39--the syndrome of celiac disease, 12--congenital lactase deficiency and 57--the syndrome of disaccharidase insufficiency . In these patients a significant decrease in the average characteristics of the main protective flora and the growth of hemolytic and lactose-negative enterobacteria were established . In all groups of patients increased amounts of Proteus were detected, which was indicative of profound dysbiosis . The content of bifidobacteria was found to be decreased in 89.5-97% of the patients and the content of lactic acid bacteria, in 15.8-33.3% . The decreased content of Escherichia coli with normal enzymatic activity (less than 10(7) colony-forming units) was noted in one-third of the patients with the syndrome of celiac disease and congenital lactase deficiency, in about a half of the patients with the syndrome of disaccharidase insufficiency and least of all in patients with celiac disease (9.1%) . The association of opportunistic microbes was detected in 15.6% of the patients, more often in those with celiac disease, the syndrome of celiac disease and congenital lactase deficiency . The severity of disturbances in intestinal eubiosis was found to depend on the gravity of the patients' state.

J Med Microbiol, 2001 Sep, 50(9), 833 - 42
Modulation of genotoxic enzyme activities by non-digestible oligosaccharide metabolism in in-vitro human gut bacterial ecosystems; McBain AJ et al.; Supplementation of the human diet with prebiotic substances such as inulin and non-digestible oligosaccharides (NDO), e.g., galacto-oligosaccharides (GOS), has been associated with various health benefits . However, little information is available regarding the spatial location of their metabolism in human gut bacterial ecosystems . Therefore, the present study investigated the metabolism of inulin and GOS with respect to bacterial growth, bifidobacterial stimulatory properties and anti-mutagenicity potential, in a three-stage continuous culture model of the colon which reproduces the physicochemical characteristics of the proximal (V1) and distal (V2, V3) colons . Fermentation of both carbohydrates was rapid, and occurred primarily in V1, as evidenced by acid formation . Inulin metabolism was associated with 10-fold stimulation of lactobacillus populations, together with smaller increases in bifidobacterial cell counts in V1 . However, peptostreptococci, enterococci and Clostridium perfringens also increased in this fermentation vessel . In contrast, GOS was only weakly bifidogenic in V1, although these bacteria did proliferate in V2 . GOS also increased lactobacilli by an order of magnitude in V1 . However, overall changes in microbial populations resulting from inulin or GOS addition were minimal in V2 and V3 . Potential beneficial effects of inulin metabolism included minor reductions in beta-glucosidase and beta-glucuronidase, whereas GOS strongly suppressed these enzymes, together with arylsulphatase (AS) . Growth of putatively health promoting micro-organisms was not only associated with reductions in enzymes linked to genotoxicity . For example, both carbohydrates stimulated synthesis of nitroreductase and azoreductase, throughout the fermentation system, while inulin increased AS . Colonic transit time is an important factor in bacterial metabolism in the large bowel, and these data suggest that, in some circumstances, NDO fermentation will occurprincipally in the proximal colon.

Zh Mikrobiol Epidemiol Immunobiol, 2001 Mar-Apr, (2), 91 - 4
{Vaginal microflora in girls of prepuberal age}; Boiko AV et al.; The bacteriological study of vaginal microflora in 24 clinically healthy girls aged 7-8 years was carried out . Two criteria of normal vaginal biocenosis in girls of prepuberal age were proposed . The proportion of hemolytic bacteria in the vagina of a girl of prepuberal age could reach 100% of all microflora, provided the total bacterial count (TBC) exceed average values . A high proportion of hemolytic bacteria in the vaginal microflora of girls of prepuberal age in combination with a high level of TBC (even with TBC not exceeding the upper limit of its normal value) could probably by regarded as a border state . No fungi of the genus Candida should be isolated from vagina in girls of prepuberal age . Facultative anaerobic microflora was represented mainly by Staphylococcus epidermidis and S . saprophyticus . In 64.3 +/- 12.8% of cases bifidobacteria were detected in the girls vagina.

Zh Mikrobiol Epidemiol Immunobiol, 2001 Mar-Apr, (2), 114 - 8
{Biological activity of Spirulina}; Blinkova LP et al.; In this review information of Spirulina platensis (SP), a blue-green alga (photosynthesizing cyanobacterium) having diverse biological activity is presented . Due to high content of highly valuable proteins, indispensable amino acids, vitamins, beta-carotene and other pigments, mineral substances, indispensable fatty acids and polysaccharides, PS has been found suitable for use as bioactive additive . SP produces an immunostimulating effect by enhancing the resistance of humans, mammals, chickens and fish to infections, the capacity of influencing hemopoiesis, stimulating the production of antibodies and cytokines . Under the influence of SP macrophages, T and B cells are activated . SP sulfolipids have proved to be effective against HIV . Preparations obtained from SP biomass have also been found active against herpesvirus, cytomegalovirus, influenza virus, etc . SP extracts are capable in inhibiting cancerogenesis . SP preparations are regarded as functional products contributing to the preservation of the resident intestinal microflora, especially lactic acid bacilli and bifidobacteria, and to a decrease in the level of Candida albicans . The biological activity of SP with respect to microorganisms holds good promise for using these microalgae as components of culture media.

Appl Environ Microbiol, 2001 Sep, 67(9), 4256 - 63
Molecular and biochemical analysis of two beta-galactosidases from Bifidobacterium infantis HL96; Hung MN et al.; Two genes encoding beta-galactosidase isoenzymes, beta-galI and beta-galIII, from Bifidobacterium infantis HL96 were revealed on 3.6- and 2.4-kb DNA fragments, respectively, by nucleotide sequence analysis of the two fragments . beta-galI (3,069 bp) encodes a 1,022-amino-acid (aa) polypeptide with a predicted molecular mass of 113 kDa . A putative ribosome binding site and a promoter sequence were recognized at the 5' flanking region of beta-galI . Further upstream a partial sequence of an open reading frame revealed a putative lactose permease gene transcribing divergently from beta-galI . The beta-galIII gene (2,076 bp) encodes a 691-aa polypeptide with a calculated molecular mass of 76 kDa . A rho-independent transcription terminator-like sequence was found 25 bp downstream of the termination codon . The amino acid sequences of beta-GalI and beta-GalIII are homologous to those found in the LacZ and the LacG families, respectively . The acid-base, nucleophilic, and substrate recognition sites conserved in the LacZ family were found in beta-GalI, and a possible acid-base site proposed for the LacG family was located in beta-GalIII, which featured a glutamate at residue 160 . The coding regions of the beta-galI and beta-galIII genes were each cloned downstream of a T7 promoter for overexpression in Escherichia coli . The molecular masses of the overexpressed proteins, as estimated by polyacrylamide gel electrophoresis on sodium dodecyl sulfate-polyacrylamide gels, agree with their predicted molecular weights . beta-GalI and beta-GalIII were specific for beta-D-anomer-linked galactoside substrates . Both are more active in response to ONPG (o-nitrophenyl-beta-D-galactopyranoside) than in response to lactose, particularly beta-GalIII . The galacto-oligosaccharide yield in the reaction catalyzed by beta-GalI at 37 degrees C in 20% (wt/vol) lactose solution was 130 mg/ml, which is more than six times higher than the maximum yield obtained with beta-GalIII . The structure of the major trisaccharide produced by beta-GalI catalysis was characterized as O-beta-D-galactopyranosyl-(1-3)-O-beta-D-galactopyranosyl-(1-4)-D-glucopyranose (3'-galactosyl-lactose).

Int J Food Microbiol, 2001 Aug 5, 67(3), 207 - 16
In vitro adherence properties of Lactobacillus rhamnosus DR20 and Bifidobacterium lactis DR10 strains and their antagonistic activity against an enterotoxigenic Escherichia coli; Gopal PK et al.; Adhesion and colonisation properties of three probiotic strains namely, Lactobacillus rhamnosus DR20, L . acidophilus HN017, and Bifidobacterium lactis DR10, were determined in vitro using the differentiated human intestinal cell-lines including HT-29, Caco-2, and HT29-MTX, and compared with properties of L . acidophilus LA-1 and L . rhamnosus GG (two commercial probiotic strains) . Two independent methods were employed to quantitate the "adhesiveness" of each strain . In the first method, the bacteria adhered to human cells were detected by Gram staining and counted in different fields under a microscope . Bacteria were also radio-labelled and extent of adhesion determined by scintillation counting . All three strains showed strong adhesion with the human intestinal cell lines in vitro . Adhesion indices of the three strains to two cell lines, i.e . HT-29, and Caco-2 varied between 99 +/- 17 and 219 +/- 36 . With mucus-secreting cell-line HT29-MTX, the adhesion indices of all the strains were 2-3 times higher . The adhesion indices of L . acidophilus LA-1 and L . rhamnosus GG were comparable to the other three probiotic strains . We also investigated the inhibitory effect of adhering strains against the intestinal cell monolayer colonization by a known enterotoxigenic strain of Escherichia coli (strain O157:H7) . Pre-treatment of E . coli O157:H7 with 2.5-fold concentrated cell-free culture supernatants from L . acidophilus HN017, L . rhamnosus DR20 and B . lactis DR10 reduced the culturable E . coli numbers on TSB plates and also reduced the invasiveness and cell association characteristics of this toxic strain . The inhibitory molecules secreted into the spent media by these strains were partially affected by treatments with lactate dehydrogenase, trypsin and proteinase K suggesting that overall inhibition may be due to a synergistic action of lactic acid and proteinaceous substances.

Syst Appl Microbiol, 2001 Jul, 24(2), 232 - 7
In vitro study of prebiotic properties of levan-type exopolysaccharides from Lactobacilli and non-digestible carbohydrates using denaturing gradient gel electrophoresis; Bello FD et al.; Batch cultures inoculated with human faeces were used to study the prebiotic properties of levan-type exopolysaccharides (EPS) from Lactobacillus sanfranciscensis as well as levan, inulin, and fructooligosaccharide (FOS) . Denaturing gradient gel electrophoresis of 16S rDNA fragments generated by PCR with universal primers was used to analyse the cultures . Characteristic changes were revealed in the composition of the gut bacteria during fermentation of the carbohydrates . An enrichment of Bifidobacterium spp . was found for the EPS and inulin but not for levan and FOS . The bifidogenic effect of the EPS was confirmed by culturing on selective medium . In addition, the use of EPS and FOS resulted in enhanced growth of Eubacterium biforme and Clostridium perfringens, respectively.

Int J Tissue React, 2001, 23(3), 81 - 7
The synergistic effect of lactic acid bacteria and alkylglycerols on humoral immunity in mice; Chorostowska-Wynimko J et al.; Investigations on immune suppression and reconstitution of immune functions dependent on the presence of physiological microflora allow us to conclude that symbiotic microorganisms such as Lactobacillus sp . are essential for adequate activity of the defense system in humans . In addition to their beneficial influence on the intestinal microbial balance, these microorganisms exert a variety of immunomodulatory effects on the host immune system . On the other hand, immunostimulatory animal-derived substances rich in alkylglycerols have been shown to enhance lactic acid bacteria proliferation . Therefore, the aim of the present study was to evaluate the effects on murine humoral response of the combined administration of lyophilized combination of three lactic acid bacteria: L . acidophilus, L . bulgaricus and Bifidobacterium bifidum together with alkylglycerol-rich shark liver oil . The lactic acid bacteria mixture induced markedly stronger enhancement of the humoral response than alkylglycerols did . A significant synergistic stimulatory effect of lactic acid bacteria and alkylglycerols was observed in both treatment schedules: post- as well as in preimmunization with sheep red blood cells . However, their concomitant administration exerted stronger immunomodulatory effect than did the alternative route of treatment.

Indian J Exp Biol, 2001 May, 39(5), 441 - 6
Sub chronic toxicity studies of lactulose in rats; Baskaran V et al.; Lactulose has profound health benefits by way of increasing bifidobacterial flora in the intestine of infants thereby protecting them against enteric infection, constipation and systemic encephalopathy . In the present study to assess the sub chronic toxicity of lactulose syrup, the rats were fed on a basal feed supplemented with lactulose syrup at 0.5, 1.0, 2.0 and 5.0% for a period of 21 weeks . Monitoring of food consumption, gain in body weight and physical observations did not reveal any treatment-related toxicity in any of the group of rats . Terminal autopsy also did not reveal any signs of toxicity . Further, no significant alterations in relative organ weight, serum biochemistry and urinalysis were observed up to 1% lactulose supplementation level . The results suggest that supplementation of lactulose in the diet does not produce any toxicity at the doses tested.

J Clin Immunol, 2001 Jul, 21(4), 264 - 71
Dietary probiotic supplementation enhances natural killer cell activity in the elderly: an investigation of age-related immunological changes; Gill HS et al.; Many elderly subjects are at increased risk of infectious and noninfectious diseases due to an age-related decline in lymphoid cell activity (immunosenescence) . Noninvasive means of enhancing cellular immunity are therefore desirable in the elderly . Previous reports have suggested that dietary supplementation could represent an effective means of enhancing the activity of circulating natural killer (NK) cells in the elderly . In the present study, we have conducted a pre-post intervention trial to determine the impact of dietary supplementation with probiotic lactic acid bacteria (LAB) on peripheral blood NK cell activity in healthy elderly subjects . Twenty-seven volunteers consumed low-fat/low-lactose milk supplemented with known immunostimulatory LAB strains (Lactobacillus rhamnosus HN001 or Bifidobacterium lactis HN019) for a period of 3 weeks . A dietary run-in of milk alone was shown to have no significant effect on NK cells . In contrast, the proportion of CD56-positive lymphocytes in peripheral circulation was higher following consumption of either LAB strain, and ex vivo PBMC tumoricidal activity against K562 cells was also increased . Supplementation with HN001 or HN019 increased tumoricidal activity by an average of 101 and 62%, respectively; these increases were significantly correlated with age, with subjects older than 70 years experiencing significantly greater improvements than those under 70 years . These results demonstrate that dietary consumption of probiotic LAB in a milk-based diet may offer benefit to elderly consumers to combat some of the deleterious effects of immunosenescence on cellular immunity.

Folia Microbiol (Praha), 2001, 46(1), 73 - 5
Enrichment of bifidobacteria in the hen caeca by dietary inulin; Rada V et al.; Caecal bifidobacterial concentration was increased more than 3-fold in inulin-treated laying hens . The counts of bifidobacteria in birds fed as the control were 9.64, in inulin-diet fed ones 10.17 log CFU/g of caecal content, respectively . Dietary inulin had no effect on caecal microbial metabolite concentration . The proportion of inulin-fermenting bifidobacteria in the total bifidobacteria increased 2-fold in inulin-treated birds.

Probl Tuberk, 2001, (2), 13 - 4
{Pathogenetic mechanisms of impaired intestinal microflora in patients with destructive pulmonary tuberculosis}; Lineva ZE et al.; Following 2-3 months of combined chemotherapy, most (97.5%) patients with destructive pulmonary tuberculosis were found to develop pathological changes in the intestinal microflora . Of them 92.3% had bifidobacterium deficiency, which creates conditions for opportunistic microorganisms to colonize the bowel, promoting the development of an inflammatory process in the gastroduodenal area, Bifidobacterium deficiency being a pathogenetic factor and opportunistic microbes acting as an etiological factor of the impaired intestinal microflora . Assessing of dysbiotic impairments makes it possible to specify the mechanisms of development of toxic reactions and inflammatory processes in the digestive tract.

Int J Food Microbiol, 2001 Jul 20, 67(1-2), 97 - 106
Inhibition of the binding of enterotoxigenic Escherichia coli Pb176 to human intestinal epithelial cell line HCT-8 by an extracellular protein fraction containing BIF of Bifidobacterium longum SBT2928: suggestive evidence of blocking of the binding receptor gangliotetraosylceramide on the cell surface; Fujiwara S et al.; The extracellular protein fraction (P100V) containing the protein BIF produced by Bifidobacterium longum SBT2928 (BL2928), which inhibits the binding of enterotoxigenic Escherichia coli Pb176 (ETEC) to the glycolipid binding receptor gangliotetraosylceramide (GA1) also inhibited the binding of ETEC to the human intestinal epithelial cell line HCT-8 (ATCC CCL 244) in a dose-dependent manner . ETEC-binding inhibitory experiments using crude colonization factor antigen (CFA)-II prepared from ETEC, rabbit anti-GA1 antiserum, medium containing GA1 and media containing lectins, as the binding-inhibitors, suggest that the interaction between the CFA-II antigen present on the cell surface of ETEC and GA1 expressed on HCT-8 cells plays a significant role in the adherence between them . It is strongly suggested that the P100V fraction works as a blocker for the ETEC receptor GA1 on HCT-8 cells.

Zhongguo Zhong Xi Yi Jie He Za Zhi, 1998 Apr, 18(4), 236 - 8
{An experimental study on curative effect of Chinese medicine qing yi tang in acute necrotizing pancreatitis}; Wu C et al.; OBJECTIVE: To observe the curative effect of the traditional Chinese medicine (TCM) Qing Yi Tang (QYT) in acute necrotizing pancreatitis (ANP) . METHODS: Twenty three dogs were randomly divided into 3 groups . In control group (n = 7), animals underwent laparotomy only . In ANP group (n = 8), acute necrotizing pancreatitis was induced by injection of 0.5 ml/kg 5% sodium taurocholate with 300 u/kg trypsin into the pancreatic duct . While in TCM group (n = 8) were fed everyday with QYT after onset of ANP . All animals were sacrified 7 days later and organs were gathered and cultured . Mucosal and luminal floras of the intestine were analysed . Pancreas and ileal mucosa were examined histologically and ultra-microscopically, the levels of lipopolysaccharide (LPS) and amylase in blood were determined . RESULTS: In TCM groups, histologic and ultra-structural damages in pancreas and ileal mucosa were much milder as compared with those of ANP group . In ANP group, there was a significant increase of E . coli and bacterocoids, and a significant decrease of bifidobacteria, lactobacilli and enterococci in the intestinal mucosa, while in TCM groups, these changes were alleviated significantly (P < 0.05 or P < 0.01) . As compared with ANP group, the bacterial translocation (BT) rate was reduced from 100% to 50%, and the counts of translocated bacteria were decreased 10-40 times, the levels of LPS and amylase reduced 2 to 3 times . CONCLUSION: TCM recipe QYT showed their protective effects on gut barrier function by alleviating the damage of intestinal mucosa and microecologic disturbance following acute pancreatitis . As a result, the chances of BT and enterogenic infection declined . These preparation might be promising in the prophylaxis and treatment of infection complicating ANP.

Curr Issues Mol Biol, 1999, 1(1-2), 53 - 64
Identification of lactobacilli and bifidobacteria; Tannock GW; Selective culture media and phenotypic tests enable lactobacilli to be differentiated from morphologically similar bacteria . The accurate identification of Lactobacillus species can be accomplished by reference to 16S rRNA gene sequences . Species-specific, PCR primers that target the 16S-23S rRNA spacer region are available for a limited number of Lactobacillus species . Molecular methods for the comprehensive identification of Bifidobacterium species are not yet available . Only DNA-DNA reassociation provides a reliable means of species identification for this genus at present . Bifidobacteria can be differentiated from morphologically similar bacteria by the use of genus-specific, PCR primers or oligonucleotide probes.

Appl Environ Microbiol, 2001 Aug, 67(8), 3469 - 75
Adhesion of bifidobacteria to granular starch and its implications in probiotic technologies; Crittenden R et al.; Adhesion of 19 Bifidobacterium strains to native maize, potato, oat, and barley starch granules was examined to investigate links between adhesion and substrate utilization and to determine if adhesion to starch could be exploited in probiotic food technologies . Starch adhesion was not characteristic of all the bifidobacteria tested . Adherent bacteria bound similarly to the different types of starch, and the binding capacity of the starch (number of bacteria per gram) correlated to the surface area of the granules . Highly adherent strains were able to hydrolyze the granular starches, but not all amylolytic strains were adherent, indicating that starch adhesion is not a prerequisite for efficient substrate utilization for all bifidobacteria . Adhesion was mediated by a cell surface protein(s) . For the model organisms tested (Bifidobacterium adolescentis VTT E-001561 and Bifidobacterium pseudolongum ATCC 25526), adhesion appeared to be specific for alpha-1,4-linked glucose sugars, since adhesion was inhibited by maltose, maltodextrin, amylose, and soluble starch but not by trehalose, cellobiose, or lactose . In an in vitro gastric model, adhesion was inhibited both by the action of protease and at pH values of < or =3 . Adhesion was not affected by bile, but the binding capacity of the starch was reduced by exposure to pancreatin . It may be possible to exploit adhesion of probiotic bifidobacteria to starch granules in microencapsulation technology and for synbiotic food applications.

Aliment Pharmacol Ther, 2001 Aug, 15(8), 1139 - 45
A study of fructo oligosaccharides in the prevention of travellers' diarrhoea; Cummings JH et al.; BACKGROUND: Prebiotic carbohydrates selectively stimulate the growth of bifidobacteria and lactobacilli in the human colon . These bacteria form part of the gut's inherent defence against invading pathogens . AIM: To test the effectiveness of fructo oligosaccharides in preventing travellers' diarrhoea . METHODS: A total of 244 healthy subjects, travelling to high and medium risk destinations for travellers' diarrhoea, took part in a randomized, double-blind, placebo-controlled study . The protocol comprised a preliminary week for recording bowel habit by diary, a 2-week pre-holiday period with the diary and consumption of 10 g of fructo oligosaccharides or placebo daily, followed by a 2-week holiday with continuation of treatment and diary . A post-study questionnaire was completed by all subjects on their return to the UK . RESULTS: The consumption of fructo oligosaccharides led to a small (6%; P < 0.02) increase in stool frequency in the pre-holiday period and gave a significantly better sense of 'well-being' during the holiday, although subjects reported more flatulence . There were non-significant decreases in episodes of diarrhoea with 20% on placebo and 11% on fructo oligosaccharides recording episodes in the post-study questionnaire (P=0.08) and 46% placebo, 38% fructo oligosaccharides recording episodes in the diary (P > 0.1) . No change in bowel frequency, consistency or stool size was recorded . CONCLUSION: Travel to high risk areas increases diarrhoea . Fructo oligosaccharides alone are not sufficient to prevent this, although do have some benefits for the subjects.

Curr Issues Mol Biol, 2000 Apr, 2(2), 41 - 50
Genetic modification of intestinal lactobacilli and bifidobacteria; Kullen MJ et al.; Lactobacilli and bifidobacteria are important members of the gastrointestinal microflora of man and animals . There is a substantial and growing body of evidence that these microbes provide benefits to the host in which they reside . Understanding the roles of these two groups of bacteria in the intestine continues to be a significant challenge . To this end, genetic characterisation and manipulation of intestinal lactobacilli and bifidobacteria is essential to define their contributions to the intestinal microflora, and to potentially exploit any beneficial or unique properties . This review will describe the tools and strategies currently available for the genetic manipulation of lactobacilli and bifidobacteria . Additionally, the ramifications and opportunities that may arise as a result of the genetic manipulation of probiotic lactobacilli and bifidobacteria will be addressed.

J Dairy Sci, 2001 Jul, 84(7), 1583 - 9
Suppressive effects of bifidobacteria on lipid peroxidation in the colonic mucosa of iron-overloaded mice; Ito M et al.; The antioxidative effects of live bifidobacteria on lipid peroxidation in the colonic mucosa were investigated . Bifidobacterium bifidum strain Yakult, which has been used for production of fermented milk, most effectively inhibited lipid peroxidation catalyzed by ferrous iron in liposomes among 10 species of bifidobacteria from human intestinal flora . Oral administration of B . bifidum strain Yakult for 2 wk significantly decreased the level of lipid peroxide (thiobarbituric acid reactive substance) in the colonic mucosa of iron-overload mice (Fe 0.07% in diet) . The iron concentrations in plasma and cecum contents were not affected by administration of B . bifidum strain Yakult . Bifidobacterium bifidum strain Yakult had no chelating or incorporating activity for ferrous iron in vitro . Therefore, the antioxidative effect of B . bifidum strain Yakult in the colonic mucosa was not thought to be based on the removal of ferrous iron from the reaction system of lipid peroxidation . These results suggested that B . bifidum strain Yakult protected the colonic mucosa from oxidative injury without inhibiting iron absorption.

FEMS Microbiol Ecol, 2001 Jul, 36(2-3), 113 - 121
Molecular microbial analysis of Bifidobacterium isolates from different environments by the species-specific amplified ribosomal DNA restriction analysis (ARDRA); Ventura M et al.; One hundred and six isolates of the genus Bifidobacterium, isolated from different environments (mainly gastrointestinal), were identified and classified taxonomically to species level by amplified ribosomal DNA restriction analysis . Two restriction endonucleases (Sau3AI and BamHI) were chosen for aligning the 16S rRNA sequences of 16 bifidobacterial species retrieved from various databases, to obtain species-specific restriction patterns . A rapid and accurate identification scheme was obtained by comparing the resulting 16S rDNA digestion profiles of 16 Bifidobacterium type-strains and 90 strains of various origins . All of the investigated strains were previously confirmed at the species level as belonging to the genus Bifidobacterium by fluorescence in-situ hybridisation and by polymerase chain reaction amplification with genus- and species-specific primers . The present work demonstrates that species-specific detection of Bifidobacterium adolescentis, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium coryneforme, Bifidobacterium cuniculi, Bifidobacterium dentium, Bifidobacterium infantis, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium suis, Bifidobacterium magnum, Bifidobacterium pseudolongum, Bifidobacterium pseudocatenulatum and Bifidobacterium pullorum present in different micro-ecological environments (e.g . gastrointestinal tract) can be accomplished in a reliable, rapid and accurate manner, circumventing the recognised deficiencies of traditional identification techniques.

Am J Health Syst Pharm, 2001 Jun 15, 58(12), 1101 - 9
Probiotics: "living drugs"; Elmer GW; The uses, mechanisms of action, and safety of probiotics are discussed . Probiotics are live microorganisms or microbial mixtures administered to improve the patient's microbial balance, particularly the environment of the gastrointestinal tract and the vagina . The yeast Saccharomyces boulardii and the bacterium Lactobacillus rhamnosus, strain GG, have shown efficacy in clinical trials for the prevention of antimicrobial-associated diarrhea . Other probiotics that have demonstrated at least some promise as prophylaxis for this type of diarrhea are Lactobacillus acidophilus, Bifidobacterium longum, and Enterococcus faecium . The use of S . boulardii as an adjunctive treatment to therapy with metronidazole or vancomycin has been found in controlled studies to decrease further recurrences of Clostridium difficile-associated disease . Other gastrointestinal disorders for which probiotics have been studied include traveler's diarrhea, acute infantile diarrhea, and acute diarrhea in adults . Several Lactobacillus species given in yogurt or in tablet or suppository form have shown clinical efficacy as a treatment for vaginal infections . Lactobacillus strains have also been examined as a treatment for urinary-tract infections . Putative mechanisms of action of probiotics include production of pathogen-inhibitory substances, inhibition of pathogen attachment, inhibition of the action of microbial toxins, stimulation of immunoglobulin A, and trophic effects on intestinal mucosa . The available probiotics are considered nonpathogenic, but even benign microorganisms can be infective when a patient is severely debilitated or immunosuppressed . Probiotics have demonstrated an ability to prevent and treat some infections . Effective use of probiotics could decrease patients' exposure to antimicrobials . Additional controlled studies are needed to clearly define the safety and efficacy of these agents.

Prikl Biokhim Mikrobiol, 2001 May-Jun, 37(3), 317 - 25
{Microelements preparations obtained during processing of natural flint affect the physiology and biochemistry of bifidobacteria}; Novik GI et al.; Microelement preparations obtained in the course of processing of flint powder stimulate the biological activity of Bifidobacterium adolescentis 94 BIM, grown on complex and synthetic nutritive media . The composition of the microelement preparations differed in the content of cations and anions . Introduction of the preparations into the cultures of physiologically active or anabiotic forms of bifidobacteria changed the parameters of exponential growth: compared to controls, the cultures were characterized by increased specific growth rate and decreased generation time . In the presence of microelements, the development of populations of bifidobacteria was associated with more pronounced accumulation of metabolic products (acetate, lactate, and ethanol) . Introduction of microelement preparations increased the rate of synthesis of the extracellular proteinase (maximum content of the enzyme was observed after 3 h, whereas control cultures attained this level only after 6 h).

Scand J Immunol, 2001 Jul-Aug, 54(1-2), 171 - 9
Experimental chronic arthritis and granulomatous inflammation induced by bifidobacterium cell walls; Zhang X et al.; Effects of cell walls (CWs) from two almost identical strains of Bifidobacterium adolescentis were studied in rats, using three different doses . A single i.p . injection of both CWs triggered a long-lasting arthritis with CW degradation products present in the joint tissue . Histologically, the arthritis was characterized by inflammatory cells, synovial hyperplasia, pannus formation and bone erosion, closely resembling human rheumatoid arthritis (RA) . In addition, CWs of the other strain induced a remarkable granuloma formation in the spleen and liver . Both CWs have the same peptidoglycan (PG) type A4alpha/beta, but differ from each other in three aspects . CW of the granuloma inducing strain: firstly has more lysine and less ornithine in PG stem peptides; secondly is more resistant to lysozyme degradation, and thirdly is better retained in the spleen . All these in comparison to the other strain used . Such characteristics are associated with the capacity to induce chronic arthritis, but it remains open how crucial they are for the granuloma formation.

Breast Cancer Res Treat, 2001 Mar, 66(2), 165 - 70
Bifidobacterium longum as a delivery system for gene therapy of chemically induced rat mammary tumors; Yazawa K et al.; A fundamental obstacle in cancer gene therapy is the specific targeting of therapy directly to a solid tumor, and no systemic delivery system yet exists . A strain of domestic bacteria, Bifidobacterium longum, which is nonpathogenic and anaerobic, selectively localized to and proliferated in 7,12-dimethylbenz{a}anthracene-induced rat mammary tumors after systemic application . We further ascertained the tumor specificity of genetically engineered, as well as wild-type, Bifidobacterium longum . This is the first demonstration that Bifidobacterium longum can be utilized as a specific gene delivery vector for gene therapy on solid breast tumors.

Int Arch Allergy Immunol, 2001 Jun, 125(2), 112 - 9
Can immunoregulatory lactic acid bacteria be used as dietary supplements to limit allergies?
Cross ML, Gill HS.
Studies in gnotobiotic animals have suggested that the intestinal bacterial flora may play an important role in priming the immune system during ontogeny to limit dysfunctional responses, including allergy . Prospective clinical studies have identified a higher incidence of allergy expression in early childhood among children who have low enteric populations of lactic acid bacteria (LAB), such as lactobacilli and bifidobacteria, further supporting a role for gut-colonizing bacteria in regulating immunological atopy . There is some evidence to suggest that supplementing the human diet with probiotic LAB might combat both allergy development and expression of atopy in allergy sufferers; however, definitive information, in the form of controlled intervention trials, remains scant . Recent immunological evidence has indicated that certain strains of LAB can stimulate the production of type I and II interferons and pro-interferon monokines (IL-12 and IL-18), following contact with the immune system; therefore, probiotic forms of immunoregulatory LAB could be used as dietary supplements to modify the gut microflora and provide pro-T helper cell 1 (Th1) STAT-activating signals sufficient to deviate the immune phenotype and correct the Th2-type bias which promotes allergy . This review outlines the clinical and laboratory evidence of a role for LAB in combating allergies, and attempts to explain this phenomenon in terms of our current understanding of immunoregulatory signals produced by gut-colonizing microbes .

Clin Diagn Lab Immunol, 2001 Jul, 8(4), 762 - 7
Enhanced antigen-specific delayed-type hypersensitivity and immunoglobulin G2b responses after oral administration of viable Lactobacillus casei YIT9029 in Wistar and Brown Norway rats; de Waard R et al.; In this study, the effects of orally administered viable Lactobacillus casei Shirota strain YIT9029 on the immunity parameters of Wistar and Brown Norway rats were examined . For this purpose, we used the Trichinella spiralis host resistance model . Two weeks before and during T . spiralis infection, rats were fed 10(9) viable L . casei bacteria 5 days per week . The T . spiralis-specific delayed-type hypersensitivity (DTH) response was significantly enhanced in both Wistar and Brown Norway rats given L . casei . In both rat strains fed L . casei, serum T . spiralis-specific immunoglobulin G2b (IgG2b) concentrations were also significantly increased . In the model, no significant effects of L . casei on larval counts or inflammatory reactions in the tongue musculature, body weights, or lymphoid organ weights were observed . Serum specific antibody responses, other than IgG2b, were not changed by feeding of L . casei . In contrast to L . casei, it was shown that orally administered Bifidobacterium breve or Bifidobacterium bifidum had no influence on the measured infection and immunity indices in the rat infection model . Since the rat DTH response is considered to be a manifestation of Th1 cell-mediated immunity and the IgG2b isotype has been associated with Th1 activity, it was concluded that Th1 cells could play an active role in the immunomodulatory effects of orally administered L . casei . Furthermore, our data do not indicate that the effect of oral supplementation with L . casei is dependent on the genetic background of the host.

J Appl Microbiol, 2001 Jun, 90(6), 859 - 65
Fermentations of fructo-oligosaccharides and their components by Bifidobacterium infantis ATCC 15697 on batch culture in semi-synthetic medium; Perrin S et al.; AIMS: To compare the physiological behaviour of Bifidobacterium infantis ATCC 15697 growing on synthetic oligofructose or its components . METHODS AND RESULTS: The studies were carried out in regulated or non-regulated batch cultures on semi-synthetic media . Differences between the carbohydrate utilization patterns with glucose, fructose, sucrose and fructo-oligosaccharides (FOS) were determined . Glucose was the preferred substrate for growth and biomass production, whereas fructose was the best for lactate and acetate production . With sucrose, biomass production reached the level obtained with glucose, whereas with FOS, more metabolites were produced, as with fructose . In a mixture of FOS, the shorter saccharides were used first and fructose was released in the medium . Fructofuranosidase, an enzyme necessary to hydrolyse FOS, was inducible by fructose . CONCLUSION: Glucose contained in FOS and sucrose might sustain growth and cell production, while fructose might enable the production of major metabolites . SIGNIFICANCE AND IMPACT OF THE STUDY: A better understanding of the bifidogenic nature of oligofructose has been gained.

Drug Dev Ind Pharm, 2001 Apr, 27(4), 309 - 19
Preparation and physical characterization of alginate microparticles using air atomization method; Cui JH et al.; Alginate microparticles were prepared using an air atomization method and varying formulation and processing conditions . Thereafter, the size and surface morphology of alginate microparticles were characterized . The trapping efficiencies of the ketoconazole, acetaminophen, vitamin C, and Bifidobacteria bifidum as model core materials were then determined . The air atomization process produced free-flowing and small-size microparticles after the freeze-drying process . The size distribution and surface morphology varied depending on the concentration of wall-forming materials and processing conditions . Generally, the geometric mean size increased as the concentration of alginate and poly-1-lysine and the delivery rate increased, but the air pressure decreased . Most of all, the ratio of delivery rate of alginate solution and air pressure could affect the size and surface morphology of alginate microparticles . However, the geometric mean size of alginate poly-1-lysine microparticles reproducibly ranged from about 80 to 130 microm . The microparticles were irregularly spherical or elliptical . The trapping efficiencies of ketoconazole, acetaminophen, vitamin C, and bifidobacteria were determined to be 71.5%, 60.1%, 1.6%, and 31%, respectively, when alginate concentration (1.5%), poly-1-lysine concentration (0.02%), air pressure (0.75 bar), delivery rate (8 ml/min), and spraying distance (45 cm) were applied . The current microencapsulation process using the air atomization method provides an alternative to entrapping small molecules and macromolecules without using harmful organic solvents . In addition, the small-size and free-flowing alginate microparticles containing active substances can be used as an intermediate in pharmaceutical applications.

J Vet Med B Infect Dis Vet Public Health, 2001 Apr, 48(3), 227 - 33
Bifidobacteria are obligate inhabitants of the crop of adult laying hens; Petr J et al.; Modified TPY agar (MTPY; with addition of glacial acetic acid and mupirocin) was used in the enumeration of bifidobacteria from hen crop and faeces . The colonies on MTPY medium inoculated with crop and faeces samples were Gram-stained, screened for presence of fructose-6-phosphate phosphoketolase activity and tested for fermentation patterns using ANAEROtest kits . It was revealed that bifidobacteria are obligate inhabitants of the hen crop, reaching counts of 10(7)/g of crop content . The occurrence of bifidobacteria in the hen crop was evidently not the consequence of ingestion of faeces as all the bifidobacteria strains isolated from the hen crop fermented glucose and fructose, while most strains isolated from hen faeces did not ferment either glucose or fructose . The results suggest that bifidobacteria are common and may even be one of the predominant parts of the hen crop flora.

Med Microbiol Immunol (Berl), 2001 Apr, 189(3), 147 - 52
A dietary probiotic (Bifidobacterium lactis HN019) reduces the severity of Escherichia coli O157:H7 infection in mice; Shu Q et al.; The protective effects of the probiotic Bifidobacterium lactis HN019 against Escherichia coli O157:H7 were investigated in murine challenge infection models . BALB/c or C57BL/6 mice were fed milk-based diets supplemented with B . lactis HN019 (3 x 10(8) cfu/g) for 7 days prior to and following oral challenge with E . coli O157:H7 . Behavioral parameters (morbidity, feed intake) were measured for 7 days following challenge; immunological responses (phagocytosis, antibody) and pathogen translocation were measured in a sub-sample of ostensibly healthy animals 1 week post-challenge . Results showed that HN019-fed mice maintained significantly higher post-challenge feed intake and exhibited a lower cumulative morbidity rate, compared to control mice which did not receive the probiotic . Significantly higher proportions of phagocytically active cells in the blood and peritoneum, and higher intestinal tract IgA anti-E . coli antibody responses, were recorded among HN019-fed mice compared to controls . Among HN019-fed mice, pathogen translocation was identified in one of five BALB/c and one of five C57 mice; the comparative figures in control mice were two of five and three of five, respectively, and the mean bacterial burdens in these mice were over 100-fold higher than in HN019-fed mice . These results demonstrate that HN019 can reduce the severity of infection due to the enterohemolytic pathogen E . coli O157:H7, and suggest that this reduction may be associated with enhanced immune protection conferred by the probiotic.

Exp Anim, 2001 Apr, 50(2), 105 - 13
Establishment of specific pathogen-free guinea-pig colonies using limited-flora guinea-pigs associated with conventional guinea-pig flora, and monitoring of their cecal flora; Yanabe M et al.; Six groups of limited flora (LF) Hartley guinea-pigs were produced by inoculation of hysterectomy-derived GF guinea-pigs with various combinations of cecal bacteria of conventional (CV) guinea-pigs to determine the effective bacterial cocktails for the establishment of a specific pathogen free (SPF) colony . Bifidobacterium magnum (Bif) isolated from CV guinea-pigs was used for pretreatment . The mortality of LF guinea-pigs inoculated with only Bif was 75%, and that of those inoculated with Bif plus chloroform-treated cecal suspension (CHF) or Bif plus CHF plus 32 isolates from CV guinea-pigs was 40 to 66.7% . These three groups were in an unhealthy condition with mucoid enteritis-like diarrhea . However, the mortality of LF guinea-pigs inoculated with the anaerobic growth on EG plates injected with 10(-5) dilution of cecal contents (CF) or inoculated with Bif plus CF was 6.3 and 15%, respectively . These latter two groups of LF guinea-pigs were transferred to separate barrier rooms and some of the LF guinea-pigs were maintained in isolators as a source of intestinal flora for SPF guinea-pigs . The composition of cecal flora of LF guinea-pigs was stable for a long time, and bacteroidaceae and peptococcaceae were maintained as predominant components . The basic composition of the cecal flora of SPF guinea-pigs originated from LF guinea-pigs, which consists mainly of the anaerobic bacteria, was not changed over a long period, and the flora composition became similar to that in CV guinea-pigs . Guinea-pig-specific pathogens from the SPF colonies were not detected during experiments.

Appl Environ Microbiol, 2001 Jun, 67(6), 2760 - 5
Specific identification and targeted characterization of Bifidobacterium lactis from different environmental isolates by a combined multiplex-PCR approach; Ventura M et al.; The species Bifidobacterium lactis, with its main representative strain Bb12 (DSM 10140), is a yoghurt isolate used as a probiotic strain and is commercially applied in different types of yoghurts and infant formulas . In order to ensure the genetic identity and safety of this bacterial isolate, species- and strain-specific molecular tools for genetic fingerprinting must be available to identify isolated bifidobacteria or lactic acid bacteria from, e.g., various clinical environments of relevance in medical microbiology . Two opposing rRNA gene-targeted primers have been developed for specific detection of this microorganism by PCR . The specificity of this approach was evaluated and verified with DNA samples isolated from single and mixed cultures of bifidobacteria and lactobacilli (48 isolates, including the type strains of 29 Bifidobacterium and 9 Lactobacillus species) . Furthermore, we performed a Multiplex-PCR using oligonucleotide primers targeting a specific region of the 16S rRNA gene for the genus Bifidobacterium and a conserved eubacterial 16S rDNA sequence . The specificity and sensitivity of this detection with a pure culture of B . lactis were, respectively, 100 bacteria/ml after 25 cycles of PCR and 1 to 10 bacteria/ml after a 50-cycle nested-PCR approach.

Appl Environ Microbiol, 2001 Jun, 67(6), 2526 - 30
Synthesis and fermentation properties of novel galacto-oligosaccharides by beta-galactosidases from Bifidobacterium species; Rabiu BA et al.; beta-Galactosidase enzymes were extracted from pure cultures of Bifidobacterium angulatum, B . bifidum BB-12, B . adolescentis ANB-7, B . infantis DSM-20088, and B . pseudolongum DSM-20099 and used in glycosyl transfer reactions to synthesize oligosaccharides from lactose . At a lactose concentration of 30% (wt/wt) oligosaccharide yields of 24.7 to 47.6% occurred within 7 h . Examination of the products by thin-layer chromatography and methylation analysis revealed distinct product derived spectra from each enzyme . These were found to be different to that of Oligomate 55, a commercial prebiotic galacto-oligosaccharide . Fermentation testing of the oligosaccharides showed an increase in growth rate, compared to Oligomate 55, with products derived from B . angulatum, B . bifidum, B . infantis, and B . pseudolongum . However B . adolescentis had a lower growth rates on its oligosaccharide compared with Oligomate 55 . Mixed culture testing of the B . bifidum BS-4 oligosaccharide showed that the overall prebiotic effect was equivalent to that of Oligomate 55.

Appl Environ Microbiol, 2001 Jun, 67(6), 2430 - 5
Characterization of the properties of human- and dairy-derived probiotics for prevention of infectious diseases in fish; Nikoskelainen S et al.; The present study aimed to investigate the potential probiotic properties of six lactic acid bacteria (LAB) intended for human use, Lactobacillus rhamnosus ATCC 53103, Lactobacillus casei Shirota, Lactobacillus bulgaricus, L . rhamnosus LC 705, Bifidobacterium lactis Bb12, and Lactobacillus johnsonii La1, and one for animal use, Enterococcus faecium Tehobak, for use as a fish probiotic . The strains for human use were specifically chosen since they are known to be safe for human use, which is of major importance because the fish are meant for human consumption . The selection was carried out by five different methods: mucosal adhesion, mucosal penetration, inhibition of pathogen growth and adhesion, and resistance to fish bile . The adhesion abilities of the seven LAB and three fish pathogens, Vibrio anguillarum, Aeromonas salmonicida, and Flavobacterium psychrophilum, were determined to mucus from five different sites on the surface or in the gut of rainbow trout . Five of the tested LAB strains showed considerable adhesion to different fish mucus types (14 to 26% of the added bacteria) . Despite their adhesive character, the LAB strains were not able to inhibit the mucus binding of A . salmonicida . Coculture experiments showed significant inhibition of growth of A . salmonicida, which was mediated by competition for nutrients rather than secretion of inhibitory substances by the probiotic bacteria as measured in spent culture liquid . All LAB except L . casei Shirota showed tolerance against fish bile . L . rhamnosus ATCC 53103 and L . bulgaricus were found to penetrate fish mucus better than other probiotic bacteria . Based on bile resistance, mucus adhesion, mucus penetration, and suppression of fish pathogen growth, L . rhamnosus ATCC 53103 and L . bulgaricus can be considered for future in vivo challenge studies in fish as a novel and safe treatment in aquaculture.

J Appl Microbiol, 2001 May, 90(5), 749 - 54
Characterization of factors affecting attachment of Bifidobacterium species to amylomaize starch granules; O'Riordan K et al.; AIMS: Two human-derived Bifidobacterium strains, PL1 and PL2, were tested for their ability to attach to amylomaize starch granules, and factors affecting binding were assessed . METHODS AND RESULTS: Good binding to granules was observed when the strains were grown on maltose or amylomaize starch, but not on glucose . Binding activity was localized to cell wall components and was sensitive to treatment with proteolytic enzymes . Several methodologies were employed to confirm these observations, including studies using radiolabelled cells, dot blot assays and scanning electron microscopy (SEM) analysis . CONCLUSION: Results from this study indicated that binding of strains PL1 and PL2 to amylomaize starch granules was mediated by a cell wall-associated proteinaceous factor that was induced when the strains were grown on starch or a related substrate, but not glucose . SIGNIFICANCE AND IMPACT OF THE STUDY: Attachment of probiotic strains to starch or other dietary fibres is believed to offer a selective advantage in the host intestine and may even prolong viability in adverse food environments . Therefore, characterizing the mechanisms of attachment has commercial implications in the design of synbiotic products.

Vopr Pitan, 2001, 70(1), 29 - 32
{Functional constipation syndrome: experience in the use of sour milk products and biologically active food supplements in the complex treatment of patients}; Blokhina LV et al.; Sour milk product "Bifeedock" and biologically-active food additives of "Litovite" and "Pektsecom" type are used in the overall treatment of functional constipation and accompanying colon disbacteriosis of I and II degree . They help to improve the clinical symptomatics and the functional state of the gastrointestinal tract . According to the data analysis, the elimination of the colon stasis results in the improved microbiocenosis expressed in the suppression of growth of conditionally pathogenic microflora . It also stimulates the growth of the symbiotic elements of microbiota (bifidobacteria, lactobacteria and the intestinal E . coli).

Vopr Pitan, 2001, 70(1), 15 - 20
{Comparative study of the effects of different sour milk products on the intestinal microflora of healthy infants}; Safronova AI et al.; Our study was aimed to investigate in comparable conditions the composition of intestinal microflora of healthy infants who were given 9 types of fermented milk products . There were observed 103 practically healthy infants aged from 4 month to 1,5 years old . The obtained data have shown that studied products improve intestinal microflora in infants . Significant increase of bifidobacteria in infants intestines was promoted by fermented milk products containing alive bifidobacteria with rather high acidity . The most of products which contain bifidobacteria decreased the level of hemolisinproducing Escherichia coli and coagulase-positive staphylococcus . The products which not contain bifidobacteria decreased the level of Candida albicans and protei.

Int J Food Microbiol, 2001 Apr 11, 65(1-2), 45 - 54
Dynamics of the microbial community responsible for traditional sour cassava starch fermentation studied by denaturing gradient gel electrophoresis and quantitative rRNA hybridization; Ampe F et al.; The microbial community developing during the spontaneous fermentation of sour cassava starch was investigated by cultivation-independent methods . Denaturing gradient gel electrophoresis (DGGE) of partially amplified 16S rDNA followed by sequencing of the most intense bands showed that the dominant organisms were all lactic acid bacteria (LAB), mainly close relatives of Bifidobacterium minimum, Lactococcus lactis, Streptococcus sp., Enterococcus saccharolyticus and Lactobacillus plantarum., Close relatives of Lb . panis, Leuconostoc mesenteroides and Ln . citreum were also found . A complementary analysis using hybridization of 16S rRNA with phylogenetic probes was necessary to detect the presence of the recently discovered species Lb . manihotivorans . Although it represented up to 13% of the total lactic acid bacteria of sour cassava starch, this species could not be detected by DGGE as the PCR product migrated to the same position as Lc . lactis . In addition, it was shown that a strong pH decrease in the time course of fermentation was most probably responsible for the competitive selection of acid-resistant LAB vs . both homo and heterofermentative acid-sensitive LAB.

Br J Nutr, 2001 Mar, 85 Suppl 1, S47 - 53
Non-polyol low-digestible carbohydrates: food applications and functional benefits; Murphy O; Many LDCs currently on the market are not digested in the upper gastrointestinal tract and become fermented in the large intestine . They possess physiological benefits similar to those of dietary fibre . For some of these materials the fermentation process is highly specialised and leads to the selective stimulation and growth of beneficial gut bacteria, e.g . bifidobacteria . These materials are described as prebiotics, which are defined as nutrients fermented in the large bowel that favour the growth of desirable large bowel microflora . This activity has been demonstrated for inulin and oligofructose . Two other carbohydrates with low digestibility that offer desirable physiological properties are resistant starch (RS) and polydextrose (PD) . These 'functional benefits have led to considerable interest from the food industry leading to the use of these ingredients in the development of new 'healthy' products . This paper describes the use of these materials in the development of 'healthy' products, some of their functional properties, and the benefits they confer on different food systems.

Appl Environ Microbiol, 2001 May, 67(5), 2276 - 83
Intra- and extracellular beta-galactosidases from Bifidobacterium bifidum and B . infantis: molecular cloning, heterologous expression, and comparative characterization; Moller PL et al.; Three beta-galactosidase genes from Bifidobacterium bifidum DSM20215 and one beta-galactosidase gene from Bifidobacterium infantis DSM20088 were isolated and characterized . The three B . bifidum beta-galactosidases exhibited a low degree of amino acid sequence similarity to each other and to previously published beta-galactosidases classified as family 2 glycosyl hydrolases . Likewise, the B . infantis beta-galactosidase was distantly related to enzymes classified as family 42 glycosyl hydrolases . One of the enzymes from B . bifidum, termed BIF3, is most probably an extracellular enzyme, since it contained a signal sequence which was cleaved off during heterologous expression of the enzyme in Escherichia coli . Other exceptional features of the BIF3 beta-galactosidase were (i) the monomeric structure of the active enzyme, comprising 1,752 amino acid residues (188 kDa) and (ii) the molecular organization into an N-terminal beta-galactosidase domain and a C-terminal galactose binding domain . The other two B . bifidum beta-galactosidases and the enzyme from B . infantis were multimeric, intracellular enzymes with molecular masses similar to typical family 2 and family 42 glycosyl hydrolases, respectively . Despite the differences in size, molecular composition, and amino acid sequence, all four beta-galactosidases were highly specific for hydrolysis of beta-D-galactosidic linkages, and all four enzymes were able to transgalactosylate with lactose as a substrate.

Biosci Biotechnol Biochem, 2001 Feb, 65(2), 466 - 9
Dietary curdlan increases proliferation of bifidobacteria in the cecum of rats; Shimizu J et al.; Significant increases in the amounts of short-chain fatty acids and lactate, and in numbers of bifidobacteria were observed in the cecum of curdlan (CD) -fed rats as compared with those of cellulose-fed ones . The in vitro proliferation of 5 species of bifidobacteria was markedly increased in the cultures containing the supernatant obtained from the cecal contents of CD-fed rats . These findings suggest that bifidus factors have been produced in the cecum of CD-fed rats.

Biosci Biotechnol Biochem, 2001 Feb, 65(2), 264 - 9
Effect of dietary short-chain fructooligosaccharides on the cecal microflora in gastrectomized rats; Sakai K et al.; Total gastric resection is known to lead to changes in the microflora in the whole gastrointestinal tract . Dietary short-chain fructooligosaccharides (Sc-FOS) have been shown to also induce a change in the microflora in the large bowel by promoting an increase in the numbers of Bifidobacterium and Lactobacillus which have beneficial effects on the host . In the present study, 4-week-old male Sprague-Dawley rats received total gastrectomy or laparotomy, and each of these surgically treated groups was randomly divided into two experimental diet groups and given a 7.5% Sc-FOS diet or control diet . Enumeration and identification of the cecal bacteria was performed by using selective and non-selective media . In the gastrectomized rats, the total bacterial count, and the counts of Bacteroidaceae and Enterobacteriaceae were higher than those in the sham-operated rats . Sc-FOS promoted an increase in the numbers of Bifidobacterium and Lactobacillus, In the rats fed on the Sc-FOS diet, the predominant type of bacteria was Lactobacillus and in the rats fed on the control diet, it was Bacteroidaceae irrespective of gastrectomy . We confirmed that both gastrectomy and dietary Sc-FOS changed the composition of cecal microflora in the rats . Dietary Sc-FOS in the gastrectomized rats increased the proportions of Lactobacillus relative to other types of bacteria to levels similar to those seen in healthy normal rats, and decreased the proportion of Bacteroidaceae.

Lett Appl Microbiol, 2001 Apr, 32(4), 235 - 9
Purification and N-terminal amino acid sequence of fructose-6-phosphate phosphoketolase from Bifidobacterium longum BB536; Fandi KG et al.; AIMS: The key enzyme in the fructose-6-phosphate shunt in bifidobacteria, Fructose-6-phosphate phosphoketolase (F6PPK; E.C . 4.1.2.22.), was purified to electrophoretic homogeneity for the first time from Bifidobacterium longum (BB536) . METHODS AND RESULTS: A three-step procedure comprising acetone fractionation followed by fast protein liquid chromatography (FPLC) resulted in a 30-fold purification . The purified enzyme had a molecular mass of 300 +/- 5 kDa as determined by gel filtration . It is probably a tetramer containing two different subunits with molecular masses of 93 +/- 1 kDa and 59 +/- 0.5 kDa, as determined by SDS-PAGE . CONCLUSION: The deduced N-terminal amino acid sequences of the two subunits revealed no significant similarity between them and other proteins when compared to the data bases of EMBL and SWISS-PROT, indicating that this could be the first report on N-terminal amino acid sequence of F6PPK . SIGNIFICANCE AND IMPACT OF THE STUDY: The data from this study will be used to design oligonucleotide probe specific for bifidobacteria and to study the gene encoded F6PPK.

J Bacteriol, 2001 May, 183(9), 2929 - 36
Characterization of the D-xylulose 5-phosphate/D-fructose 6-phosphate phosphoketolase gene (xfp) from Bifidobacterium lactis; Meile L et al.; A D-xylulose 5-phosphate/D-fructose 6-phosphate phosphoketolase (Xfp) from the probiotic Bifidobacterium lactis was purified to homogeneity . The specific activity of the purified enzyme with D-fructose 6-phosphate as a substrate is 4.28 Units per mg of enzyme . K(m) values for D-xylulose 5-phosphate and D-fructose 6-phosphate are 45 and 10 mM, respectively . The native enzyme has a molecular mass of 550,000 Da . The subunit size upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis (90,000 Da) corresponds with the size (92,529 Da) calculated from the amino acid sequence of the isolated gene (named xfp) encoding 825 amino acids . The xfp gene was identified on the chromosome of B . lactis with the help of degenerated nucleotide probes deduced from the common N-terminal amino acid sequence of both the native and denatured enzyme . Comparison of the deduced amino acid sequence of the cloned gene with sequences in public databases revealed high homologies with hypothetical proteins (26 to 55% identity) in 20 microbial genomes . The amino acid sequence derived from the xfp gene contains typical thiamine diphosphate (ThDP) binding sites reported for other ThDP-dependent enzymes . Two truncated putative genes, pta and guaA, were localized adjacent to xfp on the B . lactis chromosome coding for a phosphotransacetylase and a guanosine monophosphate synthetase homologous to products of genes in Mycobacterium tuberculosis . However, xfp is transcribed in B . lactis as a monocistronic operon . It is the first reported and sequenced gene of a phosphoketolase.

Acta Paediatr Taiwan, 2001 Jan-Feb, 42(1), 11 - 7
Sialic acid in human milk: composition and functions; Nakano T et al.; Breast milk is the best nutrient source for infants . It contains all elements needed for a normal growth and development of infants . Human milk contains a large amount of sialic acid compared with bovine milk . Sialic acid contained in oligosaccharides, glycolipids and glycoproteins in milk is considered to play important roles in physiological functions in infancy . Thus, we have investigated the sialic acid composition and the functions of sialylated compounds in human milk . Sialic acids comprise a family of neuraminic acid derivatives present in secretions, fluids and tissues of mammals . In milk, sialic acid is present in different sialoglycoconjugate compounds such as oligosaccharides, glycolipids and glycoproteins, not in a free form . Human milk contains 0.3-1.5 mg/ml of sialic acid . Sialic acid bound to oligosaccharides accounts for about 75% of the total sialic acid contained in human milk . Most of the sialic acid contained in human milk is found in the form of sialyllactose, an oligosaccharide formed from lactose and sialic acid . In milk, gangliosides, sialic acid-containing glycolipid, occur mainly as monosialoganglioside 3 (GM3) and disialoganglioside 3 (GD3) . The concentration of GM3 in human milk increases, while that of GD3 concentration decreases during lactation . Because the brain and central nervous system contain considerable level of sialic acid in infancy, it is considered to play important roles on the expression and development of their functions . Moreover, we found that some sialylated compounds had inhibited the adhesion of toxins, bacteria and viruses to the receptors on the surface of epithelial cells . Additionally, we found that some sialylated compounds had growth-promoting effects on bifidobacteria and lactobacilli, predominantly present in the intestinal flora of infants fed with human milk . The results suggested that sialylated compounds in human milk possibly behaved as a physiological component in the intestinal tract of infants to protect them against enteric infections.

Arch Inst Pasteur Alger, 1998, 62, 63 - 76
{In vitro study of antagonistic activity of bifidobacteria against Campylobacter and Escherichia coli causing gastroenteritis in children}; Loucif F et al.; Bifidobacteria represents the major constituents of the human intestinal microflora . It's a non mobile Gram (+) bacteria, which interest almost of the research about the relationships between the intestinal flora and human health . Bifidobacteria has an inhibition activity on the growth of the Campylobacter and E . coli G.E.I., in case of children gastro-intestinal infections by mixed cultivation in reconstituted infant milk adapted for the first age . This activity is the result of a group of mechanisms acting at the same time, it's probably the following: Competition of the nutrients indispensable for bacterial growth, action of the produced acid do not favorite the growth of the active pathogens germs (bacteriocine) with acid pH.

Br J Nutr, 2000 Nov, 84 Suppl 1, S39 - 46
Biological activities of bovine glycomacropeptide; Brody EP; Biological activity of bovine kappa-caseino glycomacropeptide (GMP) has received much attention in recent years . Research has focused on the ability of GMP to bind cholera and Escherichia coli enterotoxins, inhibit bacterial and viral adhesion, suppress gastric secretions, promote bifidobacterial growth and modulate immune system responses . Of these, protection against toxins, bacteria, and viruses and modulation of the immune system are the most promising applications.

Zh Mikrobiol Epidemiol Immunobiol, 2001 Jan-Feb, (1), 80 - 1
{Multiplication of Listeria in milk products}; Karpova TI et al.; The results of the evaluation of the multiplication dynamics of Listeria cells in milk and Bifidok, a lactic acid product, are presented . The samples were inoculated on thioglycol agar and studied at different exposure time after incubation at 37 degrees C, 20 degrees C and 4 degrees C . The study revealed the intensive multiplication of Listeria cells in milk, also during storage in a household refrigerator . The presence of bifidobacteria mixed with kefir-producing culture in dairy products was shown to essentially inhibit the growth of Listeria cells which were not detected by bacteriological techniques on day 7.

Zh Mikrobiol Epidemiol Immunobiol, 2000 Sep-Oct, (5), 69 - 72
{The use of biological preparations for correction of microbial status in pregnant women}; Korshunova OV et al.; 15 pregnant women with pregnancy lasting 28-32 weeks, whose medico-laboratory data (the positive result of the amino test, high pH value and the detection of "key" cells) suggested the presence of bacterial vaginosis, were placed under observation . The bacteriological study of vaginal microflora in all these women revealed pronounced disturbances simultaneously with the development of intestinal dysbacteriosis . The local application of the bacterial preparation "Zhlemik" containing freeze-dried live lactic acid bacteria of vaginal origin and the oral administration of lactic acid bifidumbacterin containing live bifidobacteria of intestinal origin permitted the successful restoration of vaginal and intestinal microbiocenosis . The capacity of biotherapeutic preparations for inducing non-specific immunostimulation led to a significant rise in the levels of IgA, IgM and IgG in vaginal secretions.

Biofactors, 2000, 12(1-4), 289 - 97
When is dietary fiber considered a functional food?
Prosky L.
Before answering the question of when dietary fiber can be considered a functional food we must first decide what can be called a dietary fiber . The generally accepted definition of dietary fiber is that of Trowell that dietary fiber consists of the remnants of edible plant cells polysaccharides, lignin, and associated substances resistant to (hydrolysis) digestion by the alimentary enzymes of humans . In Japan the food tables list the dietary fiber content of animal as well as plant tissues, while many countries accept saccharides of less than DP-10 as dietary fiber (inulin, oligofructose, Fibersol-2, polydextrose, fructooligosaccharides, galactooligosaccharides etc.) . These shorter chain oligosaccharides do not precipitate as dietary fiber in the standard Association of Official Analytical Chemists (AOAC) method, which is accepted by the US Food & Drug Administration, the US Department of Agriculture and the Food & Agriculture Organization of the World Health Organization for nutrition labeling purposes . In the United Kingdom the term dietary fiber has been replaced in nutrition labeling by nonstarch polysaccharides . Therefore the American Association of Cereal Chemists (AACC) commissioned an ad hoc committee of scientists to evaluate continuing validity of the currently used definition, and if appropriate, to modify and update that definition . Obtaining scientific input from the community of analysts, health professionals, and dietary fiber researchers was considered a high priority . To this end three meetings were held in the space of six months to assure input from all persons knowledgeable in the field with the answer expected sometime before 2000 . Dietary fiber can be considered a functional food when it imparts a special function to that food aside from the normal expected function and similarly when the dietary fiber is used as an additive to foods . For example, dietary fiber contributes to colonic health, bifidobacterial or lactobacillus stimulation in the gut, coronary artery health, cholesterol reduction, glucose metabolism, insulin response, blood lipids, cancer etc . The author discusses in detail the functional food properties of dietary fiber.

Zh Mikrobiol Epidemiol Immunobiol, 2000 Nov-Dec, (6), 80 - 2
{Improving a process of cultivation of bifidobacteria in a bioreactor to obtain lyophilized seed culture}; Murashova AO et al.; The parameters of the cultivation process and the characteristics of the culture growth have been analyzed . The phases of the population growth have been determined . The existence of short stages differing in the physiological state of the cells, their resistance to damaging factors and metabolic activity within the deceleration phase of the growth of bifidobacteria has been experimentally substantiated . To harvest the biomass, the period of the completion of the cultivation process has been chosen . The lyophilized seed culture in which a high percentage of viable cells is preserved for one year has been obtained.

Zh Mikrobiol Epidemiol Immunobiol, 2000 Nov-Dec, (6), 55 - 8
{Effect of bifidobacteria and lactobacillus and antibiotics in combination with common gnotobiological isolation on the survival of mice with acute radiation sickness}; Pikina AP et al.; The influence of the combined use of bacterial preparations (Bifidobacterium and Lactobacillus) and antibiotics (ciprofloxacin, lomefloxacin and amikacin) on the survival rate of irradiated mice placed under the conditions of general gnotobiological isolation was studied . Bacterial strains used in combination with quinolones (ciprofloxacin, lomefloxacin) significantly increased the mean survival time of the animals (p < 0.05) when introduced in a dose of 1.0 x 10(9) microbial cells per mouse on days 1, 3, 5, 7, 9, 11, 13 and 15 after irradiation . At the same time a short course of treatment with bacterial preparations (two injections on days 5 and 7 after irradiation) proved to be insufficient for increasing the survival rate of the animals . The mean survival time of the irradiated mice was higher after the use of bacterial preparations in combination with lomefloxacin or ciprofloxacin than after their use with amikacin.

New Microbiol, 2001 Jan, 24(1), 23 - 33
Behaviour of the pharmaceutical probiotic preparation VSL#3 in human ileostomy effluent containing its own natural elements; Bianchi-Salvadori B et al.; The pharmaceutical probiotic VSL#3 (300 billion cfu/g lactic acid bacteria & bifidobacteria) was inoculated into human ileostomy effluent (HIE) to assess its behaviour vs the ileo-caecal tract . Separately, yogurt bacteria (yog) and bifidobacteria (Bif) present in VSL#3 were also inoculated into HIE . During 37 degrees C incubation (anaerobic condition) at zero, six and 24 hours, both cell growth in control HIE and indigenous Bif growth in HIE+yog were observed . Cells remained viable and metabolically active as shown by the increase in L(+) lactic acid in HIE+VSL#3 and HIE+yog and the pH decreased (approx . 5.5 compared with the 6.2 of control HIE) . Total SCFA Short Chain Fatty Acids decreased in HIE+yog and HIE+VSL#3 at 6h and in all cultures at 24h; butyric acid decreased in HIE+Bif and HIE+VSL#3 . Generally in vivo the bacteria remain in the ileo-caecal tract no longer than six h, therefore VSL#3 bacteria seem able to pass this barrier viably, colonizing the large bowel.

Int J Food Microbiol, 2001 Jan 22, 63(1-2), 81 - 90
Potential probiotic lactic acid bacteria Lactobacillus rhamnosus (HN001), Lactobacillus acidophilus (HN017) and Bifidobacterium lactis (HN019) do not degrade gastric mucin in vitro; Zhou JS et al.; The mucus layer (mucin) coating the surface of the gastrointestinal tract (GIT) plays an important role in the mucosal barrier system . Any damage or disturbance of this mucin layer will compromise the host's mucosal defence function . In the present study, the ability of three potential probiotic lactic acid bacteria (LAB) strains (Lactobacillus rhamnosus HN001, Lactobacillus acidophilus HN017, Bifidobacterium lactis HN019) to degrade mucin in vitro was evaluated, in order to assess their potential pathogenicity and local toxicity . The LAB strains were incubated in medium containing hog gastric mucin (HGM, 0.3%) at 37 degrees C for 48 h, following which any decrease in carbohydrate and protein concentration in the ethanol-precipitated portion of the culture medium was determined, using phenol-sulphuric acid and bicinchonic acid (BCA) protein assays, respectively . The change in molecular weight of mucin glycoproteins, following incubation with the test strains, was monitored by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) . In order to expose any ability of the test strains to degrade mucin visually and more directly, the test strains were also cultured on agarose containing 0.3% HGM and incubated anaerobically for 72 h at 37 degrees C . No significant change in the carbohydrate or protein concentration in mucin substrates was found following incubation with the test strains . No mucin fragments were derived from the mucin suspension incubated with test strains, and no mucinolysis zone was identified on agarose . These results demonstrate that the potential probiotic LAB strains tested here were unable to degrade gastrointestinal mucin in vitro, which suggests that these novel probiotic candidates are likely to be non-invasive and non-toxic at the mucosal interface.

Immunobiology, 2000 Nov, 202(5), 429 - 41
Bifidobacterium adolescentis modulates the specific immune response to another human gut bacterium, Bacteroides thetaiotaomicron, in gnotobiotic rats; Scharek L et al.; In order to investigate the capability of an autochthonous bacterium to modulate the host's immune response against the indigenous microfiora, the immunogenicity of two selected bacterial species of the human gut was investigated in a gnotobiotic rat model . Germ-free (GF) rats were monoassociated with either Bifidobacterium (B.) adolescentis or Bacteroides (B.) thetaiotaomicron and the development of bacteria-specific IgG and IgA in serum and specific secretory IgA (sIgA) in feces of the animals were measured . Knowing the antibody levels in gnotobiotic rats induced by monoassociation, we subsequently diassociated two groups of rats in order to investigate the impact of B . adolescentis on the immune reaction against B . thetaiotaomicron . One group was diassociated simultaneously with B . adolescentis and B . thetaiotaomicron, the second group was diassociated with these bacteria in sequence . In contrast to B . thetaiotaomicron, B . adolescentis was not able to induce a systemic immune response in monoassociated animals as evident from serum IgG and IgA . However, both bacterial species challenged the mucosal immune system as indicated by an increase in sIgA in the feces . The specific immune response to B . thetaiotaomicron was significantly lower in diassociated animals than in animals monoassociated with B . thetaiotaomicron . This effect was more pronounced in the rats, that had been associated sequentially . The presence of B . adolescentis down-regulated the humoral immunity to B . thetaiotaomicron.

Arerugi, 2000 Dec, 49(12), 1146 - 55
{Intestinal flora of infants with cow milk hypersensitivity fed on casein-hydrolyzed formula supplemented raffinose}; Hattori K et al.; We studied the intestinal flora of infants with cow milk hypersensitivity fed on casein-hydrolyzed formula (MA-1) and the influence of that supplemented with Raffinose (MA-1{R}) . Infants with cow milk hypersensitivity were fed with MA-1 for 2 weeks, after which the formula was changed to MA-1{R} . Fourteen subjects were enrolled in this study and divided into two groups; three who fed with breast or conventional milk in addition to MA-1 or MA-1{R}(BM group) and 11 mainly fed with MA-1 or MA-1{R} (TF group) . Intestinal flora was investigated at two weeks after MA-1 feeding and at two weeks after MA-1{R} feeding, respectively . Bifidobacterium was detected as the most predominant bacteria in all examples in the BM group, and that count and the ratio in all bacteria remained high even after changing MA-1 to MA-1{R} . On the other hand, bacteria count and ratios of Bifidobacterium in all bacteria were conspicuously low in the TF group as compared with the BM group . And with the change from MA-1 to MA-1{R} in the TF group, the bacterial number and the occupation ratio of Bifidobacterium were increased, and Enterobacteriaceae bacterial count and the occupation ratio were decreased . The change of the intestinal flora with MA-1{R} feeding was mainly caused by the breeding action of Raffinose on bifidobacteria . Further studies are needed from a viewpoint of clinical effectiveness about the influence of normalization of the intestinal flora for the treatment of food hypersensitivity.

Mikrobiologiia, 2000 Nov-Dec, 69(6), 774 - 7
{Production of proteinases associated with Bifidobacterium adolescentis 94-BIM cell wall}; Samartsev AA et al.; Bifidobacterium adolescentis 94-BIM was found to produce cell-wall bound proteolytic enzymes active at acidic, neutral, and alkaline pH values . The solubilization of proteinases with 0.5% Triton X-100 substantially improved the yield of the enzymes . The most active accumulation of cell-bound proteinases was observed in the third hour of cultivation at rates of 156.7, 179.5, and 111.1 U/(mg h), measured at pH 2.5, 7.0, and 9.0, respectively . It is suggested that the cell-wall bound proteinases of B . adolescentis 94-BIM are the precursors of the enzymes secreted into the medium.

Digestion, 2001, 63 Suppl 1, 60 - 7
Germinated barley foodstuff feeding . A novel neutraceutical therapeutic strategy for ulcerative colitis; Kanauchi O et al.; A germinated barley foodstuff (GBF) contained glutamine-rich protein and the hemicellulose-rich fiber was made from brewer's spent grain by physical isolation (milling and sieving) . Both in vivo and in vitro studies demonstrated that the fiber fraction of GBF supports maintenance of epithelial cell populations, facilitates epithelial repair, and suppresses epithelial nuclear factor kappa B-DNA binding activity through generating increased short-chain fatty acid (especially butyrate) production by luminal microflora which includes Bifidobacterium and Eubacterium, thereby preventing experimental colonic injury . The fiber fraction also modulates stool water content by its high water-holding capacity . The protein fraction which contains larger glutamine prevents experimental small bowel injury . Based on these observations, clinical studies were initiated in patients with mild to moderate active ulcerative colitis . The patients who had been unresponsive to or intolerant of standard treatment received 30 grams of GBF feeding daily in a nonrandomized, open-label fashion . At 4 weeks, this treatment resulted in a significant clinical and endoscopic improvement independent of disease extent . The improvement was associated with an increase in stool butyrate concentrations and in luminal Bifidobacterium and Eubacterium levels . After the end of GBF treatment the patients had an exacerbation of the disease . GBF was safe and well tolerated . These results indicate that GBF feeding is a potentially attractive treatment in patients with ulcerative colitis .

Am J Physiol Gastrointest Liver Physiol, 2001 Mar, 280(3), G368 - 80
Influence of microbial species on small intestinal myoelectric activity and transit in germ-free rats; Husebye E et al.; The effect of an intestinal microflora consisting of selected microbial species on myoelectric activity of small intestine was studied using germ-free rat models, with recording before and after specific intestinal colonization, in the unanesthetized state . Intestinal transit, neuropeptides in blood (RIA), and neuromessengers in the intestinal wall were determined . Clostridium tabificum vp 04 promoted regular spike burst activity, shown by a reduction of the migrating myoelectric complex (MMC) period from 30.5 +/- 3.9 min in the germ-free state to 21.2 +/- 0.14 min (P < 0.01) . Lactobacillus acidophilus A10 and Bifidobacterium bifidum B11 reduced the MMC period from 27.9 +/- 4.5 to 21.5 +/- 2.1 min (P < 0.02) and accelerated small intestinal transit (P < 0.05) . Micrococcus luteus showed an inhibitory effect, with an MMC period of 35.9 +/- 9.3 min compared with 27.7 +/- 6.3 min in germ-free rats (P < 0.01) . Inhibition was indicated also for Escherichia coli X7 gnotobiotic rats . No consistent changes in slow wave frequency were observed . The concentration of neuropeptide Y in blood decreased after introduction of conventional intestinal microflora, suggesting reduced inhibitory control . Intestinal bacteria promote or suppress the initiation and aboral migration of the MMC depending on the species involved . Bacteria with primitive fermenting metabolism (anaerobes) emerge as important promoters of regular spike burst activity in small intestine.

J Agric Food Chem, 2001 Jan, 49(1), 378 - 83
Bifidobacteria: genetic modification and the study of their role in the colon; van der Werf MJ et al.; Bifidobacteria are among the most common bacteria in the human intestine and are thought to have a positive effect on human health . Therefore, there is an increasing interest in using these microorganisms as probiotics, either in fermented dairy products or formulated as tablets . However, convincing scientific data supporting their health claims are scarce . The study of the role of bifidobacteria in the colon is complicated by the fact that they are part of a complex ecosystem also interacting with the human host and by the fact that their in vivo study encounters many ethical constraints . Several tools have been developed at TNO with which the role of bifidobacteria can be studied . These include (i) an efficient transformation protocol for the introduction of foreign DNA into Bifidobacterium strains and (ii) in vitro models of the stomach/small intestine (TIM-1) and large intestine (TIM-2), creating an environment closely resembling that of the in vivo situation . With these tools, biomarkers from bifidobacteria quantifying their positive effect on gut health can be identified.

J Appl Microbiol, 2001 Feb, 90(2), 268 - 78
Selection of a Bifidobacterium strain to complement resistant starch in a synbiotic yoghurt; Crittenden RG et al.; AIMS: To employ an in vitro screening regime to select a probiotic Bifidobacterium strain to complement resistant starch (Hi-maizetrade mark) in a synbiotic yoghurt . METHODS AND RESULTS: Of 40 Bifidobacterium isolates examined, only B . lactis Laftitrade mark B94 possessed all of the required characteristics . This isolate hydrolysed Hi-maizetrade mark, survived well in conditions simulating passage through the gastrointestinal tract and possessed technological properties suitable for yoghurt manufacture . It grew well at temperatures up to 45 degrees C, and grew to a high cell yield in an industrial growth medium . In addition to resistant starch, the organism was able to utilize a range of prebiotics including inulin, and fructo-, galacto-, soybean- and xylo-oligosaccharides . Pulse field gel electrophoresis of restriction enzyme cut chromosomal DNA revealed that B . lactis Laftitrade mark B94 was very closely related to the B . lactis Type Strain (DSM 10140), and to the commercial strains B . lactis Bb-12 and B . lactis DS 920 . However, B . lactis Laftitrade mark B94 was the only one of these isolates that could hydrolyse Hi-maizetrade mark . This phenotypic difference did not appear to be due to the presence of plasmid encoded amylase . Bifidobacterium lactis Laftitrade mark B94 survived without substantial loss of viability in synbiotic yoghurt containing Hi-maizetrade mark during storage at 4 degrees C for six weeks . CONCLUSION: Bifidobacterium lactis Laftitrade mark B94 is a promising new yoghurt culture that warrants further investigation to assess its probiotic potential . SIGNIFICANCE AND IMPACT OF THE STUDY: In vitro screening procedures can be used to integrate complementary probiotic and prebiotic ingredients for new synbiotic functional food products.

Int J Pharm, 2000 Dec 4, 210(1-2), 51 - 9
Survival and stability of bifidobacteria loaded in alginate poly-l-lysine microparticles; Cui JH et al.; Bifidobacteria-loaded alginate microparticles were prepared by spraying a mixture of alginate and bifidobacteria culture using an air atomization method . Survival and stability of bifidobacteria loaded in microparticles were then evaluated . Survival of bifidobacteria from alginate poly-l-lysine microparticles was significantly increased when MRS broth or yeast extract was added in simulated intestinal fluid (pH 6.8) . The number of bifidobacteria gradually increased for 8 h (10(8) cfu/g) and then reached about 10(9)-10(10) cfu/g when incubated over 12 h in intestinal fluid containing 0.5% yeast extract and 0.05% L-cysteine . The survival of bifidobacteria was highly dependent on the pH of the exposing media . When the bifidobacteria was immobilized with alginate or even poly-l-lysine treatment, the survival of bifidobacteria was highly enhanced in the low pH conditions (ca . > 10(8) vs . < 10(3) cfu/g) . The stability of free flowing bifidobacteria-loaded alginate poly-l-lysine microparticles was significantly improved during storage at 4 degrees C in a refrigerator when compared to bifidobacteria cultures . The bifidobacteria-loaded alginate poly-l-lysine microparticles could be applied to various dairy products.

Am J Clin Nutr, 2001 Feb, 73(2 Suppl), 476S - 483S
Market potential for probiotics; Stanton C et al.; "Functional foods" as a marketing term was initiated in Japan in the late 1980s and is used to describe foods fortified with ingredients capable of producing health benefits . This concept is becoming increasingly popular with consumers because of a heightened awareness of the link between health, nutrition, and diet . Food manufacturers are enthusiastic about developing such products because the added ingredients give increased value to food . The global market for functional foods in the coming years is predicted to grow rapidly . Although Japan currently accounts for about one-half of this market, the fastest rate of growth is expected to be in the United States . Probiotic products represent a strong growth area within the functional foods group and intense research efforts are under way to develop dairy products into which probiotic organisms such as Lactobacillus and Bifidobacterium species are incorporated . Such probiotic foods may modulate gut microbial composition, thereby leading to improved gut health, for example, through improved tolerance to lactose in lactose-intolerant individuals or improved resistance to pathogenic bacteria . Large numbers of viable microorganisms are likely to be required in the food product, which should be consumed regularly to experience the health effect . The probiotic market, especially dairy products such as yogurts and fermented milks, has experienced rapid growth in Europe . The long-term exploitation of probiotics as health promoters is dependent on several factors, including sound, scientifically proven clinical evidence of health-promoting activity; accurate consumer information; effective marketing strategies; and, above all, a quality product that fulfills consumer expectations.

Am J Clin Nutr, 2001 Feb, 73(2 Suppl), 465S - 470S
Probiotics and safety; Ishibashi N et al.; Bacterial species that have traditionally been regarded as safe are used in probiotics; the main strains used include lactic acid bacteria and bifidobacteria that inhabit the intestinal tracts of humans and animals . However, reports of frequent isolation of bacteria used in probiotics from infection sources in recent years have raised much debate over the safety of probiotics . This article describes the status quo of isolation of probiotic bacteria from infections and reviews each of the factors that have to be addressed in assessing the safety of probiotics, namely pathogenicity, infectivity, toxicity, and intrinsic properties of the bacteria . Monoassociation with Bifidobacterium longum in gnotobiotic mice as a method to assess safety with respect to infection, and translocation and immune responses as a result of the monoassociation are also described.

Am J Clin Nutr, 2001 Feb, 73(2 Suppl), 415S - 420S
Prebiotic digestion and fermentation; Cummings JH et al.; Prebiotics, as currently conceived of, are all carbohydrates of relatively short chain length . To be effective they must reach the cecum . Present evidence concerning the 2 most studied prebiotics, fructooligosaccharides and inulin, is consistent with their resisting digestion by gastric acid and pancreatic enzymes in vivo . However, the wide variety of new candidate prebiotics becoming available for human use requires that a manageable set of in vitro tests be agreed on so that their nondigestibility and fermentability can be established without recourse to human studies in every case . In the large intestine, prebiotics, in addition to their selective effects on bifidobacteria and lactobacilli, influence many aspects of bowel function through fermentation . Short-chain fatty acids are a major product of prebiotic breakdown, but as yet, no characteristic pattern of fermentation acids has been identified . Through stimulation of bacterial growth and fermentation, prebiotics affect bowel habit and are mildly laxative . Perhaps more importantly, some are a potent source of hydrogen in the gut . Mild flatulence is frequently observed by subjects being fed prebiotics; in a significant number of subjects it is severe enough to be unacceptable and to discourage consumption . Prebiotics are like other carbohydrates that reach the cecum, such as nonstarch polysaccharides, sugar alcohols, and resistant starch, in being substrates for fermentation . They are, however, distinctive in their selective effect on the microflora and their propensity to produce flatulence.

Am J Clin Nutr, 2001 Feb, 73(2 Suppl), 410S - 414S
Molecular assessment of intestinal microflora; Tannock GW; The application of molecular methodologies to intestinal microflora analysis should enable the development of a detailed knowledge of the microbial ecology of the human colon . This knowledge is essential to derive scientifically valid probiotics . Molecular typing (genetic fingerprinting) methods, eg, ribotyping and pulsed field gel electrophoresis of DNA digests, provide a means of distinguishing bacterial strains inhabiting the intestinal tract . Analysis of lactobacillus, bifidobacterial, and enterobacterial populations with the use of these methods has shown that human and porcine subjects harbor a characteristic collection of bacterial strains . Additionally, perturbations and transitions that occur in these populations and are caused by antibiotic administration or by autogenic or allogenic factors can be detected by molecular analysis of the intestinal microflora . In future studies, molecular typing methods could be used to analyze the composition of bacterial populations before, during, and after the administration of the probiotic product . This experimental approach would provide information on the effect of the probiotic on indigenous strains inhabiting the intestinal tract of humans and other animals.

Am J Clin Nutr, 2001 Feb, 73(2 Suppl), 406S - 409S
Prebiotics: preferential substrates for specific germs?
Roberfroid MB.
A prebiotic is "a non-digestible food ingredient that beneficially affects the host by selectively stimulating the growth and/or the activity of one or a limited number of bacteria in the colon." The premise is based on the hypothesis that the large gut in humans contains bacteria that are beneficial or detrimental to health . Although this generalization probably gives too simplistic a view of gut microbiology, it is a feasible working concept . Currently, food components that seem to exert the best prebiotic effects are inulin-type fructans . In pure culture, most species of bifidobacteria are adapted to the utilization of these nondigestible oligosaccharides but many other bacteria are also capable of metabolizing them . Clearly, these studies of pure bacteria are of limited use unless their results are supported by the results of studies using mixed cultures . Indeed, as many components of the gut microbiota as possible should be measured to indicate a true prebiotic effect . Simple stimulation of bifidobacteria is insufficient to demonstrate an effect; the effects on other gut microorganisms in vivo with human volunteers is necessary . Adjustment of the composition and activities of the colonic microflora so that health-promoting activities are optimized remains key in functional food development . New methods are being applied extensively to human gut microbiology and promise the degree of reliability required to detect subtle changes in colonic microflora composition and to correlate such changes with health benefits . This is a review of the present state of knowledge concerning prebiotics, with emphasis on the criteria used for classification, mechanisms of selective growth stimulation, and physiologic effects.

Am J Clin Nutr, 2001 Feb, 73(2 Suppl), 399S - 405S
Probiotics: determinants of survival and growth in the gut; Bezkorovainy A; Bifidobacteria and lactobacilli are purportedly beneficial to human health and are called probiotics . Their survival during passage through the human gut, when administered in fermented milk products, has been investigated intensely in recent years . Well-controlled, small-scale studies on diarrhea in both adults and infants have shown that probiotics are beneficial and that they survive in sufficient numbers to affect gut microbial metabolism . Survival rates have been estimated at 20-40% for selected strains, the main obstacles to survival being gastric acidity and the action of bile salts . Although it is believed that the maximum probiotic effect can be achieved if the organisms adhere to intestinal mucosal cells, there is no evidence that exogenously administered probiotics do adhere to the mucosal cells . Instead, they seem to pass into the feces without having adhered or multiplied . Thus, to obtain a continuous exogenous probiotic effect, the probiotic culture must be ingested continually . Certain exogenously administered substances enhance the action of both exogenous and endogenous probiotics . Human milk contains many substances that stimulate the growth of bifidobacteria in vitro and also in the small intestine of infants; however, it is unlikely that they function in the colon . However, lactulose and certain fructose-containing compounds, called prebiotics, are not digested in the small intestine but pass into the cecum unchanged, where they are selectively utilized by probiotics . Beneficial effects may thus accrue from exogenously administered probiotics, often administered with prebiotics, or by endogenous bifidobacteria and lactobacilli, whose metabolic activity and growth may also be enhanced by the administration of prebiotics.

Am J Clin Nutr, 2001 Feb, 73(2 Suppl), 365S - 373S
Taxonomy and important features of probiotic microorganisms in food and nutrition; Holzapfel WH et al.; Lactic acid bacteria are among the most important probiotic microorganisms typically associated with the human gastrointestinal tract . Traditionally, lactic acid bacteria have been classified on the basis of phenotypic properties, eg, morphology, mode of glucose fermentation, growth at different temperatures, lactic acid configuration, and fermentation of various carbohydrates . Studies based on comparative 16S ribosomal RNA sequencing analysis, however, showed that some taxa generated on the basis of phenotypic features do not correspond with the suggested phylogenetic relations . Thus, some species are not readily distinguishable by phenotypic characteristics . This is especially true for the so-called Lactobacillus acidophilus group, the Lactobacillus casei and Lactobacillus paracasei group, and some bifidobacteria, strains of which have been introduced in many probiotic foods, eg, the novel yogurt-like commodities . Consequently, modern molecular techniques, including polymerase chain reaction-based and other genotyping methods, have become increasingly important for species identification or for the differentiation of probiotic strains . Probiotic strains are selected for potential application on the basis of particular physiologic and functional properties, some of which may be determined in vitro . The classification and identification of a probiotic strain may give a strong indication of its typical habitat and origin . The species, or even genus name, may also indicate the strain's safety and technical applicability for use in probiotic products . Molecular typing methods such as pulsed-field gel electrophoresis, repetitive polymerase chain reaction, and restriction fragment length polymorphism are extremely valuable for specific characterization and detection of such strains selected for application as probiotics.

J Agric Food Chem, 1998 Jan 19, 46(1), 8 - 12
Growth-Inhibiting Effects of Cinnamomum cassia Bark-Derived Materials on Human Intestinal Bacteria; Lee HS et al.; The growth-inhibiting activity of Cinnamomum cassia (Blume) bark-derived materials toward five intestinal bacteria was examined using an impregnated paper disk method and compared with that of tetracycline and chloramphenicol, as well as four commercially available compounds (cinnamyl alcohol, trans-cinnamic acid, eugenol, and salicylaldehyde) . The biologically active component of C . cassia bark was characterized as cinnamaldehyde by spectral analysis . The growth responses varied with each bacterial strain tested . In a test using 1 and 0.5 mg/disks, cinnamaldehyde revealed potent inhibition against Clostridium perfringensand Bacteroides fragilis . At 1 and 0.5 mg/disk, growth of Bifidobacterium bifidum was significantly inhibited, whereas weak or no inhibitory activity was obtained against Bifidobacterium longum or Lactobacillus acidophilus . The inhibitory effect was much more pronounced in Cl . perfringens, B . fragilis, and Bi . bifidum, compared to Bi . longum or L . acidophilus . Salicylaldehyde exhibited moderate growth-inhibiting activity, but little or no inhibition was observed from treatments with cinnamyl alcohol, trans-cinnamic acid, and eugenol . In contrast, tetracycline and chloramphenicol significantly inhibited growth of all test bacteria as low as 0.01 mg/disk . These results may be an indication of at least one of the pharmacological actions of C . cassia bark.

FEMS Microbiol Lett, 2001 Jan 1, 194(1), 33 - 7
Rapid CE microbial assays for consumer products that contain active bacteria; Armstrong DW et al.; Recent advances in high efficiency separation methods of bacteria allow their rapid identification and quantitation in some cases . A specific capillary electrophoresis (CE) technique is used to identify and quantitate Lactobacillus acidophilus in both pill and syrup health products as well as Bifidobacterium infantis in a powdered formula supplement . Cell viability can be evaluated as well . In some cases, both the living and dead bacterial cells as well as the molecular excipients can be evaluated in a single run.

Biol Pharm Bull, 2000 Dec, 23(12), 1481 - 5
Constitutive beta-glucosidases hydrolyzing ginsenoside Rb1 and Rb2 from human intestinal bacteria; Bae EA et al.; When ginsenoside Rb1 and Rb2 were anaerobically incubated with human intestinal microflora, these ginsenosides were metabolized to 20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol (compound K) and 20(S)-protopanaxadiol . Several kinds of intestinal bacteria hydrolyzed these ginsenosides . Eubacterium sp., Streptococcus sp . and Bifidobacterium sp., which more potently hydrolyzed gentiobiose than sophorose, metabolized ginsenoside Rb1 to compound K via ginsenoside Rd rather than gypenoside XVII . However, Fusobacterium K-60, which more potently hydrolyzed sophorose than gentiobiose, metabolized to compound K via gypenoside XVII . Ginsenoside Rb2 was also metabolized to compound K via ginsenoside Rd or compound O by human intestinal microflora . Eubacterium sp., Streptococcus sp . and Bifidobacterium sp . metabolized ginsenoside Rb2 to compound K via ginsenoside Rd rather than compound O . Fusobacterium K-60 metabolized ginsenoside Rb2 to compound K via compound O.

Int J Food Microbiol, 2000 Dec 5, 62(1-2), 47 - 55
Encapsulation of probiotic bacteria with alginate-starch and evaluation of survival in simulated gastrointestinal conditions and in yoghurt; Sultana K et al.; A modified method using calcium alginate for the microencapsulation of probiotic bacteria is reported in this study . Incorporation of Hi-Maize starch (a prebiotic) improved encapsulation of viable bacteria as compared to when the bacteria were encapsulated without the starch . Inclusion of glycerol (a cryo-protectant) with alginate mix increased the survival of bacteria when frozen at -20 degrees C . The acidification kinetics of encapsulated bacteria showed that the rate of acid produced was lower than that of free cultures . The encapsulated bacteria, however, did not demonstrate a significant increase in survival when subjected to in vitro high acid and bile salt conditions . A preliminary study was carried out in order to monitor the effects of encapsulation on the survival of Lactobacillus acidophilus and Bifidobacterium spp . in yoghurt over a period of 8 weeks . This study showed that the survival of encapsulated cultures of L . acidophilus and Bifidobacterium spp . showed a decline in viable count of about 0.5 log over a period of 8 weeks while there was a decline of about 1 log in cultures which were incorporated as free cells in yoghurt . The encapsulation method used in this study did not result in uniform bead size, and hence additional experiments need to be designed using uniform bead size in order to assess the role of different encapsulation parameters, such as bead size and alginate concentration, in providing protection to the bacteria.

Appl Environ Microbiol, 2001 Jan, 67(1), 420 - 5
Direct in situ viability assessment of bacteria in probiotic dairy products using viability staining in conjunction with confocal scanning laser microscopy; Auty MA et al.; The viability of the human probiotic strains Lactobacillus paracasei NFBC 338 and Bifidobacterium sp . strain UCC 35612 in reconstituted skim milk was assessed by confocal scanning laser microscopy using the LIVE/DEAD BacLight viability stain . The technique was rapid (<30 min) and clearly differentiated live from heat-killed bacteria . The microscopic enumeration of various proportions of viable to heat-killed bacteria was then compared with conventional plating on nutrient agar . Direct microscopic enumeration of bacteria indicated that plate counting led to an underestimation of bacterial numbers, which was most likely related to clumping . Similarly, LIVE/DEAD BacLight staining yielded bacterial counts that were higher than cell numbers obtained by plate counting (CFU) in milk and fermented milk . These results indicate the value of the microscopic approach for rapid viability testing of such probiotic products . In contrast, the numbers obtained by direct microscopic counting for Cheddar cheese and spray-dried probiotic milk powder were lower than those obtained by plate counting . These results highlight the limitations of LIVE/DEAD BacLight staining and the need to optimize the technique for different strain-product combinations . The minimum detection limit for in situ viability staining in conjunction with confocal scanning laser microscopy enumeration was approximately 10(8) bacteria/ml (equivalent to approximately 10(7) CFU/ml), based on Bifidobacterium sp . strain UCC 35612 counts in maximum-recovery diluent.

Biosci Biotechnol Biochem, 2000 Oct, 64(10), 2083 - 8
Purification and identification of a growth-stimulating peptide for Bifidobacterium bifidum from natural rubber serum powder; Etoh S et al.; Natural rubber serum powder, which is a by-product obtained in the production of latex rubber, has a strong growth-stimulating activity for Bifidobacterium bifidum JCM 1254 . The retained fraction obtained by ultrafiltration (molecular weight cutoff 1000) showed a growth-stimulating activity in a dose-dependent manner on B12 assay medium with ammonium sulfate . One of the growth stimulators was purified from the retained fraction by acetone precipitation, solid-phase extraction with a hydrophobic pretreatment column, and multistage reversed-phase HPLC . An increase of 53-fold in the specific activity, and a recovery of 1.3% were obtained . The amino acid composition and N-terminal sequence analysis of this growth stimulator provided the structure of Ala-Thr-Pro-Glu-Lys-Glu-Glu-Pro-Thr-Ala . The molecular mass was 1075 by MALDI-TOF MS analysis . These results showed that this growth stimulator was a decapeptide with the sequence shown above . This is the first report that clarified the structure of an active peptide for the growth of Bifidobacterium.

Microbiol Immunol, 2000, 44(10), 815 - 20
Bifidobacterium lactis Meile et al . 1997 is a subjective synonym of Bifidobacterium animalis (Mitsuoka 1969) Scardovi and Trovatelli 1974; Cai Y et al.; Bifidobacterium lactis JCM 10602T (T = type strain) and Bifidobacterium animalis JCM 1190T were found to be phenotypically similar . These strains were subjected to investigation of their genetic relationships . The 16S rRNA sequence of B . animalis JCM 1190T was aligned with that of other Bifidobacterium species . B . animalis and B . lactis were the most closely related species in the phylogenetic tree and showed a high similarity in sequences (98.8%) . The levels of DNA-DNA hybridization between the type strains of B . lactis and B . animalis ranged from 85.5 to 92.3%, showing that they represent a single species . It is proposed that B . lactis should be considered as a junior subjective synonym of B . animalis.

Lett Appl Microbiol, 2000 Dec, 31(6), 438 - 42
Adhesion, autoaggregation and hydrophobicity of 13 strains of Bifidobacterium longum; Del Re B et al.; To identify bacterial traits related to adhesion ability in human bifidobacteria, 13 strains of Bifidobacterium longum isolated from human gastric juice and intestine were studied . Strains were tested for their capability to adhere to Caco-2 cells and classified as adhesive (Adh+) or non-adhesive (Adh-) . Adh+ and Adh- strains were then investigated for their autoaggregation ability and surface hydrophobicity . Comparing the properties of Adh+ and Adh-, we observed that strains were able to adhere to cell monolayers if they autoaggregate and manifest a good degree of hydrophobicity as determined by microbial adhesion to hydrocarbons . These two traits could be used for preliminary screening to identify potentially adherent isolates.

J Microbiol Methods, 2000 Dec 15, 43(2), 127 - 32
A new selective medium for the isolation of glucose non-fermenting bifidobacteria from hen caeca; Rada V et al.; Avian caeca contain a large and diverse population of bacteria . Certain genera, including Bifidobacterium, are thought to exert health-promoting effects . Two media were evaluated to determine their sensitivity and selectivity for bifidobacteria in the hen caecal samples: modified Wilkins-Chalgren agar (MW; Oxoid) with the addition of glacial acetic acid (1 ml/l) and mupirocin (100 mg/l) and modified TPY agar (MTPY; ADSA, Spain) with glacial acetic acid (1 ml/l) and mupirocin (100 mg/l) . The colonies arising on the plates inoculated with caecal samples were Gram stained, screened for the presence of fructose-6-phosphate phosphoketolase activity, and tested for fermentation patterns using ANAEROtest (Lachema, Czech Republic) and API 50 CHL (BioMerieux, France) kits . Both agars were selective for bifidobacteria, however, MTPY agar showed higher cfu/g than MW agar . Bifidobacterial counts were higher than 10(10) cfu/g of caecal contents using MTPY agar . Most of strains isolated from this medium fermented melibiose, sucrose, and raffinose, but not glucose . Soya peptone (5 g/l; Oxoid) stimulated the growth of glucose non-fermenting strains in complex liquid media . The results suggest that the media for selective enumeration and isolation of bifidobacteria in poultry caecal samples should not contain glucose as the sole carbon source . It can be concluded that MTPY medium is highly selective and permits the growth of both glucose fermenting and glucose non-fermenting bifidobacteria.

Int J Antimicrob Agents, 2000 Dec, 16(4), 521 - 6
Treatment and prevention of antibiotic associated diarrhea; Bergogne-Berezin E; Mild or severe episodes of antibiotic-associated diarrhea (AAD) are common side effects of antibiotic therapy . The incidence of AAD differs with the antibiotic and varies from 5 to 25% . The major form of intestinal disorders is the pseudomembranous colitis associated with Clostridium difficile which occurs in 10-20% of all AAD . In most cases of AAD discontinuation or replacement of the inciting antibiotic by another drug with lower AAD risk can be effective . For more severe cases involving C . difficile, the treatment of diarrhea requires an antibiotic treatment, with glycopeptides (vancomycin) or metronidazole . Another approach to AAD treatment or prevention is based on the use of non-pathogenic living organisms, capable of re-establishing the equilibrium of the intestinal ecosystem . Several organisms have been used in treatment or prophylaxis of AAD such as selected strains of Lactobacillus acidophilus, L . bulgaricus, Bifidobacterium longum, and Enterococcus faecium . Another biotherapeutic agent, a non-pathogenic yeast, Saccharomyces boulardii has been used . In animal models of C . difficile colitis initiated by clindamycin, animals treated with S . boulardii (at end of vancomycin therapy) had a significant decrease in C . difficile colony-forming units, and of toxin B production . In several clinical randomised trials (versus placebo), S . boulardii has demonstrated its effectiveness by decreasing significantly the occurrence of C . difficile colitis and preventing the pathogenic effects of toxins A and B of C . difficile . It has been shown to be a safe and effective therapy in relapses of C . difficile colitis . A good response has been seen in children with AAD, treated by S . boulardii only . In ICUs prevention of AAD remains based on limitation of antibiotic overuse and spread of C . difficile or other agents of AAD should be prevented by improved hygiene measures (single rooms, private bathrooms for patients, use of gloves and hand washing for personnel) . In addition the increasing use of biotherapeutic agents such as S . boulardii should permit the prevention of the major side effect of antibiotics, i.e . AAD in at risk patients.

Exp Anim, 2000 Oct, 49(4), 305 - 7
Inhibitory effect of apple pectin and culture condensate of Bifidobacterium longum on colorectal tumors induced by 1,2-dimethylhydrazine in transgenic mice harboring human prototype c-Ha-ras genes; Ohno K et al.; The number and tumor score of colorectal tumors induced by 1,2-dymethylhydrazine in transgenic (Tg) mice carrying human c-Ha-ras genes were significantly reduced by ingestion of apple pectin (AP) or a culture condensate of Bifidobacterium longum (MB) when compared with a control diet . There was no statistical difference in the incidence of colorectal tumors in Tg mice between the AP or MB diet and the control diet . This study demonstrated that Tg mice are a useful tool for screening inhibition of colorectal tumors by functional foods.

Syst Appl Microbiol, 2000 Oct, 23(3), 386 - 90
Simultaneous identification of five bifidobacterium species isolated from human beings using multiple PCR primers; Dong X et al.; On the basis of 16S rRNA sequences, 5 species-specific forward primers were designed for the identification of 5 Bifidobacterium species isolated from human intestine, namely B . bifidum, B . adolescentis, B . infantis, B . breve and B . longum . As the 5 primers targeted at different sites of 16S rDNA, by using their mixture and a genus-specific reversed primer, the 5 Bifidobacterium species can be simultaneously identified in individual or in mixed culture through PCR amplification . The specificity of the primers was confirmed by the use of genomic DNAs from type strains of all 32 Bifidobacterium species and 6 other relatives . The 5-primer mixture was also applied to the identification of Bifidobacterium strains used commercially . The results turned out to be in accordance with those from conventional identification . This multiple-primer method provides a useful tool for rapid identification of the 5 Bifidobacterium species indicated.

Alcohol Alcohol, 2000 Nov-Dec, 35(6), 561 - 8
Acetaldehyde production and metabolism by human indigenous and probiotic Lactobacillus and Bifidobacterium strains; Nosova T et al.; Many human gastrointestinal facultative anaerobic and aerobic bacteria possess alcohol dehydrogenase (ADH) activity and are therefore capable of oxidizing ethanol to acetaldehyde . We examined whether human gastrointestinal lactobacilli (three strains), bifidobacteria (five strains) and probiotic Lactobacillus GG ATCC 53103 are also able to metabolize ethanol and acetaldehyde in vitro . Acetaldehyde production by bacterial suspensions was determined by gas chromatography after a 1-h incubation with 22 mM ethanol . To determine the acetaldehyde consumption, the suspensions were incubated with 50 microM or 500 microM acetaldehyde as well as with 500 microM acetaldehyde and 22 mM ethanol, i.e . under conditions resembling those in the human colon after alcohol intake . The influence of growth media and bacterial concentration on the ability of lactobacilli to metabolize acetaldehyde and to produce acetate from acetaldehyde were determined . ADH and aldehyde dehydrogenase (ALDH) activities were determined spectrophotometrically . Neither measurable ADH nor ALDH activities were found in aerobically grown Lactobacillus GG ATCC 53103 and Lactobacillus acidophilus ATCC 4356 strains . All the lactobacilli and bifidobacteria strains revealed a very limited capacity to oxidize ethanol to acetaldehyde in vitro . Lactobacillus GG ATCC 53103 had the highest acetaldehyde-metabolizing capacity, which increased significantly with increasing bacterial concentrations . This was associated with a marked production of acetate from acetaldehyde . The type of the growth media had no effect on acetaldehyde consumption . Addition of ethanol to the incubation media diminished the acetaldehyde-metabolizing capacity of all strains . However, in the presence of ethanol, Lactobacillus GG ATCC 53103 still demonstrated the highest capacity for acetaldehyde metabolism of all strains . These data suggest a beneficial impact of Lactobacillus GG ATCC 53103 on high gastrointestinal acetaldehyde levels following alcohol intake . The possible clinical implications of this finding remain to be established in in vitro studies.

J Agric Food Chem, 2000 Nov, 48(11), 5643 - 8
Ascorbate regeneration by the reduced form of 2-amino-3-carboxy-1, 4-naphthoquinone, a strong growth stimulator for bifidobacteria; Yamazaki S et al.; Nonenzymatic reduction of dehydroascorbate into ascorbate by the reduced form (quinol form) of 2-amino-3-carboxy-1,4-naphthoquinone, a strong growth stimulator for bifidobacteria, has been found . The bimolecular reaction rate constant was evaluated as 9 M(-)(1) s(-)(1) at pH 7.0 . This reaction has been successfully coupled with enzymatic regeneration of the naphthoquinol by NAD(P)H in cell-free extracts of Bifidobacterium longum 6001 . The overall reaction is a regeneration of NAD(P)(+) by dehydroascorbate {or a regeneration of ascorbate by NAD(P)H}, in which the naphthoquinone/quinol redox couple functions as an electron transfer mediator . Kinetic study of the reduction of dehydroascorbate with related quinol compounds suggested the significance of the amino substituent of the naphthoquinol . A mechanism of the electron transfer from the quinol to dehydroascorbate is proposed, where the first step of the reaction is a nucleophilic addition of the C(2)-amino substituent of the naphthoquinol to the C(2)-position of dehydroascorbate to form a Schiff base intermediate.

Clin Exp Allergy, 2000 Nov, 30(11), 1604 - 10
Probiotics in the management of atopic eczema; Isolauri E et al.; BACKGROUND: Over the last two decades the incidence of allergic diseases has increased in industrialized countries, and consequently new approaches have to be explored . OBJECTIVE: The potential of probiotics to control allergic inflammation at an early age was assessed in a randomized double-blind placebo-controlled study . METHODS: A total of 27 infants, mean age 4.6 months, who manifested atopic eczema during exclusive breast-feeding and who have had no exposure to any infant or substitute formula were weaned to probiotic-supplemented, Bifidobacterium lactis Bb-12 or Lactobacillus strain GG (ATCC 53103), extensively hydrolysed whey formulas or to the same formula without probiotics . The extent and severity of atopic eczema, the growth and nutrition of infants, and concentrations of circulating cytokines/chemokines and soluble cell surface adhesion molecules in serum and methyl-histamine and eosinophilic protein X in urine were determined . RESULTS: The SCORAD score reflecting the extent and severity of atopic eczema was 16 (7-25) during breast-feeding, median (interquartile range) . After 2 months, a significant improvement in skin condition occurred in patients given probiotic-supplemented formulas, as compared to the unsupplemented group; chi(2) = 12.27, P = 0.002 . SCORAD decreased in the Bifidobacterium lactis Bb-12 group to 0 (0-3.8), and in the Lactobacillus GG group to 1 (0.1-8.7), vs unsupplemented 13.4 (4.5-18.2), median (interquartile range), in parallel with a reduction in the concentration of soluble CD4 in serum and eosinophilic protein X in urine . CONCLUSION: The results provide the first clinical demonstration of specific probiotic strains modifying the changes related to allergic inflammation . The data further indicate that probiotics may counteract inflammatory responses beyond the intestinal milieu . The combined effects of these probiotic strains will guide infants through the weaning period, when sensitization to newly encountered antigens is initiated . The probiotic approach may thus offer a new direction in the search for future foods for allergy treatment and prevention strategies.

Can J Microbiol, 2000 Oct, 46(10), 878 - 84
Bile salt toxicity to some bifidobacteria strains: role of conjugated bile salt hydrolase and pH; Grill JP et al.; The purpose of this work was to study some aspects of bile salt toxicity towards bifidobacteria . A strain (Bifidobacterium coryneforme ATCC 25911) was selected for its lack of conjugated bile salt hydrolase activity (CBSH-), and was used with three deconjugating strains (CBSH+), for study of their growth and viability in the presence of two dihydroxylated conjugated bile salts (tauro- and glyco-deoxycholic acids) . The presence of the glycoconjugate induced a more significant growth inhibition for the four strains than the tauroconjugate . The viability of the strains was measured at several pH levels . Glycodeoxycholic acid, but not taurodeoxycholic acid, exerted a lethal effect, which increased at low pH . This phenomenon was more pronounced for the CBSH- strain . We explain some of these results using an hypothesis based on the consequence of dissociation of conjugated and deconjugated bile salts, and the value of their pKa.

Arch Pharm Res, 2000 Oct, 23(5), 482 - 7
Antitumor activity of Bifidobacterium spp . isolated from a healthy Korean; Rhee YK et al.; The antitumor activity of Bifidobacterium breve K-110, and K-111, and B . infantis K-525 was investigated . These Bifidobacterial cells and their cell wall preparations (WPG) significantly increased the survival rate of mice who had been intraperitoneally implanted with sarcoma 180 cells . Solid tumor growth was inhibited even when the sarcoma 180 cells were implanted into the groins of the mice . However, the Bifidobacterial cells did not show in vitro cytotoxicity against tumor cell lines . Cell kinetic studies revealed that these WPGs induced neutrophils, which were followed by macrophages, at the site of peritoneal injection . The WPGs directly activated these cells to inhibit the growth of tumor cells in in vitro assays . Our results suggest that Bifidobacterial WPGs induce and activate nonspecific phagocytes in situ to reject growing tumor cells in the mouse peritoneal cavity.

J Appl Microbiol, 2000 Oct, 89(4), 668 - 77
Evaluation of 16s rRNA and cellular fatty acid profiles as markers of human intestinal bacterial growth in the chemostat; Hopkins MJ et al.; Chemostats were used to study the effects of carbon and nitrogen limitation and specific growth rate on 16S rRNA synthesis and cellular fatty acid (CFA) profiles in four human intestinal bacteria (Bacteroides thetaiotaomicron, Bifidobacterium adolescentis, Clostridium bifermentans and Cl . difficile) . Cellular fatty acid synthesis varied with dilution rate and nutrient availability in different species, but these cellular constituents were relatively stable phenotypic characteristics in Bact . thetaiotaomicron, where branched chain and hydroxy CFA were good taxonomic markers . Conversely, CFA in the Gram-positive bacteria varied markedly with changes in growth environment . For example, in chemostats, cyclopropane CFA were only synthesized in Cl . bifermentans and Cl . difficile under N-limited conditions . Similarly, Dimethyl acetal (DMA) fatty acids in Bif . adolescentis were primarily produced during N-limited growth, and this was inversely related to dilution rate . At low growth rates, 16S rRNA concentrations (microg rRNA per ml culture) correlated with viable bacterial counts, but were more closely related to specific growth rate when expressed as a function of cell mass (microg rRNA per mg dry weight bacteria) . However, this did not reveal differences in bacterial population size and rRNA concentration in C-limited cultures . Thus, at low dilution rates, C limitation strongly reduced rRNA synthesis in Cl . bifermentans, despite viable cell counts being similar to those in N-limited cultures . These results indicate that, while 16S rRNA is a useful indicator of microbial activity, cell growth rate does not necessarily relate to rRNA concentration under all nutritional conditions . Consequently, bowel habit and diet will affect both CFA and rRNA content in bacteria isolated from intestinal samples, and this should be taken into consideration when interpreting such data measurements.

J Food Prot, 2000 Oct, 63(10), 1369 - 76
Effect of conjugated bile salts on antibiotic susceptibility of bile salt-tolerant Lactobacillus and Bifidobacterium isolates; Charteris WP et al.; Virtually every antibiotic may cause in vivo alterations in the number, level, and composition of the indigenous microbiotae . The degree to which the microbiotae are disturbed depends on many factors . Although bile may augment antibiotic activity, studies on the effect of bile on the antibiotic susceptibility of indigenous and exogenous probiotic microorganisms are lacking . It was against this background that the antibiotic susceptibility of 37 bile salt-tolerant Lactobacillus and 11 Bifidobacterium isolates from human and other sources was determined in the presence of 0.5% wt/wt oxgall (conjugated bile salts) . Oxgall did not affect the intrinsic resistance of lactobacilli to metronidazole (5 microg), vancomycin (30 microg), and cotrimoxazole (25 microg), whereas it resulted in a complete loss of resistance to polymyxin B (300 microg) and the aminoglycosides gentamicin (10 microg), kanamycin (30 microg), and streptomycin (10 microg) for most strains studied (P < 0.001) . Oxgall did not affect the intrinsic resistance of bifidobacteria to metronidazole and vancomycin, whereas polymyxin B and co-trimoxazole resistance was diminished (P < 0.05) and aminoglycoside resistance was lost (P < 0.001) . Seven lactobacilli, but no bifidobacteria strain, showed unaltered intrinsic antibiotic resistance profiles in the presence of oxgall . Oxgall affected the extrinsic susceptibility of lactobacilli and bifidobacteria to penicillin G (10 microg), ampicillin (10 microg), tetracycline (30 microg), chloramphenicol (30 microg), erythromycin (15 microg), and rifampicin (5 microg) in a source- and strain-dependent manner . Human strain-drug combinations of lactobacilli (P < 0.05) and bifidobacteria (P < 0.01) were more likely to show no change or decreased susceptibility compared with other strain-drug combinations . The antimicrobial activity spectra of polymyxin B and the aminoglycosides should not be considered limited to gram-negative bacteria but extended to include gram-positive genera of the indigenous and transiting microbiotae in the presence of conjugated bile salts . Those lactobacilli (7 of 37) that show unaltered intrinsic and diminished extrinsic antibiotic susceptibility in the presence of oxgall may possess greater upper gastrointestinal tract transit tolerance in the presence of antibiotics.

Gut, 2000 Nov, 47(5), 646 - 52
Bifidobacterium strains from resident infant human gastrointestinal microflora exert antimicrobial activity; Lievin V et al.; BACKGROUND AND AIMS: The gastrointestinal microflora exerts a barrier effect against enteropathogens . The aim of this study was to examine if bifidobacteria, a major species of the human colonic microflora, participates in the barrier effect by developing antimicrobial activity against enterovirulent bacteria . METHODS: Antibacterial activity was examined in vitro against a wide range of Gram negative and Gram positive pathogens . Inhibition of Salmonella typhimurium SL1334 cell association and cell invasion was investigated in vitro using Caco-2 cells . Colonisation of the gastrointestinal tract in vivo by bifidobacteria was examined in axenic C3/He/Oujco mice . Antimicrobial activity was examined in vivo in axenic C3/He/Oujco mice infected by the lethal S typhimurium C5 strain . RESULTS: Fourteen human bifidobacterium strains isolated from infant stools were examined for antimicrobial activity . Two strains (CA1 and F9) expressed antagonistic activity against pathogens in vitro, inhibited cell entry, and killed intracellular S typhimurium SL1344 in Caco-2 cells . An antibacterial component(s) produced by CA1 and F9 was found to be a lipophilic molecule(s) with a molecular weight of less than 3500 . In the axenic C3/He/Oujco mice, CA1 and F9 strains colonised the intestinal tract and protected mice against S typhimurium C5 lethal infection . CONCLUSION: Several bifidobacterium strains from resident infant human gastrointestinal microflora exert antimicrobial activity, suggesting that they could participate in the "barrier effect" produced by the indigenous microflora.

Int J Food Microbiol, 2000 Oct 1, 61(1), 17 - 25
Survival of bifidobacteria in yogurt and simulated gastric juice following immobilization in gellan-xanthan beads; Sun W et al.; A novel acid-stable bead made of gellan gum and xanthan gum was used to immobilize Bifidobacteria . The beads (0.75% gellan and 1% xanthan gum) had an average diameter of 3 mm and did not shrink in 25% lactic acid solution, pH 1.5, or 20% acetic acid solution, pH 1.5, after storage at 4 degrees C for 4 weeks . Bifidobacterium infantis ATCC 15697, the most acid-tolerant strain tested, was immobilized in gellan-xanthan beads and its survival in peptone water, pH 4, pasteurized yogurt, and simulated gastric juice was monitored . In peptone water, pH 4, the reduction in cell count of immobilized cells of B . infantis ATCC 15697 was not significantly different from that obtained with free cells during 6 weeks of storage at 4 degrees C . However, counts of immobilized cells of B . infantis ATCC 15697 remained significantly higher than free cells (P < 0.0001) when both were exposed to simulated gastric juices at pH 2.5, 2.0 and 1.5 . At pH 2.5, the viable count of free cells dropped from 1.23 x 10(9) CFU/ml to an undetectable level (< 10 CFU/ml) in 30 min, while the viable count of immobilized cells decreased by only 0.67 log cycle in the same time period . Immobilized cells also survived significantly better than free cells (P <0.05) in pasteurized yogurt after refrigerated storage for 5 weeks.

Dig Dis Sci, 2000 Aug, 45(8), 1617 - 22
Antioxidative effect of intestinal bacteria Bifidobacterium longum ATCC 15708 and Lactobacillus acidophilus ATCC 4356; Lin MY et al.; The antioxidative effect of intact cells and intracellular cell-free extracts of intestinal lactic acid bacteria B . longum (ATCC 15708) and L . acidophilus (ATCC 4356) was investigated . Both intact cells and intracellular cell-free extracts of 10(9)cells of B . longum and L . acidophilus demonstrated antioxidative activity, inhibiting linoleic acid peroxidation by 28-48% . This indicated that these two strains demonstrated excellent antioxidative activity . B . longum and L . acidophilus also showed the ability to scavenge alpha,alpha-diphenyl-beta-picrylhydrazyl (DPPH) free radical, scavenging 21-52% . The intact cells of these two intestinal bacteria demonstrated a high inhibitory effect on the cytotoxicity of 4-nitroquinoline-N-oxide (4NQO) . Cytotoxicity of 4NQO was reduced by L . acidophilus by approximately half and by almost 90% by B . longum . Nevertheless, no inhibition of cytoxicity observed for intracellular cell-free extracts of 10(9) cells of B . longum and L . acidophilus . The effect of B . longum and L . acidophilus on inhibiting plasma lipid peroxidation was also evaluated . The results showed that both intestinal strains were able to protect plasma lipid from oxidation at different degrees . The inhibition rates on plasma lipid peroxidation ranged from 11 to 29% for 10(9) cells of B . longum and L . acidophilus . Generally speaking, B . longum demonstrated better antioxidative ability than L . acidophilus in this study.

FEMS Microbiol Lett, 2000 Oct 1, 191(1), 17 - 24
Molecular differentiation of Bifidobacterium species with amplified ribosomal DNA restriction analysis and alignment of short regions of the ldh gene; Roy D et al.; The differentiation of Bifidobacterium species was performed with specific primers using the PCR technique, the amplified ribosomal DNA restriction analysis (ARDRA) technique based on reports on the sequence of the 16S rRNA gene and speciation based on a short region of the ldh gene . Four specific primer sets were developed for each of the Bifidobacterium species, B . animalis, B . infantis and B . longum . The use of the ARDRA method made it possible to discriminate between B . infantis, B . longum and B . animalis with the combination of BamHI, TaqI and Sau3AI restriction enzymes . The ldh gene sequences of 309-312 bp were determined for 19 Bifidobacterium strains . Alignment of these short regions of the ldh gene confirmed that it is possible to distinguish between B . longum and B . infantis but not between B . lactis and B . animalis.

J Dairy Sci, 2000 Sep, 83(9), 1946 - 51
Viability of microencapsulated bifidobacteria in set yogurt during refrigerated storage; Adhikari K et al.; Bifidobacteria are probiotic organisms that improve the microbial balance in the human gut . They can be incorporated as live cultures in fermented dairy foods, including yogurt, for transmission to humans . Because bifidobacteria are sensitive to high acidity, their viability in yogurt is limited . The objective of the present study was to investigate the effect of microencapsulation on the viability of bifidobacteria in yogurt during refrigerated storage for 30 d . Live bifidobacterial cells were encapsulated in kappa-carrageenan . Cell enumeration, determination of titratable acidity and pH, quantitation of lactic and acetic acids, and sensory evaluation (consumer test) were carried out on the yogurt samples . Microbiological results showed a decline of 78 and 70.5% in the population of Bifidobacterium longum B6 and B . longum ATCC 15708, respectively, for the treatments containing nonencapsulated cells . No difference in bifidobacterial population was observed in the encapsulated treatments . The acetic acid content in the yogurt with nonencapsulated bifidobacteria was higher than that in the plain yogurt (control) and encapsulated treatments . The increase in lactic acid content during storage was not different among the various treatments for B . longum B6, but was greater for nonencapsulated than encapsulated B . longum 15708 and the control . Consumers judged the nonencapsulated treatment as the most sour, which was likely due to the higher acetic acid content . Consumers preferred the control and nonencapsulated treatments over the encapsulated treatment . Microencapsulation appears to increase the viability of bifidobacteria in yogurt . This technique can be used to transmit bifidobacteria via fermented products provided that sensory characteristics of the product are improved or maintained.

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