Carbohydr Res, 2000 Aug 7, 327(4), 377 - 83 The efficient enzymatic synthesis of N-acetyllactosamine in an organic co-solvent; Yoon JH et al.; In the presence of beta-galactosidase from Bifidobacterium bifidum, N-acetyllactosamine was synthesized in significantly enhanced yield compared with earlier routes . Different proportions of the (1-->4)- and (1-->6)-linked forms were obtained depending on the choice of enzyme and reaction conditions, viz . the nature of added organic co-solvent (20-80% of 2-ethoxy ethyl ether, trimethyl phosphate, or acetone) . The beta-(1-->4)-linked disaccharide was the major product and the beta-(1-->6)-linked disaccharide was the minor product . With beta-galactosidases from P . multicolor, A . oryzae, B . longum the beta-(1-->6) linkage was exclusively synthesized . Procedures for optimising the yield of N-acetyllactosamine are discussed . An immobilized enzyme on a nylon powder column was used for the efficient recycling of enzyme and synthesizing the disaccharide. Lett Appl Microbiol, 2000 Aug, 31(2), 154 - 6 Effects of Lactobacillus amylovorus and Bifidobacterium breve on cholesterol; Grill JP et al.; To determine the validity of the hypothesis of assimilation and/or precipitation of cholesterol by Lactobacillus and Bifidobacterium species, culture were undertaken in TPY medium containing oxgall or taurocholic acid . In the case of growing cells, both strains were able to remove cholesterol in the presence of bile salts . Nevertheless, the behaviour was different according to the kind of bile salt . In the presence of taurocholic acid, the removal of cholesterol was due to both bacterial uptake and precipitation . In the presence of Oxgall, bacterial uptake and precipitation were observed for Lactobacillus but only precipitation occurred for Bifidobacterium. J Food Prot, 2000 Aug, 63(8), 1133 - 6 Binding of aflatoxin B1 to bifidobacteria in vitro; Oatley JT et al.; Aflatoxins are mycotoxins that cause health and economic problems when they contaminate food and feed . One potential method for reducing human health effects due to aflatoxin ingestion is to block uptake via binding by bacteria that either make up the normal gut flora or are present in fermented foods in our diet . These bacteria would bind aflatoxin and make it unavailable for absorption in the intestinal tract . Bifidobacteria comprise a large fraction of the normal gut flora, are thought to provide many probiotic effects and are increasingly used in fermented dairy products . These qualities targeted bifidobacteria for studies to determine if various strains of heat-killed bifidobacteria can bind aflatoxin B1 (AFB1) in vitro . The AFB1 binding affinities of various strains of bifidobacteria, Staphylococcus aureus, and Escherichia coli were quantitated utilizing enzyme-linked immunosorbent and {3H}AFB1 binding assays . The bacteria analyzed were found to bind significant quantities of AFB1 ranging from 25% to nearly 60% of the added toxin . The data also suggest that there are reproducible strain differences in AFB1 binding capacity. Drugs Exp Clin Res, 2000, 26(3), 95 - 111 Bifidobacteria and lactobacilli in human health; Orrhage K et al.; The gastrointestinal microflora is a complex ecological system, normally characterized by a flexible equilibrium . The most important role of the microflora, from the point of view of the host, is probably to act in colonization resistance against exogenous, potentially pathogenic, microorganisms . Bifidobacteria and lactobacilli are Gram-positive lactic acid-producing bacteria constituting a major part of the intestinal microflora in humans and other mammals . Administration of antimicrobial agents may cause disturbances in the ecological balance of the gastrointestinal microflora with several unwanted effects such as colonization by potential pathogens . To maintain or reestablish the balance in the flora, supplements of intestinal microorganisms, mainly bifidobacteria and lactobacilli, sometimes called probiotics, have been successfully used . This article reviews the role of bifidobacteria and lactobacilli in human health. Rev Latinoam Microbiol, 1998 Jul-Dec, 40(3-4), 166 - 72 A review on the use of microorganisms as probiotics; Gomez-Gil B et al.; A literature review on the use of live microorganisms as probiotics is presented . Topics discussed are the definition of probiotic; the normal microflora of the digestive system of mammals, including bacterial interactions in the gut, colonization, modification of metabolic processes, and immunostimulation . Probiotics studies in humans and in farmed animals, with special emphasis on the use of Lactobacillus spp . and Bifidobacterium spp . are also discussed. Zh Mikrobiol Epidemiol Immunobiol, 2000 May-Jun, (3), 20 - 4 {The action of a preparation of Escherichia coli M-17 growth autostimulants (Actoflor) on the growth of pure and mixed bacterial cultures}; Vakhitov TIa et al.; The effect of the preparation of E . coli M-17 low-molecular exometabolites (Actoflor), containing growth autostimulators, on the growth of pure cultures of E . coli M-17 E . coli K-12, Salmonella enteritidis, Serratia marcescens and Bifidobacterium adolescentis MC-42 was studied . This preparation was shown to stimulate the growth of all above-mentioned bacteria . The addition of Actoflor also led to the acceleration of growth in the cultivation of mixed cultures of E . coli M-17 with E . coli K-12 (or S . enteritidis), the producer strain (E . coli M-17) showing the highest degree of acceleration . Moreover, the action of Actoflor led to the elimination of competitor strains and to the increase of the antagonistic activity of E . coli M-17 . Actoflor may be supposedly used as a therapeutic or prophylactic remedy. DNA Seq, 2000, 11(1-2), 101 - 7 Isolation and sequence analysis of repA from the incurable 90 MDA plasmid of Azospirillum brasilense; Vande Broek A et al.; An open reading frame encoding a protein of 312 amino acids was identified in an essential region of the 90 MDa plasmid of Azospirillum brasilense . The putative protein shows similarity with RepA proteins of plasmids of Sphingomonas aromacitovorans, Rhodotermus marinus, Chlorobium limicola and Bifidobacterium asteroides and with the RepA protein of the broad host range plasmid pSa . A putative helix-turn-helix motif was identified in the central part of the A . brasilense RepA protein . Analysis of the upstream and downstream region of the A . brasilense repA gene revealed the presence of eight 15-bp repeats, which can possibly function as iterons. J Agric Food Chem, 2000 Jul, 48(7), 2744 - 8 Cordycepin: selective growth inhibitor derived from liquid culture of Cordyceps militaris against Clostridium spp; Ahn YJ et al.; The growth responses of nine human intestinal bacteria to liquid culture of Cordyceps militaris Link . Pt . (Ascomycotina: Clavicipitaceae) collected from a pupa of Bombyx mori L . (Lepidoptera: Bombycidae) were examined using spectrophotometric and impregnated paper disk methods and compared to those of tetracycline and chloramphenicol, as well as those of Coptis japonica root-derived berberine chloride . The biologically active constituent of the cultures was characterized as cordycepin (3'-deoxyadenosine) by spectroscopic analysis . This compound revealed potent growth-inhibiting activity toward Clostridium paraputrificum and Clostridium perfringens at 10 microgram/disk without adverse effects on the growth of Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium adolescentis, Lactobacillus acidophilus, and Lactobacillus casei, whereas tetracycline and chloramphenicol inhibited the growth of these lactic acid-producing bacteria, clostridia and Escherichia coli . However, C . militaris-derived materials revealed no growth stimulation on the bifidobacteria and lactobacilli . These results may be an indication of at least one of the pharmacological actions of C . militaris . As a naturally occurring antibacterial agent, cordycepin could be useful as a new preventive agent against various diseases caused by clostridia. Lett Appl Microbiol, 2000 Jul, 31(1), 57 - 62 Antimicrobial susceptibility of bifidobacteria; Yazid AM et al.; Eighteen Bifidobacterium strains were tested for their susceptibility to a range of antimicrobial agents . All the strains tested, including the reference culture Lactobacillus acidophilus CH2, were susceptible to several groups of antimicrobial agents, they were cephalosporin (cefamandole, cefazolin, cefaperazone, cefoxitin), polypeptide (bacitracin), macrolide (erythromycin), penicillin (amoxicillin), phenicol (chloramphenicol) and beta-lactam (imipenem) . Fourteen strains were resistant to more than 10 antibiotics . The reference culture was resistant to only three antibiotics . The results showed that bifidobacteria are resistant to a wide range of antimicrobial agents. J Nutr Sci Vitaminol (Tokyo), 2000 Apr, 46(2), 105 - 8 Effect of soy milk and bifidobacterium-fermented soy milk on plasma and liver lipids in ovariectomized Syrian hamsters; Kikuchi-Hayakawa H et al.; The effects of soy milk and fermented soy milk on lipid metabolism were studied in ovariectomized Syrian hamsters . Five mo-old Syrian hamsters were randomly assigned to four treatment groups: ovariectomized (OVX)+control diet (OVX-C); OVX+soy milk diet (OVX-SM); OVX+fermented soy milk diet (OVX-FSM); and sham-operated+control diet (Sham-C) . The hamsters were fed on these diets for 4 wk . The atherogenic index value of the OVX-FSM group was lower than that of the OVX-C group . The plasma triglyceride level of the OVX-FSM group was significantly lower than that of the OVX-C group . The liver total cholesterol contents in the OVX-SM and OVX-FSM groups were significantly lower than that in the OVX-C group . Thus, these results demonstrate that bifidobacterium-fermented soy milk had a hypolipidemic effect in ovariectomized hamsters. Br J Nutr, 2000 Mar, 83(3), 247 - 55 In vitro fermentability of dextran, oligodextran and maltodextrin by human gut bacteria; Olano-Martin E et al.; Anaerobic batch culture fermenters were used for a preliminary screening of the in vitro utilization by human gut microflora of dextran and novel oligodextrans (I, II and III) produced in the University of Reading (UK) . Glucose and fructooligosaccharides (FOS) were used as reference carbohydrates . As expected, FOS acted as a good prebiotic in that it selectively increased numbers of bifidobacteria in the early stages of the fermentation . Dextran and oligodextrans each resulted in an enrichment of bifidobacteria in the batch cultures, with high levels of persistence up to 48 h . They also produced elevated levels of butyrate ranging from 5 to 14.85 mmol/l . To more effectively simulate conditions that prevail in different regions of the large intestine, a three-stage continuous culture cascade system was used to study further the fermentation of dextran, a low-molecular-mass oligodextran (IV) and maltodextrin . Oligodextran IV was shown to be the best substrate for bifidobacteria and lactobacilli with steady-state populations of bifidobacteria and lactobacilli being higher in all three vessels of the gut model than the respective populations resulting from dextran and maltodextrin . A maximum difference of 1.9 log was observed in vessel 1 for both bifidobacteria and lactobacilli in the case of dextran fermentation, with 1.4 log and 0.8 log in vessel 3 were the maximum differences for bifidobacteria and lactobacilli when maltodextrin was used as the carbohydrate source . Moreover, dextran and oligodextran appeared to stimulate butyrate production, with a maximum production up to 25.39 mmol/l in vessel 3 when fermenting dextran, followed by 21.70 mmol/l in the case of oligodextran IV and only 12.64 mmol/l in the case of maltodextrin. Zh Mikrobiol Epidemiol Immunobiol, 2000 Jan-Feb, (1), 88 - 90 {The use of the biotherapeutic agent Zhlemik for correcting the microflora in bacterial vaginosis}; Sarkisov SE et al.; The vaginal microflora in 11 women suffering with bacterial vaginosis was corrected by BTA "Zlemik" contained highly adhesive strain Lactobacillus . BTA was administered intravaginal in dose 1 suppositoria at night during 15 days . It was established in majority cases that clinical symptoms and discharges have disappeared . The value of pH has decreased from 5.6-6.2 to 4.6-4.8 . After biotherapeutic therapy significantly increased of the levels of Lactobacillus and percentage of the presence Bifidobacterium . The number of non-spore-forming obligatory anaerobic bacteria, Streptococcus and Gardnerella decreased in 10 and 100 times respectively. Int J Food Microbiol, 2000 May 25, 56(1), 87 - 96 Safety assessment of potential probiotic lactic acid bacterial strains Lactobacillus rhamnosus HN001, Lb . acidophilus HN017, and Bifidobacterium lactis HN019 in BALB/c mice; Zhou JS et al.; The general safety of immune-enhancing lactic acid bacteria (LAB) strains Lactobacillus rhamnosus HN001 (DR20), Lb . acidophilus HN017, and Bifidobacterium lactis HN019 (DR10) was investigated in a feeding trial . Groups of BALB/c mice were orally administered test LAB strains or the commercial reference strain Lb . acidophilus LA-1 at 2.5 x 10(9), 5 x 10(10) or 2.5 x 10(12) colony forming units (CFU)/kg body weight/day for 4 weeks . Throughout this time, their feed intake, water intake, and live body weight were monitored . At the end of the 4 week observation period, samples of blood, liver, spleen, kidney, mesenteric lymph nodes, and gut tissues (ileum, caecum, and colon) were collected to determine: haematological parameters (red blood cell and platelet counts, haemoglobin concentration, mean corpuscular volume, mean corpuscular haemoglobin, and mean corpuscular haemoglobin concentration); differential leukocyte counts; blood biochemistry (plasma total protein, albumin, cholesterol, and glucose); mucosal histology (epithelial cell height, mucosal thickness, and villus height); and bacterial translocation to extra-gut tissues (blood, liver, spleen, kidney and mesenteric lymph nodes) . DNA finger printing techniques were used to identify any viable bacterial strains recovered from these tissues . The results demonstrated that 4 weeks consumption of these LAB strains had no adverse effects on animals' general health status, haematology, blood biochemistry, gut mucosal histology parameters, or the incidence of bacterial translocation . A few viable LAB cells were recovered from the tissues of animals in both control and test groups, but DNA fingerprinting did not identify any of these as the inoculated strains . The results obtained in this study suggest that the potentially probiotic LAB strains HN001, HN017, and HN019 are non-toxic for mice and are therefore likely to be safe for human use. Int J Food Microbiol, 2000 Jun 1, 56(2-3), 113 - 21 Growth of bifidobacteria in soymilk and their survival in the fermented soymilk drink during storage; Chou CC et al.; Growth of Bifidobacterium infantis CCRC 14633 and B . longum B6, in soymilk was investigated in the present study . It was found that soymilk could support the growth of both organisms tested . B . infantis grew better than B . longum in soymilk . Supplementation of bifitose (isomaltooligosccharie), glucose, lactose or galactose to soymilk increased the growth of B . infantis and B . longum as determined after 48 h of fermentation . On the other hand, addition of yeast extract, peptone, tryptone, casitone or N-Z-Case plus to soymilk enabled B . infantis to reach its maximum population in a shorter cultivation time of 24 h . Acid production by B . longum and B . infantis in soymilk was mainly non-growth associated, while in the yeast extract-supplemented soymilk, acid produced by B . infantis was found to be growth-associated . Populations of B . longum reduced more than did B . infantis in the prepared fermented soymilk drink during storage period . Viable population of both test organisms reduced less in the fermented drink held at 5 degrees C than at 25 degrees C . After a 10-day storage at 5 degrees C, viable B . infantis and B . longum reduced by 0.44 and 3.18 log CFU/ml, respectively, in the fermented drink . Addition of sucrose to the fermented drink resulted in an increased reduction of viable bifidobacteria during the storage period . This phenomenon was most prominent with B . infantis in the fermented drink held at 25 degrees C. Zh Mikrobiol Epidemiol Immunobiol, 1999 Jul-Aug, (4), 7 - 10 {The subcellular mechanisms of the action of disinfectants . The effect of iodine and chloramine-B on bacterial ribosomes}; Shandala MG et al.; The composition of ribosomal material in bacterial cells of 5 families, subjected to the minimal bactericidal action of iodine and chloramine-B, was studied with the use of electrophoresis in polyacrylamide gel . The study demonstrated that under these conditions protein-synthesizing organelles of vibrios and bifidobacteria did not undergo perceptible changes, while Escherichia, Pseudomonas and bacilli essential ribosomal degradation was found to occur, which was manifested by a decrease in the intensity of bands 23S and 16S of ribosomal RNA . The conclusion was made that the in vivo destruction of ribosomes was not the direct result of the action of disinfectants, but mediated by some cellular mechanisms. Zh Mikrobiol Epidemiol Immunobiol, 1999 Sep-Oct, (5), 56 - 61 {Bifidogenic factors as drug preparations}; Murashova AO et al.; The review of new data on the study of bifidobacterial factors of different origin and the probable mechanisms of their favorable action on the microflora of the intestinal tract if presented . The main emphasis is made on the analysis of data on the use of oligosaccharides, including fructo-oligosaccharides, as compounds stimulating the growth and development of bifidobacteria both in pure cultures and in intestinal microflora . Methods for the treatment of natural compounds with a view to enhancing their bifidogenic effect are presented . The possibilities and/or advantages of using bifidogenic factors in vivo and in vitro as medicinal preparations either alone or incorporated in probiotic compositions are evaluated . Suggestion has been made that the choice of the method for using bifidogenic factors may depend on the kind and severity of disturbances in indigenous microflora. J Appl Microbiol, 2000 Jun, 88(6), 968 - 74 Effects of fructooligosaccharides and their monomeric components on bile salt resistance in three species of bifidobacteria; Perrin S et al.; The influence of fructooligosaccharides (FOS) and their monomeric components on bile salt resistance of Bifidobacterium breve ATCC 15700, Bif . longum ATCC 15707 and Bif . animalis ATCC 25527 was examined . The neosugars induced fructofuranosidase activities for the degradation of these saccharides . For the three strains tested the growth was identical and bile salts had the same inhibitory effect on growth whatever the carbohydrate used . The survival of Bif . breve and Bif . longum, in the presence of glycodeoxycholic acid depended, however, on carbohydrates: the toxic effects of the bile salt could be partly alleviated by the addition of a metabolizable C-source . For Bif . animalis, the presence of any carbohydrate in the incubation medium did not enhance the viability of the strain . But in the three deconjugating strains of bifidobacteria studied, the presence of neosugar during the growth led to improved resistance to the bactericidal effect of the bile salt compared with the monomeric components of these neosugars (glucose and fructose). Microbiol Immunol, 2000, 44(4), 213 - 22 Dietary Bifidobacterium lactis (HN019) enhances resistance to oral Salmonella typhimurium infection in mice; Shu Q et al.; The ability of a newly identified probiotic lactic acid bacterial strain, Bifidobacterium lactis (HN019), to confer protection against Salmonella typhimurium was investigated in BALB/c mice . Feeding mice with B . lactis conferred a significant degree of protection against single or multiple oral challenge with virulent S . typhimurium, in comparison to control mice that did not receive B . lactis . Protection included a ten-fold increase in survival rate, significantly higher post-challenge food intake and weight gain, and reduced pathogen translocation to visceral tissues (spleen and liver) . Furthermore, the degree of pathogen translocation showed a significant inverse correlation with splenic lymphocyte proliferative responses to mitogens, blood and peritoneal cell phagocytic activity and intestinal mucosal anti-S . typhimurium antibody titers in infected mice; all of these immune parameters were enhanced in mice fed B . lactis . Together, these results suggest that dietary B . lactis can provide a significant degree of protection against Salmonella infection by enhancing various parameters of immune function that are relevant to the immunological control of salmonellosis . Thus dietary supplementation with B . lactis provides a unique opportunity for developing immune-enhancing probiotic dairy food products with proven health benefits. Appl Environ Microbiol, 2000 Jun, 66(6), 2682 - 4 Fermentation of fructooligosaccharides by lactic acid bacteria and bifidobacteria; Kaplan H et al.; Lactic acid bacteria and bifidobacteria were screened of their ability to ferment fructooligosaccharides (FOS) on MRS agar . Of 28 strains of lactic acid bacteria and bifidobacteria examined, 12 of 16 Lactobacillus strains and 7 of 8 Bifidobacterium strains fermented FOS . Only strains that gave a positive reaction by the agar method reached high cell densities in broth containing FOS. Appl Environ Microbiol, 2000 Jun, 66(6), 2502 - 12 Bile salt hydrolase of Bifidobacterium longum-biochemical and genetic characterization; Tanaka H et al.; A bile salt hydrolase (BSH) was isolated from Bifidobacterium longum SBT2928, purified, and characterized . Furthermore, we describe for the first time cloning and analysis of the gene encoding BSH (bsh) in a member of the genus Bifidobacterium . The enzyme has a native molecular weight of 125,000 to 130,000 and a subunit molecular weight of 35,024, as determined from the deduced amino acid sequence, indicating that the enzyme is a tetramer . The pH optimum of B . longum BSH is between 5 and 7, and the temperature optimum is 40 degrees C . The enzyme is strongly inhibited by thiol enzyme inhibitors, indicating that a Cys residue is likely to be involved in the catalytic reaction . The BSH of B . longum can hydrolyze all six major human bile salts and at least two animal bile salts . A slight preference for glycine-conjugated bile acids was detected based on both the specificity and the K(m) values . The nucleotide sequence of bsh was determined and used for homology studies, transcript analysis, and construction and analysis of various mutants . The levels of homology with BSH of other bacteria and with penicillin V acylase (PVA) of Bacillus sphaericus were high . On the basis of the similarity of BSH and PVA, whose crystal structure has been elucidated, BSH can be classified as an N-terminal nucleophile hydrolase with Cys as the N-terminal amino acid . This classification was confirmed by the fact that a Cys1Ala exchange by site-directed mutagenesis resulted in an inactive protein . Reverse transcription-PCR experiments revealed that bsh is part of an operon containing at least two genes, bsh and glnE (GlnE is glutamine synthetase adenylyltransferase) . Two UV-induced BSH-negative mutants and one spontaneous BSH-negative mutant were isolated from B . longum SBT2928 cultures and characterized . These mutants had point mutations that inactivated bsh by premature termination, frameshift, or amino acid exchange. Poult Sci, 2000 May, 79(5), 689 - 97 Preventive effect of partially hydrolyzed guar gum on infection of Salmonella enteritidis in young and laying hens; Ishihara N et al.; The preventive effect of partially hydrolyzed guar gum (PHGG) on the colonization of Salmonella enteritidis (SE) in young and laying hens was investigated . The effects of feed supplemented with 0.025, 0.05, and 0.1% PHGG was examined on young hens orally infected with SE . The incidence of SE in organs was decreased, the excretion of SE into feces was increased, and the agglutinating antibody titer to SE in serum was decreased by the administration of PHGG to young hens . In particular, feed supplemented with 0.025% PHGG was the most effective . It was also shown that feed supplemented with 0.025% PHGG increased the number of Bifidobacterium spp . and Lactobacillus spp., the most numerous intestinal bacteria in the cecum of young hen . The effect of the excretion of SE via feces was also observed in an experiment using laying hens . The incidence of SE on the surface of the eggshell and in egg white and egg yolk was also decreased when the feed of laying hens was supplemented with 0.025% PHGG . These results show that the administration of feed supplemented with PHGG can prevent the colonization of SE in young and laying hens, which, in turn, could be related to improvement in the balance of intestinal microflora. J Agric Food Chem, 2000 May, 48(5), 1644 - 52 Fermentation of plant cell wall derived polysaccharides and their corresponding oligosaccharides by intestinal bacteria; Van Laere KM et al.; New types of nondigestible oligosaccharides were produced from plant cell wall polysaccharides, and the fermentation of these oligosaccharides and their parental polysaccharides by relevant individual intestinal species of bacteria was studied . Oligosaccharides were produced from soy arabinogalactan, sugar beet arabinan, wheat flour arabinoxylan, polygalacturonan, and rhamnogalacturonan fraction from apple . All of the tested substrates were fermented to some extent by one or more of the individual species of bacteria tested . Bacteroides spp . are able to utilize plant cell wall derived oligosaccharides besides their reported activity toward plant polysaccharides . Bifidobacterium spp . are also able to utilize the rather complex plant cell wall derived oligosaccharides in addition to the bifidogenic fructooligosaccharides . Clostridium spp., Klebsiella spp., and Escherichia coli fermented some of the selected substrates in vitro . These studies do not allow prediction of the fermentation in vivo but give valuable information on the fermentative capability of the tested intestinal strains. Zh Mikrobiol Epidemiol Immunobiol, 2000 Mar-Apr, (2), 63 - 6 {The antibacterial resistance of the strains making up the components of probiotic preparations}; Lykova EA; The spectrum of the antibiotic resistance of 21 strains of normal microflora, mainly forming constituents of widely used probiotics to 25 antibacterial preparations . Lactobacillus spp . were prevalent in the strains under test . The spectrum of the antibiotic resistance of lactobacilli varied, the gradation of resistance being more pronounced with respect to strains and not species . Both highly sensitive and highly resistant Lactobacillus strains were found: L . acidophilus (a component of biopreparation "Linex"), L . plantarum 8RA3, L . fermentum 90T4C (probiotic "Lactobacterin"), L . fermentum BL96 . Bacteria used as the components of combined preparation "Linex" exhibited the highest resistance to a number of modern antibiotics . Strains of bifidobacteria were found to be highly sensitive to antibiotics. Am J Respir Crit Care Med, 2000 May, 161(5), 1750 - 3 Bacteriology of infected cavitating lung tumor; Liao WY et al.; Differentiation between in situ infection and simple tumor necrosis in cavitating lung tumors by means of imaging studies is difficult . In this study, we prospectively investigated the role of ultrasound (US)-guided transthoracic aspiration for bacteriologic examination of infected cavitating lung tumors, and the influence of the culture results on the treatment of patients . Twenty-two patients (18 men and four women) with cavitating lung tumors treated from January 1996 to October 1998 were included . All patients underwent US-guided transthoracic aspiration for bacterial, fungal, and mycobacterial cultures . Microorganisms were isolated from six of seven febrile patients and one of 15 nonfebrile patients . A total of nine pathogens were isolated from seven patients: Klebsiella pneumoniae (n = 3); Haemophilus influenzae (n = 2); Enterococcus faecium (n = 1); Bifidobacterium (n = 1); Shewanella putrefaciens (n = 1); and Mycobacterium tuberculosis (n = 1) . Two pathogens were isolated from the aspirate cultures in two patients, while the others had monomicrobial infection . The six febrile patients who had positive lung aspirate cultures were treated with empiric antimicrobial agents before the culture results were available, and the culture results led to adjustment of the antibiotic regimen in five of these . The clinical conditions of the six patients with infected cavitating lung tumors improved after the initiation of individualized antimicrobial treatment . Pneumothorax occurred in one patient, and was the sole procedure-related complication . In conclusion, US-guided transthoracic aspiration is helpful for differentiating infected cavitating lung tumors from simple tumor necrosis . Infection in cavitating lung tumors is common among febrile patients, and the culture results can guide modification of the antimicrobial therapy. J Microbiol Methods, 2000 May, 40(3), 221 - 4 Modification of the phosphoketolase assay for rapid identification of bifidobacteria; Orban JI et al.; The phosphoketolase assay is commonly used as a definitive criterion for identification of bifidobacteria . A limitation of the assay is the time-consuming process of cell disruption, either by use of the French Pressure Cell or by sonication . We have replaced the time consuming cell disruption process with a more rapid cell membrane disruption process by pretreating cells with the detergent hexadecyltrimethylammonium bromide (cetrimonium bromide, CTAB) . The effect of no pretreatment, sonication or the addition of CTAB (0.45 mg/ml) on color development in the phosphoketolase assay was tested using pure cultures of bifidobacteria and lactobacilli . No phosphoketolase activity was observed with bifidobacterial cultures without cell disruption or with lactobicilli that had undergone cell disruption . All bifidobacterial cultures gave a similar color formation whether sonication or CTAB addition was used to disrupt cells . Use of CTAB to disrupt cell membranes is an effective alternative to the time consuming traditional cell disruption procedures and increases the number of cultures that can be simultaneously assayed and presumptively identified using the phosphoketolase assay. J Med Microbiol, 2000 May, 49(5), 431 - 9 Translocation of indigenous microflora in an experimental model of sepsis; Naaber P et al.; Translocation of viable bacteria from gut to bloodstream and other sterile body sites during shock has been demonstrated in several experimental and clinical studies . The factors causing translocation and its incidence at different stages of shock are not known . The aim of the study was to evaluate the importance of several factors causing translocation of indigenous microflora in an experimental model of septic shock based on intraperitoneal Escherichia coli sepsis in rats . Counts of inoculated E . coli and translocated bacteria in different locations, gut morphology and haematological values were evaluated at different stages of sepsis . Sepsis developed in all animals and E . coli achieved the highest counts in blood 6 h after inoculation . Translocation was commonest at 6 and 12 h after inoculation . Frequently translocating bacteria were lactobacilli, bifidobacteria, bacteroides and peptostreptococci . In early sepsis, translocation was associated with high E . coli counts in blood, yet in late sepsis the opposite correlation was present . Low infiltration by neutrophils in the ileum and decreased mitotic activity in the colon were associated with a high translocation rate . In early sepsis, translocation was associated with low lymphocyte counts, but in late sepsis, with low neutrophil counts . Translocation of bacteria (including anaerobes) that colonise the gut in high counts takes place during sepsis . Putative influencing factors such as activity of the primary disease (bacterial counts in blood), gut morphology or haematological values seem to have different impacts on translocation, depending on the stage of the disease. J Dairy Sci, 2000 Apr, 83(4), 894 - 907 Probiotic bacteria: selective enumeration and survival in dairy foods; Shah NP; A number of health benefits have been claimed for probiotic bacteria such as Lactobacillus acidophilus, Bifidobacterium spp., and Lactobacillus casei . Because of the potential health benefits, these organisms are increasingly incorporated into dairy foods . However, studies have shown low viability of probiotics in market preparations . In order to assess viability of probiotic bacteria, it is important to have a working method for selective enumeration of these probiotic bacteria . Viability of probiotic bacteria is important in order to provide health benefits . Viability of probiotic bacteria can be improved by appropriate selection of acid and bile resistant strains, use of oxygen impermeable containers, two-step fermentation, micro-encapsulation, stress adaptation, incorporation of micronutrients such as peptides and amino acids and by sonication of yogurt bacteria . This review will cover selective enumeration and survival of probiotic bacteria in dairy foods. Int J Food Microbiol, 2000 Apr 10, 55(1-3), 41 - 5 Basic features of the stress response in three species of bifidobacteria: B . longum, B . adolescentis, and B . breve; Schmidt G et al.; The presence of the dnaK heat shock gene could be demonstrated for B . longum NCC481, B . longum NCC490, B . longum NCC585, B . adolescentis NCC251, and B . breve NCC298 . Induction of dnaK on the transcriptional level was shown for NCC251 and NCC481 by increasing temperatures . NCC251 showed an additional chaperone-induction after salt or bile-salt treatment . In both strains preconditioning with bile-salts protected against otherwise lethal concentrations thereof . NCC251 when subjected to a heat stress was able to survive an otherwise lethal temperature (55 degrees C) . Cross-protection was demonstrated for NCC251 since salt pretreatment resulted in increased tolerance after freeze-thawing cycles or lethal heat stress. Appl Environ Microbiol, 2000 May, 66(5), 2227 - 31 Use of a single, triplicate arbitrarily primed-PCR procedure for molecular fingerprinting of lactic acid bacteria; Cusick SM et al.; Arbitrarily primed (AP)-PCR can be used to generate characteristic DNA fingerprint patterns . However, small changes in reaction conditions can cause band irreproducibility . In this study, a single methodology encompassing triplicate reactions, which were intentionally exposed to three different annealing temperatures, enabled bands that were reproducibly generated to be recognized . A single triplicate AP-PCR (TAP-PCR) procedure, using an 18-mer primer, was developed and used to fingerprint representative isolates from the major genera of lactic acid bacteria and Bifidobacterium to the strain level. Carcinogenesis, 2000 May, 21(5), 937 - 41 Inhibitory effects of Bifidobacterium-fermented soy milk on 2-amino-1-methyl-6-phenylimidazo{4,5-b}pyridine-induced rat mammary carcinogenesis, with a partial contribution of its component isoflavones; Ohta T et al.; High consumption of soybean and soybean-related products is hypothesized to contribute to protection against breast cancer . Soybean is a rich source of genistein, a putative cancer chemopreventive agent . Fermented soy milk (FSM), which is made of soy milk fermented with the Bifidobacterium breve strain Yakult, contains larger amounts of the isoflavone aglycones genistein and daidzein than unfermented soy milk . In the present study, we examined the effects of FSM and its component isoflavone mixture (genistein:daidzein 4:1) on 2-amino-1-methyl-6-phenylimidazo{4, 5-b}pyridine (PhIP)-induced mammary carcinogenesis in rats . Starting at 7 weeks of age, female Sprague-Dawley rats were given PhIP at a dose of 85 mg/kg body wt by intragastric administration four times a week for 2 weeks . They were fed control high fat basal diet or experimental high fat diet containing 10% FSM or 0.02 or 0.04% isoflavone mixture during and after carcinogen exposure . The incidences (percentage of rats with tumors) of mammary gland tumors were 71% in the control diet group, 51% in the FSM group and 68 and 61% in the groups treated with isoflavone mixture at 0.02 and 0.04%, respectively . Mammary tumor multiplicities (number of tumors per rat) were 1.2 +/- 0.2 for 10% FSM, 2.2 +/- 0.4 for 0.02% isoflavone mixture and 1.5 +/- 0.3 for 0.04% isoflavone mixture, being clearly smaller than the control diet value (2.6 +/- 0.5) . Furthermore, feeding of FSM and the isoflavone mixture at both doses reduced the sizes of mammary tumors . Since the amounts of isoflavones in 10% FSM are approximately equivalent to those in the 0.02% isoflavone mixture, the chemopreventive activity of FSM could be partly attributable to the presence of isoflavones such as genistein and daidzein. Cancer Gene Ther, 2000 Feb, 7(2), 269 - 74 Bifidobacterium longum as a delivery system for cancer gene therapy: selective localization and growth in hypoxic tumors; Yazawa K et al.; A fundamental obstacle in gene therapy for cancer is the specific delivery of an anticancer gene product to a solid tumor, and yet no systemic delivery system that specifically targets solid tumors currently exists . A strain of domestic bacteria, Bifidobacterium longum, which is nonpathogenic and anaerobic, selectively localized and proliferated in several types of mouse solid tumors after systemic application . In this report, we further describe a novel approach to cancer gene therapy in which genetically engineered Bifidobacterium is used as a tumor-specific vector . Similarly to wild-type B . longum, genetically engineered B . longum could be detected in tumor tissue only and was not found in a large survey of normal mouse tissues after intravenous injection . This finding strongly suggests that obligate anaerobic bacteria such as Bifidobacterium can be used as highly specific gene delivery vectors for cancer gene therapy. Prikl Biokhim Mikrobiol, 2000 Jan-Feb, 36(1), 104 - 10 {Production of the therapeutic and prophylactic preparation enterobifidin on the basis of Bifidobacterium adolescentis MC-42}; Novik GI et al.; Production of Enterobifidin comprises preparation of culture media, reparation of lyophilized Bifidobacterium adolescentis MS-42 culture, preparation of starters, cultivation of bacteria in fermenters, biomass conservation, and its biological control . The preparation contains physiologically active bifidobacterium cells with high activities of growth (mu = 0.7 h-1, g = 1.0 h) and acid formation (titratable acidity is approximately 120-140 degrees T; acetate concentration, 0.50-0.75%; and lactate concentration, 0.33-0.50%) . The antagonistic activity of these bacteria towards Escherichia coli 08, E . coli 086, E . coli 015, E . coli 0115, and E . coli 0101 amounts to 98.2;, to Proteus vulgaris 102, to 87.2; and Staphylococcus aureus 209p, to 83.2% . The bifidobacteria (with a titer of 10(9) CFU/ml) remained viable for two to five months. Biol Pharm Bull, 1999 Dec, 22(12), 1314 - 8 Metabolism of 6"-O-xylosyltectoridin and tectoridin by human intestinal bacteria and their hypoglycemic and in vitro cytotoxic activities; Bae EA et al.; 6"-O-Xylosyltectoridin and tectoridin isolated from the flowers of Pueraria thunbergiana (Leguminosae), are metabolized to tectorigenin by human intestinal bacteria . Although tectoridin is metabolized to tectorigenin by most intestinal bacteria, 6"-O-xylosyltectoridin is metabolized to tectorigenin via tectoridin by only a few intestinal bacteria, such as Bifidobacterium breve K-110 and Eubacterium A-44 . The metabolite, tectorigenin, had more potent hypoglycemic activity as well as in vitro cytotoxic activity against tumor cell lines than 6"-O-xylosyltectoridin and tectoridin . These results suggest that 6"-O-xylosyltectoridin and tectoridin are prodrugs which can be transformed to the active agents by human intestinal bacteria. Br J Nutr, 2000 Feb, 83(2), 167 - 76 Enhancement of natural and acquired immunity by Lactobacillus rhamnosus (HN001), Lactobacillus acidophilus (HN017) and Bifidobacterium lactis (HN019); Gill HS et al.; Consumption of lactic acid bacteria (LAB) has been suggested to confer a range of health benefits including stimulation of the immune system and increased resistance to malignancy and infectious illness . In the present study, the effects of feeding Lactobacillus rhamnosus (HN001, DR20), Lactobacillus acidophilus (HN017) and Bifidobacterium lactis (HN019, DR10) on in vivo and in vitro indices of natural and acquired immunity in healthy mice were examined . Mice were fed daily with L . rhamnosus, L . acidophilus or B . lactis (10(9) colony forming units) and their immune function was assessed on day 10 or day 28 . Supplementation with L . rhamnosus, L . acidophilus or B . lactis resulted in a significant increase in the phagocytic activity of peripheral blood leucocytes and peritoneal macrophages compared with the control mice . The proliferative responses of spleen cells to concanavalin A (a T-cell mitogen) and lipopolysaccharide (a B-cell mitogen) were also significantly enhanced in mice given different LAB . Spleen cells from mice given L . rhamnosus, L . acidophilus or B . lactis also produced significantly higher amounts of interferon-gamma in response to stimulation with concanavalin A than cells from the control mice . LAB feeding had no significant effect on interleukin-4 production by spleen cells or on the percentages of CD4+, CD8+ and CD40+ cells in the blood . The serum antibody responses to orally and systemically administered antigens were also significantly enhanced by supplementation with L . rhamnosus, L . acidophilus or B . lactis . Together, these results suggest that supplementation of the diet with L . rhamnosus (HN001), L . acidophilus (HN017) or B . lactis (HN019) is able to enhance several indices of natural and acquired immunity in healthy mice. Appl Environ Microbiol, 2000 Apr, 66(4), 1379 - 84 Characterization of a novel beta-galactosidase from Bifidobacterium adolescentis DSM 20083 active towards transgalactooligosaccharides; Van Laere KM et al.; This paper reports on the effects of both reducing and nonreducing transgalactooligosaccharides (TOS) comprising 2 to 8 residues on the growth of Bifidobacterium adolescentis DSM 20083 and on the production of a novel beta-galactosidase (beta-Gal II) . In cells grown on TOS, in addition to the lactose-degrading beta-Gal (beta-Gal I), another beta-Gal (beta-Gal II) was detected and it showed activity towards TOS but not towards lactose . beta-Gal II activity was at least 20-fold higher when cells were grown on TOS than when cells were grown on galactose, glucose, and lactose . Subsequently, the enzyme was purified from the cell extract of TOS-grown B . adolescentis by anion-exchange chromatography, adsorption chromatography, and size-exclusion chromatography . Beta-Gal II has apparent molecular masses of 350 and 89 kDa as judged by size-exclusion chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, respectively, indicating that the enzyme is active in vivo as a tetramer . Beta-Gal II had an optimal activity at pH 6 and was not active below pH 5 . Its optimum temperature was 35 degrees C . The enzyme showed highest V(max) values towards galactooligosaccharides with a low degree of polymerization . This result is in agreement with the observation that during fermentation of TOS, the di- and trisaccharides were fermented first . Beta-Gal II was active towards beta-galactosyl residues that were 1-->4, 1-->6, 1-->3, and 1 <--> 1 linked, signifying its role in the metabolism of galactooligosaccharides by B . adolescentis. Microbios, 2000, 101(399), 105 - 14 A new factor from Bacillus mesentericus which promotes the growth of Bifidobacterium; Seo G et al.; It was reported previously that supernatants of cultures of Bacillus mesentericus TO-A promote the growth of Bifidobacterium species . In this study, a new growth-promoting factor, BM-1, was purified from the supernatant of such a culture and its chemical structure was determined . BM-1 was identified as 3,3-dihydroxyazetidine, and it promoted the growth of several strains of Bifidobacterium. Lett Appl Microbiol, 2000 Jan, 30(1), 10 - 3 The mucus binding of Bifidobacterium lactis Bb12 is enhanced in the presence of Lactobacillus GG and Lact . delbrueckii subsp . bulgaricus; Ouwehand AC et al.; The ability to adhere to mucosal surfaces is related to many probiotic health effects . In the presence of Lactobacillus GG or Lact . bulgaricus, the adhesion of Bifidobacterium lactis Bb12 to a mucus model was more than doubled . Other tested lactobacilli did not affect the adhesion, nor was the adhesion of the lactobacilli influenced by the bifidobacteria . Co-aggregation between Bif . lactis Bb12 and the tested lactobacilli was insignificant and does not explain the observed effect . The results suggest that combinations of probiotics strains may have synergistic adhesion effects . Such specific strain combinations should also be assessed in clinical studies. J Nutr, 2000 Feb, 130(2S Suppl), 432S - 436S Interactions mediating bacterial translocation in the immature intestine; Duffy LC; Systemic disease caused by transmucosal passage of enterovirulent bacteria and toxins from the gut lumen into the mesenteric lymph nodes (MLN) is reviewed, with particular concern for bacterial interactions in the developing gut of premature newborns . Anaerobic bacteria are rarely observed to translocate to the MLN . Bifidobacterial strains have been tested for their abilities to adhere to enterocyte-like Caco-2 cells in culture . We have investigated the inhibitory effect of adherent human bifidobacterial strains against colonization by a number of diarrheagenic bacteria (Escherichia coli O157; Salmonella typhimurium) and viruses (murine and rhesus rotavirus), in various in vitro and in vivo models . The phagocytic cell (macrophage) may be a key factor in bacterial translocation (BT) . Human breast milk contains abundant bioactive substances (immunologic, nutritional) that provide protective effects through inhibition of bacterial overgrowth and BT . New biotherapeutic therapies that stimulate beneficial anaerobic microflora (Lactobacillus, Bifidobacterium) are promising avenues of research to combat BT in disease treatment. J Nutr, 2000 Feb, 130(2S Suppl), 396S - 402S The role of probiotic cultures in the control of gastrointestinal health; Rolfe RD; The use of probiotics to enhance intestinal health has been proposed for many years . Probiotics are traditionally defined as viable microorganisms that have a beneficial effect in the prevention and treatment of specific pathologic conditions when they are ingested . There is a relatively large volume of literature that supports the use of probiotics to prevent or treat intestinal disorders . However, the scientific basis of probiotic use has been firmly established only recently, and sound clinical studies have begun to be published . Currently, the best-studied probiotics are the lactic acid bacteria, particularly Lactobacillus sp . and Bifidobacterium sp . However, other organisms used as probiotics in humans include Escherichia coli, Streptococcus sp., Enterococcus sp., Bacteroides sp., Bacillus sp., Propionibacterium sp . and various fungi . Some probiotic preparations contain mixtures of more than one bacterial strain . Probiotics have been examined for their effectiveness in the prevention and treatment of a diverse spectrum of gastrointestinal disorders such as antibiotic-associated diarrhea (including Clostridium difficile-associated intestinal disease), infectious bacterial and viral diarrhea (including diarrhea caused by rotavirus, Shigella, Salmonella, enterotoxigenic E . coli, Vibrio cholerae and human immunodeficiency virus/acquired immunodeficiency disorder, enteral feeding diarrhea, Helicobacter pylori gastroenteritis, sucrase maltase deficiency, inflammatory bowel disease, irritable bowel syndrome, small bowel bacterial overgrowth and lactose intolerance . Probiotics have been found to inhibit intestinal bacterial enzymes involved in the synthesis of colonic carcinogens . There are many mechanisms by which probiotics enhance intestinal health, including stimulation of immunity, competition for limited nutrients, inhibition of epithelial and mucosal adherence, inhibition of epithelial invasion and production of antimicrobial substances . Probiotics represent an exciting prophylactic and therapeutic advance, although additional investigations must be undertaken before their role in intestinal health can be delineated clearly. J Nutr, 2000 Feb, 130(2S Suppl), 391S - 395S Aspects of in vitro and in vivo research approaches directed toward identifying probiotics and prebiotics for human use; Gibson GR et al.; The microbiota of the human gastrointestinal tract plays a key role in nutrition and health . Through the process of fermentation, gut bacteria metabolize various substrates (principally dietary components) to end products such as short-chain fatty acids and gases . This anaerobic metabolism is thought to contribute positively toward host daily energy requirements . However, under certain circumstances, the fermentative process may produce undesirable metabolites . This may cause the onset of gut disorders that can be manifest through both acute and chronic conditions . Moreover, the gut flora may become contaminated by transient pathogens that serve further to upset the normal community structure . There has been a recent increase in the use of dietary components that help to maintain, or even improve, the gut microflora "balance." Probiotics are live microbial feed supplements added to appropriate food vehicles (usually fermented milks), whereas prebiotics are dietary carbohydrates that have a selective metabolism in the colon and serve to increase numbers of bacteria seen as desirable . Because of their purported health-promoting properties, lactic acid-producing bacteria, including bifidobacteria, are the usual target organisms . The market value and biological potential of both approaches are enormous . This article will summarize how efficacious types can be identified. Food Chem Toxicol, 2000 Feb-Mar, 38(2-3), 153 - 61 Acute oral toxicity and bacterial translocation studies on potentially probiotic strains of lactic acid bacteria; Zhou JS et al.; Three potentially probiotic lactic acid bacteria (LAB) strains, Lactobacillus rhamnosus HN001 (DR20(TM)), Lb . acidophilus HN017 and Bifidobacterium lactis HN019 (DR10()), have recently been identified and characterized . The present study was designed to evaluate the acute oral toxicity of these strains to mice, and also to investigate bacterial translocation and gut mucosal pathology in BALB/c mice fed HN019, HN001 or HN017 for 8 consecutive days at a high dose of 10(11)cfu/mouse/day . Results showed that these probiotic strains had no adverse effect on general health status, feed intake, body weight gain and intestinal mucosal morphology (villus height, crypt depth, epithelial cell height and mucosal thickness) . No viable bacteria were recovered from blood and tissue samples (mesenteric lymph nodes, liver and spleen) of mice, and no treatment-associated illness or death was observed . According to these results, the oral LD(50) of HN019, HN001 and HN017 is more than 50g/kg/day for mice, and their acceptable daily intake (ADI) value is 35g dry bacteria per day for a 70-kg person . This suggests that the probiotic strains HN019, HN001 and HN017 are non-pathogenic and likely to be safe for human consumption. J Food Prot, 2000 Mar, 63(3), 327 - 31 Viability of bifidobacteria in commercial dairy products during refrigerated storage; Shin HS et al.; Commercial milk and two brands of yogurt containing bifidobacteria were obtained from retail outlets . All products were evaluated for viability of bifidobacteria and lactic acid bacteria during refrigerated storage at 4 degrees C . Milk was evaluated at 9, 6, and 3 days prior and past its expiration date . The yogurts were evaluated at 3, 2, and 1 week prior and past their expiration . Viability of bifidobacteria and lactic acid bacteria in milk and yogurt remained above 10(6) CFU/ml or g until the expiration date of the respective products . This microbial concentration is the recommended minimum dose to receive the health benefits of these organisms. J Food Prot, 2000 Mar, 63(3), 322 - 6 An enzymatic-colorimetric assay for the quantification of Bifidobacterium; Bibiloni R et al.; An enzymatic-colorimetric assay for the quantification of Bifidobacterium was developed . The method, based upon the standard detection of fructose-6-phosphate phosphoketolase activity, was optimized with respect to bacterial cell pretreatment, time of incubation, and substrate concentration . The relationship between bacterial biomass and phosphoketolase activity was linear in a wide spectrum of bacterial densities . Higher sensitivity over the standard method was achieved by using 0.25% Triton X-100 in the reaction mixture to pretreat the bacterial cells . Because autoaggregation is a frequent feature among Bifidobacterium strains, this simple and reproducible method offers good advantage over viable plate count and turbidimetric techniques . The methodology can also be applied to the assessment of adherent Bifidobacterium strains to human epithelial cells. Scand J Infect Dis, 2000, 32(1), 81 - 5 Comparative effects of moxifloxacin and clarithromycin on the normal intestinal microflora; Edlund C et al.; Twelve healthy male subjects age range 24-40 y participated in the investigation . The trial was divided into 2 35-d periods . The 2 treatment regimens were: (i) 1 x 400 mg moxifloxacin tablet in the morning and 1 placebo tablet in the evening for 7 d; and (ii) 1 x 500 mg clarithromycin tablet in the morning and 1 x 500 mg clarithromycin tablet in the evening for 7 d . Each subject received firstly I treatment regimen and secondly the other treatment regimen . The wash-out period was 6 weeks between the two treatment regimens . Moxifloxacin caused significant decreases of enterococci and enterobacteria during the administration period while the numbers of staphylococci, streptococci, Bacillus and Candida were not affected . No impact on peptostreptococci, lactobacilli, Veillonella, Bacteroides or fusobacteria was observed, while bifidobacteria and clostridia decreased during moxifloxacin administration . The microflora was normalized after 35 d . Clarithromycin caused significant reduction of Escherichia coli while the numbers of enterococci, Enterobacter, Citrobacter, Klebsiella and Pseudomonas increased markedly . No significant changes in the numbers of staphylococci, streptococci, Bacillus and Candida were noticed . In the anaerobic microflora bifidobacteria, lactobacilli and clostridia were suppressed, while no changes in peptostreptococci, Veillonella, Bacteroides and fusobacteria were found . The microflora was normalized in all volunteers after 35 d. Eur J Clin Nutr, 2000 Mar, 54(3), 263 - 7 Enhancement of natural immune function by dietary consumption of Bifidobacterium lactis (HN019); Arunachalam K et al.; OBJECTIVE: To determine the effects of dietary consumption of Bifidobacterium lactis (strain HN019, DR10TM) on natural immunity . DESIGN: A randomized, double blind, placebo-controlled clinical trial . SETTING: Janeway Medical Centre, Memorial University, St Johns, Newfoundland . SUBJECTS: Twenty-five healthy elderly volunteers (median age 69 y; range 60-83 y) . INTERVENTIONS: Twelve control subjects consumed 180 ml low-fat/low-lactose milk twice daily for a period of 6 weeks; 13 test subjects consumed milk supplemented with 1.5x1011 colony-forming units of B . lactis twice daily . Indices of natural immunity, including interferon production, phagocytic capacity and phagocyte-mediated bactericidal activity, were determined via peripheral blood at 0, 3, 6 and 12 weeks post-trial commencement . RESULTS: Subjects who consumed milk containing B . lactis for 6 weeks produced significantly enhanced levels of interferon-alpha, upon stimulation of their peripheral blood mononuclear cells in culture, in comparison to the placebo control group who received milk alone . There were also significant increases in polymorphonuclear cell phagocytic capacity among test group subjects, following consumption of milk supplemented with B . lactis, while individuals who consumed B . lactis-supplemented milk or milk alone showed enhanced phagocyte-mediated bactericidal activity . CONCLUSIONS: The results demonstrate that dietary consumption of B . lactis HN019 can enhance natural immunity in healthy elderly subjects, and that a relatively short-term dietary regime (6 weeks) is sufficient to impart measurable improvements in immunity that may offer significant health benefits to consumers . SPONSORS: Financial support for this project was provided by the New Zealand Dairy Board. Appl Microbiol Biotechnol, 2000 Feb, 53(2), 219 - 23 Influence of a synbiotic mixture consisting of Lactobacillus acidophilus 74-2 and a fructooligosaccharide preparation on the microbial ecology sustained in a simulation of the human intestinal microbial ecosystem (SHIME reactor); Gmeiner M et al.; Lactobacillus acidophilus 74-2, which is used in probiotic products, was administered, with fructo-oligosaccharide in a milk-based product, to the second vessel (duodenum/jejunum) of the SHIME reactor, an in vitro simulation of the human intestinal microbial ecology . The main focus of this study was to monitor the changes of the population density of selected bacterial species in the intestine and the changes of metabolic activities during the supplementation of L . acidophilus and fructooligosaccharide in the SHIME reactor . Interestingly, the addition of L . acidophilus 74-2 with fructooligosaccharide gave rise to an increase of bifidobacteria . Moreover, major positive changes occurred in the production of volatile fatty acids: a strong upward trend was observed especially in the case of butyric acid and propionic acid . Furthermore a noticeable increase of beta-galactosidase activity was monitored, while the activity of beta-glucuronidase, generally considered undesirable, declined. Antimicrob Agents Chemother, 2000 Mar, 44(3), 775 - 7 Occurrence of the new tetracycline resistance gene tet(W) in bacteria from the human gut; Scott KP et al.; Members of our group recently identified a new tetracycline resistance gene, tet(W), in three genera of rumen obligate anaerobes . Here, we show that tet(W) is also present in bacteria isolated from human feces . The tet(W) genes found in human Fusobacterium prausnitzii and Bifidobacterium longum isolates were more than 99.9% identical to those from a rumen isolate of Butyrivibrio fibrisolvens. Appl Environ Microbiol, 2000 Feb, 66(2), 869 - 73 Survival of Bifidobacterium longum immobilized in calcium alginate beads in simulated gastric juices and bile salt solution; Lee KY et al.; Bifidobacterium longum KCTC 3128 and HLC 3742 were independently immobilized (entrapped) in calcium alginate beads containing 2, 3, and 4% sodium alginate . When the bifidobacteria entrapped in calcium alginate beads were exposed to simulated gastric juices and a bile salt solution, the death rate of the cells in the beads decreased proportionally with an increase in both the alginate gel concentration and bead size . The initial cell numbers in the beads affected the numbers of survivors after exposure to these solutions; however, the death rates of the viable cells were not affected . Accordingly, a mathematical model was formulated which expressed the influences of several parameters (gel concentration, bead size, and initial cell numbers) on the survival of entrapped bifidobacteria after sequential exposure to simulated gastric juices followed by a bile salt solution . The model proposed in this paper may be useful for estimating the survival of bifidobacteria in beads and establishing optimal entrapment conditions. Immunol Cell Biol, 2000 Feb, 78(1), 80 - 8 Survival and therapeutic potential of probiotic organisms with reference to Lactobacillus acidophilus and Bifidobacterium spp; Kailasapathy K et al.; The present paper provides an overview on the use of probiotic organisms as live supplements, with particular emphasis on Lactobacillus acidophilus and Bifidobacterium spp . The therapeutic potential of these bacteria in fermented dairy products is dependent on their survival during manufacture and storage . Probiotic bacteria are increasingly used in food and pharmaceutical applications to balance disturbed intestinal microflora and related dysfunction of the human gastrointestinal tract . Lactobacillus acidophilus and Bifidobacterium spp . have been reported to be beneficial probiotic organisms that provide excellent therapeutic benefits . The biological activity of probiotic bacteria is due in part to their ability to attach to enterocytes . This inhibits the binding of enteric pathogens by a process of competitive exclusion . Attachment of probiotic bacteria to cell surface receptors of enterocytes also initiates signalling events that result in the synthesis of cytokines . Probiotic bacteria also exert an influence on commensal micro-organisms by the production of lactic acid and bacteriocins . These substances inhibit growth of pathogens and also alter the ecological balance of enteric commensals . Production of butyric acid by some probiotic bacteria affects the turnover of enterocytes and neutralizes the activity of dietary carcinogens, such as nitrosamines, that are generated by the metabolic activity of commensal bacteria in subjects consuming a high-protein diet . Therefore, inclusion of probiotic bacteria in fermented dairy products enhances their value as better therapeutic functional foods . However, insufficient viability and survival of these bacteria remain a problem in commercial food products . By selecting better functional probiotic strains and adopting improved methods to enhance survival, including the use of appropriate prebiotics and the optimal combination of probiotics and prebiotics (synbiotics), an increased delivery of viable bacteria in fermented products to the consumers can be achieved. Am J Clin Nutr, 2000 Feb, 71(2), 405 - 11 Effects of probiotic bacteria on diarrhea, lipid metabolism, and carcinogenesis: a review of papers published between 1988 and 1998; de Roos NM et al.; We reviewed the evidence from human intervention studies for the health effects of probiotic bacteria, ie, live bacteria that survive passage through the gastrointestinal tract and have beneficial effects on the host . Of the 49 studies reviewed, 26 dealt with the prevention or treatment of diarrheal disease, 9 with the prevention of cancer or of the formation of carcinogens, 7 with the lowering of serum cholesterol, and 7 with the stimulation of the immune system . The most widely studied probiotic bacteria were Lactobacillus GG (22 studies), Lactobacillus acidophilus (16 studies), Bifidobacterium bifidum (6 studies), and Enterococcus faecium (7 studies) . Intake of Lactobacillus GG consistently shortened the diarrheal phase of rotavirus infection by 1 d . However, evidence for the prevention by Lactobacillus GG and other probiotics of diarrhea due to viral or bacterial infections was less strong . Effects of probiotics on the immune system are inconclusive because of the variety of outcome variables reported . Cholesterol lowering by L . acidophilus was shown in some but not all studies; cholesterol lowering by E . faecium seems to be transient . Two studies of one research group showed a smaller recurrence of bladder tumors in patients after treatment with Lactobacillus casei; these results await confirmation . The production of mutagens after a meal might be reduced by the concomitant intake of probiotics, but the relevance of this finding is unclear . In conclusion, consumption of foods containing Lactobacillus GG may shorten the course of rotavirus infection . Other health effects of probiotic bacteria have not been well established . Well-designed placebo-controlled studies with validated outcome variables are needed to determine the health effects of probiotics. Adv Pediatr, 1999, 46, 353 - 82 Protective nutrients and bacterial colonization in the immature human gut; Dai D et al.; The normal human microflora is a complex ecosystem that is in part dependent on enteric nutrients for establishing colonization . The gut microbiota are important to the host with regard to metabolic functions and resistance to bacterial infections . At birth, bacterial colonization of a previously germ-free human gut begins . Diet and environmental conditions can influence this ecosystem . A breast-fed, full-term infant has a preferred intestine microbiota in which bifidobacteria predominate over potentially harmful bacteria, whereas in formula-fed infants, coliforms, enterococci, and bacteroides predominate . The pattern of bacterial colonization in the premature neonatal gut is different from that in the healthy, full-term infant gut . Those infants requiring intensive care acquire intestinal organisms slowly, and the establishment of bifidobacterial flora is retarded . A delayed bacterial colonization of the gut with a limited number of bacterial species tends to be virulent . Bacterial overgrowth is one of the major factors that promote bacterial translocation . The aberrant colonization of the premature infant may contribute to the development of necrotizing enterocolitis . Breast-feeding protects infants against infection . Oligo-saccharides and glycoconjugates, natural components in human milk, may prevent intestinal attachment of enteropathogens by acting as receptor homologues . Probiotics and prebiotics modulate the composition of the human intestinal microflora to the benefit of the host . These beneficial effects may result in the suppression of harmful microorganisms, the stimulation of bifidobacterial growth, or both . In the future, control and manipulation of the bacterial colonization in the neonatal gut may be a new approach to the prevention and treatment of intestinal infectious diseases of various etiologies. Mikrobiol Z, 1999 Sep-Oct, 61(5), 41 - 7 {The colonization resistance of the mucous membrane of the large intestine in patients with rheumatoid arthritis in a period of exacerbation}; Pishak OV; Specific and quantitative compositions of the colon mucous microflora in 36 patients with rheumatic arthritis (RA) in the remission period were studied . The mucous membrane of healthy people is colonized by bifidobacteria, lactobacilli, Bacteroides, Escherichia and enterococci . The mucous membrane in such people is mainly colonized by aerobic opportunistic conventionally pathogenic enterobacteria (enteropathogenic Escherichia, Citrobacter {correction of cytobacter}, Enterobacter, Klebsiella, etc.), staphylococci, enterococci and anaerobic bacteria (Bacteroides, peptococci, peptostreptococci, etc.) . Taking into account significant changes of colonization resistance in the colon mucous membrane in remission period of RA, it is necessary to apply bacteriotherapy, using bacterial drugs containing bifidobacteria and lactobacteria. Am J Gastroenterol, 2000 Jan, 95(1 Suppl), S16 - 8 Probiotics and infectious diarrhea; Saavedra J; Numerous probiotic agents have been studied for the management of diarrheal disease . In particular, the prevention and management of acute viral diarrhea, the treatment of recurrent Clostridium difficile diarrhea, as well as the control of antibiotic-associated diarrhea seem to be areas of significant potential benefit . A few agents, including Lactobacillus GG, Lactobacillus reuteri, and Saccharomyces boulardii, seem to be promising agents for the amelioration of the course of acute diarrhea in children when used therapeutically . The use of other agents, particularly Bifidobacteria, supplementing the regular feed of infants may have an effect as prophylactic agents against acute diarrheal diseases . In general, in pediatric populations, the effect of probiotic agents appears to be most significant against viral (rotaviral) diarrhea, suggesting that an immunological mechanism is responsible for the beneficial effects . The numerous agents, doses, and populations used make generalization difficult . Nevertheless, it is clear probiotic agents are becoming an important part of the armamentarium against gastrointestinal problems in infants and children. Eur J Clin Nutr, 2000 Nov, 54(11), 849 - 55 Enhancing immunity by dietary consumption of a probiotic lactic acid bacterium (Bifidobacterium lactis HN019): optimization and definition of cellular immune responses; Chiang BL et al.; OBJECTIVE: To define the cellular basis for immune enhancement by a probiotic lactic acid bacteria strain (Bifidobacterium lactis HN019); and to determine whether immune enhancement can be optimized by delivery in oligosaccharide-enriched low-fat milk . DESIGN: A double-blind, three-stage before-and-after intervention trial . SETTING: Taipei Medical College Hospital, Taipei, Taiwan . SUBJECTS: Fifty healthy Taiwanese citizens (age range 41-81; median 60) randomly allocated to two groups . INTERVENTIONS: In stage 1 (run-in control stage) all subjects consumed reconstituted low-fat milk (LFM) for 3 weeks; in stage 2 (probiotic intervention) subjects consumed B . lactis in LFM (group A) or B . lactis in lactose-hydrolysed LFM (group B) for 3 weeks; in stage 3 all subjects returned to non-supplemented LFM for a further 3 weeks (washout stage) . The innate immune functions of two different leucocyte types (polymorphonuclear (PMN) cells and natural killer (NK) cells) were assessed at four time points via in vitro analyses on peripheral blood samples . RESULTS: While consumption of LFM alone had no significant effect on immune responses, stage 2 results indicated significantly enhanced PMN cell phagocytosis and NK cell tumour killing activity following consumption of milk containing B . lactis . These increases levelled off following cessation of B . lactis consumption, but remained above the pre-treatment values . Increases in PMN and NK cell activity were greatest among subjects who consumed B . lactis in lactose-hydrolysed LFM . CONCLUSIONS: Dietary consumption of the probiotic bacterium B . lactis HN019 enhanced immune function of two different types of leucocytes; the degree of enhancement was increased by consuming B . lactis in an oligosaccharide-rich substrate . SPONSORSHIP: Financial support was provided by the New Zealand Dairy Board. Reprod Nutr Dev, 1999 Sep-Dec, 39(5-6), 563 - 88 Non-digestible oligosaccharides used as prebiotic agents: mode of production and beneficial effects on animal and human health; Grizard D et al.; Prebiotic agents are food ingredients that are potentially beneficial to the health of consumers . The main commercial prebiotic agents consist of oligosaccharides and dietary fibres (mainly inulin) . They are essentially obtained by one of three processes: 1) the direct extraction of natural polysaccharides from plants; 2) the controlled hydrolysis of such natural polysaccharides; 3) enzymatic synthesis, using hydrolases and/or glycosyl transferases . Both of these enzyme types catalyse transglycosylation reactions, allowing synthesis of small molecular weight synthetic oligosaccharides from mono- and disaccharides . Presently, in Europe, inulin-type fructans, characterised by the presence of fructosyl units bound to the beta-2,1 position of sucrose, are considered as one of the carbohydrate prebiotic references . Prebiotics escape enzymatic digestion in the upper gastrointestinal tract and enter the caecum without change to their structure . None are excreted in the stools, indicating that they are fermented by colonic flora so as to give a mixture of short-chain fatty acids (acetate, propionate and butyrate), L-lactate, carbon dioxide and hydrogen . By stimulating bifidobacteria, they may have the following implications for health: 1) potential protective effects against colorectal cancer and infectious bowel diseases by inhibiting putrefactive bacteria (Clostridium perfringens ) and pathogen bacteria (Escherichia coli, Salmonella, Listeria and Shigella ), respectively; 2) improvement of glucid and lipid metabolisms; 3) fibre-like properties by decreasing the renal nitrogen excretion; 4) improvement in the bioavailability of essential minerals; and 5) low cariogenic factor . These potential beneficial effects have been largely studied in animals but have not really been proven in humans . The development of a second generation of oligosaccharides and the putative implication of a complex bacterial trophic chain in the intestinal prebiotic fermentation process are also discussed. FEMS Microbiol Lett, 2000 Jan 1, 182(1), 163 - 9 Metabolization of beta-(2,6)-linked fructose-oligosaccharides by different bifidobacteria; Marx SP et al.; Low-molecular-mass beta-(2,6)-linked fructose-oligosaccharides (beta-(2,6)-FOS) were examined as a new carbohydrate source for growth of bifidobacteria . beta-(2,6)-FOS were prepared from microbial high-molecular-mass levan by acid hydrolysis and refined by cation-exchange chromatography . (13)C-NMR spectroscopy confirmed the presence of predominantly beta-(2,6)-fructosyl linkages in the oligosaccharides . More than 80% beta-(2,6)-FOS was recovered after in vitro incubation with amylolytic and proteolytic enzymes, implying resistance to degradation in the upper intestinal tract . Bifidobacterium adolescentis, B . longum, B . breve, and B . pseudocatenulatum were studied in vitro for their ability to metabolize beta-(2,6)-FOS . Growth, decrease in pH, formation of short- chain fatty acids (lactate, acetate, formate) and degradation of beta-(2,6)-FOS were markedly different among species . B . adolescentis showed the best growth, produced the highest amounts of organic acids and metabolized both short- and long-chain beta-(2, 6)-FOS. Z Geburtshilfe Neonatol, 1999 Sep-Oct, 203(5), 213 - 7 {Effects of oral administration of bifidobacteria on intestinal microflora in premature and newborn infants}; Uhlemann M et al.; In a prospective, randomised study the effects of orally administered bifidobacteria on the intestinal microflora were investigated in 100 preterm and term neonates under intensive care conditions during the first 21 days of life . The 50 infants (group with bifidobacteria) received lyophilized bifidobacteria (Topfer Bifidus) via nasogastral tube with an initial dosage of 3 times daily 1.25 x 10(8) bifidobacteria on day 2 of life and a daily dosage of 6 times 1.25 x 10(8) bifidobacteria on day 3 until day 21 of life . The other 50 infants (control group) did not receive bifidobacteria . The preterm and term neonates were fed either with pasteurized mother's milk or milk from healthy female donors (n = 79) or with an infant formula (Alfare, n = 13) or initially with Alfare and thereafter with mother's milk (n = 8) . The intestinal microflora of preterm and term neonates under intensive care conditions could be influenced by the oral administration of bifidobacteria . The administration of bifidobacteria resulted in the group of inoculated infants in a significantly earlier colonization of bifidobacteria (8.1 +/- 3.9 days of life) than in the control group (11.3 +/- 4.7 days of life) . On day 7 a bifidobacterial dominance (> 90% of the intestinal microflora) could be found in 26% of infants with inoculation of bifidobacteria and only in 2% of the control group (p < 0.001) . These significant differences could be shown until day 21 of life . A difference in septicemia frequency between the two groups could not be demonstrated . At the beginning of the infection a bifidobacterial dominance was found in only one of 23 cases of septicemia. J Agric Food Chem, 1999 Sep, 47(9), 3661 - 4 Inhibition of lipid peroxidation by Lactobacillus acidophilus and Bifidobacterium longum; Lin MY et al.; The inhibition of lipid peroxidation by Lactobacillus acidophilus and Bifidobacterium longum was investigated using two lipid model systems . All eight strains, including six strains of L . acidophilus and two strains of B . longum, demonstrated an inhibitory effect on linoleic acid peroxidation . The inhibitory rates on linoleic acid peroxidation ranged from 33 to 46% when 1 mL of intracellular cell-free extract was tested . In the second model system, the cell membrane of osteoblast was used as the source for biological lipid . The results indicated that all strains were able to protect biological lipids from oxidation . The inhibition rates on cell membrane lipid peroxidation ranged from 22 to 37% . The effect of L . acidophilus and B . longum on inhibition of fluorescent tissue pigment accumulation was also obtained for osteoblastic cells . The inhibition rates on fluorescent tissue pigment accumulation ranged from 20 to 39% . The antioxidative effect of each milliliter of intracellular cell-free extract of L . acidophilus and B . longum was equivalent to 104-172 ppm of butylated hydroxytoluene (BHT) . These results indicated that all strains demonstrated high antioxidative activity . The scavenging ability of lipid peroxidation products, tert-butyl hydroperoxide and malondialdehyde, was also evaluated . The results showed that L . acidophilus and B . longum were not able to scavenge the tert-butyl hydroperoxide . Nevertheless, malondialdehyde was scavenged well by these strains. J Agric Food Chem, 1999 Mar, 47(3), 934 - 8 Growth-inhibiting effects of Coptis japonica root-derived isoquinoline alkaloids on human intestinal bacteria; Chae SH et al.; The growth-inhibiting activity of Coptis japonica (Makino) root-derived materials toward eight human intestinal bacteria was examined using an impregnated paper disk method and compared to that of four commercially available isoquinoline alkaloids {berberine sulfate (BS), berberine iodide (BI), palmatine chloride (PC), and palmatine sulfate(PS)}, as well as that of Thea sinensis leaf-derived epigallocatechin gallate (EGCG) . The biologically active constituents of the Coptis extract were characterized as the isoquinoline alkaloids berberine chloride (BC), palmatine iodide (PI), and coptisine chloride (CC) by spectral analysis . The growth responses varied with both chemical and bacterial strain used . In a test using 500 microg/disk, BC and PI produced a clear inhibitory effect against Bifidobacterium longum, Bifidobacterium bifidum, Clostridium perfringens, and Clostridium paraputrificum, whereas weak or no inhibition was observed in Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactobacillus casei, and Escherichia coli . At 1000 microg/ disk, CC revealed weak or no growth inhibition toward all test bacteria, whereas EGCG exhibited weak growth inhibition against only C . perfringens and C . paraputrificum . Among various isoquinoline alkaloids, BC exhibited more potent inhibitory activity toward C . perfringens than BI and BS, whereas the inhibitory effect was more pronounced in PI compared to PC and PS . The Coptis root-derived materials did not promote growth of B . longum and C . perfringens. Biosci Biotechnol Biochem, 1999 Sep, 63(9), 1569 - 75 Effects of a probiotic on the lipid metabolism of cocks fed on a cholesterol-enriched diet; Endo T et al.; The effects of a probiotic (a mixture of Bacillus, Lactobacillus, Streptococcus, Clostridium, Saccharomyces and Candida) on the lipid metabolism, and caecal flora and metabolites of cocks were studied . The cholesterol level of the liver and serum was significantly decreased in the cocks fed on the cholesterol-enriched diet containing the probiotic . The distribution and frequency of occurrence of flora, and the chemical characteristics of the metabolites in the caecal content of the cocks were also affected by the inclusion of the probiotic in the basal and cholesterol-enriched diets . The Enterobacteriaceae species were significantly decreased in number, while the Bacillus, Streptococcus, Bifidobacterium and Lactobacillus species were significantly increased . The presence of yeast was observed, and the ammonia level was significantly reduced . The pH value, however, was not affected . The concentration of short-chain fatty acids in the caecal content of the cocks fed on the cholesterol-enriched diet supplemented with the probiotic was increased . It is, therefore, suggested that the incorporation of a probiotic in the diet would improve the balance of the intestinal flora and metabolites . Furthermore, it would also suppress the serum and liver cholesterol levels of cocks fed on the cholesterol-enriched diet. J Dairy Sci, 1999 Nov, 82(11), 2308 - 14 Increase of intestinal Bifidobacterium and suppression of coliform bacteria with short-term yogurt ingestion; Chen RM et al.; To determine whether ingestion of yogurt would alter human intestinal bacterial composition and whether Bifidobacterium numbers would increase in the intestine, 34 healthy volunteers were studied . The experimental period was 26 d, including an initial 8 d without yogurt, 10 d with three bottles (230 ml each) of AB yogurt per day (President Enterprise Corporation, Tainan, Taiwan), and 8 d without yogurt . Stool samples were taken at 3- to 4-d intervals . The bacteria of each fresh stool sample were promptly analyzed by dilution and culture on blood, MacConkey, Center for Disease Control and NNLP agars, the agar contained nalidixic acid, neomycin sulfate, LiCl, and paromomycin sulfate for aerobes, coliforms, anaerobes, and bifidobacteria, respectively . The number of bacteria was determined as colony-forming units per gram of dried stool . Results indicated that ingestion of AB yogurt increased the counts of anaerobic bacteria, suppressed aerobic bacteria, and significantly elevated the bifidus to coliform ratio . Arbitrarily primed polymerase chain reaction was used to differentiate the identity of bifidobacteria in four volunteers before and after yogurt ingestion and confirmed that B . bifidum ingested from the yogurt survived and proliferated in the stool throughout the experiment . However, the elevated bifidus to coliform ratio gradually diminished and disappeared after yogurt consumption was discontinued . In conclusion, ingestion of yogurt increased the numbers of stool bifidobacteria and suppressed coliform bacteria . The ingested bifidobacteria survived for more than 8 d after yogurt consumption was discontinued. Int J Infect Dis, 1999 Summer, 3(4), 197 - 202 Reduced incidence of necrotizing enterocolitis associated with enteral administration of Lactobacillus acidophilus and Bifidobacterium infantis to neonates in an intensive care unit; Hoyos AB; OBJECTIVES: Necrotizing enterocolitis (NEC) has been associated with a wide variety of bacteria and their cytotoxins . The content and the nature of gut bacterial colonization in newborns that require intensive care hospitalization has been demonstrated to be abnormal . In the 25-bed neonatal intensive care unit in Hospital Simon Bolivar, in Bogota, Colombia, cases of NEC are common causes of morbidity and mortality . This article examines the hypothesis that oral administration of prophylactic Lactobacillus acidophilus and Bifidobacterium infantis to all neonates in an intensive care unit, would decrease the incidence of NEC . METHODS: Daily doses of 250 million live L . acidophilus and 250 million B . infantis were given to all 1237 newborns (both inpatients and transfer patients) admitted to the unit during 1 year, until they were discharged from the hospital . In this study, 1282 patients hospitalized during the previous year were used as controls . RESULTS: There were no complications attributed to the daily administration of L . acidophilus and B . infantis . The study groups were compared for place of origin, clinical, and demographic variables, and there was no statistically significant difference in those variables . In the historic control group, there were 85 NEC cases compared to 34 cases in the group that received probiotic prophylaxis (P < 0.0002) . In the historic control group, there were 35 NEC-associated fatalities compared to 14 fatalities in the group that received probiotic prophylaxis (P < 0.005) . CONCLUSIONS: The positive results in this study support the need for further investigation of bacterial colonization and its role in NEC. Appl Microbiol Biotechnol, 1999 Nov, 52(5), 681 - 8 Transglycosidase activity of Bifidobacterium adolescentis DSM 20083 alpha-galactosidase; Van Laere KM et al.; Bifidobacterium adolescentis, a gram-positive saccharolytic bacterium found in the human colon, can, alongside other bacteria, utilise stachyose in vitro thanks to the production of an alpha-galactosidase . The enzyme was purified from the cell-free extract of Bi . adolescentis DSM 20083T . It was found to act with retention of configuration (alpha-->alpha), releasing alpha-galactose from p-nitrophenyl galactoside . This hydrolysis probably operates with a double-displacement mechanism, and is consistent with the observed glycosyltransferase activity . As alpha-galactosides are interesting substrates for bifidobacteria, we focused on the production of new types of alpha-galactosides using the transgalactosylation activity of Bi . adolescentis alpha-galactosides . Starting from melibiose, raffinose and stachyose oligosaccharides could be formed . The transferase activity was highest at pH 7 and 40 degrees C . Starting from 300 mM melibiose a maximum yield of 33% oligosaccharides was obtained . The oligosaccharides formed from melibiose were purified by size-exclusion chromatography and their structure was elucidated by NMR spectroscopy in combination with enzymatic degradation and sugar linkage analysis . The trisaccharide alpha-D-Galp-(1-->6)-alpha-D-Galp-(1-->6)-D-Glcp and tetrasaccharide alpha-D-Galp-(1-->6)-alpha-D-Galp- (1-->6)-alpha-D-Galp-(1-->6)-D-Glcp were identified, and this indicates that the transgalactosylation to melibiose occurred selectively at the C-6 hydroxyl group of the galactosyl residue . The trisaccaride alpha-D-Galp-(1-->6)-alpha- D-Galp-(1-->6)-D-Glcp formed could be utilised by various intestinal bacteria, including various bifidobacteria, and might be an interesting pre- and synbiotic substrate. Acta Paediatr Suppl, 1999 Aug, 88(430), 47 - 57 Factors controlling the bacterial colonization of the intestine in breastfed infants; Orrhage K et al.; This article summarizes the published data on the intestinal microflora in breastfed infants published during the last 15 y . Enterobacteria and enterococci are found in high numbers in most infants during the first week of life . Bifidobacteria and Bacteroides spp . are found in increasing numbers at the following weeks . The intestinal microflora in breastfed infants can also be followed by different biochemical parameters . Acetic acid is found in higher concentrations in breastfed than in formula-fed infants . Degradation of mucin starts later in breastfed than in formula-fed infants . The conversion of cholesterol to coprostanol is also delayed by breastfeeding . Geographical differences in the composition of the intestinal microflora in infants have been reported, i.e . enterobacteria, enterococci, bifidobacteria, lactobacilli and bacteroides show different occurrences in developed and developing countries . There are minor differences in the infant's intestinal microflora due to breastfeeding or/and formula feeding. Eur J Gastroenterol Hepatol, 1999 Nov, 11(11), 1195 - 8 Probiotics: established effects and open questions; von Wright A et al.; Human intestinal microflora is a complex ecosystem with hundreds of bacterial species . Its metabolic functions and interactions with the host probably affect the human health and well being, but these effects are extremely difficult to study . However, for about 100 years, the idea of modifying the composition of colonic flora by consuming viable bacteria in order to improve the quality of life and to prevent and treat intestinal disorders has had some popularity . Solid clinical data have usually been lacking to support the health claims associated with these so-called probiotics . The situation, however, is rapidly changing . Competent clinical studies are accumulating, showing that specific probiotic microbes, mainly lactic acid bacteria and bifidobacteria, can alleviate or prevent diverse intestinal disorders and reduce the risk of some intestinal diseases . Some indication of the mechanisms of action can also be deduced from the data available, while rapidly developing molecular biological methods offer new tools to verify the survival of the probiotics in the gut and the subsequent adhesion to mucosae . While development of foods containing probiotic bacteria has a great potential for the food industry and can be expected to positively affect the general health of the population, safety considerations have to be taken into account while introducing new species or strains without a previous history of safe food use. Appl Environ Microbiol, 1999 Nov, 65(11), 5173 - 6 A new selective medium for Bifidobacterium spp; Nebra Y et al.; A new selective antibiotic-free medium for Bifidobacterium spp . is defined . This medium has lactulose as the main carbon source and includes methylene blue, propionic acid, and lithium chloride as inhibitors of some related bacterial species . The low pH of the medium contributes to the inhibition of the growth of Enterobacteriaceae . This new selective medium has a simple composition, and the level of recovery it yields is similar to those yielded by nonselective media for Bifidobacterium strains . It could thus be used for routine analysis in environmental or food microbiology. Appl Environ Microbiol, 1999 Nov, 65(11), 4848 - 54 In vitro utilization of amylopectin and high-amylose maize (Amylomaize) starch granules by human colonic bacteria; Wang X et al.; It has been well established that a certain amount of ingested starch can escape digestion in the human small intestine and consequently enters the large intestine, where it may serve as a carbon source for bacterial fermentation . Thirty-eight types of human colonic bacteria were screened for their capacity to utilize soluble starch, gelatinized amylopectin maize starch, and high-amylose maize starch granules by measuring the clear zones on starch agar plates . The six cultures which produced clear zones on amylopectin maize starch- containing plates were selected for further studies for utilization of amylopectin maize starch and high-amylose maize starch granules A (amylose; Sigma) and B (Culture Pro 958N) . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to detect bacterial starch-degrading enzymes . It was demonstrated that Bifidobacterium spp., Bacteroides spp., Fusobacterium spp., and strains of Eubacterium, Clostridium, Streptococcus, and Propionibacterium could hydrolyze the gelatinized amylopectin maize starch, while only Bifidobacterium spp . and Clostridium butyricum could efficiently utilize high-amylose maize starch granules . In fact, C . butyricum and Bifidobacterium spp . had higher specific growth rates in the autoclaved medium containing high-amylose maize starch granules and hydrolyzed 80 and 40% of the amylose, respectively . Starch-degrading enzymes were cell bound on Bifidobacterium and Bacteroides cells and were extracellular for C . butyricum . Active staining for starch-degrading enzymes on SDS-PAGE gels showed that the Bifidobacterium cells produced several starch-degrading enzymes with high relative molecular (M(r)) weights (>160,000), medium-sized relative molecular weights (>66,000), and low relative molecular weights (<66,000) . It was concluded that Bifidobacterium spp . and C . butyricum degraded and utilized granules of amylomaize starch. Antonie Van Leeuwenhoek, 1999 Jul-Nov, 76(1-4), 383 - 9 Immunomodulatory function of lactic acid bacteria; Yasui H et al.; Using mice, we found that oral administration of Bifidobacterium breve YIT4064 (B . breve) activated the humoral immune system, augmented anti-rotavirus IgA production or anti-influenza virus (IFV) IgG production and protected against rotavirus infection or influenza infection, respectively . Furthermore, when the B . breve was given to infants from an infant home, there was a significant reduction of the frequency of rotavirus shedding in stool samples during the administration of the bacteria . It was also found, again using mice, that oral administration of Lactobacillus casei strain Shirota (LcS) stimulated type 1 helper T (Th1) cells, activated the cellular immune system and inhibited incidence of tumors and IgE production . These results demonstrated that these two strains of lactic acid bacteria modulated the different immune systems each in its own way and prevented against various diseases. J Basic Microbiol, 1999, 39(4), 243 - 51 Production and shelf-life studies of low cost beverage with soymilk, buffalo cheese whey and cow milk fermented by mixed cultures of Lactobacillus casei ssp . shirota and Bifidobacterium adolescentis; Macedo RF et al.; A study was performed to develop a fermented milk beverage with the aim to increase the potential application of buffalo cheese whey and soymilk . A mixed substrate was prepared by selective combination, which contained buffalo cheese whey 35%, soymilk 30% and cow milk 35% . The substrate mixture was fermented by a mixed culture of Lactobacillus casei shirota and Bifidobacterium adolescentis at 37 degrees C for 8 h keeping a 1:1.5 proportion between the lactic and bifidobacteria within a 5% (v/v) inoculum size . The fermented beverage was lightly extra-flavoured with vanilla essence and subjected to chemical, microbiological and sensory evaluations during storage for 28 days at 4 degrees C . Except a slight variation in the acidity, no other properties changed even after 28 days . There were no contaminating organisms (Salmonella and coliforms), which indicated the sanitary and hygienic conditions of the processing and the viable cells of the bacterial strains was well within recommended limits (6.8 x 10(8) cells for L . casei and 2.3 x 10(7) cells for Bifidobacterium) . No negative changes were found in the sensory characteristics of the beverage allowing its good acceptability in all during the storage period. Antonie Van Leeuwenhoek, 1999 May, 75(4), 329 - 33 Sequence analysis of 16S rRNA genes amplified from two ribosomal RNA gene clusters of Bifidobacterium bifidum; Yoon JH et al.; Two rRNA gene clusters were detected in the genome of Bifidobacterium bifidum KCTC 3202T using Southern blot analysis . To analyse the sequences of the 16S rRNA genes from rrnA and rrnB, 16S rDNAs were amplified by PCR using DNA fragments purified from gel slices containing each of the rRNA gene clusters . The amplified 16S rDNAs from rrnA and rrnB were cloned into vectors and three clones of each gene sequenced . The resultant sequences were confirmed by direct sequencing of the 16S rDNAs from rrnA and rrnB . Sequence differences were not found between rrnA and rrnB in 1488 bp of the 16S rRNA genes. J Dairy Sci, 1999 Sep, 82(9), 1869 - 76 Bifidobacterium bifidum exhibits a lipopolysaccharide-like mitogenic activity for murine B lymphocytes; Ko EJ et al.; To determine the role of bifidobacteria in the systemic and mucosal antibody response, we examined the direct modulatory effect of bifidobacteria on the synthesis of antibodies by murine spleen B cells . Whole spleen B cells were cultured with Bifidobacterium bifidum or Clostridium perfringens (Welch's bacilli, negative control), and antibody synthesis was measured by ELISA and enzyme-linked immunospot assay . The B . bifidum, but not C . perfringens, substantially increased total secretion of major immunoglobulin (Ig) isotypes and the number of IgA-secreting cells . In addition, B . bifidum increased proliferation of spleen cells by threefold, and C . perfringens had little to diminishing effect on the cells . These results indicate that B . bifidum increased Ig synthesis through its mitogenic influence on B cells . Further, B . bifidum induced spleen B cells to be reactive to transforming growth factor-beta 1 and interleukin-5 and resulted in increased surface IgA expression (approximately threefold) and total IgA production (> 20-fold) but not increased production of IgM and IgG2a isotypes . Together, these studies indicate that B . bifidum can act as a lipopolysaccharide-like polyclonal activator for B cells . Furthermore, that bifidobacteria enable B cells to respond to transforming growth factor-beta 1 and interleukin-5 for the IgA production has important implications for the primary defense against pathogens in the gastrointestinal tract. Dig Dis Sci, 1999 Jul, 44(7), 1485 - 93 Probiotic preparations dose-dependently increase net production rates of organic acids and decrease that of ammonia by pig cecal bacteria in batch culture; Sakata T et al.; We tested probiotic preparations containing Bifidobacterium, Enterococcus, or Lactobacillus to see if they affect production of organic acids and ammonia by mixed cecal bacteria . Four preparations (Bibalance, Neorakuton, Yakult Seichoyaku, or Yakult Seichoyaku BL) were digested with HC and pancreatin before adding to batch cultures of 50% pig cecal contents . Concentrations of organic acids and ammonia in the culture were quantified . Two preparations stimulated the net production of short-chain fatty acids (SCFA) during the first 8 hr of incubation . From 8 to 24 hr of incubation, all preparations accelerated the net production of SCFA, succinic and lactic acids, and three preparations slowed the net production of isovaleric acid and ammonia, all dose-dependently . The above results indicate that the preparations tested accelerated the breakdown of hard-to-degrade carbohydrate(s) and, possibly thereby, decreased the breakdown of proteinous materials or increased bacterial cell body synthesis in a mixed culture of cecal bacteria. Zentralbl Veterinarmed B, 1999 Aug, 46(6), 369 - 73 Evaluation of selective media for bifidobacteria in poultry and rabbit caecal samples; Rada V et al.; Five media were evaluated to determine their selectivity for Bifidobacterium sp . in hen and rabbit caecal samples . The colonies arising on the plates inoculated with the caecal samples were Gram stained and screened for the presence of fructose-6-phosphate phosphoketolase activity . Rogosa agar modified by the addition of cysteine-hydrochloride (0.05% w/v), Beeren's agar (with 5 ml/l of propionic acid as a selective agent), BS 2 agar (containing per one litre sodium propionate 15 g, lithium chloride 3 g, paromomycin sulphate 50 mg, neomycin sulphate 200 mg), and Wilkins-Chalgren agar (MW) modified by the addition of acetic acid (1 ml/l) and mupirocin (100 mg/l) were selective for Bifidobacterium sp . from rabbit caecal samples . In contrast, only MW medium was suitable for the isolation and enumeration of bifidobacteria in hen caecal samples . In conclusion, the results suggest that MW agar showed the greatest selectivity . A further advantage of this medium is its case of preparation . Therefore this agar could contribute to the study of the effects of the ingestion both probiotics and prebiotics . Finally, it could be noted that the bifidobacteria selective media should be chosen in respect of the animal species origin of the sample tested. Rocz Panstw Zakl Hig, 1999, 50(1), 89 - 95 {Nutritional importance of oligosaccharides}; Amarowicz R; Oligosaccharides are widely distributed in higher plants, especially leguminous seeds . This review described the structure of galactooligosaccharides, fructooligosaccharides and raffinose-type oligosacchariddes . Flatulance--causes, relation to diet and composition of intenstinal gas are discussed . Emphasis is placed upon the fact that ingestion of oligosaccharides increases the bifidobacteria population in the colon, which in turn contributes to human health in many ways. Gastroenterology, 1999 Sep, 117(3), 577 - 83 Bifidobacterial supplementation reduces the incidence of necrotizing enterocolitis in a neonatal rat model; Caplan MS et al.; BACKGROUND & AIMS: Neonatal necrotizing enterocolitis (NEC) is a devastating gastrointestinal disease of premature infants partly caused by intestinal bacterial proliferation . Because bifidobacteria are thought to reduce the risk for intestinal disturbances associated with pathogenic bacterial colonization, we hypothesized that exogenous bifidobacterial supplementation to newborn rats would result in intestinal colonization and a reduction in the incidence of neonatal NEC . METHODS: Newborn rat pups were given Bifidobacterium infantis (10(9) organisms per animal daily), Escherichia coli, or saline control and exposed to the NEC protocol consisting of formula feeding (Esbilac; 200 cal . kg(-1) . day(-1)) and asphyxia (100% N(2) for 50 seconds followed by cold exposure for 10 minutes) . Outcome measures included stool and intestinal microbiological evaluation, gross and histological evidence of NEC, plasma endotoxin concentration, intestinal phospholipase A(2) expression, and estimation of intestinal mucosal permeability . RESULTS: Bifidobacterial supplementation resulted in intestinal colonization by 24 hours and appearance in stool samples by 48 hours . Bifidobacteria-supplemented animals had a significant reduction in the incidence of NEC compared with controls and E . coli-treated animals (NEC, 7/24 B . infantis vs . 19/27 control vs . 16/23 E . coli; P < 0.01) . Plasma endotoxin and intestinal phospholipase A(2) expression were lower in bifidobacteria-treated pups than in controls, supporting the role of bacterial translocation and activation of the inflammatory cascade in the pathophysiology of NEC . CONCLUSIONS: Intestinal bifidobacterial colonization reduces the risk of NEC in newborn rats. J Food Prot, 1999 Aug, 62(8), 905 - 12 Screening of bile resistance and bile precipitation in lactic acid bacteria and bifidobacteria; Kociubinski G et al.; A modification of the ecometric method was developed for a rapid screening of bile resistance in lactic acid bacteria and bifidobacteria . Validation of the MEM bile assay (modified ecometric method) was performed comparing the bile resistance index (RIbile) and the bile resistance ratio (R%) . Most Bifidobacterium strains assayed were bile sensitive (83.3%), while only 62.1% of lactobacilli showed that behavior . Some bifidobacterial strains (55.6%) showed a crystalline precipitate when grown on solid medium supplemented with 0.5% ox bile . The crystalline structures produced by B . pseudolongum CIDCA 531 were isolated and analyzed by optical and scanning electron microscopy, thin-layer chromatography, melting point, and specific cholesterol reactions . Those studies confirmed the presence of cholesterol in these crystalline structures . On the other hand, none of the lactobacilli and streptococci studied had the ability to produce crystalline precipitates. Br J Nutr, 1999 Feb, 81(2), 121 - 32 Functional food properties of non-digestible oligosaccharides: a consensus report from the ENDO project (DGXII AIRII-CT94-1095); Van Loo J et al.; This paper results from the final phase of the ENDO project (DGXII AIRII-CT94-1095), a European Commission-funded project on non-digestible oligosaccharides (NDO) . All participants in the programme met to perform a consensus exercise on the possible functional food properties of NDO . Topics studied during the project (including a workshop on probiotics and prebiotics) and related aspects, for which considerable evidence has been generated recently, were evaluated on the basis of existing published scientific evidence . There was a general consensus that: (1) there is strong evidence for a prebiotic effect of NDO in human subjects . A prebiotic effect was defined as a food-induced increase in numbers and/or activity predominantly of bifidobacteria and lactic acid bacteria in the human large intestine; (2) there is strong evidence for the impact that NDO have on bowel habit; (3) there is promising evidence that consumption of inulin-type fructans may result in increased Ca absorption in man; (4) there are preliminary indications that inulin-type fructans interact with the functioning of lipid metabolism; (5) there is preliminary evidence in experimental animals of a preventive effect against colon cancer . Human nutrition studies are needed to substantiate these findings . It was concluded that the nutritional properties of NDO may prove to be a key issue in nutritional research in the future. Biochim Biophys Acta, 1999 Aug 5, 1428(2-3), 241 - 50 Role of 2-amino-3-carboxy-1,4-naphthoquinone, a strong growth stimulator for bifidobacteria, as an electron transfer mediator for NAD(P)(+) regeneration in Bifidobacterium longum; Yamazaki S et al.; 2-Amino-3-carboxy-1,4-naphthoquinone (ACNQ) is a novel growth stimulator for bifidobacteria . The role of ACNQ as a mediator of the electron transfer from NAD(P)H to dioxygen (O(2)) and hydrogen peroxide (H(2)O(2)), proposed in our previous paper, was examined using the cell-free extract and whole cells of Bifidobacterium longum . Continuous monitoring of ACNQ, O(2) and H(2)O(2) by several amperometric techniques has revealed that ACNQ works as a good electron acceptor of NAD(P)H diaphorase and that the reduced form of ACNQ is easily autoxidized and also acts as a better electron donor of NAD(P)H peroxidase than NAD(P)H . The generation of H(2)O(2) by B . longum under aerobic conditions is effectively suppressed in the presence of ACNQ . These ACNQ-mediated reactions would play roles as NAD(P)(+)-regeneration processes . The accumulation of ACNQ in the cytosol has been also suggested . These characteristics of ACNQ seem to be responsible for the growth stimulation of bifidobacteria . Vitamin K(3), which has an extremely low growth-stimulating activity and was used as a reference compound, exhibits much lower activity as an electron transfer mediator . The difference in the activity is discussed in terms of the redox potential and partition property of the quinones. Res Microbiol, 1999 Jun, 150(5), 343 - 50 Molecular monitoring of human intestinal Bifidobacterium strain diversity; Mangin I et al.; Predominant Bifidobacterium strains belonging to the intestinal flora of four human volunteers were isolated on selective medium before and after eight days of treatment with oral amoxicillin-clavulanic acid (Augmentin) . These antimicrobial agents are known to be strongly active against the genus Bifidobacterium . A fifth volunteer did not receive the antibiotics and was considered as a control . Bifidobacteria were characterized by hybridizing a ribosomal 23S DNA probe onto their EcoRV restriction patterns, and were identified by comparing the ribosomal patterns obtained to collection strains . A total of 17 distinct ribosomal patterns and 23 distinct pattern types were revealed for the 95 isolates tested . Each type characterized was correlated with a specific ribosomal pattern associated with a specific total restriction pattern . Similar-sized molecular bands permitted isolates to be unambiguously discriminated into the species B . longum, B . bifidum, and B . adolescentis . This study enabled us to show considerable strain variability among individuals . Three months after penicillin ingestion, no significant changes were observed in Bifidobacterium flora . Each flora remained relatively stable for strain composition over time, with some slight variations also detected in our control subject. Nahrung, 1999 Jun, 43(3), 148 - 53 Probiotics in the new millennium; Vaughan EE et al.; Beneficial effects on human health by specific probiotic microorganisms such as prevention of gastrointestinal tract infections immune stimulation, and balancing of the intestinal microflora have been established in numerous clinical trials . The successful probiotic strains which are mainly members of Lactobacillus and more recently Bifidobacterium naturally found in the human intestinal tract, have been traditionally incorporated into fermented milk products but have excellent potential for further inclusion in functional foods and health-related products . While the health claims are generally accepted by both scientists and consumers, often the molecular mechanisms underlying the probiotic properties remains controversial . Further progress concerning the molecular basis of probiotic traits will give vital reinforcement to the probiotic concept and is a prerequisite for rational development into further applications . In this review, current research on the genetics of properties of the intestinal lactobacilli that may contribute to the activity and effe