J Dairy Sci, 1996 May, 79(5), 750 - 7 Improvement of lactose digestion in humans by ingestion of unfermented milk containing Bifidobacterium longum; Jiang T et al.; Fifteen lactose malabsorbers were studied to evaluate the effects of consumption of milk containing different strains of Bifidobacterium longum on lactose digestion . Influences of different growth substrates, bile sensitivity, and lactose transport on lactose digestion by bifidobacteria were also investigated . Lactose malabsorption was determined by measuring breath hydrogen excretion of subjects fed four different test milks (three of which contained 5 x 10(8) cfu/ml of B . longum) on 4 different d using a randomized, double-blinded trial . Test milks included 1) 400 ml of lowfat milk (control), 2) 400 ml of milk containing B . longum B6 that had been grown with lactose, 3) 400 ml of milk containing B . longum B6 grown with lactose plus glucose, or 4) 400 ml of milk containing B . longum ATCC 15708 grown with lactose . beta-Galactosidase activity was highest in milk containing B6 grown with lactose but was extremely low in milk containing B6 grown with lactose and glucose . Consumption of milk containing B6 grown with lactose resulted in significantly less hydrogen production and flatulence than occurring after consumption of control milk or the milk containing B6 grown with both lactose and glucose . Hydrogen production after ingestion of 15708 was also significantly lower than hydrogen production after ingestion of the control milk . We concluded that milks containing B . longum might reduce breath hydrogen response and symptoms from lactose malabsorption when the culture is grown in a medium containing only lactose to induce a higher beta-galactosidase level and increase rate of lactose uptake. Biol Pharm Bull, 1996 May, 19(5), 705 - 9 Purification and characterization of a novel sennoside-hydrolyzing beta-glucosidase from Bifidobacterium sp . strain SEN, a human intestinal anaerobe; Yang L et al.; A novel beta-glucosidase, which is inducible and capable of catalyzing the hydrolysis of sennosides, was purified from Bifidobacterium sp . strain SEN with Triton X-100 solubilization and DEAE-cellulose column chromatography, by which hydrolytic activities toward sennoside B, 4-methylumbelliferyl beta-glucoside (MUG), and p-nitrophenyl beta-glucoside (pNPG) were obtained together in the same eluted fractions . The activity was stable against detergents such as sodium dodecyl sulfate (SDS) and Triton X-100, but was denatured by SDS and beta-mercaptoethanal when heated . The final preparation was shown to be nearly homogeneous on SDS-polyacrylamide gel electrophoresis (PAGE) either after the enzyme was denatured or when it was not denatured . In the non-denaturing SDS-PAGE, a single protein band hydrolyzed MUG on the gel . In the denaturing SDS-PAGE, the subunit mass of the enzyme was estimated to be 110 kDa . The enzyme was optimally active at pH 6.0 for hydrolysis of sennoside B and MUG . Km values for sennoside B and MUG are 0.94 and 0.53 mM, respectively . The enzyme also catalyzed the hydrolysis of pNPG, amygdalin, geniposide and salicin . It was less active against methyl beta-glucoside and incapable of hydrolyzing cellobiose . The beta-glucosidase activity was inhibited by deoxynojirimycin and p-chloromercuribenzenesulfonic acid, but was less susceptible to several metals (FeSO4, ZnCl2, and CuSO4), and 5,5'-dithio-bis(2-nitrobenzoic acid). Biol Pharm Bull, 1996 May, 19(5), 701 - 4 A sennoside-hydrolyzing beta-glucosidase from Bifidobacterium sp . strain SEN is inducible; Yang L et al.; Bifidobacterium sp . strain SEN was isolated and characterized by hydrolytic conversion of sennosides to sennidins (Akao et al., Appl . Environ . Microbiol., 60, 1041 (1994)) . The sennoside-hydrolyzing capacity of the strain SEN was disappeared following the addition of glucose to the media in spite of good bacterial growth and potent activity hydrolyzing p-nitrophenyl beta-D-glucopyranoside (pNPG) . In a fructose-containing medium, no such suppressing effect was shown . Following a 10 h incubation in 50 mM potassium phosphate buffer (pH 7.4), the sennoside-hydrolyzing activity of the bacterium increased, dose-dependently, with the addition of sennoside B . Inhibition of the substrate-induced increase in sennoside-hydrolyzing activity was observed following the addition of some antibiotics (chloramphenicol, streptomycin, and rifampicin) . In particular, chloramphenicol completely inhibited the increase of sennoside-hydrolyzing activity while 38% pNPG-hydrolyzing activity remained . It is suggested that the strain SEN produces two different beta-glucosidases of which the sennoside-hydrolyzing enzyme is inducible . In addition, the glucosides pNPG, esculin, salicin, or amygdalin stimulated the induction of the sennoside beta-glucosidase, but less markedly than sennoside . Sennidin A or sugars (glucose, fructose, cellobiose, or maltose) did not induce the enzyme. Appl Microbiol Biotechnol, 1996 May, 45(4), 484 - 9 Construction of an integrative food-grade cloning vector for Lactobacillus acidophilus; Lin MY et al.; An integrative cloning vector was constructed using a randomly cloned HindIII-digested chromosomal fragment from Lactobacillus acidophilus ADH inserted into an Escherichia coli vector, pBluescript II SK+ . Southern hybridization studies demonstrated homology of the inserted fragment with one other L . acidophilus strain and one Bifidobacterium strain . Identification of a SauI site located near the middle of the 1.9-kb ADH chromosomal fragment made it possible to clone the Lactobacillus bulgaricus beta-galactosidase (EC 3.2.1.23) gene into this vector . The vector was unable to replicate in the homologous host, L . acidophilus ADH, following electroporation . The chromosomal fragment allowed the integration of the beta-galactosidase gene (beta gal) into the host chromosome via homologous recombination . The size of the two flanking L . acidophilus ADH chromosomal fragments, approximately 0.95 kb each, was sufficient to allow the double cross-over to take place . Southern hybridization demonstrated that only L . acidophilus and L . bulgaricus DNA had been integrated into the chromosome of the host strain . The beta-galactosidase activity of the transformant was increased approximately 200-fold when compared to the enzyme activity of the wild-type strain . The beta gal gene remained stable in the transformant strain after 30 transfers in growth media without selection pressure . This first-generation integrative cloning vector is constructed solely of DNA from organisms consumed by humans and could be considered a food-grade vector system. J Nutr, 1996 May, 126(5), 1505 - 11 Feeding human milk to rats increases Bifidobacterium in the cecum and colon which correlates with enhanced folate status; Krause LJ et al.; The purpose of this investigation was to determine if feeding diets containing human milk resulted in increased numbers of microorganisms implicated in increased folate production and the effect on folate availability . Following a folate-depletion period (5 wk), 30 rats were fed folate-repletion diets (4 wk) with or without 20% milk solids (human, cow or goat) and containing either 906 or 4530 nmol folic acid/kg . At the end of the test period, the cecum and colon were removed in an anaerobic chamber, homogenized, diluted (10(-2) -10(-8)), and the contents of each plated on selective and nonselective media . In addition to enumeration of the total anaerobic load, five genera of bacteria were counted (Bacteroides, Bifidobacterium, Clostridium, Escherichia and Streptococcus) . Rats fed human milk solids had at least a seven- and onefold mean increase in the Bifidobacterium concentration in the cecum (P < 0.006) and colon (P < 0.04), respectively, compared with rats fed other diets . The total anaerobic bacterial concentration in the cecum and the colon of rats fed human milk solids was also greater than that of rats fed the other diets (P < 0.05) . The single exception was the total anaerobic count in the cecum of rats consuming goat milk solids, which did not differ from that of rats consuming human milk solids . Further, rats fed human milk solids had at least a 42 and 48% higher mean plasma folate concentration and total cecal material folate content, respectively, than rats in other dietary treatments containing 906 nmol/kg folic acid . Therefore, the improved folate status of rats fed human milk-containing diets appears to be due, at least in part, to increased folate synthesis by Bifidobacteria and other folate-synthesizing microbes in the cecum and colon. J Nutr, 1996 May, 126(5), 1362 - 71 Probiotics, cecal microflora, and aberrant crypts in the rat colon; Gallaher DD et al.; Our hypothesis was that administration of bifidobacteria, Lactobacillus acidophilus or both to rats will minimize the numbers of aberrant crypts in the distal colon that develop in response to the carcinogen 1,2-dimethylhydrazine (DMH) . A series of experiments was designed to test this hypothesis where the treatments used were as follows: skim milk controls (Skim-Basal), skim milk + bifidobacteria (Bifido-Basal), skim milk + fructooligosaccharide (Skim-FOS), and skim milk + bifidobacteria + fructooligosaccharide (Bifido-FOS) . In two experiments, bifido-bacteria + FOS administration significantly decreased the number of aberrant crypts that developed, but there was no clear relationship of aberrant crypts to numbers of bifidobacteria or Clostridium perfringens . In the third experiment, the Bifido-FOS treatment led to significantly fewer aberrant crypts and aberrant crypt foci than the Bifido-Basal treatment . The Skim-FOS group had significantly more cecal bifidobacteria than the Skim-Basal group and significantly fewer C . perfringens than the Skim-Basal and Bifido-Basal . In a fourth experiment, L . acidophilus was added as an additional treatment . The number of aberrant crypts was not significantly different among the groups . However, the number of C . perfringens was significantly decreased by the addition of bifidobacteria, L . acidophilus or the combination of the two, whereas the numbers of bifidobacteria and L . acidophilus were not affected by treatment . A significant correlation (R2 = 0.84, P < 0.01) was noted between the body weight of rats at DMH administration and the magnitude of the difference in aberrant crypts between the Skim-Basal rats and the Bifido-FOS rats . The results suggest that there is variability in the effects of bifidobacteria and L . acidophilus administration on both aberrant crypt formation and C . perfringens. Biol Pharm Bull, 1996 Apr, 19(4), 647 - 48 Reduction of oxazepam to desmethyldiazepam by human intestinal bacteria; Okamura T et al.; The biotransformation of oxazepam by Bifidobacterium bifidum was studied . The major metabolite was purified by chromatographic methods and found to be desmethyldiazepam using NMR, IR and other physicochemical data. Int J Syst Bacteriol, 1996 Apr, 46(2), 564 - 71 Bifidobacterium inopinatum sp . nov . and Bifidobacterium denticolens sp . nov., two new species isolated from human dental caries; Crociani F et al.; In a previous investigation of bifidobacteria isolated from human dental caries (V . Scardovi and F . Crociani, Int . J . Syst . Bacteriol . 24:6-20, 1974), 40 strains were assigned to the new species Bifidobacterium dentium . In this study we examined 70 new strains of bifidobacteria isolated from dental caries . The morphological characteristics, biochemical reactions, fermentation patterns, end products from glucose metabolism, protein electrophoretic patterns, levels of DNA hybridization, and DNA G+C contents of these organisms revealed that they belong to three different taxa . One of these taxa was identified as B . dentium . The other two are described as the following new Bifidobacterium species in this paper: Bifidobacterium inopinatum (type strain, DSM 10107) and Bifidobacterium denticolens (type strain, DSM 10105) . The two new species differ from other Bifidobacterium species in their morphological characteristics (especially B . inopinatum, with its very small coccoid cells), in their carbohydrate fermentation patterns (most strains ferment dextran, and B . inopinatum does not ferment galactose), and in their DNA base compositions (especially B . inopinatum). Jpn J Antibiot, 1996 Apr, 49(4), 367 - 76 {In vitro activities of sulopenem, a new parenteral penem, against anaerobes}; Watanabe K et al.; In vitro activities of sulopenem, a novel parenteral penem, was compared with those of imipenem, flomoxef, cefuzonam, cefoperazone and sulbactam/ampicillin against 66 reference strains (19 genera, 61 species) and 392 recent clinical isolates of anaerobic bacteria and fastidious aerobic bacteria . Sulopenem had a very broad spectrum against anaerobic bacteria . In general, this compound was active against anaerobic reference strains with MICs of < or = 0.78 micrograms/ml, while being the least active against Bifidobacterium spp . and less active than imipenem against Lactobacillus spp . Sulopenem was more active against Bacteroides fragilis isolates than imipenem and had the highest activities against Bacteroides thetaiotaomicron, Prevotella intermedia, Porphyromonas gingivalis, Fusobacterium spp . and Peptostreptococcus spp . among the antibiotics tested . Sulopenem was not hydrolyzed by oxyiminocephalosporinase type 1 produced by B . fragilis GAI-0558, GAI-7955 and GAI-10150 and its stability was comparable to imipenem . Its susceptibilities to hydrolysis by a metallo-beta-lactamase from B . fragilis GAI-30144 was less than imipenem . Sulopenem (120 mg/kg, 3 times a day for 4 days) was as effective as imipenem/cilastatin against a mixed intraabdominal mice infection due to E . coli and B . fragilis . Sulopenem (20 mg/kg twice a day for 5 days) did not induce an overgrowth of Clostridium difficile in the caecum of mice. J Dent Res, 1996 Apr, 75(4), 1008 - 14 The final pH of bacteria comprising the predominant flora on sound and carious human root and enamel surfaces; van Houte J et al.; Acidogenesis at low pH appears to be an important bacterial cariogenic trait . However, most information in this regard pertains to only a few of the acidogenic dental plaque bacteria . Therefore, the 'final' pH in sugar broth was determined for a wide variety of oral bacteria . Their source was: (1) carious material from advanced root lesions (ARL), (2) plaque from sound root surfaces of root-caries-free subjects (SRS), (3) plaque from "white spot" coronal lesions and sound coronal surfaces of caries-active subjects, and (4) plaque from sound coronal surfaces of caries-free subjects . Strains from groups 1 and 2 (ARL, 389 strains; SRS, 358 strains) were previously identified (van Houte et al., 1994) to the genus/species level and belonged to the predominant cultivable flora (PCF) . Strains from groups 3 and 4 also belonged to the PCF but were not identified . All strains were placed in one of 4 final pH categories: < 4.2, 4.2-4.4, 4.4-4.6, and > or = 4.6 . The main findings were: (1) ARL samples contained many strains with a final pH < 4.2 (mean percentage of 25.7) . They included all strains of Lactobacillus and mutans streptococci (MS), most Bifidobacterium strains and non-mutans streptococci (non-MS), and about 20% of the Actinomyces strains . By contrast, SRS samples contained far fewer strains with a final pH < 4.2 (mean percentage of 8.4) which were nearly all non-MS . (2) Organisms with a final pH < 4.4 constituted mean percentages of 41.5 and 32.1 for the ARL and SRS samples, respectively . (3) The final pH distribution of strains in samples from coronal surfaces showed a tendency relative to caries activity (group 3 vs . group 4) similar to that for groups 1 and 2 . Our findings further support the concept that increased cariogenic conditions are associated with increased proportions of organisms capable of acidogenesis at a low pH and that this shift involves organisms other than the MS and lactobacilli. Z Ernahrungswiss, 1996 Mar, 35(1), 22 - 31 {Structural and functional aspects of oligosaccharides in human milk}; Kunz C et al.; About a century ago, pediatricians observed that in feces of breast-fed infants, compared to those of bottle-fed infants, Bifidobacterium bifidum was the predominant microorganism . It was shown thereafter that aminosugar-containing oligosaccharides are growth factors for a specific strain of Bifidobacterium . Meanwhile, more than 130 lactose-derived oligosaccharides have been identified in human milk . Some of these oligosaccharides like Lacto-N-Tetraose and Lacto-N-Fucopentaose I and II do not occur in minute amounts but in concentrations up to 1-2 g/L . As the total amount of complex oligosaccharides is between 3-6 g/L those components have to be considered as major human milk constituents . There is striking evidence that human milk oligosaccharides are potent inhibitors of bacterial adhesion to epithelial surfaces, an initial stage of infective processes . Therefore, these oligosaccharides are considered to be soluble receptor analogues of epithelial cell surfaces participating in the non-immunological defense system of human milk-fed infants. Res Microbiol, 1996 Mar-Apr, 147(3), 183 - 92 Ribosomal DNA polymorphism in the genus Bifidobacterium; Mangin I et al.; Ribosomal DNA polymorphism was studied in order to demonstrate intra- and interspecies differentiation of 42 Bifidobacterium strains . DNA from these strains was digested with the endonucleases BamHI, EcoRV, HindIII and PvuII and then analysed by Southern blotting . Ribosomal patterns using a part of an rRNA 23S gene as a probe clearly differentiated the majority of species from each other . Only B . indicum ATCC 25912T and B . infantis ATCC 15697T displayed identical ribosomal patterns, even though they are classified into two different species . Moreover, ribotypes were able to distinguish between strains belonging to the same species . Furthermore, these strains generally showed common bands, except for B . infantis strains and two strains of B . animalis. Res Microbiol, 1996 Mar-Apr, 147(3), 133 - 43 Characterization of the plasmid pMB1 from Bifidobacterium longum and its use for shuttle vector construction; Rossi M et al.; The nucleotide sequence of the 1847-bp Bifidobacterium longum B2577 cryptic plasmid pMB1 was determined . The plasmid had a G+C content of 62.0%, and contained two open reading frames, orf1 and orf2, likely arranged in an operon . The proteins encoded by orf1 and orf2 show the highest degree of similarity with similarly arranged peptide sequences translated from Corynebacterium glutamicum pXZ10142 and Mycobacterium fortuitum pAL5000 plasmids . Recombinant plasmids containing the pMB1 replicon were able to replicate in Bifidobacterium animalis MB209 . The successful transformation of this strain with pMB1-based plasmids facilitated characterization of this replicon, results of which showed that both orf1 and orf2 are necessary for plasmid replication . A family of new Escherichia coli-B . animalis shuttle plasmids, based on the pMB1 replicon and expressing a cat and an ery gene, was constructed. Brain Dev, 1996 Mar-Apr, 18(2), 160 - 2 Neonatal meningitis caused by Bifidobacterium breve; Nakazawa T et al.; We are reporting a male neonate with meningitis caused by Bifidobacterium breve . This is only the second case reported so far to our knowledge . The patient's clinical course was excellent and inflammatory indications, such as serum C-reactive protein, were weak . Although the antibiotics used for the patient were effective against this bacterium both in vitro and in vivo, two relapses occurred which might have been caused by an incomplete remission due to the low permeability of antibiotics through the blood-brain barrier under the very mild inflammation of the meninges, and also by the discrepancy between minimum bactericidal concentrations (MBC) and minimum inhibitory concentrations (MIC) . Anaerobic meningitis is very rare, but it may exist in more than the reported cases . Anaerobic culture should be performed for patients with culture-negative purulent meningitis. Zh Mikrobiol Epidemiol Immunobiol, 1996 Mar-Apr, (2), 88 - 91 {The probiotic correction of microecological and immune disorders in gastroduodenal pathology in children}; Lykova EA et al.; The state of microbiocenosis was studied and the level of immunoglobulins was determined in the gastric juice and feces of children with chronic diseases of the digestive system . In 20% of patients an increase in the contamination of the gastric biotope with opportunistic microflora was established . The isolation rate of Helicobacter pylori was 56% . The detection of H . pylori was found to be accompanied by the aggravation of the form and course of gastritis . In cases of Helicobacter-associated pathology the deficiency of Lactobacillus sp . in the stomach was established, which was accompanied by their deficiency and absence in feces . The study also revealed a decrease in the population level of Bifidobacterium sp . with a simultaneous increase in the population of opportunistic enterobacteria and changes in the state of local immunity: the decreased level of SIgA in most samples and the decreased level of IgG in the presence of H . pylori . The correction of microecological and immune disturbances with probiotic preparations, containing bifidobacteria (bifidumbacterin-forte) and lactobacilli, yielded good results. Carbohydr Res, 1996 Feb 23, 281(2), 285 - 91 Structure of a galactan from cell walls of Bifidobacterium catenulatum YIT4016; Nagaoka M et al.; A structural study was carried out on a galactose-rich polysaccharide fraction isolated from cell walls of Bifidobacterium catenulatum YIT4016 after N-acetylmuramidase digestion . The polysaccharide contained galactose and glucosamine in a molar ratio of 16.9:1.0 . Data obtained by 13C NMR spectroscopy showed that the backbone chain of this polysaccharide is composed of galactofuranose residues, while the branches consist of galactopyranosyl residues . Furthermore, the data obtained from NaIO4 oxidation, partial methanolysis and methylation analysis indicated that this polysaccharide consists of a trisaccharide repeating unit having the following structure: {sequence: see text} Biosci Biotechnol Biochem, 1996 Feb, 60(2), 188 - 93 Purification and characterization of beta-D-glucosidase (beta-D-fucosidase) from Bifidobacterium breve clb acclimated to cellobiose; Nunoura N et al.; The beta-D-glucosidase (EC . 3.2.1.21) activity of Bifidobacterium breve 203 was increased by acclimation with cellobiose, and the enzyme was purified to homogeneity from cell-free extracts of an acclimatized strain of B . breve clb, by ammonium sulfate fractionation and column chromatographies of anion-exchange, gel filtration, Gigapaite, and hydrophobic interaction . This enzyme had not only beta-D-glucosidase activity but also beta-D-fucosidase activity, which is specific to Bifidobacteria in intestinal flora . The molecular weight of the purified enzyme was estimated to be 47,000-48,000 and the enzyme was assumed to be a monomeric protein . The optimum pH and temperature of the enzyme were around 5.5 and 45 degrees C, respectively . The enzyme was stable up to 40 degrees C and between pH 5 and 8 . The isoelectric point of the enzyme was 4.3 and the Km values for p-nitrophenyl-beta-D-glucoside and p-nitrophenyl-beta-D-fucoside were 1.3 mM and 0.7 mM, respectively . This enzyme had also transferase activity for the beta-D-fucosyl group but not for the beta-D-glucosyl group . The N-terminal amino acid sequence of this enzyme was similar to those of beta-D-glucosidase from other bacteria, actinomycetes, and plants. Lett Appl Microbiol, 1996 Feb, 22(2), 145 - 8 A partially purified beta-glucosidase from Bifidobacterium adolescentis converts cycasin to a mutagenic compound; Choi YJ et al.; beta-Glucosidase was extracted from sonicated Bifidobacterium adolescentis Int-57 and partially purified by Sepharose CL-6B gel-filtration and DEAE-cellulose ion-exchange chromatography . The partially purified enzyme was confirmed to convert cycasin to a mutagen in the Ames and SOS chromotests . beta-Glucosidase negative strains were unable to activate cycasin mutagenically. J Clin Microbiol, 1996 Feb, 34(2), 376 - 84 Rapid characterization of periodontal bacterial isolates by using fluorogenic substrate tests; Maiden MF et al.; Eighty-nine species of subgingival bacteria, represented by 121 reference strains and 892 patient isolates, including gram-negative, gram-positive, aerobic, facultatively anaerobic, microaerophilic, and anaerobic species, were characterized with a panel of fluorogenic, 4-methylumbelliferyl-linked substrate tests . Identifications of all patient isolates were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of whole-cell proteins relative to reference strains . Characteristic profiles of positive fluorogenic reactions differentiated most of the species, including five Porphyromonas species, six pigmenting and five nonpigmenting Prevotella species, Bacteroides forsythus, three Capnocytophaga species, six Actinomyces species, four Propionibacterium species, and eight Streptococcus species . Two mannoside isomers differentiated Actinomyces israelii and Actinomyces gerencseriae . In addition to Porphyromonas gingivalis, B . forsythus, and Capnocytophaga species, Fusobacterium alocis, Actinomyces odontolyticus, Actinomyces meyeri, and Bifidobacterium dentium were all positive for so-called trypsin-like activity . Fusobacterium nucleatum, Eikenella corrodens, Actinobacillus actinomycetemcomitans, and Campylobacter species were nonreactive with the carbohydrate-based substrates tested . Fluorogenic substrate tests provided a sensitive and simple method for biochemical characterization that could presumptively identify to species level most subgingival isolates within 4 h . The method was ideal for rapidly obtaining presumptive identifications of isolates prior to confirming identifications by definitive methods, such as SDS-PAGE. Int J Food Microbiol, 1996 Feb, 29(1), 11 - 29 Differentiation of bifidobacteria by use of pulsed-field gel electrophoresis and polymerase chain reaction; Roy D et al.; Several different genomic fingerprints can be obtained from various commercially-important species of Bifidobacterium using pulsed-field gel electrophoresis (PFGE) following digestion of DNA with XbaI and SpeI . Four different genomic finger printings were discernible for reference strains of Bifidobacterium animalis, five for B . bifidum, three for B . breve, five for B . infantis and three for B . longum . Standard commercially-available industrial strains of B . animalis are identical to the reference strain ATCC 27536, previously isolated from chicken feces . There was more genomic heterogeneity among industrial strains of B . longum, in that only one gave profiles similar to the type strain of this species (ATCC 15707) . The other 14 commercially-available strains of B . longum (mainly isolated from Japanese commercial preparations) were divided into four new molecular types based on their PFGE patterns . The PFGE method indicated that only five distinct strains of B . longum and one strain of B . animalis are used in commercial preparations . Additionally, the use of polymerase chain reaction amplification of portions of 16S rDNA provides a highly specific technique to discriminate between the species B . breve, B . infantis and B . longum. Antibiot Khimioter, 1996, 41(11), 28 - 32 {The efficacy of the eubiotic bifacide when used in the combined antibacterial therapy of newborn infants with infectious-inflammatory diseases}; Kushnareva MV et al.; The efficacies of bifacid and bifidumbacterin were studied comparatively in the correction of intestinal biocenosis in 60 newborns with infectious inflammatory diseases and intestine disfunction treated with massive doses of antibacterial drugs . The study showed that the use of bifidumbacterin was accompanied by significant disturbances in the biocenosis and by development of the intestinal syndrome . The protective action of the drug was observed after a short-term use only of one antibiotic and when the course of the bifidum therapy was continued after discontinuation of the treatment with antibacterial drugs . The use of bifacid was accompanied by a rapid (by the 2nd or the 5th day fo the treatment) and stable normalization of the stools and a marked improvement of the patient general state . The clinical efficacy of bifacid was much higher than that of bifidumbacterin . At the background of the bifacid therapy there was observed correction of the intestinal microflora composition due to normalization of the count of Bifidobacterium, Lactobacillus and Colibacillus as well as to eradication of opportunistic pathogens. Ter Arkh, 1996, 68(2), 24 - 7 {Enterosorption in ulcerative lesions of the gastrointestinal tract with concomitant intestinal dysbacteriosis}; Krylov AA et al.; Adjuvant use of enterosorbents in combined therapy of gastroduodenal ulcers (158 patients), nonspecific ulcerative colitis with intestinal dysbacteriosis was assessed . Enterosorption in combined therapy of gastrointestinal diseases with dysbacteriosis potentiated positive effect . The highest benefit of enterosorption occurs in intestinal affections (nonspecific ulcerative colitis, irritable bowel-syndrome, chronic enteritis, colitis) . In gastroduodenal ulcer lignin sorbent (polyfepan) is preferable . Polyfepan acted positively on intestinal dysbacteriosis through eliminating Escherichia with hemolyzing properties and enterococci, by reducing lack of bifidobacteria. Vestn Ross Akad Med Nauk, 1996, (2), 8 - 11 {Role of anaerobic non spore-forming bacteria in maintaining human health}; Shenderov BA; The paper gives brief information on the fact that anaerobic non spore-forming bacteria take an active part in the metabolism of various substances and imbalance in their microscopic flora may be a trigger in the development of many human diseases . In the past 2 decades, Russia has been using various pharmaceuticals based on bifidobacteria, lactobacilli and their complexes, increasing colonization resistance, normalizing the pool of cholesterol, oxalic acid, free histamine, and making the liver function normally . Special emphasis is laid on the design of functional nutrition products by means of the above bacteria . It is emphasized that the industrial development of functional nutrition will determine human health in the twenty-first century. Vestn Ross Akad Med Nauk, 1996, (2), 60 - 5 {Rational approach to correction of intestinal microflora}; Korshunov VM et al.; This paper reviews the present notions of the mechanisms of probiotics' action and analyzes selective approaches to correcting the intestinal microflora, such as the use of antibiotic-resistant and highly-adhesive probiotics, treatment with autostrains of lactobacilli and bifidobacteria, and the application of fermented-milk probiotics . Methods for optimization of the intestinal microflora in the newborns by using the maternal strains of bifidobacteria and the drug Zlemik that contains highly-adhesive lactobacilli are discussed . It is shown that parameters of immunotropic activity and involvement in the bacteriocin-mediated interactions may be used to design new probiotics . In future, the application of gene engineering methods will aid in designing a new generation of probiotics with predicted biological properties. Microbiology, 1996 Jan, 142 ( Pt 1), 109 - 14 A convenient and reproducible method to genetically transform bacteria of the genus Bifidobacterium; Argnani A et al.; A protocol was developed for the introduction of foreign plasmid DNA into various Bifidobacterium strains . The method, which is applicable to all Bifidobacterium species tested so far, is based on electroporation of bacteria made competent by preincubation in electroporation buffer for several hours at 4 degrees C . Transformation of Bifidobacterium could be achieved with a plasmid vector originating from Bifidobacterium and with plasmid vectors from Corynebacterium, but not with vectors carrying replicons from Lactococcus or Lactobacillus. Int J Syst Bacteriol, 1996 Jan, 46(1), 102 - 11 16S rRNA and 16S to 23S internal transcribed spacer sequence analyses reveal inter- and intraspecific Bifidobacterium phylogeny; Leblond-Bourget N et al.; In the last few years many attempts have been made to differentiate more than 20 Bifidobacterium species . It has been recognized that identification of bifidobacterial species is problematic because of phenetic and genetic heterogeneities . In order to contribute to our understanding of Bifidobacterium taxonomy, we studied Bifidobacterium phylogeny by performing both 16S rRNA and 16S to 23S (16S-23S) internally transcribed spacer (ITS) sequence analyses . In this study, we determined 16S rRNA sequences of five Bifidobacterium strains representing four species, and compared them with the sequences available in the GenBank database, and used them to construct a distance tree and for a bootstrap analysis . Moreover, we determined the ITS sequences of 29 bifidobacterial strains representing 18 species and compared these sequences with each other . We constructed a phylogenetic tree based on these sequence data and compared this tree with the tree based on 16S rRNA sequence data . We found that the two trees were similar topologically, suggesting that the two types of molecules provided the same kind of phylogenetic information . However, while 16S rRNA sequences are a good tool to infer interspecific links, the 16S-23S rDNA spacer data allowed us to determine intraspecific relationships . Each of the strains was characterized by its own ITS sequence; hence, 16S-23S rRNA sequences are a good tool for strain identification . Moreover, a comparison of the ITS sequences allowed us to estimate that the maximum level of ITS divergence between strains belonging to the same species was 13% . Our data allowed us to confirm the validity of most of the Bifidobacterium species which we studied and to identify some classification errors . Finally, our results showed that Bifidobacterium strains have no tRNA genes in the 16S-23S spacer region. J Bacteriol, 1996 Jan, 178(1), 317 - 20 Isolation and structural analysis of polysaccharide containing galactofuranose from the cell walls of Bifidobacterium infantis; Tone-Shimokawa Y et al.; We isolated cell wall polysaccharides (PS-1 and PS-2) from Bifidobacterium infantis Reuter ATCC 15697 and found that the backbone of PS-2 is-->3)-beta-D-Galf-(1-->3)-alpha-D-Galp- (1-->in which beta-D-Galf and alpha-D-Galp are partially substituted at O-6 with beta-D-Glcp . This is the first report of the presence of this disaccharide backbone in a gram-positive bacterium; it resembles the O antigen of some bacteria. Appl Microbiol Biotechnol, 1995 Nov, 43(6), 995 - 1000 Isolation and characterization of exocellular polysaccharides produced by Bifidobacterium longum; Abbad Andaloussi S et al.; When grown anaerobically at pH values above 5.0, on ultrafiltered complex media containing excess lactose, Bifidobacterium longum formed up to 140 mg l-1 (glucose equiv.) exopolysaccharides . The highest yield was obtained when the cells were cultivated in a peptone/yeast extract medium with pH controlled by additions of NH4OH . Whatever the conditions under study, exopolysaccharides represented about 30% of the polysaccharides produced by B . longum after 48 h of culture . Crude pronase-treated exopolysaccharide preparations were adsorbed on ion-exchange chromatographic resin to yield an anionic heteropolysaccharide fraction . Two subfractions with apparent molecular masses of 1.2 MDa and 0.36 MDa respectively were subsequently recovered after gel filtration on Sepharose 4B . In both subfractions, glucose, galactose and small amounts of uronic acids and hexosamines were present in similar molar proportions, suggesting that the excreted polymers may be synthesized from the same base unit and may have a structure resulting from repeating subunits. Mol Gen Mikrobiol Virusol, 1995 Oct-Dec, (4), 27 - 9 {Approaches to classifying bifidobacteria based on DNA structural data}; Blokhina IN et al.; A brief historical review of classifications of Bifidobacteria demonstrates the difficulties in this problem, for the present-day classifications are far from being satisfactory . Based on the genotypical heterogeneity of the genus Bifidobacterium, the authors validate the possibility of raising it to the status of the family Bidobacteriaceae fam . nov . The principle of forming new genera within the frames of this hypothetical family, discussed in the paper, originates from the genomic similarity of representatives of some species of this group of microorganisms. J Dairy Sci, 1995 Oct, 78(10), 2108 - 12 A selective enumeration medium for bifidobacteria in fermented dairy products; Lim KS et al.; A selective medium, blood-glucose-liver agar containing oxgall (.2 mg/ml) and gentamicin (30 micrograms/ml), was formulated for the selective enumeration of bifidobacteria in fermented dairy products containing both lactobacilli and streptococci . Recovery rates of bifidobacteria on this selective medium were around 90% of recovery on blood-glucose-liver agar . Strains of lactobacilli and streptococci were mostly inhibited with higher dilutions on this selective medium. J Appl Bacteriol, 1995 Oct, 79(4), 475 - 9 In vitro inhibition of Helicobacter pylori NCTC 11637 by organic acids and lactic acid bacteria; Midolo PD et al.; In this study the effects of both pH and organic acids on Helicobacter pylori NCTC 11637 were tested . Lactobacillus acidophilus, Lact . casei, Lact . bulgaricus, Pediococcus pentosaceus and Bifidobacterium bifidus were assayed for their lactic acid production, pH and inhibition of H . pylori growth . A standard antimicrobial plate well diffusion assay was employed to examine inhibitory effects . Lactic, acetic and hydrochloric acids demonstrated inhibition of H . pylori growth in a concentration-dependent manner with the lactic acid demonstrating the greatest inhibition . This inhibition was due both to the pH of the solution and its concentration . Six strains of Lact . acidophilus and one strain of Lact . casei subsp . rhamnosus inhibited H . pylori growth where as Bifidobacterium bifidus, Ped . pentosaceus and Lact . bulgaricus did not . Concentrations of lactic acid produced by these strains ranged from 50 to 156 mmol l-1 and correlated with H . pylori inhibition . The role of probiotic organisms and their metabolic by-products in the eradication of H . pylori in vivo remains to be determined. J Nutr, 1995 Oct, 125(10), 2604 - 9 Dietary fructooligosaccharide, xylooligosaccharide and gum arabic have variable effects on cecal and colonic microbiota and epithelial cell proliferation in mice and rats; Howard MD et al.; Two experiments were conducted to determine if supplementing soluble fiber (fructooligosaccharide, xylooligosaccharide or gum arabic) to a semi-elemental diet would beneficially change cecal and colonic microbiota populations and enhance epithelial cell proliferation . Experiments 1 and 2 used identical dietary regimens; mice and rats were given free access to a powdered semi-elemental diet . Animals were assigned to one of the four following treatment groups: control, no supplemental dietary fiber, fructooligosaccharide, xylooligosaccharide and gum arabic . Dietary fiber was supplied via drinking water at 30 g/L . In Experiment 1 populations of Bifidobacteria and total anaerobic flora were enumerated from the contents of the cecum and colon of weanling mice . Consumption of fructooligosaccharide increased (P < 0.05) the concentrations of Bifidobacteria and the ratio of Bifidobacteria to total anaerobic flora . In Experiment 2 tissue from the cecum and distal colon of weanling rats was examined for morphological changes of the mucosa . Consumption of xylooligosaccharide increased (P < 0.05) cecal crypt depth and labeling index relative to the other three treatments . Consumption of gum arabic and the control diet increased (P < 0.01) cecal proliferation zone . Consumption of xylooligosaccharide and the control diet increased (P < 0.01) cecal cell density (number of cells in a vertical-half of the crypt) . Distal colonic crypt depth was greatest (P < 0.05) in controls and rats fed fructooligosaccharide, intermediate in those fed gum arabic, and smallest in those fed xylooligosaccharide . These results suggest that fructooligosaccharide effectively stimulates growth of Bifidobacteria and xylooligosaccharide supports a modest enhancement of cecal epithelial cell proliferation. Carbohydr Res, 1995 Sep 8, 274, 245 - 9 Structural studies on a cell wall polysaccharide from Bifidobacterium longum YIT4028; Nagaoka M et al.; The major fraction of rhamnogalactan was isolated from the cell wall of Bifidobacterium longum YIT4028 by treatment with N-acetylmuramidase . This polysaccharide was composed of rhamnose and galactose in a molar ratio of about 2:3 . The 13C NMR spectrum indicated that it contained a pentasaccharides repeating unit . This observation, and the results of Smith degradation, partial acid hydrolysis and methylation analysis led to the conclusion that its structure is {formula: see text} Antibiot Khimioter, 1995 Sep, 40(9), 20 - 5 {Ribosomal RNA degradation in gram-negative and gram-positive bacteria under the action of minimal bactericidal doses of chlorhexidine and heating}; Lishchenko NN et al.; The mechanism of the minimal bactericidal action of chlorhexidine, an antiseptic, and heating was studied . The bactericidal doses (BD99) of the above factors were determined with respect to the representatives of 5 families: Enterobacteriaceae, Pseudomonas, Vibrio, Bacillus and Bifidobacterium . Their effect on the ribosomes in the cells of the exponentially growing cultures was estimated . It was shown that these factors induced selective damage of the 30S subunits in the gram-negative bacteria at the account of a specific degradation rRNA in them while in the gram-positive bacteria there was observed no rRNA degradation under such conditions . Differences in the mechanisms of the ribosome damage in the cells of the gram-negative bacteria of the various families were detected . Since the above factors at the levels used had no effect on the bacterial ribosomes in vitro, it was suggested that the primary link in the damages was the cell membrane while the ribosome degradation was secondary, though biologically the process leading to the bacteria death was of great importance. Int Endod J, 1995 Sep, 28(5), 244 - 8 Studies into the microbial spectrum of apical periodontitis; Brauner AW et al.; This study examined the variety of obligate and facultative anaerobic bacterial species recovered from cases of acute apical periodontitis . A total of 19 root canal samples and 24 periapical granuloma samples were taken from patients suffering pain and discomfort . Bacteria were identified by applying the following techniques: culturing on various media, Gram-staining and using commercially available biochemical test strips . In addition, Prevotella intermedia and Porphyromonas endodontalis were differentiated on a molecular genetic level using species-specific oligodeoxynucleotide probes . The most frequently identified bacteria were Prevotella intermedia, Bifidobacterium spp., Streptococcus sanguis, Streptococcus milleri-group and Bacteroides spp . Obligate anaerobes occurred at a rate of 82.3%, and the average number of isolates was 6.4 per sample. Mutat Res, 1995 Sep, 331(1), 133 - 41 Antimutagenicity of components of dairy products; Abdelali H et al.; This paper reports the potential antimutagenicity of some components of dairy products . For both milk and fermented milk, Bifidobacterium sp . Bio (strain Danone 173010), casein and calcium showed a dose-dependent antimutagenic activity against benzo{a}pyrene mutagenicity in the Ames test using Salmonella typhimurium TA98 . Fatty compounds present in milk had no antimutagenic effect . At the level occurring in cultured or uninoculated milks, and even more, the bifidobacteria, casein and calcium showed less antimutagenic activity than fermented and uninoculated milks . So, the mix of all these components must contribute to the total protective effect of dairy products against induced mutagenicity, and this does not rule out the possibility of contribution of other unknown substances . The total antimutagenicity of uninoculated skim milk corresponds to the additional activity of casein and calcium . The observed antimutagenic activity of fermented skim milk remains lower than expected. Lett Appl Microbiol, 1995 Sep, 21(3), 149 - 51 Effects of three strains of bifidobacteria on cholesterol; Tahri K et al.; To determine the validity of the hypothesis of assimilation or precipitation of cholesterol by Bifidobacterium species, resting cell assays and cultures were undertaken in TPY medium containing oxgall . With resting cell assays (pH 5), cholesterol was precipitated and redissolved in phosphate buffer (pH 7) . At the end of the cultures, only part of the removed cholesterol from the culture medium was found in the phosphate buffer, while the missing cholesterol was in cell extracts . It appeared that removal of cholesterol during culturing was not solely due to its precipitation . It is concluded that growing bifidobacteria cells are able to remove cholesterol both by precipitation and assimilation. Lett Appl Microbiol, 1995 Sep, 21(3), 146 - 8 Adhesion of different bifidobacteria strains to human enterocyte-like Caco-2 cells and comparison with in vivo study; Crociani J et al.; The validity of the in vitro adhesion tests performed with cultured cell lines, was determined in this study by comparison with results obtained in vivo, in a previous study . To make this experiment the in vitro adhesion tests were performed during a long period by utilization of an appropriate medium, to determine the capacity of the adhered strain to colonize the intestinal tract . It was demonstrated that the ability of the strain to adhere and colonize the intestinal cell in vivo or the cultured intestinal cells in intro was similar. Poult Sci, 1995 Sep, 74(9), 1418 - 25 In vitro fructooligosaccharide utilization and inhibition of Salmonella spp . by selected bacteria; Oyarzabal OA et al.; In vitro experiments were conducted to determine: 1) inhibitory capacities of potential direct-fed microbial bacteria against Salmonella serotypes; and 2) the ability of Bifidobacterium bifidum, Enterococcus faecium, Lactobacillus casei, Lactococcus lactis, Pediococcus sp., and Salmonella spp . to grow in media containing fructooligosaccharides (FOS-50 or FOS pure formulation) as the only carbohydrate source . Thirteen bacteria (two strains of Bacillus coagulans, Bacillus licheniformis, Bacillus subtilis, B . bifidum, E . faecium, two strains of Lactobacillus acidophilus, L . casei, Pediococcus sp., Propionibacterium acidopropionici, P . jensenii, and Propionibacterium sp.) were tested for inhibition of six Salmonella serotypes (S . california, S . enteritidis, S . heidelberg, S . mission, S . senftenberg, and S . typhimurium) using a spot-the-lawn technique . Bifidobacterium bifidum, E . faecium, all lactobacilli, and Pediococcus sp . clearly inhibited growth of all Salmonella serotypes . In the growth experiments, E . faecium, L . lactis, and Pediococcus sp . grew in media with either FOS-50 or the pure formulation of FOS as the sole carbohydrate source . All tested Salmonella serotypes utilized FOS-50 for growth; however growth varied among the serotypes . In contrast, none of the Salmonella serotypes grew in media containing the pure formulation of FOS as the only carbohydrate source. J Infect Dis, 1995 Aug, 172(2), 403 - 9 Passive protection against rotavirus-induced diarrhea of mouse pups born to and nursed by dams fed Bifidobacterium breve YIT4064; Yasui H et al.; Mouse pups born to and nursed by dams fed Bifidobacterium breve YIT4064 and immunized orally with rotavirus were more strongly protected against rotavirus-induced diarrhea than those born to and nursed by dams immunized with rotavirus only . The level of antirotavirus IgA in milk of dams fed B . breve YIT4064 and immunized orally with rotavirus was significantly higher than that of dams immunized with rotavirus only . The antirotavirus IgA level in feces of dams immunized orally with rotavirus was also increased by oral administration of B . breve YIT4064 . These findings demonstrate that oral administration of B . breve YIT4064 enhanced antigen-specific IgA antibody in the mammary gland and in the intestine . Oral administration of B . breve YIT4064 may enhance antigen-specific IgA antibody against various pathogenic antigens taken orally and induce protection against infections in various mucosal tissues. J Appl Bacteriol, 1995 Aug, 79(2), 117 - 27 Degradation and fermentation of alpha-gluco-oligosaccharides by bacterial strains from human colon: in vitro and in vivo studies in gnotobiotic rats; Djouzi Z et al.; The ability of several human gut bacteria to break down alpha-1,2 and alpha-1,6 glycosidic linkages in alpha-gluco-oligosaccharides (GOS) was investigated in vitro in substrate utilization tests . Bacteroides thetaiotaomicron, Bifidobacterium breve and Clostridium butyricum, which are usually found in the infant gut and have been associated with both beneficial and deleterious effects on health, were studied . Alpha-Gluco-oligosaccharide degradation was compared in vitro and in vivo in gnotobiotic rats associated with these organisms, inoculated alone or in combination . Oligomer breakdown and short chain fatty acid and gas production indicated hydrolysis and fermentation of the substrate . In vitro and in vivo, Cl . butyricum was the least efficient in utilizing GOS, whereas Bact . thetaiotaomicron was the most efficient . Kinetic studies on GOS hydrolysis in pH-regulated fermenters showed that alpha-1,2 glucosidic bonds, which characterize the substrate, were more resistant than alpha-1,6 linkages . Adaptation of gnotobiotic rats to a diet containing 2% (w/w) GOS significantly increased the hydrolysis of alpha-1,2 glucosidic bonds . Combination of bacteria in trixenic rats improved GOS degradation and inhibited Cl . butyricum metabolism . This inhibition was confirmed in pH-regulated fermenters containing GOS as the principal carbon source . The association of beneficial bacteria and GOS may therefore have a potential health-promoting effect in human neonates. J Pediatr Gastroenterol Nutr, 1995 Jul, 21(1), 54 - 8 Enhancement of lysozyme trypsin-mediated decay of intestinal bifidobacteria and lactobacilli; Heine W et al.; Lysozyme-mediated lysis of Bifidobacteria and Lactobacilli was studied in in vitro tests using the agar gel plate and turbidometric Micrococcus luteus (lysodeikticus) procedure as a standard . Suspensions of the strains Bifidobacterium infantis, B . infantis liberorum, B . breve, B . longum, B . ssp, and Lactobacillus acidophilus proved to be resistant to egg white lysozyme and human milk lysozyme when incubated at 37 degrees C in concentrations of 5, 50, and 500 mg lysozyme/L, respectively, through 30 and 60 min . Heat treatment at 100 degrees C for 1 h and pretreatment with ether, acetone, ascorbic acid, and hydrogen peroxide failed to incline the bacteria to the lytic effects of lysozyme . Consecutive incubation of the lysozyme-pretreated bacteria with trypsin resulted in a significantly enhanced bacteriolysis in all strains of bacteria, with the exception of B . longum . The mode of action of lysozyme and proteolytic enzymes on Bifidobacteria and Lactobacilli offers an explanation for the release of microbial building blocks and their colonic absorption and retention in the breast-fed baby. Curr Microbiol, 1995 Jul, 31(1), 49 - 54 Characterization of fructose 6 phosphate phosphoketolases purified from Bifidobacterium species; Grill JP et al.; Fructose 6 phosphate phosphoketolases (F6PPKs) were purified from Bifidobacterium longum BB536, B . dentium ATCC 27534, B . globosum ATCC 25864, and Bifidobacterium animalis ATCC 25527 . Concerning ions (Cu++, Zn++, Ca++, Mg++, Fe++, Co++, Mn++) and common enzyme inhibitors (fructose, ammonium sulfate, iodoacetate, and parachloromercuribenzoic acid), no difference appeared between the enzymes . Cu++, parachloromercuribenzoic acid (pCMB), and mercuric acetate induced high enzymatic inhibition . The study of pCMB demonstrated a noncompetitive inhibition . Additional results showed that the sulfhydryl group was not involved in catalytic reaction . Photooxidation experiments and determination of ionizable group pKas (5.16-7.17) suggested the presence of one or more histidines necessary for the catalytic reaction and explained the inhibition observed with pCMB . In light of the noncompetitive inhibition, this group was not directly involved in substrate binding . Determination of Km demonstrated that the affinities for fructose 6 phosphate in the case of animal and human origin strains were close . In addition, the same enzymatic efficiency (Kcat/Km) was obtained for each strain . The F6PPK activity was regulated by sodium pyrophosphate, ATP, and especially by ADP. Curr Microbiol, 1995 Jul, 31(1), 23 - 7 Bifidobacteria and probiotic effects: action of Bifidobacterium species on conjugated bile salts; Grill JP et al.; The effect of six different conjugated bile salts (two trihydroxyconjugated bile salts: tauro and glycocholic acids; and four dihydroxyconjugated bile salts: tauro- and glycochenodeoxycholic, tauro- and glycodeoxycholic acids) on eight bifidobacteria strains were studied . A strong growth-inhibitory effect was observed (80% at 0.95 mM) for each bile salt and strain . This phenomenon was explained by the production of deconjugated bile salt during bifidobacteria growth . The deconjugation phenomenon was concurrent with biomass production, and deconjugated bile salts were the sole compound produced during bifidobacteria biotransformation . In resting cell experiments, differences appeared between the strains and the kind of bile salts, particularly concerning taurocholic acid . The Bifidobacterium longum strains were the most efficient among the bacteria tested. Scand J Gastroenterol, 1995 Jul, 30(7), 675 - 80 Inability of Lactobacillus casei strain GG, L . acidophilus, and Bifidobacterium bifidum to degrade intestinal mucus glycoproteins; Ruseler-van Embden JG et al.; BACKGROUND: Lactic acid bacteria have been suggested for use in the prevention of relapse of ulcerative colitis and of recurrent pouchitis . These strains may not damage the protective intestinal mucus glycoproteins . METHODS: Lactobacillus casei strain GG and strains isolated from a commercial fermented product (Lactobacillus acidophilus, Bifidobacterium bifidum, and a mesophylic lactic culture) were cultured in vitro on hog gastric mucin and human intestinal glycoproteins . Furthermore, germ-free rats were mono-associated with Lactobacillus GG and poly-associated with the other strains . Glycoproteins were isolated from rat distal ileum, cecum, and colon . Mucus degradation was established by assaying carbohydrates (hexosamines, hexoses, pentoses), proteins, and blood group antigenicity . RESULTS: All strains colonized the intestinal mucus but were not found in the deep crypts . Degradation of mucus glycoproteins was observed neither in vitro nor in vivo . CONCLUSION: The tested strains do not break down intestinal mucus glycoproteins and thus far are safe to use for therapy. J Nutr, 1995 Jun, 125(6), 1401 - 12 Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics; Gibson GR et al.; Because the human gut microbiota can play a major role in host health, there is currently some interest in the manipulation of the composition of the gut flora towards a potentially more remedial community . Attempts have been made to increase bacterial groups such as Bifidobacterium and Lactobacillus that are perceived as exerting health-promoting properties . Probiotics, defined as microbial food supplements that beneficially affect the host by improving its intestinal microbial balance, have been used to change the composition of colonic microbiota . However, such changes may be transient, and the implantation of exogenous bacteria therefore becomes limited . In contrast, prebiotics are nondigestible food ingredients that beneficially affect the host by selectively stimulating the growth and/or activity of one or a limited number of bacterial species already resident in the colon, and thus attempt to improve host health . Intake of prebiotics can significantly modulate the colonic microbiota by increasing the number of specific bacteria and thus changing the composition of the microbiota . Nondigestible oligosaccharides in general, and fructooligosaccharides in particular, are prebiotics . They have been shown to stimulate the growth of endogenous bifidobacteria, which, after a short feeding period, become predominant in human feces . Moreover, these prebiotics modulate lipid metabolism, most likely via fermentation products . By combining the rationale of pro- and prebiotics, the concept of synbiotics is proposed to characterize some colonic foods with interesting nutritional properties that make these compounds candidates for classification as health-enhancing functional food ingredients. Biosci Biotechnol Biochem, 1995 Jun, 59(6), 1150 - 1 Natural rubber serum that contains a special growth promoter for Bifidobacterium; Ishizaki A; Natural rubber serum (NRS), was found to have a remarkable growth-promoting effect on various kinds of microorganisms, in particular, anaerobes . NRS stimulated the growth of all the species of Bifidobacterium tried and synergism was noted between the effects of NRS and nutrients in media that contained yeast extract, meat extract, and the casein as nutrients. Biosci Biotechnol Biochem, 1995 Jun, 59(6), 1021 - 6 Formation of oligosaccharides from lactose by Bacillus circulans beta-galactosidase; Yanahira S et al.; Eleven oligosaccharides formed by a transglycosylation reaction during lactose hydrolysis with Bacillus circulans beta-galactosidase were purified by gel permeation chromatography, charcoal chromatography, and HPLC . From the results of methylation analysis, and MS and NMR studies, it was concluded that these oligosaccharides were beta-D-Galp-(1-->3)-D-Glc, beta-D-Galp-(1-->6)-D-Glc, beta-D-Galp-(1-->2)-D-Glc, beta-D-Galp-(1-->4)-beta-D-Galp-(1-->4)-D-Glc, beta-D-Galp-(1-->6)-{beta-D-Galp-(1-->2)}-D-Glc, beta-D-Galp-(1-->6)-{beta-D-Galp-(1-->4)}-D-Glc, beta-D-Galp-(1-->4)-beta-D-Galp-(1-->3)-D-Glc, beta-D-Galp-(1-->4)-beta-D-Galp-(1-->2)-D-Glc, beta-D-Galp-(1-->4)-{beta-D-Galp-(1-->2)}-D-Glc, beta-D-Galp-(1-->4)-beta-D-Galp-(1-->6)-D-Glc, beta-D-Galp-(1-->6){beta-D-Galp-(1-->3)}-D-Glc . The last five are newly observed oligosaccharides . The results of a use test (in vitro) by human intestinal bacteria showed that the oligosaccharides containing lactose units were predominantly used by human intestinal bifidobacteria. Mol Cell Probes, 1995 Jun, 9(3), 167 - 74 Development of a species-specific polymerase chain reaction assay for Gardnerella vaginalis; van Belkum A et al.; The nucleotide sequence of the region between the 16S and 23S rRNA genes of the facultative anaerobic bacterium Gardnerella vaginalis has been determined, together with the 5' proximal 500 nucleotides of the 23S rRNA gene . Regions suited for the development of specific, probe-confirmable polymerase chain reaction (PCR) assays were selected . PCR assays were evaluated with respect to sensitivity and specificity, the latter in comparison with a number of G . vaginalis reference strains and closely related species like Bifidobacterium spp . In an initial diagnostic study it appeared that the PCR test detected G . vaginalis in 40% of women irrespective of their clinical status . Ten out of 11 patients suffering from bacterial vaginosis as defined on the basis of clinical parameters were carrying G . vaginalis. Biosci Biotechnol Biochem, 1995 May, 59(5), 860 - 3 Effects of the lipid-saccharide complex and unsaponifiable matter from sunflowers on liver lipid metabolism and intestinal flora in rats; Fukushima M et al.; The effects of the flower lipid-saccharide complex and unsaponifiable matter (1 g/kg of diet) from the sunflower on liver lipid metabolism and intestinal flora was studied in rats given cholesterol-enriched diets . After six weeks of feeding, the microsomal cholesterol concentration in the liver had been significantly reduced with the sunflower diet . The ratio of cholesterol/phospholipid was also reduced by the sunflower diet . The 3-hydroxy-3-methylglutaryl coenzyme A reductase activity of the sunflower groups was significantly lower than that of the control group . There was no significant difference in the cholesterol 7 alpha-hydroxylase activity, although this tended to increase with dietary sunflower consumption . The number of Bacillus in the cecum flora was significantly higher in the lipid-saccharide complex group than in the other groups, while Bifidobacterium and Eubacterium in the cecum flora was significantly higher in the unsaponifiable matter group when compared to the control group . These results suggest that the lipid-saccharide complex and unsaponifiable matter in the sunflower are related to liver cholesterol synthesis and intestinal flora. Lett Appl Microbiol, 1995 May, 20(5), 328 - 30 Effect of bifidobacteria on nitrites and nitrosamines; Grill JP et al.; The effects of six different bifidobacteria strains were studied on two procarcinogens: nitrite and nitrosamines . Growth of bifidobacteria was not affected by nitrite concentrations below 50 mumol l-1 . At nitrite concentrations greater than 2000 mumol l-1, total growth inhibition was observed . Nitrite elimination by a non-enzymic mechanism was noted for six strains of bifidobacteria . Acids produced by the bacteria seem to be involved in nitrite elimination . Nitrosamines tested had no effect on growth of bifidobacteria . Only one strain (Bifidobacterium longum BB 536) was able to metabolize nitrosamines by an intracellular mechanism. Zh Mikrobiol Epidemiol Immunobiol, 1995 May-Jun, (3), 113 - 6 {The genital tract microflora in patients with a papillomavirus infection}; Bagirova MSh et al.; In 70 patients of reproductive age (20-30 years) with the papilloma virus infection of the uterine neck the microflora of vaginal contents was studied . The study revealed the specific diversity of bacteria colonizing the vagina and the uterine neck . High occurrence of Chlamydia and Gardnerella was established . The detected dysbiotic disturbances in patients with condylomatosis of the uterine neck were manifested by a decrease in the isolation rate of lactobacteria and bifidobacteria and by an increase in the isolation rate of opportunistic bacteria . The most pronounced dysbiosis in the microflora of the vagina and the uterine neck was characteristic of patients with papilloma virus infection in association with cervical intraepithelial neoplasia of the III degree. Carbohydr Res, 1995 Apr 18, 270(1), 33 - 42 Structure determination of galacto-oligosaccharides by pyridylamination and NMR spectroscopy; Kimura K et al.; Galacto-oligosaccharides formed from lactose by the action of some beta-galactosidases were subjected to gel chromatography on Bio-Gel P-2, and the resulting oligosaccharide fractions were converted into pyridylamino (PA) derivatives . Each PA-oligosaccharide fraction, which consisted of several isomers in a given size-class, was then subjected to HPLC on an ODS column . Twenty-one individual galacto-oligosaccharide components were isolated in this way . The structures of most of these compounds, namely six disaccharides, five trisaccharides, two tetrasaccharides, and a pentasaccharide, were determined by 13C-NMR spectroscopy . The results obtained will be useful for the study of the activity of various galacto-oligosaccharides on the growth of Bifidobacterium species. Mikrobiol Z, 1995 Mar-Apr, 57(2), 54 - 60 {A microflora study of the gastrointestinal tract of mink housed within and outside of the area of the Chernobyl Atomic Electric Power Station}; Sudenko VI et al.; Quantitative differences in the content of lactic acid bacteria isolated from the content of the stomach, small and large intestine have been established when studying microflora of the gastrointestinal tract of minks kept in the 30-kilometer zone of the Chernobyl NPP (experimental animals) and at the Cherkassy fur farm (control animals) . Obligate heterofermentative species of lactic acid bacteria related to Lactobacillus fermentum and L . reuteri prevailed in the stomach of experimental minks . Species composition of lactic acid bacteria isolated from the stomach of the Cherkassy minks is characterized by the availability of both obligate and facultative heterofermentative species of bacteria--L . bavaricus, L . coryniformis, L . reuteri and of obligate homofermentative bacteria--L . salivarius and L . jensenii . In limiting dilutions (10(-9)-10(-10)) of the content of small intestine of the control minks one could find bacteria of L . coryniformis species and representatives of obligate heterofermentative bacteria--L . confusus and L . fermentum that is 1-2 orders higher then in the experimental minks . Both lactic acid bacteria and bifidobacteria (the latter up to 10(+9) cells/g of the content) were isolated from the lower departments of small and large intestine of the Chernobyl and Cherkassy minks . Among the species of lactic acid bacteria isolated from the experimental animals homofermentative species (L . acidophilus, L . sharpeae) and, heterofermentative (L . confusus, L . fermentum) in the control were found.(ABSTRACT TRUNCATED AT 250 WORDS) Mikrobiologiia, 1995 Mar-Apr, 64(2), 222 - 7 {Architectonics of Bifidobacteria populations: submicroscopic aspect of cell cohesion in Bifidobacterium adolescentis and Bifidobacterium bifidum}; Novik GI et al.; Populations (cultures) of Bifidobacterium adolescents and B . bifidum, growing on artificial liquid and agar media, are presented by highly ordered mycelial structures . The topography of them depends on mutual arrangement of polymorphic cells and the way of their daughter cells separation after division . Evidences obtained by scanning electron microscopy (SEM) of total preparations and by transmission electron microscopy (TEM) of ultrathin sections correlate well . These data showed the existence of morphologically varied intercellular contacts (coherence) that ensure the stability of such microbial consortia during adaptation to ambient conditions . Intercellular contacts with the aid of different extracellular structures--microfibrillae, knob-like juts, cell wall evaginations, and capsuleform stuff(glycocalix)--are the result of genetically determined self-regulating development of microbial populations as multicellular systems. J Dairy Sci, 1995 Feb, 78(2), 268 - 76 Viability and enzymatic activity of bifidobacteria in milk; Hughes DB et al.; Bifidobacterium breve NCFB 2258, Bifidobacterium bifidum NCFB 2715, Bifidobacterium longum ATCC 15707, Bifidobacterium angulatum ATCC 27535, and Lactobacillus acidophilus N2 were evaluated for viability and beta-galactosidase and alpha-galactosidase activities under conditions of refrigerated and frozen storage in reconstituted NDM . beta- and alpha-Galactosidase activities were variable . Bifidobacterium angulatum 27535 exhibited much greater enzymatic activities than those of the other strains . All strains demonstrated culture and enzyme stability upon refrigerated storage in fermented and unfermented reconstituted NDM . Bifidobacteria were significantly less tolerant of low temperature storage than was L . acidophilus N2. J Periodontol, 1995 Feb, 66(2), 102 - 8 Detection and incidence of the tetracycline resistance determinant tet(M) in the microflora associated with adult periodontitis; Lacroix JM et al.; Subgingival plaque samples were collected from 68 patients with adult periodontitis, enumerated on Trypticase-soy blood agar plates, with and without tetracycline at 4 micrograms/ml, and incubated anaerobically for 5 days . Each different colony morphotype was enumerated, and a representative colony was subcultured for identification and examined for the tetracycline resistance gene tet(M) . Both PCR amplification and DNA hybridization, using a fragment of tet(M) from Tn1545, were used to detect tet(M) . The PCR primers (5'-GACACGCCAGGACATATGG-3' and 5'-TGCTTTCCTCTTGTTCGAG-3') were chosen to amplify a 397 bp region of tet(M) . Tetracycline-resistant bacteria represented approximately 12% of the total viable count . The percentage of tet(M)-positive bacteria in the tetracycline resistant microflora varied from < or = 0.05 to 83% (mean of 10%) . tet(M) was detected in 60% of 204 tetracycline-resistant strains subcultured and identified . The tet(M) containing strains consisted of streptococci (55%, mainly S . intermedius, S . oralis, S . sanguis, and Streptococcus SM4), Actinomyces D01 (14%), Bifidobacterium D05 (11%), and Veillonella spp . (10%) . Tetracycline-resistant strains in which tet(M) was not detected included the Prevotella and Bacteroides species (41%, mainly Bacteroides D28, P . intermedia, P . nigrescens, and P . oris) . These results suggest that tet(M) is widely spread in the adult periodontal microflora, but it appears, with the exception of S . intermedius, to be mainly associated with microorganisms not considered to be periodontopathogens . Assessment of other tetracycline-resistant genes in oral organisms is needed to fully evaluate the nature of resistance to this antibiotic in the oral flora. Nutr Cancer, 1995, 24(2), 99 - 109 The effect of dietary fermented milk products and lactic acid bacteria on the initiation and promotion stages of mammary carcinogenesis; Rice LJ et al.; The effects of spray-dried yogurt powder product (YPP), bifidobacteria, and Lactobacillus acidophilus were studied during the initiation and promotion phases of carcinogenesis using the 7,12-dimethylbenz{a}anthracene (DMBA)-induced mouse mammary carcinogenesis model . In two separate studies, Sencar mice were fed a diet consisting of 86%, 43%, or 0% YPP or 0% YPP, but with added cultures of bifidobacteria or L . acidophilus . When the animals were 55-63 days old, DMBA was administered by intragastric gavage at 1 mg/mouse and continued once a week for six weeks . During the initiation study, the test diets were fed for four weeks before and during DMBA administration . One week after the final DMBA treatment, all animals were switched to a basal diet based on the AIN-76 formulation . For the promotion study, the diets were introduced one week after the final dose of DMBA and fed for the remainder of the study . Palpable tumor development was monitored weekly throughout the studies . For the initiation study, mice fed 86%, 43%, or 0% YPP or 0% YPP supplemented with bifidobacteria or L . acidophilus had a histologically verified mammary tumor incidence of 15%, 35%, 19%, 30%, and 20%, respectively . The histologically verified tumor incidence for the promotion study was 48%, 58%, 36%, 59%, and 43% in the mice fed diets consisting of 86%, 43%, or 0% YPP or 0% YPP supplemented with bifidobacteria or L . acidophilus, respectively . The data indicate that neither the initiation nor the promotion phase of carcinogenesis is significantly affected by diets composed of 86% YPP, 43% YPP, 0% YPP, or 0% YPP supplemented with bifidobacteria or L . acidophilus. Nutr Cancer, 1995, 24(2), 121 - 32 Effect of dairy products on initiation of precursor lesions of colon cancer in rats; Abdelali H et al.; This study reports the modulating effect of some dairy products on initiation of putative preneoplasic lesions in rat colon (aberrant crypts) by 1,2-dimethylhydrazine dihydrochloride . Uninoculated skim milk, skim milk fermented with Bifidobacterium sp Bio (Danone strain 173010), and a suspension of the same lactic acid bacteria were incorporated in the animals' diet . The tested diets significantly reduced the incidence of aberrant crypts compared with the control diet by 51%, 49%, and 61%, respectively . The effects of the diets on cecal pH, hepatic UDP-glucuronyltransferase activity, and cecal microflora enzyme beta-glucuronidase were also studied . There was no significant difference in cecal pH between rats fed experimental diets and control rat . The diet supplemented with the Bifidobacterium strain suspension significantly decreased only the cecal beta-glucuronidase activity . Both enzyme activities were reduced in rats fed fermented skim milk- or uninoculated skim milk-supplemented diets compared with control animals. Res Microbiol, 1995 Jan, 146(1), 59 - 71 Selection of species-specific DNA probes which detect strain restriction polymorphism in four Bifidobacterium species; Mangin I et al.; Randomly cloned fragments (in a size range 1 to 2.5 kb) of DNA from Bifidobacterium longum ATCC 15707, B . adolescentis CIP 64.59T, B . bifidum CIP 64.65 and B . animalis ATCC 25527 were used as hybridization probes to characterize strains of these species and distinguish them from closely related Bifidobacterium species . The fragments were screened for hybridization with native DNA from 41 different Bifidobacterium strains . For each species, a fragment hybridizing specifically with DNA from strains of the same species was isolated . Each fragment was then hybridized with restriction digests in order to study the genome polymorphism . In some of the tested B . longum strains including strain ATCC 15707, the species-specific fragment L6/45 hybridized with 2 fragments instead of one as expected . Sequence of the fragment revealed the presence of an ORF which had an amino acid sequence similar to the site-specific recombinases of lambda integrase family . Moreover, Southern analysis demonstrated that at least 3 copies of this fragment are present in the chromosome of B . longum ATCC 15707 and in some other B . longum strains . The species-specific fragment A6/17 of B . adolescentis hybridized with the same restriction fragment on the eight strains of this species tested . The B . bifidum-specific fragment hybridized with different DNA restriction fragments according to the strain . The restriction fragment an1 from B . animalis ATCC 25527 hybridized with the same restriction fragment from strain B . animalis ATCC 27536 . However, these two strains could be differentiated by another restriction pattern . Thus, hybridization results highlight the genetic polymorphism which exists among Bifidobacterium strains of the same species. Biol Pharm Bull, 1995 Jan, 18(1), 148 - 53 Analysis of antitumor properties of effector cells stimulated with a cell wall preparation (WPG) of Bifidobacterium infantis; Sekine K et al.; Intestinal Bifidobacterium species are thought to be beneficial in animal and human intestines . We studied the mechanisms of Bifidobacteria in antitumor activity using a cell wall preparation (WPG) of B . infantis (Cancer Res., 45, 1300, (1985)) . WPG enhanced the in vitro antitumor activities of mouse peritoneal exudate cells elicited with proteose-peptone (P-PEC) and thioglycollate broth (TG-PEC), determined by cytostatic ({3H}thymidine uptake inhibition) and cytolytic ({3H}uridine release) assays . Tumor necrosis factor-alpha (TNF-alpha) and reactive nitrogen intermediates (RNI) play a role in such augmented cytotoxicity, because anti-TNF-alpha antibody almost completely blocked the increased cytolytic activity of P-PEC in the presence of WPG . Moreover, WPG induced RNI in the supernatant of TG-PEC in a dose-dependent manner . The mRNA expression of several cytokines (IL-1 beta, IL-6, IL-10, IFN-alpha and TNF-alpha) was induced in BALB/c mouse peritoneal cells 3 h after an intraperitoneal injection of WPG (3 h WPG-PEC) . However, this expression disappeared from 24 h WPG-PEC, except for that of IFN-alpha . IFN-gamma was not induced . Kinetic studies of the tumor neutralizing activities of the WPG-PECs by means of the in vivo Winn assay revealed that the activity emerged at 1.5 h, became maximal at 3 h and disappeared at 24h . These results indicated that Bifidobacterial WPG is a Biological Response Modifier (BRM) with characteristics similar to those of other bacterial BRMs. Int J Food Microbiol, 1994 Dec, 24(1-2), 199 - 210 Degradation of complex carbohydrates by Bifidobacterium spp; Crociani F et al.; Two hundred and ninety strains of 29 species of bifidobacteria from human and animal origin were surveyed for their ability to ferment complex carbohydrates . The substrates fermented by the largest number of species were D-galactosamine, D-glucosamine, amylose and amylopectin . Many of the species isolated from animal habitats showed reduced fermentation activity . Bifidobacterium dentium strains fermented gum guar and gum locust bean; porcine gastric mucin was fermented only by B . bifidum, B . infantis was the only species to ferment D-glucuronic acid; strains of B . longum fermented arabinogalactan and the gums arabic, ghatti and tragacanth; alpha-L-fucose was fermented by strains of B . breve, B . infantis and B . pseudocatenulatum . A key to the differentiation of Bifidobacterium species of human origin is provided. Dig Dis Sci, 1994 Nov, 39(11), 2334 - 40 Reduction of virus shedding by B . bifidum in experimentally induced MRV infection . Statistical application for ELISA; Duffy LC et al.; The protective effect of a human strain of Bifidobacterium bifidum (B . bifidum) against murine group A rotavirus (MRV) was examined in the intestines of BALB/c infected mice . In experiments designed to determine whether B . bifidum mediated MRV shedding during diarrheal disease, pregnant dams (and their expected litters) were randomly assigned to the following groups: (1) mice infected with MRV alone; (2) B . bifidum-treated + MRV-infected mice; (3) B . bifidum-treated controls; and (4) saline control animals . An enzyme-linked immunosorbent assay (ELISA) for the detection of group A rotavirus was used to measure virus protein . The sensitivity of the MRV antigen detector ELISA was determined by serially diluting the rotavirus antigen in test samples . Antigen was detected in dilution ranges of 1:256-1:4096 during the acute phase and 1:16-1:512 in the recovery phase of MRV clinical disease, in the samples tested . Treatment with B . bifidum significantly reduced shedding of MRV antigen (P < 0.009) on days 2-10 postinoculation . The reduction in shedding of virus protein corresponded well with delayed onset of acute diarrhea (P < 0.02) . Closer examination of tissue cross sections under electron microscopy revealed that the B . bifidum-ingested strain adhered to the epithelium of the small intestine . These results suggest that priming the intestine with B . bifidum is effective against experimental MRV challenge and confirmed the potential usefulness of this detector ELISA for studying the kinetics of group A rotavirus infection in animals and humans. Mikrobiol Z, 1994 Nov-Dec, 56(6), 51 - 7 {The microbiocenosis of the large intestine in animals with constant radiation exposure in the area of the Chernobyl Atomic Electric Power Station}; Zagoruiko EE et al.; State of microbiocenosis of large intestine in rats and furo subjected to constant combined irradiation has been studied in full-scale experiments in the emergency rooms of the Chernobyl NPP and in the nearest zone of its effect . The higher diversity of microflora (as compared to initial one), sharp increase of the amount of Candida genus fungi, a decrease of the number of bifidobacteria and increase of the amount of conditionally pathogenic flora were observed in rats with the dose of external irradiation of 1.43 Gr absorbed for 14 days . The expressed associative character of disbioses is distinctly traced . In furo affected by the radion exceeding the background level feeding with products contaminated by radionuclides leads to changes whose individual character corresponds to the picture of disbiosis under radiation injury. Immunopharmacol Immunotoxicol, 1994 Nov, 16(4), 589 - 609 Adjuvant activity of the cell wall of Bifidobacterium infantis for in vivo immune responses in mice; Sekine K et al.; We examined the adjuvant activity of the Bifidobacterial Cell Wall preparation (WPG) for in vivo immune responses in mice . We studied three classical immune responses, which are thought to be T-cell mediated responses, to evaluate the adjuvant activity of WPG . The delayed type hypersensitivity (DTH) responses of sheep blood red cell (SRBC)-sensitized mice were significantly augmented by WPG, although the enhancement varied with the timing, route and dosage of injection . The adjuvant activity of WPG was also confirmed by using a glutaraldehyde treated- and Concanavalin A associated- tumor vaccine (G-Con A tumor vaccine) system . BALB/c mice sensitized with G-Con A tumor vaccine and WPG improved synergistically in survival time and cure rate compared with those given G-Con A vaccine alone . Spleen cells of Meth A tumor-bearing mice induced antitumor neutralizing activity with the growth of tumor but the activity declined and disappeared at the late stage of tumor growth (over 28 days after tumor transplantation) . On the other hand, antitumor neutralizing immunity was prolonged for as long as 33 days in mice inoculated with Meth A tumor and WPG . The requirement of a T-cell subpopulation in the spleen cells of tumor plus WPG treated mice was confirmed using anti-Thy 1.2 antiserum + complement to deplete them . The adjuvant activities of the Bifidobacterial cell wall demonstrated by the in vivo immune responses predict that Bifidobacteria may play a role as an immunomodulator in human and animal intestines. Poult Sci, 1994 Nov, 73(11), 1663 - 72 Effects of dietary supplementation with lactosucrose (4G-beta-D-galactosylsucrose) on cecal flora, cecal metabolites, and performance in broiler chickens; Terada A et al.; The effects of dietary lactosucrose on cecal flora, cecal metabolites, and performance were studied in eight 20-d-old and eight 62-d-old broiler chickens fed a basal diet (control) or a diet with .15% lactosucrose added . On Day 20 of age, the frequency of occurrence of lecithinase-negative clostridia were decreased (P < .05) by lactosucrose consumption . On Day 62 of age, the numbers of bifidobacteria were increased (P < .05) by lactosucrose consumption, but the counts of lecithinase-positive clostridia, including Clostridium perfringens, bacteriodaceae, and staphylococci, total anaerobic bacteria, and the frequency of occurrence of pseudomonads were decreased (P < .05) . No detectable change was observed in counts of other organisms throughout the experimental period . Cecal concentration of ammonia (P < .01), phenol (P < .05), and cresol (P < .05) were decreased on Day 62 of lactosucrose consumption . Acetic acid and butyric acid were increased (P < .01 and P < .05, respectively) on Day 62 of lactosucrose consumption . Environmental ammonia and odor of chicken ceca were greatly reduced by lactosucrose consumption. J Dairy Res, 1994 Nov, 61(4), 545 - 52 Inhibitory effect of dairy products on the mutagenicities of chemicals and dietary mutagens; Cassand P et al.; The antimutagenic effects of uninoculated milk and milks cultured with Bifidobacterium or Lactobacillus strains towards the mutagenicity induced by two direct mutagens, 4-nitroquinoline N-oxide and 2-nitrofluorene, and three dietary indirect mutagens, aflatoxin B1, benzo(a)pyrene and quercetin, were investigated using the in vitro Salmonella typhimurium test . Each cultured milk sample and control milk had a significant antimutagenic effect, to an extent varying with the mutagen used . Uninoculated milk had a greater inhibitory effect than cultured milks towards dietary indirect mutagens. J Appl Bacteriol, 1994 Oct, 77(4), 412 - 20 Regulatory effects of bifidobacteria on the growth of other colonic bacteria; Gibson GR et al.; In the human large intestine bifidobacteria are a numerically important group of micro-organisms which are considered to exert a range of biological activities related to host health . One aspect is the inhibitory effect of these bacteria on other species, possibly excluding long term colonization by invasive pathogens . It has been suggested that the mechanism of inhibition carried out by bifidobacteria is related to the fermentative production of acids such as acetate and lactate . Experiments reported in this paper attempted to address this theory . Co-culture experiments whereby Bifidobacterium infantis was incubated with Escherichia coli and Clostridium perfringens, in a variety of fermentation systems, indicated that the bifidobacterium was able to exert an inhibitory effect not necessarily related to acid production . Further studies showed that eight species of bifidobacteria could variously excrete an anti-microbial substance with a broad spectrum of activity . Species belonging to the genera Salmonella, Listeria, Campylobacter and Shigella, as well as Vibrio cholerae, were all affected . These results show that bifidobacteria are able to exert more than one mechanism of inhibition, which may be of some importance with regard to protection against gastroenteritis. Vet Q, 1994 Oct, 16(3), 152 - 6 Establishment of a microbiologically acceptable daily intake of antimicrobial drug residues; Nouws JF et al.; A model is presented to calculate the microbiologically acceptable daily intake (ADIm) of antibiotic residues in food products . The ADIm calculation is based on MIC values for indicator bacteria Escherichia coli, Bacteroides fragilis, Bifidobacterium spp . and Eubacterium spp., established under gut-like conditions in an in vitro simulation model . The maximum residue level (MRL) for residues in food products can be derived from the ADIm . Four phases can be distinguished in this gastro-intestinal simulation model, namely: 1 . In vitro determination of the MIC for each bacterial strain by a standard method . 2 . Incorporation of the drug into food (meat, milk) followed by testing of the stability of the antibiotic under gut-like conditions . 3 . Adjustment of the 'gastric' fluid to the duodenal situation, inoculation with the test bacteria and anaerobic incubation at 37 degrees C for at least 18 h . 4 . MIC reading confirmed by counting bacteria growing on specific solidified media . In this study the method for calculation of ADIm and MRL is given for flumequine as model drug . On the basis of MIC50 values for E . coli strains, a MRL for flumequine of 1.0 microgram/g meat or 0.25 microgram/ml milk was calculated . It is suggested that, depending on the antibacterial spectrum of the antibiotic involved, the ADIm can be determined with selected indicator bacteria, incubated under simulated gastrointestinal conditions. New Microbiol, 1994 Oct, 17(4), 327 - 31 Characterization of the plasmid pVS809 from Bifidobacterium globosum; Mattarelli P et al.; A plasmid from a B . globosum strain was cut with 38 restriction enzymes and a physical map was constructed . Out of a total of 121 clones from curing experiments, plasmid was lost in 58% and 100% for acridine orange and ethidium bromide curing agent respectively . The plasmid does not exist as a chromosomal integrated form . An attempt to determine phenotypic characters encoded by the plasmid was made by electrophoretic analyses of the total proteins. J Dairy Sci, 1994 Oct, 77(10), 2854 - 64 Growth and viability of Bifidobacterium bifidum in cheddar cheese; Dinakar P et al.; Bifidobacterium bifidum (ATCC 15696) was grown in MRS broth containing cysteine.HCl at 37 degrees C, and the cells were harvested by centrifuging at 1300 x g for 15 min at 4 degrees C . Equal volumes of the cell slurry and a 2.5% solution of kappa-carrageenan were mixed and transferred by drops into a solution of .3 M KCl at 20 degrees C under an atmosphere of nitrogen . The gelled beads were separated, frozen, and lyophilized immediately . This preparation and a commercial powder preparation were added to Cheddar cheese curd at milling as two treatments . Treatments did not affect cheese composition . Soluble protein increased during ripening at 7 degrees C but without differences between treatments; SDS-PAGE patterns of proteolysis were also similar . Lactic acid content of cheeses increased during ripening, but differences between treatments were minor . Acetic acid and ethanol, common metabolites of bifidobacteria, were not detected during ripening . Bifidobacteria remained viable and increased in numbers in cheese during this 24-wk study but did not affect the flavor, flavor intensity, texture, or appearance of the cheese compared with that of the control. Zh Mikrobiol Epidemiol Immunobiol, 1994 Sep-Oct, (5), 13 - 7 {The effect of SolcoTrichovac on the vaginal microflora of patients with a papillomavirus infection associated with a cervical intraepithelial neoplasm}; Korshunov VM et al.; The microbiological study of vaginal secretions of 39 female patients of reproductive age (20-30 years) with papilloma virus infection associated with cervical intraepithelial neoplasia (CIN) was carried out . Of these patients, 28 with papilloma virus infection associated with CINI-II made up group I and II having this infection associated with CINII made up group 2 . Dysbiotic disturbances in vaginal bacterial flora, found in these patients, were manifested by a decrease in the isolation rate and number of the lacto- and bifidobacteria simultaneously with the excessive growth of opportunistic bacteria . The results of the oral administration of Solco-Trichovac are indicative of the effectiveness of this preparation, which was confirmed by the data of clinical and bacteriological studies . Together with an increase in the isolation rate of lacto- and bifidobacteria, the level of the contamination of the cervicovaginal niche with opportunistic and pathogenic bacterial strains decreased . The results thus obtained make it possible to recommend Solco-Trichovac for the complex treatment of with papilloma virus infection associated with CIN. Plasmid, 1994 Sep, 32(2), 208 - 11 Transformation of Bifidobacterium longum with pRM2, a constructed Escherichia coli-B . longum shuttle vector; Missich R et al.; An Escherichia coli-Bifidobacterium longum shuttle vector, designated pRM2, was constructed by cloning a B . longum plasmid and an enterococcal spectinomycin resistance gene into a commercial E . coli vector . The plasmid was successfully introduced into B . longum cells by electroporation and into E . coli cells by both electroporation and chemical transformation. Int J Food Microbiol, 1994 Sep, 23(1), 55 - 70 Characterization of dairy-related Bifidobacterium spp . based on their beta-galactosidase electrophoretic patterns; Roy D et al.; Numerical analysis of phenotypic characteristics based on enzymatic activity and carbohydrate fermentation allowed the discrimination of most strains of bifidobacteria of animal origin from those of human origin . Strains of bifidobacteria studied were separated into nine groups based on numerical analysis . Three groups contained most strains of animal origin, three groups comprised both strains of animal and human origin, and three groups were strictly composed of strains of human origin . The results indicate that one group of animal origin (group II) contained all reference strains of Bifidobacterium animals and 10 strains isolated from fermented milks or commercial preparations . Although numerical analysis of enzymatic activities and carbohydrate fermentation patterns allowed the differentiation of 'wild' strains of B . animalis and B . longum isolated from commercial preparations, this method failed to confirm the species . In the present study, the determination of electrophoretic patterns of beta-galactosidases resulted in the development of a new technique for the differentiation of Bifidobacterium species . Several isoenzymes of beta-galactosidase were detected among strains of bifidobacteria . Each species had a specific electrophoretic pattern . The detection of beta-galactosidase by electrophoresis is a new tool for distinguishing between dairy- and non-dairy-related bifidobacteria . Dairy-related bifidobacteria (B . bifidum, B . breve, B . infantis and B . longum) as well as B . animalis could be better differentiated from other bifidobacteria by comparison of their beta-galactosidase electrophoretic patterns, rather than by numerical analysis of their phenotypic characteristics. Biol Pharm Bull, 1994 Aug, 17(8), 1012 - 17 Anti-ulcer effects of lactic acid bacteria and their cell wall polysaccharides; Nagaoka M et al.; The anti-ulcer effects of bifidobacteria, lactobacilli and streptococci were examined using the acetic acid-induced gastric ulcer and ethanol-induced erosion models in rats . Bifidobacterium breve YIT4014 and 4043, and Bifidobacterium bifidum YIT4007 were administered orally, and anti-ulcer effects were confirmed for not only these organisms but also their polysaccharide fractions (PSFs) . The major component of these anti-ulcer polysaccharides was rhamnose . In particular, polysaccharides in which the rhamnose content exceeded 60% were more effective in healing gastric ulcers . After administration of the PSF from B . bifidum YIT4007, the levels of epidermal growth factor and basic fibroblast growth factor increased in gastric tissues . Similar results were observed for the culture supernatant of gastric epithelial cells cultured with PSF . Furthermore, the production of 6-ketoprostaglandin F1 alpha by macrophages was also enhanced by PSF . These results indicated that these bacteria and their polysaccharides induced host repair and protective systems in the gastric ulcer model. Zh Mikrobiol Epidemiol Immunobiol, 1994 Jul-Aug, (4), 22 - 4 {The intestinal microflora of patients with inflammatory diseases of the maxillofacial area}; Bevz NI et al.; The microflora of the large intestine in patients with odontogenic phlegmons of different localization were studied . 80.8% of such patients were found to have microecological disturbances, characterized by a decrease in the number of bacteria belonging to the genera Bifidobacterium, Lactobacillus, Enterococcus and Bacteroides and by an increase in the number of opportunistic microorganisms. J Mol Biol, 1994 May 13, 238(4), 615 - 25 Allosteric activation in Bacillus stearothermophilus lactate dehydrogenase investigated by an X-ray crystallographic analysis of a mutant designed to prevent tetramerization of the enzyme; Cameron AD et al.; The crystal structure of a mutant Bacillus stearothermophilus lactate dehydrogenase, into which an additional loop has been engineered in order to prevent tetramerization of the enzyme, has been solved and refined at 2.4 A . The minimal repeat unit in the crystal is a dimer and the tetramer cannot be generated by any of the crystallographic symmetry operations in P2(1) . The loop protrudes out into the solvent, stabilized by a good hydrogen bonding arrangement, and clearly sterically hinders tetramer formation . This is the first structure of B . stearothermophilus lactate dehydrogenase (bsLDH) in which the allosteric activator fructose, 1,6-bisphosphate (FBP) is not present . To investigate the mechanism of allosteric activation in this enzyme we have compared the structure with a ternary complex of B . stearothermophilus lactate dehydrogenase . Many of our observations confirm those reported from a comparison of FBP-bound ternary bsLDH complex with an FBP free LDH from another bacterial source, Bifidobacterium longum . Our results suggest that quaternary structural alterations may have less influence on the mechanism than previously reported . The differences in the quaternary structural behaviour of these two enzymes is discussed. Gut, 1994 May, 35(5), 658 - 64 Pouchitis: result of microbial imbalance? Ruseler-van Embden JG, Schouten WR, van Lieshout LM. To elucidate the role of microbiological factors in pouchitis, this study investigated the composition of ileal reservoir microflora, the mucus degrading capacity of bacterial enzymes as well as the pH and the proteolytic activity of pouch effluent . Stool samples were collected from five patients with pouchitis and nine patients without pouchitis . The flora of patients with pouchitis had an increased number of aerobes, a decreased ratio anaerobes to aerobes, less bifidobacteria and anaerobic lactobacilli, more Clostridium perfringens, and several species that were not found in control patients (for example, fungi) . Furthermore the pH was significantly higher in patients with pouchitis (median value 6.5) than in control patients (5.4) . To find out if the pH might influence the breakdown of intestinal mucus glycoproteins, the activity of glycosidases and proteases, and the degradation of hog gastric mucin by the pouch flora was tested at pH 5.2-7.6 . Some glycosidases were inhibited, others were stimulated by a low pH, however, in each sample the proteolytic activity was inhibited for 75% at pH 5.2 compared with pH 6.8 and 7.6 . Degradation of hog gastric mucin by the pouch flora was an active process at pH 7.2: within two to four hours of incubation more than half of the mucin was degraded . At pH 5.2 it took twice as long . It is concluded that pouchitis possibly results from instability of the flora in the pouch, which causes homeostasis to disappear (dysbiosis), and the protection of the pouch epithelium by the mucus layer becomes affected by increased activity of bacterial and host derived enzymes. Mikrobiologiia, 1994 May-Jun, 63(3), 515 - 22 {Cellular ultrastructure of various species of the genus Bifidobacterium}; Novik GI et al.; Morphologic heterogeneity of cells from developing populations of bifidobacteria correlates with ultrastructure peculiarities . Active proliferating cells in exponential phase are characterized by formation of intracytoplasmatic membrane complex represented by lamellar, myelinoform, vesicular structures . Nucleoid is localized as the central polybranched or disperse osmophobic zone . Nucleoid distribution is determined by morphogenesis processes--exobudding, branching or multiseptation . Electronograms reveal multiple polyphosphate and polysaccharide inclusions . Ageing of bifidobacterial populations is accompanied with ultrastructural changes: cell wall hypersynthesis, reorganization and increased size of intracytoplasmatic membrane complex, altered morphology and compactness of nuclei, formation and dissimilation of inclusions. Zh Mikrobiol Epidemiol Immunobiol, 1994 May-Jun, (3), 21 - 3 {The exonuclease activity of strictly anaerobic bacteria isolated from patients and healthy persons}; Polikarpov NA et al.; The comparative study of the frequency of DNAase and RNAase and the activity of their formation in 59 strains of anaerobic opportunistic bacteria isolated from traumatic and orthopedic patients and in 154 strains of bifidobacteria isolated from feces of healthy persons has been made . The study has revealed that cultures with exonuclear activity occur among opportunistic bacteria and among bifidobacteria, the representatives of human obligate anaerobic automicroflora . However, in opportunistic bacteria the frequency of the above-mentioned enzymes and the activity of their formation have proved to be significantly higher than in bifidobacteria. Biol Pharm Bull, 1994 May, 17(5), 596 - 602 Purification and characterization of beta-fructofuranosidase from Bifidobacterium infantis; Imamura L et al.; beta-Fructofuranosidase activities of eight strains of Bifidobacteria, intestinal bacteria, were assayed and Bifidobacterium infantis was selected for purification of the enzyme . beta-Fructofuranosidase activity was recovered in the supernatant fraction after disruption of B . infantis cells with sonication and was purified to homogeneity by ammonium sulfate fractionation, and DEAE-cellulose, butyl-Toyopearl and Sephacryl S-300 column chromatographies . The enzyme (molecular weight (M.W.) 232000) was composed of three identical subunits (M.W . 75000) whose NH2-terminal amino acids were threonine . The enzyme was stable at pH 6-8, having the optimum activity at pH 6.0-6.2 . The enzyme activity was stable under 40 degrees C and the optimal temperature was 55 degrees C . This enzyme catalyzed the hydrolysis of sucrose, 1-kestose, nystose, inulin and raffinose at the relative velocities of 100, 297, 365, 140 and 3.8, respectively, but did not catalyze the hydrolysis of maltose or cellobiose . These results indicated that this fructooligosaccharide hydrolyzing enzyme is a novel type of beta-fructofuranosidase. Appl Environ Microbiol, 1994 May, 60(5), 1451 - 8 Identification of Bifidobacterium strains by rRNA gene restriction patterns; Mangin I et al.; Total DNA from 21 collection or industrial Bifidobacterium strains was cleaved with various restriction endonucleases . Following electrophoresis, the fragments were subjected to Southern blot hybridization with a heterologous {alpha-32P}dCTP-labeled rDNA (genes coding for rRNA) 23S gene probe . The ribosomal patterns allowed all tested strains to be differentiated and previous classifications to be confirmed . The same method was used to characterize DNA from 121 Bifidobacterium isolates collected from the intestinal flora of five human volunteers after the induction of colonic bacterial imbalance by antibiotics and absorption of a resistant exogenous Bifidobacterium strain . Hybridizations with the ribosomal probe revealed 11 different ribosomal patterns in addition to that of the exogenous strain . They permitted the Bifidobacterium populations belonging to the dominant colonic flora to be monitored over time . This experiment revealed significant and sustained alterations of the endogenous intestinal flora; indeed, some strains were eliminated, while others, probably belonging to subdominant flora, replaced them . Furthermore, even 2 months after the end of antibiotic treatment, the colonic flora remained different from that observed before treatment . Finally, our results showed that antibiotherapy did not allow colonic colonization by the exogenous strain. Lipids, 1994 Apr, 29(4), 289 - 96 Delta 22-beta-muricholic acid in monoassociated rats and conventional rats; Kayahara T et al.; Bile acids were analyzed in the bile, small and large intestines, and feces of germ-free rats after a single inoculation with one of six intestinal bacteria that had been originally isolated from human feces . Bacteroides vulgatus and Bifidobacterium longum preferentially deconjugated tauro-beta-muricholic acid and taurocholic acid, respectively . Clostridium ramosum, Peptostreptococcus productus and Lactobacillus gasseri deconjugated both bile acids, but Escherichia coli did not deconjugate either one . Rats inoculated with bacteria that deconjugated tauro-beta-muricholic acid produced delta 22-beta-muricholic acid in the feces . In contrast, delta 22-cholic acid could not be detected in rats inoculated with bacteria that deconjugated taurocholic acid. New Microbiol, 1994 Apr, 17(2), 159 - 62 Murein types in Bifidobacterium ruminantium, Bifidobacterium merycicum and Bifidobacterium saeculare; Weiss N et al.; The murein types of strains belonging to Bifidobacterium ruminantium, Bifidobacterium merycicum and Bifidobacterium saeculare were determined . B . ruminantium was found to posses a different type of murein from B . adolescentis . B . merycicum and B . saeculare have the same type of murein that is shared by many other species of the genus. J Nutr Sci Vitaminol (Tokyo), 1994 Apr, 40(2), 181 - 8 Biotin production by bifidobacteria; Noda H et al.; Biotin production and the growth of the strains of Bifidobacterium adolescentis, B . bifidum, B . breve, B . infantis, and B . longum were studied . These five strains showed heavy growth on BL medium . But when yeast extract medium (carbon source, glucose) was used, the extent of their growth was significantly decreased, one-half or less than that of the growth on BL medium . B . bifidum grew well on yeast extract medium containing oligosaccharides, such as isomaltooligosaccharide, and produced biotin extracellularly . The utilization of oligosaccharides in biotin production by these five strains was investigated. Biosci Biotechnol Biochem, 1994 Apr, 58(4), 691 - 4 Purification and characterization of D-xylose isomerase from Bifidobacterium adolescentis; Kawai Y et al.; D-Xylose isomerase was purified to homogeneity from cell-free extracts of Bifidobacterium adolescentis by ammonium sulfate fractionation and chromatographies on DEAE-cellulose and Butyl-Toyopearl . The molecular weight of the purified enzyme was estimated to be 168,000 by gel filtration on TSKgel G-3000SW, and 53,000 on SDS-polyacrylamide gel electrophoresis . The optimum pH was around 7 and the enzyme was stable at pH 7-8 . The enzyme required bivalent cations, Mg2+, Co2+, or Mn2+ for the activity, particularly Mn2+ to be best . The enzyme had a pI of 4.3, and the Km for D-xylose was 4 mM . The N-terminal amino acid sequence of the enzyme was not similar to those of D-xylose isomerases from other sources such as Clostridium thermosulfurogenes, Escherichia coli, or Bacillus subtilis. Appl Environ Microbiol, 1994 Mar, 60(3), 1041 - 3 Isolation of a human intestinal anaerobe, Bifidobacterium sp . strain SEN, capable of hydrolyzing sennosides to sennidins; Akao T et al.; A strictly anaerobic bacterium capable of metabolizing sennosides was isolated from human feces and identified as Bifidobacterium sp., named strain SEN . The bacterium hydrolyzed sennosides A and B to sennidins A and B via sennidin A and B 8-monoglucosides, respectively . Among nine species of Bifidobacterium having beta-glucosidase activity, only Bifidobacterium dentium and B . adolescentis metabolized sennoside B to sennidin B, suggesting that the sennoside-metabolizing bacteria produce a novel type of beta-glucosidase capable of hydrolyzing sennosides to sennidins. Nat Struct Biol, 1994 Mar, 1(3), 176 - 85 T and R states in the crystals of bacterial L-lactate dehydrogenase reveal the mechanism for allosteric control; Iwata S et al.; The crystal structure of L-lactate dehydrogenase from Bifidobacterium longum, determined to 2.5 A resolution, contains a regular 1:1 complex of T- and R-state tetramers . A comparison of these two structures within the same crystal lattice and kinetical characterization of the T-R transition in solution provide an explanation for the molecular mechanism of allosteric activation . Substrate affinity is controlled by helix sliding between subunits which is triggered by the binding of the activator, fructose 1,6-bisphosphate . The proposed mechanism can explain activation by chemical modification and mutagenesis, as well as suggesting why vertebrate counterparts are not allosteric. Clin Diagn Lab Immunol, 1994 Mar, 1(2), 244 - 6 Augmentation of anti-influenza virus hemagglutinin antibody production by Peyer's patch cells with Bifidobacterium breve YIT4064; Yasui H et al.; Bifidobacterium breve YIT4064 was tested an an adjuvant for oral influenza vaccine by the murine Peyer's patch cell culture method . The organism augmented production of anti-influenza virus hemagglutinin immunoglobulin A antibody by Peyer's patch cells in response to addition of hemagglutinin . These antibodies may be disseminated to the respiratory mucosal tissue and prevent influenza virus infection. J Mol Biol, 1994 Feb 25, 236(3), 958 - 9 A regular 1:1 complex of two allosteric states in the single crystal of L-lactate dehydrogenase from Bifidobacterium longum; Iwata S et al.; Single crystals of the regular 1:1 complex of T and R-state tetramers of L-lactate dehydrogenase from