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TRUE POSITIVES:
0 ('>T', 'TI  - Murein hydrolase activity of surface layer proteins from Lactobacillus acidophilus against Escherichia coli.', 'AB  - The aim of this study was to investigate the murein hydrolase activities of the surface layer proteins (SLPs) from two strains of Lactobacillus acidophilus using zymography. The influence of these hydrolase activities on Escherichia coli ATCC 43893 was also evaluated by analysing their growth curve, cell morphology and physiological state. After the incubation of E. coli with SLPs, growth was inhibited, the number of viable cells was significantly reduced, examination by transmission electron microscopy showed that the cell wall was damaged and flow cytometry results indicated that the majority of the cells were sublethally injured. All of these results suggested that the SLPs of both L. acidophilus strains possessed murein hydrolase activities that were sublethal to E. coli cells.')

4 ('>T', 'TI  - Cocktails of probiotics pre-adapted to multiple stress factors are more robust under simulated gastrointestinal conditions than their parental counterparts and exhibit enhanced antagonistic capabilities against Escherichia coli and Staphylococcus aureus.', 'AB  - BACKGROUND: The success of the probiotics in delivery of health benefits depends  on their ability to withstand the technological and gastrointestinal conditions; hence development of robust cultures is critical to the probiotic industry. Combinations of probiotic cultures have proven to be more effective than the use of single cultures for treatment and prevention of heterogeneous diseases. We investigated the effect of pre- adaptation of probiotics to multiple stresses on their stability under simulated gastrointestinal conditions and the effect of their singular as well as their synergistic antagonistic effect against selected enteric pathogens. METHODS: Probiotic cultures were inoculated into MRS broth adjusted to pH 2 and incubated for 2 h at 37 degrees C. Survivors of pH 2 were subcultured into 2% bile acid for 1 h at 37 degrees C. Cells that showed growth after exposure to 2% bile acid for 1 h were finally inoculated in fresh MRS broth and incubated at 55 degrees C for 2 h. The cells surviving were then used as stress adapted cultures. The adapted cultures were exposed to simulated gastrointestinal conditions and their non- adapted counterparts were used to compare the effects of stress adaptation. The combination cultures were tested for their antipathogenic effects on Escherichia coli and Staphylococcus aureus. RESULTS: Acid and bile tolerances of most of the stress-adapted cells were higher than of the non-adapted cells. Viable counts of all the stress-adapted lactobacilli and Bifidobacterium longum LMG 13197 were higher after sequential exposure to simulated gastric and intestinal fluids. However, for B. longum Bb46 and B. bifidum LMG 13197, viability of non-adapted cells was higher than for adapted cells after exposure to these fluids. A cocktail containing L. plantarum + B. longum Bb46 + B. longum LMG 13197 best inhibited S. aureus while E. coli was best inhibited by a combination containing L. acidophilus La14 150B + B. longum Bb46 + B. bifidum LMG 11041. A cocktail containing the six non- adapted cultures was the least effective in inhibiting the pathogens. CONCLUSION: Multi-stress pre-adaptation enhances viability of probiotics under simulated gastrointestinal conditions; and formulations containing a mixture of multi stress-adapted cells exhibits enhanced synergistic effects against foodborne pathogens.')

5 ('>T', 'TI  - Preparation and application characteristics of microencapsulated Lactobacillus acidophilus as probiotics for dogs.', 'AB  - In this article, preparation and application characteristics of microencapsulated Lactobacillus acidophilus were investigated. Results indicated that the optimum condition for preparation of micro encapsulation were 10% (w/v) wall material and the temperature of 20 degrees C, respectively. Many micropores in the porous starch micro particles was also observed by Scanning Electron Microscope. Furthermore, the released cell counts were increase from 2.43 log cfu/g to 9.17 log cfu/g for the time prolong to 3h in the simulated colonic pH solution. On the other hand, the visible cells of Lactobacillus acidophilus in the dog feces on the 10th day after the probiotics feeding was improve about 34.8% compare to the before feeding, which was decrease about 24.6%for Escherichia coli. Furthermore, the content of is ovaleric acid, indole and 3-methylindole, putrefactive substances in dog feces, before feeding were reduce 24%, 16% and 45% in dog feces on the 10th day after feeding compared to that before feeding, respectively. Micro encapsulation can be considered a useful technology to provide the protection for Lactobacillus acidophilus and better application effective.')

7 ('>T', 'TI  - Effects of Lactobacillus acidophilus dietary supplementation on the performance,  intestinal barrier function, rectal microflora and serum immune function in weaned piglets challenged with Escherichia coli lipopolysaccharide.', 'AB  - This study was conducted with a lipopolysaccharide (LPS)-challenged piglet model  to determine the effects of diets containing Lactobacillus acidophilus on the performance, intestinal barrier function, rectal microflora and serum immune function. A total of 150 piglets (initial body weight (BW) 7.53 +/- 0.21 kg) were allotted to one of the following diets, including a basal diet, a basal diet supplemented with 250 mg/kg Flavomycin, or basal diet plus 0.05, 0.1 or 0.2 % L. acidophilus. On day 28 of the trial, the pigs were given an intraperitoneal injection of LPS (200 mug/kg body weight) followed by blood collection 3 h later. Diets with either antibiotics, 0.1 or 0.2 % Lactobacillus increased (P < 0.05) the final BW and decreased (P < 0.05) feed gain ratio (F/G) compared with the control group. Pigs fed diets containing antibiotic or Lactobacillus had greater average daily gain (ADG) (P < 0.05) than pigs fed the control diet. The rectal content Lactobacillus counts for pigs fed diet containing Lactobacillus were significant higher (P < 0.01) than those fed antibiotic or control diet. Feeding the Lactobacillus diets decreased the Escherichia coli counts of rectal content (P < 0.01). Pigs fed diets containing 0.1 or 0.2 % Lactobacillus decreased serum DAO activity (P < 0.05) compared with pigs fed the control diet. Serum IL-10 concentration was enhanced in pigs fed the diet with Lactobacillus compared to pigs fed the control diet and antibiotic diet. Feeding a diet with Lactobacillus reduced (P < 0.05) IFN-gamma concentration compared to the control diet. Inclusion of Lactobacillus in diets fed to pigs reduced TNF-alpha concentration compared with pigs fed no Lactobacillus (P < 0.05). These results indicate that feeding with L. acidophilus improved growth performance and protected against LPS-induced inflammatory status.')

12 ('>T', 'TI  - Functional characterization of probiotic surface layer protein-carrying Lactobacillus amylovorus strains.', 'AB  - BACKGROUND: Adhesiveness to intestinal epithelium, beneficial immunomodulating effects and the production of pathogen-inhibitory compounds are generally considered as beneficial characteristics of probiotic organisms. We showed the potential health-promoting properties and the mechanisms of probiotic action of seven swine intestinal Lactobacillus amylovorus isolates plus the type strain (DSM 20531T) by investigating their adherence to porcine intestinal epithelial cells (IPEC-1) and mucus as well as the capacities of the strains to i) inhibit the adherence of Escherichia coli to IPEC-1 cells, ii) to produce soluble inhibitors against intestinal pathogens and iii) to induce immune signaling in dendritic cells (DCs). Moreover, the role of the L. amylovorus surface (S) -layers - symmetric, porous arrays of identical protein subunits present as the outermost layer of the cell envelope - in adherence to IPEC-1 cells was assessed using a novel approach which utilized purified cell wall fragments of the strains as carriers for the recombinantly produced S-layer proteins. RESULTS: Three of the L. amylovorus strains studied adhered to IPEC-1 cells, while four strains inhibited the adherence of E. coli, indicating additional mechanisms other than competition for binding sites being involved in the inhibition. None of the strains bound to porcine mucus. The culture supernatants of all of the strains exerted inhibitory effects on the growth of E. coli, Salmonella, Listeria and Yersinia, and a variable, strain-dependent induction was observed of both pro- and anti-inflammatory cytokines in human DCs. L. amylovorus DSM 16698 was shown to carry two S-layer-like proteins on its surface in addition to the major S-layer protein SlpA. In contrast to expectations, none of the major S-layer proteins of the IPEC-1 -adhering strains mediated bacterial adherence. CONCLUSIONS: We demonstrated adhesive and significant pathogen inhibitory efficacies among the swine intestinal L. amylovorus strains studied, pointing to their potential use as probiotic feed supplements, but no independent role could be demonstrated for the major S-layer proteins in adherence to epithelial cells. The results indicate that many intestinal bacteria may coexist with and confer benefits to the host by mechanisms not attributable to adhesion to epithelial cells or mucus.')

16 ('>T', 'TI  - Influence of bovine lactoferrin on the growth of selected probiotic bacteria under aerobic conditions.', 'AB  - Bovine lactoferrin (bLf) is a natural glycoprotein, and it shows broad-spectrum antimicrobial activity. However, reports on the influences of bLf on probiotic bacteria have been mixed. We examined the effects of apo-bLf (between 0.25 and 128 mg/mL) on both aerobic and anaerobic cultures of probiotics. We found that bLf had similar effects on the growth of probiotics under aerobic or anaerobic conditions, and that it actively and significantly (at concentrations of >0.25 mg/mL) retarded the growth rate of Bifidobacterium bifidum (ATCC 29521), B. longum (ATCC 15707), B. lactis (BCRC 17394), B. infantis (ATCC 15697), Lactobacillus reuteri (ATCC 23272), L. rhamnosus (ATCC 53103), and L. coryniformis (ATCC 25602) in a dose-dependent manner. Otherwise, minimal inhibitory concentrations (MICs) were 128 or >128 mg/mL against B. bifidum, B. longum, B. lactis, L. reuteri, and L. rhamnosus (ATCC 53103). With regard to MICs, bLf showed at least four-fold lower inhibitory effect on probiotics than on pathogens. Intriguingly, bLf (>0.25 mg/mL) significantly enhanced the growth of Rhamnosus (ATCC 7469) and L. acidophilus (BCRC 14065) by approximately 40-200 %, during their late periods of growth. Supernatants produced from aerobic but not anaerobic cultures of L. acidophilus reduced the growth of Escherichia coli by about 20 %. Thus, bLf displayed a dose-dependent inhibitory effect on the growth of most probiotic strains under either aerobic or anaerobic conditions. An antibacterial supernatant prepared from the aerobic cultures may have significant practical use.')

17 ('>T', 'TI  - Probiotic assessment of Enterococcus durans 6HL and Lactococcus lactis 2HL isolated from vaginal microflora.', 'AB  - Forty-five lactic acid bacteria (LAB) were isolated from the vaginal specimens of healthy fertile women, and the identities of the bacteria were confirmed by sequencing of their 16S rDNA genes. Among these bacteria, only four isolates were able to resist and survive in low pH, bile salts and simulated in vitro digestion conditions. Lactococcus lactis 2HL, Enterococcus durans 6HL, Lactobacillus acidophilus 36YL and Lactobacillus plantarum 5BL showed the best resistance to these conditions. These strains were evaluated further to assess their ability to adhere to human intestinal Caco-2 cells. Lactococcus lactis 2HL and E. durans 6HL were the most adherent strains. In vitro tests under neutralized pH proved the antimicrobial activity of both strains. Results revealed that the growth of Escherichia coli O26, Staphylococcus aureus and Shigella flexneri was suppressed by both LAB strains. The antibiotic susceptibility tests showed that these strains were sensitive to all nine antibiotics: vancomycin, tetracycline, ampicillin, penicillin, gentamicin, erythromycin, clindamycin, sulfamethoxazole and chloramphenicol. These data suggest that E. durans 6HL and Lactococcus lactis 2HL could be examined further for their useful properties and could be developed as new probiotics.')

20 ('>T', 'TI  - The use of direct-fed microbials to reduce shedding of Escherichia coli O157 in beef cattle: a systematic review and meta-analysis.', 'AB  - Human illness due to infections with Escherichia coli O157 is a serious health concern. Infection occurs through direct contact with infected animals or their faeces, through contaminated food or water and/or through person-to-person transmission. A reduction in faecal E. coli O157 shedding in cattle might reduce the burden of human infections. We used systematic review and meta-analysis to assess the efficacy of direct-fed microbials (DFM), compared with placebo or no treatment, fed during the pre-harvest stage of production in reducing faecal E. coli O157 shedding in beef cattle during field trials. Four electronic databases, Nebraska Beef Reports and review article reference lists were searched. A total of 16 publications assessing faecal shedding at the end of the trial and/or throughout the trial period were included. The majority of publicly disseminated trials evaluated the prevalence of E. coli O157 faecal shedding; only two evaluated the concentration of organisms in faeces. The prevalence of faecal E. coli O157 shedding in cattle is significantly reduced by DFM treatments (summary effect size for all DFM - OR = 0.46; CI = 0.36-0.60). The DFM combination Lactobacillus acidophilus (NP51) and Propionibacterium freudenreichii (NP24) was more efficacious in reducing the prevalence of faecal E. coli O157 shedding at the time of harvest and throughout the trial period compared with the group of other DFM, although this difference was not statistically significant. Furthermore, we found that the combination [NP51 and NP24] treatment was more efficacious in reducing the prevalence of faecal E. coli O157 shedding at the time of harvest and throughout the trial period when fed at the dose of 10(9) CFU/animal/day than any lesser amount, although this difference was not statistically significant. Feeding beef cattle DFM during the pre-harvest stage of production reduces the prevalence of E. coli O157 faecal shedding and might effectively reduce human infections.')

26 ('>T', 'TI  - Effects of multistrain probiotics on growth performance, apparent ileal nutrient  digestibility, blood characteristics, cecal microbial shedding, and excreta odor contents in broilers.', 'AB  - This experiment was conducted to investigate the efficacy of Lactobacillus acidophilus, Bacillus subtilis, and Clostridium butyricum supplementation in broilers. A total of 400 one-day-old mixed sex Ross 308 broilers with an initial average BW of 46 +/- 0.5 g were randomly allotted into 4 treatments with 5 replicate pens per treatment and 20 broilers in each pen for 35 d. Dietary treatments were (1) an antibiotic-free diet (CON), (2) CON + 5 mg/kg of avilamycin, (3) CON + 1 x 10(5) cfu of multistrain probiotics/kg of diet (P1), and (4) CON + 2 x 10(5) cfu of multistrain probiotics/kg of diet (P2). Broilers fed the P1 and P2 diets had greater BW gain than broilers fed the CON diet during d 22 to 35 (P = 0.01) and overall (P = 0.02). Feed conversion ratios in P1 and P2 were decreased (P = 0.03) compared with that in CON from d 22 to 35. Ileal digestibility of most essential amino acids, with the exception of His and Phe, were increased (P < 0.05) in P1 and P2 compared with CON. Serum IgA and IgM concentrations in P2 were higher (P < 0.05) than those in CON. The cecal Lactobacillus numbers were increased (P = 0.02), and the counts of Escherichia coli were decreased (P = 0.03) in P1 and P2 compared with CON. Dietary supplementation with multistrain probiotics decreased (P < 0.05) the excreta NH3 content compared with the CON. In conclusion, dietary supplementation with multistrain probiotics improved broiler growth performance, ileal amino acids digestibility, and humoral immunity. Furthermore, the probiotics decreased the cecal numbers of E. coli and decreased the NH3 content of excreta.')

27 ('?T', 'TI  - Characterization of surface layer proteins and its role in probiotic properties of three Lactobacillus strains.', 'AB  - The objective of this study was the characterization of the surface layer proteins (SLPs) and their functional role in the probiotic activity of Lactobacillus helveticus fb213, L. acidophilus fb116 and L. acidophilus fb214. SLPs were extracted and identified by SDS-PAGE, circular dichroism spectra and LC-MS analysis. The results revealed that the molecular masses of the three proteins were 49.7 kDa, 46.0 kDa and 44.6 kDa, respectively. The secondary structures and amino acid compositions of the three proteins were found to be similar. After removing SLPs, the survival of the three lactobacilli in simulated gastric and intestinal juices was reduced by 2-3log as compared with survival of the intact cells. And the adhesion ability of the three strains to HT-29 cells decreased by 61%, 65% and 92%, respectively. SLPs also inhibited the adhesion and invasion of Escherichia coli ATCC 43893 to HT-29 cells. These results suggest that SLPs are advantageous barriers for lactobacilli in the gastrointestinal tract, and these proteins help make it possible for lactobacilli to serve their probiotic functions.')

29 ('>T', 'TI  - Potentialities of newly isolated Bacillus subtilis and Lactobacillus sp for curd  preparation and a comparative study of its physico-chemical parameters with other marketed curds.', 'AB  - Two Bacillus sp. were isolated from the local fermented milk and identified on the basis 16S rRNA sequence profile as Bacillus subtilis AKL1 and by biochemical process as Lactobacillus acidophilus AKL2. These isolates were used as fresh inoculums for curd preparation individually and in combinations. Different physico-chemical and therapeutic properties of the newly prepared curd were examined and compared with marketed local (sweet and sour) and branded (Mother Dairy and Thackar) curds. The total hydrolyzed peptides, free amino acids, lactic acid were significantly higher, whereas, total solid, ash content, syneresis and free reducing sugar were lower in the curd prepared by a mixture of AKL1 and AKL2 (0.5:0.5, v/v). The antioxidant activity against ABTS+, DPPH8, OH* and Fe3+ were also higher in the newly formulated curd. Polyphenols (85.5 microg/g), flavonoids (12.5 microg/g) and free aromatic amino acids contents were also higher in AKL1+AKL2. All these components prevent excess protein oxidation that was revealed by SDS-PAGE. The curd also exhibited potent antimicrobial activity against some entero-pathogens like Clostridium perfringens, Escherichia coli, Shigella dysentery, Vibrio cholerae and Staphylococcus aureus. It can be concluded that the combination of these Lactobacillus sp. will be a fruitful inoculum for the preparation of curd having better health promoting effects.')

31 ('>T', 'TI  - [Effect of metabolites of H2O2-producing lactobacilli on functional activity of lysozyme].', 'AB  - AIM: Study the effect of metabolites of H2O2-producing lactobacilli on enzymatic  and bactericidal activity of lysozyme. MATERIALS AND METHODS: 9 H2O2-producing vaginal lactobacilli, Micrococcus luteus NCTC 2665, Escherichia coli State Institute of Standardization and Control No 240367, Lactobacillus acidophilus Institute of Cellular and Intracellular Symbiosis No 37 were used. Ability of lactobacilli to produce H2O2 was evaluated by oxidation of tetramethylbenzidine by peroxidase. Lysozyme was modified by mixing with equal volumes of lactobacilli metabolites, metabolites of H2O2-producing vaginal lactobacilli previously treated with catalase were used in control. Lysozyme enzymatic activity was determined by speed of M. luteus lysis, bactericidal--by survivability of E. coli in Endo medium and L. acidophilus--in MRS medium. RESULTS: Decrease of enzymatic activity of lysozyme due to its contact with H2O2-producing lactobacilli metabolites was detected. This effect is accompanied by growth of bactericidal activity of lysozyme against E. coli and decrease against L. acidophilus. The degree of changes of enzymatic and bactericidal activity of lysozyme by lactobacilli metabolites depended on concentration of hydrogen peroxide in them. CONCLUSION: Modification of lysozyme by H2O2-producing lactobacilli metabolites resulting in opposite changes of its activity against autochthonous and allchthonous bacteria is one of the mechanisms of formation of stable microbial biocenosis in human organism.')

33 ('>T', 'TI  - Effect of Lactobacillus acidophilus and Bifidobacterium bifidum supplementation to standard triple therapy on Helicobacter pylori eradication and dynamic changes in intestinal flora.', 'AB  - To investigate Lactobacillus acidophilus (L. acidophilus) and Bifidobacterium bifidum (B. bifidum) supplementation to triple therapy for Helicobacter pylori (H. pylori) eradication and dynamic changes in intestinal flora in children with H. pylori infection. One hundred H. pylori-infected children were randomly assigned to two groups: treatment group (n = 43), standard triple anti-H. pylori therapy plus probiotics of L. acidophilus and B. bifidum for 2 weeks followed by taking probiotics for another 4 weeks; control group (n = 45), standard triple anti-H. pylori therapy for 6 weeks. After 6-week treatment, (1)(3)C-urease breath test was performed and side effects were monitored during the observation period. Quantitative PCR with 16S rRNA-gene-targeted species-specific primers was carried out for the analysis of human intestinal B. bifidum, L. acidophilus, and Escherichia coli (E. coli). As expected, treatment group could significantly enhance the H. pylori eradication rate (83.7 vs. 64.4 %, P < 0.05). B. bifidum, L. acidophilus, and E. coli showed no statistical difference before or after therapy in the treatment group. The number of B. bifidum and L. acidophilus was significantly decreased after 2-week treatment in the control group, but after 6-week treatment it significantly increased and nearly returned to the level before treatment. The number of E. coli increased significantly after 2-week treatment, while after 6-week treatment, it nearly decreased to the level before treatment. L. acidophilus and B. bifidum supplementation is effective for H. pylori eradication compared with triple therapy alone.')

47 ('>T', 'TI  - Comparative in vitro inhibition of urinary tract pathogens by single- and multi-strain probiotics.', 'AB  - PURPOSE: Multi-species probiotic preparations have been suggested as having a wide spectrum of application, although few studies have compared their efficacy with that of individual component strains at equal concentrations. We therefore tested the ability of 4 single probiotics and 4 probiotic mixtures to inhibit the urinary tract pathogens Escherichia coli NCTC 9001 and Enterococcus faecalis NCTC 00775. METHODS: We used an agar spot test to test the ability of viable cells to inhibit pathogens, while a broth inhibition assay was used to assess inhibition by cell-free probiotic supernatants in both pH-neutralised and non-neutralised forms. RESULTS: In the agar spot test, all probiotic treatments showed inhibition, L. acidophilus was the most inhibitory single strain against E. faecalis, L. fermentum the most inhibitory against E. coli. A commercially available mixture of 14 strains (Bio-Kult((R))) was the most effective mixture, against E. faecalis, the 3-lactobacillus mixture the most inhibitory against E. coli. Mixtures were not significantly more inhibitory than single strains. In the broth inhibition assays, all probiotic supernatants inhibited both pathogens when pH was not controlled, with only 2 treatments causing inhibition at a neutral pH. CONCLUSIONS: Both viable cells of probiotics and supernatants of probiotic cultures were able to inhibit growth of two urinary tract pathogens. Probiotic mixtures prevented the growth of urinary tract pathogens but were not significantly more inhibitory than single strains. Probiotics appear to produce metabolites that are inhibitory towards urinary tract pathogens. Probiotics display potential to reduce the incidence of urinary tract infections via inhibition of colonisation.')

50 ('?T', 'TI  - Evaluation of Lactobacillus strains for selected probiotic properties.', 'AB  - Eleven strains of Lactobacillus collected in the Culture Collection of Dairy Microorganisms (CCDM) were evaluated for selected probiotic properties such as survival in gastrointestinal fluids, antimicrobial activity, and competition with non-toxigenic Escherichia coli O157:H7 for adhesion on Caco-2 cells. The viable count of lactobacilli was reduced during 3-h incubation in gastric fluid followed by 3-h incubation in intestinal fluid. All strains showed antimicrobial activity and the three most effective strains inhibited the growth of at least 16 indicator strains. Antimicrobial metabolites of seven strains active against Lactobacillus and Clostridium indicator strains were found to be sensitive to proteinase K and trypsin, which indicates their proteinaceous nature. The degree of competitive inhibition of non-toxigenic E. coli O157:H7 adhesion on the surface of Caco-2 cells was strain-dependent. A significant decrease (P < 0.05) in the number of non-toxigenic E. coli O157:H7 adhering to Caco-2 cells was observed with all lactobacilli. Three strains were selected for additional studies of antimicrobial activity, i.e., Lactobacillus gasseri CCDM 215, Lactobacillus acidophilus CCDM 149, and Lactobacillus helveticus CCDM 82.')

53 ('>T', 'TI  - Yogurt containing bioactive molecules produced by Lactobacillus acidophilus La-5  exerts a protective effect against enterohemorrhagic Escherichia coli in mice.', 'AB  - An active fraction extracted from Lactobacillus acidophilus La5 cell-free spent medium (LAla-5AF) was incorporated in a dairy matrix and tested to assess its antivirulent effect against enterohemorrhagic Escherichia coli (EHEC). Mice in experimental groups were fed for 4 days with yogurt supplemented with LAla-5AF. On the fifth day, mice were challenged with a single dose (10(7) CFU per mouse) of E. coli O157:H7. The clinical manifestations of the infection were significantly less severe in mice fed the yogurt supplemented with LAla-5AF. EHEC attachment and colonization was attenuated by LAla-5AF. Tumor necrosis factor alpha production was down-regulated, which might indicate a protective effect in the kidney during EHEC infection. To investigate the mechanisms associated with the in vivo effects observed, LAla-5AF was tested by reverse transcription real-time PCR to confirm its effects on the expression of several virulence genes of EHEC O157. The results showed that these fractions were able to down-regulate several virulence genes of EHEC, including stxB2, qseA, luxS, tir, ler, eaeA, and hlyB.')

65 ('>T', 'TI  - Spoilage characteristics of traditionally packaged ground beef with added lactic  acid bacteria displayed at abusive temperatures.', 'AB  - Growth of pathogenic organisms such as Escherichia coli O157:H7 and Salmonella spp. can be inhibited in ground beef through the addition of certain lactic acid-producing bacteria (LAB; Lactobacillus acidophilus NP51, Lactobacillus crispatus NP35, Pediococcus acidilactici, and Lactococcus lactis ssp. lactis). This study evaluated the effects of LAB inclusion on the organoleptic and biochemical properties typically associated with spoilage in traditionally packaged ground beef displayed at abusive (10 degrees C) temperatures for 36 h. Trained and untrained panelist evaluations of lean color and off-odor, as well as instrumental color analyses, did not indicate an effect on spoilage traits due to LAB utilization (P > 0.05). However, display length affected each variable independently and was indicative of decreased stability and acceptability as display time (h) increased (P < 0.05). Thiobarbituric acid values were decreased for ground beef with added LAB (P < 0.05), but likely can be related to bacterial degradation of lipid oxidation by-products because no reduction in organoleptic traits due to oxidation was noted between treatments. Overall, LAB did not adversely influence the spoilage characteristics of traditionally packaged ground beef displayed at abusive temperatures for up to 36 h. Furthermore, biochemical and sensory indicators of spoilage were present for all treatments at the conclusion of display. Therefore, LAB can be added to ground beef in traditional packaging as a processing intervention without masking or delaying the expected spoilage characteristics.')

67 ('>T', 'TI  - Effect of Lactobacillus acidophilus KFRI342 on the development of chemically induced precancerous growths in the rat colon.', 'AB  - Lactobacillus acidophilus KFRI342, isolated from the Korean traditional food kimchi, was investigated for its suitability as a dietary probiotic. The effects of L. acidophilus KFRI342 on the development of chemically induced (1,2-dimethylhydrazine; DMH) precancerous cytological changes of the colon were investigated in rats. Forty-five male F344 rats were randomly divided into three dietary groups. The control group received a high-fat diet (HF), a second group received a high-fat diet containing the carcinogen (HFC), and a final group received a high-fat diet containing the carcinogen and L. acidophilus KFRI342 (HFCL). L. acidophilus KFRI342 was administered orally three times per week at 2x10(9) c.f.u. ml(-1). L. acidophilus KFRI342 treatments decreased the number of Escherichia coli in faecal samples, the enzyme activities of beta-glucuronidase and beta-glucosidase, and plasma triglyceride concentration compared to the HF and HFC treatments (P<0.05). L. acidophilus KFRI342 consumption also decreased the ratio of aberrant crypts to aberrant crypt foci incidence and the number of aberrant crypts in HFCL rats. Therefore, L. acidophilus showed potential probiotic activity as an inhibitor of DMH-induced symptoms in live rats. Our in vivo studies indicate that L. acidophilus from kimchi may be suitable as a probiotic for human use.')

77 ('>T', 'TI  - Therapeutic efficacy of Lactobacillus acidophilus against bacterial isolates from burn wounds.', 'AB  - BACKGROUND: Probiotics are live microorganisms which are mainly strains of Lactobacillus spp., Bifidobacterium spp. When administered in adequate amounts, these microorganisms offer a health benefit for the host. Probiotic organisms are also available commercially in milk, sour milk, ice cream and other foods. AIMS: To identify bacterial species isolated from burn wounds, and also to evaluate (In-vitro) the therapeutic efficacy of Lacto. acidophilus against these bacterial isolates. To compare this activity to other antibacterial agents which are used medically in the treatment of burn wound cases. MATERIALS AND METHODS: Burn wound swabs were obtained from 50 patients who had been admitted to hospitals in Baghdad during August to November 2009. These swabs were inoculated onto enriched and differential culture media. Subcultures were performed on selective media. The necessary biochemical tests were conducted and the organisms identified using standard procedures. Susceptibility of isolated pathogens to local isolates Lacto. Acidophilus (with 1x10(8) cells/mL) and 10 commonly used burn wounds antibiotics was examined using standard susceptibility testing. RESULTS: Ninety different organisms were isolated. Gram-positive cocci accounted for 16 (17.7%) and gram-negative bacilli for 74 (82.2%) bacterial isolates. Pseudomonas aeruginosa 30(33.3%) were the most commonly isolated organisms, followed by Escherichia coli, Enterobacter spp., Klebsiella spp., Proteus spp.(22.2,20,4.4,2.2%), respectively. Staphylococcus aureus isolates were performed in 8(8.8%). However, the incidence of Staphylococcus epidermidis was 2 (2.2%), while ss-haemolytic Streptococci was 4(4.4%). In susceptibility testing, Lacto. acidophilus had coverage against 90 (100%) of 74 gram-negative and 16 of gram-positive bacteria tested. The coverage of the remaining 10 antibacterial agents used was different in their activity (resistance or sensitivity), which ranged between 50-100%. CONCLUSION: The results of the study concluded that lactobacillus acidophilus concentration of 1x10(8) cells/mL had a high activity to inhibit the growth in-vitro of all pathogenic gram-positive and gram-negative bacteria, which cause burn wound infections. This indicated the therapeutic efficacy of lactobacillus acidophilus bacteria.')

78 ('>T', 'TI  - Probiotic yogurt effects on intestinal flora of patients with chronic liver disease.', 'AB  - BACKGROUND: Patients with chronic liver disease generally have intestinal flora imbalance that is related to the development and worsening of the disease. OBJECTIVE: The purpose of this study was to evaluate the effects of probiotic yogurt on intestinal flora of patients with chronic liver disease. METHODS: A randomized controlled trial, pretest-posttest control group design, was used. Patients were randomized to an experimental group (41 patients) or a control group (40 patients). Patients in the experimental group were given probiotic yogurt (one cup each time, three times per day for 14 days) containing Bacillus bifidus, Lactobacillus acidophilus, Lactobacillus bulgaricus, and Streptococcus thermophilus within 2 hours after meals. Levels of fecal flora, symptoms and signs, and laboratory examination indexes were collected. RESULTS: After intervention, the experimental group had a lower Escherichia coli count and reduced intestinal flora imbalance (p < .05). Comparison of the experimental and control groups after the intervention showed that the former had improved symptoms and signs, including significant improvement in debilitation, food intake, appetite, abdominal distension, and ascitic fluid (p < .05). CONCLUSION: Probiotic yogurt reduces the levels of intestinal flora imbalance and has an additional therapeutic effect on patients with chronic liver disease.')

79 ('>T', 'TI  - Probiotic properties of Lactobacillus strains isolated from the feces of breast-fed infants and Taiwanese pickled cabbage.', 'AB  - This study assessed potential probiotic Lactobacillus strains isolated from the feces of breast-fed infants and from Taiwanese pickled cabbage for their possible use in probiotic fermented foods by evaluating their (i) in vitro adhesive ability, resistance to biotic stress, resistance to pathogenic bacteria, and production of beta-galactosidase; (ii) milk technological properties; and (iii) in vivo adhesive ability, intestinal survival and microbial changes during and after treatment. Five Lactobacillus isolates identified as Lactobacillus reuteri F03, Lactobacillus paracasei F08, Lactobacillus rhamnosus F14, Lactobacillus plantarum C06, and Lactobacillus acidophilus C11 that showed resistance to gastric juice and bile salts were selected for further evaluation of their probiotic properties. All the strains demonstrated the ability to adhere to Caco-2 cells, particularly, strain L. plantarum C06 and L. reuteri F03 showed satisfactory abilities, which were similar to that of the reference strain L. rhamnosus GG. The strains L. paracasei F08 and L. acidophilus C11 had the highest beta-galactosidase activity. Most of the strains were resistant to aminoglycosides and vancomycin but sensitive to ampicillin, erythromycin, and penicillin. All the 5 strains elicited antibacterial activity against both Gram-positive (Bacillus cereus, Listeria monocytogenes and Staphylococcus aureus) and -negative (Escherichia coli and Salmonella enterica) pathogens. Moreover, the strains L. reuteri F03, L. paracasei F08, and L. plantarum C06 could grow rapidly in milk without nutrient supplementation and reached 10(8) cfu/mL after 24 h of fermentation at 37 degrees C. The viable cell counts of the 3 strains remained above 10(7) cfu/mL after 21 d of storage at 4 degrees C. In the animal feeding trial, the number of intestinal lactobacilli increased significantly after administration of milk fermented with the 3 strains, and the counts of fecal coliforms and Clostridium perfringens were markedly reduced. Lactobacillus strains could also survive in the ileal intestinal tissue of the treated rats. Technologically interesting Lactobacillus isolates may be used in the future as probiotic starter cultures for manufacturing novel fermented foods.')

85 ('>T', 'TI  - Bacteriocins produced by L. Fermentum and L. Acidophilus can inhibit cephalosporin resistant e. Coli.', 'AB  - Reemerging infections occur due to resistant bacteria. Such infections create restrictions for clinicians and microbiologists in drug selection. Such problems demand new strategies for solution. Use of bacteriocins for this purpose may be fruitful. In the present research work, the inhibitory effects of bactericins on cephalosporin resistant Escherichia coli are used as model system for the control of antibiotic resistant pathogenic bacteria. Cephalosporin resistant Escherichia coli strain was isolated from pus by using conventional methodology. For bacteriocin production, Lactobacilli strains were selected by using selective media. Out of seventy two strains isolated from yogurt, fecal materials of human, chick, parrot and cat, only two strains (strain 45 and strain 52) were found to produce bacteriocins having antimicrobial potential against cephalosporin resistant Escherichia coli. Biochemical characterization showed that strain 45 belonged to group of Lactobacillus fermentum and strain 52 to Lactobacillus acidophilus. Both strains showed maximum growth at 25 degrees C and 35 degrees C respectively. Suitable pH was 5.5 and 6.0 for Lactobacillus fermentum and Lactobacillus acidophilus respectively. Bacteriocins produced by both strains were found stable at 50, 75 and 100 degrees C for 60min. Function of bacteriocin was also not disturbed due to change in pH. These findings suggest that bacteriocin produced by Lactobacillus fermentum and Lactobacillus acidophilus can be used for the infection control of cephalosporin resistant Escherichia coli.')

88 ('>T#GUTSPECIFIC', "TI  - Association of Lactobacillus acidophilus with mice Peyer's patches.", "AB  - OBJECTIVE: To clarify the adhesion mechanism of Lactobacillus acidophilus to Peyer's patches. METHODS: Adhesion of L. acidophilus FN001 to mice Peyer's patches was studied in vitro using a fluorescent quantization method. The nature of adhesion mediator was studied by the differing effects of physical, chemical, and enzymatic pre-treatments of the bacteria and the inhibitory effects of sugars on the adhesion. The presence of lectin-like proteins on the cell surface was determined by hemagglutination assay. The effect of L. acidophilus FN001 on the inhibition of adhesion of pathogens to Peyer's patches was also studied. RESULTS: The adhesion of L. acidophilus FN001 was strongly inhibited in the presence of D-mannose and methyl-alpha-D-mannoside. Pretreatment of L. acidophilus FN001 with pepsin and trypsin decreased the adhesive capacity indicating that some cell surface proteins might be involved in the adhesion. L. acidophilus FN001 showed agglutinating activity toward the rabbit red cells in a mannose specific manner, which was decreased after protease pretreatment, suggesting possible occurrence of mannose specific lectin(s) on the L. acidophilus FN001 surface. In adhesion inhibition assay, L. acidophilus NF001, when applied to Peyer's patches first or at the same time with pathogen, significantly inhibited adhesion of Escherichia coli ATCC25922 to Peyer's patches. CONCLUSION: L. acidophilus FN001 contains some mannose-specific protein(s) on its surface that mediates its adhesion to the Peyer's patches. FN001 inhibits the adhesion of E. coli, which also contains mannose specific lectin.")

91 ('>T', 'TI  - Fates of acid-resistant and non-acid-resistant Shiga toxin-producing Escherichia  coli strains in ruminant digestive contents in the absence and presence of probiotics.', 'AB  - Healthy ruminants are the main reservoir of Shiga toxin-producing Escherichia coli (STEC). During their transit through the ruminant gastrointestinal tract, STEC encounters a number of acidic environments. As all STEC strains are not equally resistant to acidic conditions, the purpose of this study was to investigate whether acid resistance confers an ecological advantage to STEC strains in ruminant digestive contents and whether acid resistance mechanisms are induced in the rumen compartment. We found that acid-resistant STEC survived at higher rates during prolonged incubation in rumen fluid than acid-sensitive STEC and that they resisted the highly acidic conditions of the abomasum fluid, whereas acid-sensitive strains were killed. However, transit through the rumen contents allowed acid-sensitive strains to survive in the abomasum fluid at levels similar to those of acid-resistant STEC. The acid resistance status of the strains had little influence on STEC growth in jejunal and cecal contents. Supplementation with the probiotic Saccharomyces cerevisiae CNCM I-1077 or Lactobacillus acidophilus BT-1386 led to killing of all of the strains tested during prolonged incubation in the rumen contents, but it did not have any influence in the other digestive compartments. In addition, S. cerevisiae did not limit the induction of acid resistance in the rumen fluid. Our results indicate that the rumen compartment could be a relevant target for intervention strategies that could both limit STEC survival and eliminate induction of acid resistance mechanisms in order to decrease the number of viable STEC cells reaching the hindgut and thus STEC shedding and food contamination.')

96 ('>T', 'TI  - Antagonistic activity of Lactobacillus acidophilus RY2 isolated from healthy infancy feces on the growth and adhesion characteristics of enteroaggregative Escherichia coli.', 'AB  - Enteroaggregative Escherichia coli (EAggEC) infection is an important cause of acute diarrhea, affecting children in developing countries and travelers visiting tropical or subtropical areas. Three probiotics can exert bacteriostatic or bactericidal effects on human and animal intestinal pathogens, the efficiency of probiotics on EAggEC infection remains unclear. In this study, the antagonistic activity of probiotic bacteria isolated from infant faeces was examined against several EAggEC stains. While three isolates, Lactobacillus acidophilus RY2, Lactobacillus salivarius MM1 and Lactobacillus paracasei En4 were shown to significantly inhibit the growth of EAggEC. In addition, the antagonistic activity of the Lactobacillus species was maintained despite heating (100 degrees C, 15 min) of cell free culture supernatant (CFCS). The antagonistic activity of the CFCS however, could be reduced following lactate dehydrogenase treatment and at pH 7.2. Furthermore, in an adhesion-inhibition assay, L. acidophilus RY2 was shown to be more effective than L. salivarius MM1 and L. paracasei EN4. This study suggests that L. acidophilus RY2 could be used as a probiotic organism against EAggEC.')

98 ('>T#GUTSPECIFIC', 'TI  - Live and heat-inactivated lactobacilli from feces inhibit Salmonella typhi and Escherichia coli adherence to Caco-2 cells.', 'AB  - A quantitative approach has been proposed to evaluate the competitive inhibition  of Escherichia coli and Salmonella typhi by live and heat-inactivated laboratory isolated Lactobacillus sp. on adhesion to monolayer of Caco-2 cells. Three species of Lactobacillus (L. casei, L. acidophilus, L. agilis) isolated from human neonate feces and two commercial probiotic strains (L. casei, L. acidophilus) have been compared for probiotic activity. All lactobacilli were able to attach to the Caco-2 cells, however, the degree of adhesion was bacterial strain-dependent. The adhesion indices of the two commercial probiotic strains were not significantly different from the values obtained for the other two similar fecal strains (p > 0.01). The inhibition of attachment of the pathogenic bacteria by inactivated cells of fecal L. acidophilus was examined and compared to the results of live bacteria. The inhibition pattern was similar for live and heat-inactivated L. acidophilus (p > 0.01). The number of attached pathogenic bacteria to the Caco-2 cells decreased when the number of L. acidophilus increased from 10(6) to 10(9) CFU/mL. The heat-inactivated L. acidophilus displayed similar probiotic activity compared to the live bacteria.')

100 ('>T', 'TI  - Evaluation of an antimicrobial ingredient prepared from a Lactobacillus acidophilus casein fermentate against Enterobacter sakazakii.', 'AB  - Previously two antimicrobial peptides, IKHQGLPQE (caseicin A) and VLNENLLR (caseicin B), were identified following the fermentation of sodium caseinate with the proteolytic strain Lactobacillus acidophilus DPC 6026. This study evaluated the ability of these peptides to kill Enterobacter sakazakii ATCC 12868 spiked in reconstituted infant formula. The survival of E. sakazakii populations in reconstituted infant formula containing a sodium caseinate fermentate was compared with survival in formula containing positive (monocaprylin) and negative controls. The L. acidophilus DPC 6026 sodium caseinate fermentate reduced pathogen numbers by >4 log CFU/ml at 37 degrees C, comparing favorably with the activity of monocaprylin. Additionally, E. sakazakii NCTC 8155 was inoculated into pasteurized, reconstituted infant formula (6 log CFUlml) followed by the addition of increasing concentrations of the fermentate (0.21 to 6.7% [wt/vol]). At a concentration of 0.21% (wt/vol), pathogen viability was maintained over 4 h at 6.0 log CFU/ml. In contrast, pathogen numbers increased approximately 100-fold in the control formula in the same time frame. At higher final fermentate concentrations (-3.33% [wt/vol]), numbers were reduced to 0 log CFU/ml over 60 min. The spectrum of activity of the fermentate against other foodborne pathogens was also determined and shown to be effective against Escherichia coli O157:H7 and Listeria innocua. Results indicate the potential of this fermentate as a built-in protection mechanism against E. sakazakii strains in reconstituted infant formula.')

104 ('>T', 'TI  - Antagonistic potential against pathogenic microorganisms and hydrogen peroxide production of indigenous lactobacilli isolated from vagina of Chinese pregnant women.', "AB  - OBJECTIVE: To investigate the indigenous lactobacilli from the vagina of pregnant women and to screen the isolates with antagonistic potential against pathogenic microorganisms. METHODS: The strains were isolated from pregnant women's vagina and identified using the API50CH system. The ability of the isolates to produce hydrogen peroxide was analyzed semi-quantitatively using the TMB-HRP-MRS agar. The antagonistic effects of the isolates on pathogenic microorganisms were determined with a double layer agar plate. RESULTS: One hundred and three lactobacilli strains were isolated from 60 samples of vaginal secretion from healthy pregnant women. Among them, 78 strains could produce hydrogen peroxide, in which 68%, 80%, 80%, and 88% had antagonistic effects against Candida albicans CMCC98001, Staphylococcus aureus CMCC26003, Escherichia coli CMCC44113, and Pseudomonas aeruginosa CMCC10110, respectively. CONCLUSION: The recovery of hydrogen peroxide-producing lactobacilli decreases with the increasing pregnant age and time. The most commonly isolated species from vagina of Chinese pregnant women are Lactobacillus acidophilus and Lactobacillus crispatus. Most of L. acidophilus and L. crispatus produce a high H2O2 level.")

107 ('>T', 'TI  - Released exopolysaccharide (r-EPS) produced from probiotic bacteria reduce biofilm formation of enterohemorrhagic Escherichia coli O157:H7.', 'AB  - Here, we characterized released-exopolysaccharides (r-EPS) from Lactobacillus acidophilus A4 with the goal of identifying natural compounds that represses biofilm formation. In plastic 96-well microplates that contained 1.0 mg/ml of r-EPS, enterohemorrhagic Escherichia coli (EHEC) biofilms were dramatically decreased by 87% and 94% on polystyrene and polyvinyl chloride (PVC) surfaces, respectively. In the presence of r-EPS, neither their growth rate nor their autoinducer-2-like activity was affected on the EHEC O157:H7. Importantly, consistent reduction in biofilm formation was also observed when r-EPS was applied to the continuous-flow chamber models. In addition, we found that adding r-EPS significantly repressed biofilm formation by affecting genes related to curli production (crl, csgA, and csgB) and chemotaxis (cheY) in transcriptome analysis. Furthermore, these r-EPS could prevent biofilm formation by a wide range of Gram-negative and -positive pathogens. This property may lead to the development of novel food-grade adjuncts for microbial biofilm control.')

108 ('>T', 'TI  - Effect of molecules secreted by Lactobacillus acidophilus strain La-5 on Escherichia coli O157:H7 colonization.', "AB  - The probiotic bacterium Lactobacillus acidophilus strain La-5 is a gut-colonizing microorganism that, when established, becomes an important part of the gastrointestinal (GI) tract microbiota. It has been shown to be effective against enterohemorrhagic Escherichia coli (EHEC) O157:H7 infection. We have previously shown that molecules released by probiotic strain La-5 influence the transcription of EHEC genes involved in colonization and quorum sensing. In this work, we report on the ability of these molecules to prevent the adherence of EHEC to epithelial cells and on its capacity to concentrate F-actin at adhesion sites. With a fluorescein-labeled phallotoxin, it was shown that La-5 cell-free spent medium (CFSM) fractions remarkably reduced attaching and effacing lesions in HeLa cells. We also observed a significant inhibition of bacterial adhesion to Hep-2 cells when they were treated with the same La-5 CFSM fractions. In order to observe if La-5 CFSM fractions exhibited the same effect in vivo, we studied the ability of luminescent EHEC constructs (LEE1::luxCDABE) to adhere to intestinal epithelial cells of specific-pathogen-free ICR mice following intragastric inoculation. Colonization of the GI tract by luminescent EHEC O157:H7 was monitored in real time with a slow-scan charge-coupled device camera. At the same time, fecal shedding of EHEC was studied. Following oral gavage of the La-5 active fraction, we observed a reduced amount of bioluminescence signal along with a decrease in fecal shedding by mice, indicating an effect on the ability of the organism to colonize the GI tract. Our results confirm past evidence of the possibility of blocking or interfering with EHEC's virulence by active molecules contained in the probiotic CFSM and identify novel therapeutic alternatives to antibiotic treatments in the fight against food-borne pathogens.")

112 ('>T', 'TI  - Isolation and characterization of potential probiotic lactobacilli from pig feces.', 'AB  - This study examined four lactobacilli isolated from pig feces. Two hundred lactic acid bacteria strains were obtained from pig feces using selective culture media (with vancomycin and bromocresol green; termed LAMVAB agar). Microscopy, the catalase test, Gram-staining, and RAPD-PCR analysis were used to group the bacteria into 20 related clusters. Phenotypic analysis using the API 50 CH test and genotypic analysis of 16S rDNA sequences identified these clusters as representing single strains of each of Lactobacillus fermentum, Lactobacillus salivarius, Lactobacillus plantarum, and Lactobacillus reuteri. Bacterial survival under the conditions of low pH (2.0) and high concentration (5.0%, w/v) of bile salt was much better than that of the reference strain (Lactobacillus acidophilus ATCC 33199). The isolated bacteria were quite capable of inhibiting the growth of two pathogens, Escherichia coli K88 and Salmonella typhimurium. The high acid-resistance, bile resistance and antagonism against pathogens, suggest that the four lactic acid bacteria isolated from pig feces could prove useful as piglet probiotics.')

115 ('>T#GUTSPECIFIC', 'TI  - Inhibition of Escherichia coli O157:H7 attachment by interactions between lactic  acid bacteria and intestinal epithelial cells.', 'AB  - The intestinal epithelial cell (IEC) layer of the intestinal tract makes direct contact with a number of microbiota communities, including bacteria known to have deleterious health effects. IECs possess innate protective strategies against pathogenic challenge, which primarily involve the formation of a physicochemical barrier. Intestinal tract mucins are principal components of the mucus layer on epithelial surfaces, and perform a protective function against microbial damage. However, little is currently known regarding the interactions between probiotics/pathogens and epithelial cell mucins. The principal objective of this study was to determine the effects of Lactobacillus on the upregulation of MUC2 mucin and the subsequent inhibition of E. coli O157:H7 attachment to epithelial cells. In the current study, the attachment of E. coli O157:H7 to HT-29 intestinal epithelial cells was inhibited significantly by L. acidophilus A4 and its cell extracts. It is also important to note that the expression of MUC2 mucin was increased as the result of the addition of L. acidophilus A4 cell extracts (10.0 mg/ml), which also induced a significant reduction in the degree to which E. coli O157:H7 attached to epithelial cells. In addition, the mRNA levels of IL-8, IL-1beta, and TNF-alpha in HT-29 cells were significantly induced by treatment with L. acidophilus A4 extracts. These results indicate that MUC2 mucin and cytokines are important regulatory factors in the immune systems of the gut, and that selected lactobacilli may be able to induce the upregulation of MUC2 mucin and specific cytokines, thereby inhibiting the attachment of E. coli O157:H7.')

116 ('>T', 'TI  - Therapeutic potential of two probiotics in inflammatory bowel disease as observed in the trinitrobenzene sulfonic acid model of colitis.', 'AB  - PURPOSE: The pathogenesis of inflammatory bowel disease is thought to be a multifactorial process. One of the leading hypotheses is that an imbalance in normal gut flora induces an excessive immune response and contributes to inflammation in the gastrointestinal tract. Administration of probiotic bacteria reduces symptoms in patients suffering from inflammatory bowel diseases, probably via both manipulation of the microflora and stimulation of the intestinal immune system. In the current study the therapeutic potential of two different probiotics-Lactobacillus GG and a mixture of Streptococcus thermophilus, Lactobacillus acidophilus, and Bifidobacterium longum (YO-MIX Y 109 FRO 1000)--in a rat model of colitis were evaluated. METHODS: Male Wistar rats were administered probiotics for three days simultaneously with colitis induction. Colonic damage was evaluated histologically and biochemically and colonic tissues, as well as fecal samples, were used for bacterial studies using 16S rRNA gene primers. RESULTS: Probiotics administration reduced the relative amounts of the pathogenic bacteria Aeromonas and Escherichia coli in the colonic tissue. However, whereas both probiotics affected colon morphology, only Lactobacillus GG administration reduced myeloperoxidase activity. CONCLUSIONS: We report the therapeutic rather than preventive potential of two different probiotics in an animal model of colitis.')

120 ('>T#GUTSPECIFIC', 'TI  - Interaction of probiotic Lactobacillus and Bifidobacterium strains with human intestinal epithelial cells: adhesion properties, competition against enteropathogens and modulation of IL-8 production.', 'AB  - The human intestinal microbiota plays a pivotal role in human nutrition and health by promoting the supply of nutrients, preventing pathogen colonization and shaping and maintaining normal mucosal immunity. The depletion of the individual microbiota can result in a higher susceptibility to enteropathogenic bacteria infection. In order to reduce this risk, the use of food supplements containing probiotic bacteria has been recently addressed. In this paper, we investigate the protective role toward enteropathogen infection of probiotic strains belonging to Lactobacillus and Bifidobacterium. According to our experimental data, Lactobacillus acidophilus Bar13, L. plantarum Bar10, Bifidobacterium longum Bar33 and B. lactis Bar30 were effective in displacing the enteropathogens Salmonella typhimurium and Escherichia coli H10407 from a Caco-2 cell layer. Moreover, L. acidophilus Bar13 and B. longum Bar33 have been assessed for their immunomodulatory activity on IL-8 production by HT29 cells. Both strains showed the potential to protect enterocytes from an acute inflammatory response. These probiotic strains are potential candidates for the development of new functional foods helpful in counteracting enteropathogen infections.')

123 ('>T#NOTSOLID', 'TI  - Functional properties of selected starter cultures for sour maize bread.', 'AB  - This paper focuses on the functional properties of maize sour-dough microflora selected and tested for their use as starter cultures for sour maize bread. Lactic acid bacteria and yeasts isolated from spontaneously fermented maize dough were selected based on dominance during fermentation and presence at the end of fermentation. Functional properties examined included acidification, leavening and production of some antimicrobial compounds in the fermenting matrix. The organisms previously identified as Lactobacillus plantarum, Lb. brevis, Lb. fermentum, Lb. acidophilus, Pediococcus acidilactici, Leuconostoc mesenteroides and Leuconostoc dextranicum and Saccharomyces cerevisiae were used singly and as mixed cultures in the fermentation (fermentation time: 12h at 28+/-2 degrees C) of maize meal (particle size >0.2mm). The pH fell from an initial value of 5.62-3.05 in maize meals fermented with Lb. plantarum; 4.37 in L. dextranicum+S. cerevisiae compared with the value for the control (no starter) of 4.54. Significant differences (P <or =0.05) were observed in values obtained for the functional properties tested when starters were inoculated compared with the control (no starter) except for leavening. Bivariate correlations at 0.01 levels (two-tailed) showed that significant correlations existed among pH and production of antimicrobial compounds in the fermenting meals, the highest correlation being between production of diacetyl and acid (0.694), a positive correlation indicating that production of both antimicrobial compounds increase together with time. Antimicrobial activities of the fermented maize dough were confirmed by their abilities to inhibit the growth of Salmonella typhi, Escherichia coli, Staphylococcus aureus and Aspergillus flavus from an initial inoculum concentration of 7 log cfu ml(-1)) for test bacteria and zone of inhibition of up to 1.33 cm for aflatoxigenic A. flavus. The findings of this study form a database for further studies on the development of starter cultures for sour maize bread production as an alternative bread specialty.')

131 ('>T', 'TI  - Bactericidal activity of culture fluid components of Lactobacillus fermentum strain 90 TS-4 (21) clone 3, and their capacity to modulate adhesion of Candida albicans yeast-like fungi to vaginal epithelial cells.', 'AB  - Antagonistic activities of L. fermentum strain 90 TS-4 (21), L. casei ATCC 27216, and L. acidophilus ATCC 4356 and bactericidal activity of lactobacillus culture fluid towards E. coli strain K12, S. aureus, and S. epidermidis test cultures were studied. The bactericidal effect of L. fermentum strain 90 TS-4 (21) clone 3 culture fluid preparation (pH 6.0) on the test cultures was dose-dependent. Adhesion of C. albicans yeast-like fungi to vaginal epitheliocytes was more pronounced for strains isolated from women with asymptomatic infection than for strains isolated from women with manifest forms. L. fermentum strain 90 TS-4 (21) clone 3 culture fluid preparation modulated adhesion of yeast-like fungi only if the fungal strain was initially highly adherent.')

132 ('>T', 'TI  - Three Lactobacillus strains from healthy infant stool inhibit enterotoxigenic Escherichia coli grown in vitro.', 'AB  - Enterotoxigenic Escherichia coli (ETEC) are a major cause of sporadic diarrhea disease in humans, affecting mainly infants in developing countries and travelers from industrialized countries visiting tropical or subtropical areas. In this study, we screen the antagonistic activity by inoculating wells among ETEC agar cultures to assess inhibition zones created by the lactobacilli spent culture supernatant (SCS) from healthy infant stool. Only three isolates possessed antagonistic activity, acid and bile tolerance and could adhere to the cultured human intestinal C2BBel (Caco-2) cell line. Isolate identification using API 50CHL strips showed that they belonged to different Lactobacillus species, i.e., Lactobacillus acidophilus RY2, Lactobacillus salivarius MM1 and Lactobacillus paracasei En4. The SCS still had an inhibitory effect on ETEC after heating (100 degrees C, 15 min). The lactate dehydrogenase treatment or the pH of SCS was adjusted to neutral (pH 7.2) to reduce the SCS inhibitory effect. Antimicrobial activity was performed by incubating the lactic acid bacteria (LAB)-SCS with ETEC suspension. After 4h of co-culture, ETEC growth was inhibited. This study suggests that L. acidophilus RY2, L. salivarius MM1 and L. paracasei En4 could be used as an effective control for ETEC.')

134 ('>T', 'TI  - Reduction of Escherichia coli O157 and Salmonella in feces and on hides of feedlot cattle using various doses of a direct-fed microbial.', 'AB  - In this study, the effectiveness of direct-fed microbials at reducing Escherichia coli O157 and Salmonella in beef cattle was evaluated. Steers (n=240) received one of the following four treatment concentrations: control = lactose carrier only; low = 1 X 10(7) CFU per steer daily Lactobacillus acidophilus NP51; medium = 5 x 10(8) CFU per steer daily L. acidophilus NP51; and high = 1 x 10(9) CFU per steer daily L. acidophilus NP51. Low, medium, and high diets also included 1 x 10(9) CFU per steer Propionibacterium freudenreichii NP24. Feces were collected from each animal at allocation of treatment and found to have no variation (P = 0.54) between cohorts concerning E. coli O157 recovery. Feces and hide swabs were collected at harvest and analyzed for the presence of E. coli O157 by immunomagnetic separation and Salmonella by PCR. No significant dosing effects were detected for E. coli O157 recovery from feces at the medium dose or from hides at the medium and high doses. E. coli O157 was 74% (P < 0.01) and 69% (P < 0.01) less likely to be recovered in feces from animals receiving the high and low diets, respectively, compared with controls. Compared with controls, E. coli O157 was 74% (P = 0.05) less likely to be isolated on hides of cattle receiving the low dose. No significant dosing effects were detected for Salmonella recovery from feces at the medium and low doses or from hides at any doses. Compared with controls, Salmonella was 48% (P = 0.09) less likely to be shed in feces of cattle receiving the high dose. No obvious dose-response of L. acidophilus NP51 on recovery of E. coli O157 or Salmonella was detected in our study.')

136 ('>T', 'TI  - [Evaluation of the effect of Lactobacillus rhamnosus probiotic culture added to yogurt over Staphylococcus aureus, Escherichia coli O157:H7, Listeria monocytogenes and Salmonella enteritidis populations].', 'AB  - The effect of different types of probiotics present in yogurt over known populations of Staphylococcus aureus, Escherichia coli O157:H7, Listeria monocytogenes and Salmonella enteritidis was evaluated. The three types of yogurt used were: without added probiotics, with added probiotics (Lactobacillus casei CRL_431 and L. acidophilus CRL_730 CHR HANSEN) and another one with the same probiotics mentioned above and Lactobacillus rhamnosus (LR-35) culture. About 10(9) CFU/ mL of each potentially pathogenic bacteria was added to each type of yogurt tested, and kept in refrigeration at 4 degrees C during its shelf life, about 30 days. Bacterial count was done the initial day and every four days. Results obtained show that there is a difference in the inhibition between yogurts without added probiotics and the commercial yogurt with added probiotics; there is a clear inhibitory effect of the last one over S. aureus, E. coli O157:H7 and Listeria monocytogenes. The yogurt with added probiotics and L. rhamnosus did not show any additional inhibitory effect over the bacteria tested when compared with the yogurt with added probiotics. S. enteritidis could not be evaluated because it was not detectable in any yogurt samples evaluated four days after its inoculation. This study confirms the antagonic effect of probiotic cultures over potentially pathogenic bacteria for human beings and animals that may be present in food. Nevertheless, the use of L. rhamnosus did not produce any additional inhibitory effect.')

142 ('>T', 'TI  - Prevalence and enumeration of Escherichia coli O157 in steers receiving various strains of Lactobacillus-based direct-fed microbials.', 'AB  - The objective of this research was to evaluate the effect of daily dietary inclusion of specific strains of Lactobacillus acidophilus on prevalence and concentration of Escherichia coli O157 in harvest-ready feedlot cattle. Five hundred yearling steers were housed in pens of 10 animals each. At arrival, steers were randomly allocated to one of five cohorts. Four of the cohorts were fed various strains and dosages of Lactobacillus-based direct-fed microbials throughout the feeding period. Fecal samples were collected from the rectum of each animal immediately prior to shipment to the abattoir. E. coli O157 was detected using selective enrichment and immunomagnetic separation techniques. For positive samples, E. coli O157 concentration was estimated using a most-probable-number (MPN) technique that included immunomagnetic separation. Prevalence varied among the cohorts (P < 0.01). The prevalence in the controls (26.3%) was greater (P < 0.05) than that in cattle supplemented with L. acidophilus strains NP51, NP28, or NP51-NP35 (13.0, 11.0, and 11.0%, respectively). The greatest E. coli O157 concentration was also observed in the controls (3.2 log MPN/g of feces); this concentration was greater (P < 0.05) than that observed in positive animals receiving NP51, NP28, or NP51-NP35 (0.9, 1.1, 1.7 log MPN/g of feces, respectively). Specific strains of Lactobacillus-based direct-fed microbials effectively reduced the prevalence and concentration of E. coli O157 in harvest-ready cattle, whereas others did not. When using direct-fed microbials to reduce carriage of E. coli O157 in cattle, it is important to select appropriately validated products.')

148 ('>T', 'TI  - In vitro growth control of selected pathogens by Lactobacillus acidophilus- and Lactobacillus casei-fermented milk.', 'AB  - AIMS: Food-borne pathogen inhibition was tested in the presence of a mixture of Lactobacillus acidophilus and Lactobacillus casei during fermentation under controlled pH conditions. METHODS AND RESULTS: The growth of Escherichia coli O157:H7, Salmonella serotype Typhimurium, Staphylococcus aureus, Listeria innocua, Enterococcus faecium and Enterococcus faecalis was evaluated for 48 h at 37 degrees C. In the presence of the lactic acid bacteria (LAB), an increase of the generation time was observed for all the gram-positive bacteria evaluated. Staphylococcus aureus was the most sensitive strain showing an increase of the generation time by 210%. However, for all the gram-negative bacteria evaluated, no inhibition occurred after 8 h of fermentation. The soluble portion of Lact. acidophilus- and Lact. casei-fermented milk was recuperated and tested for its antimicrobial activity. Listeria innocua and Staph. aureus were the most sensitive to the presence of fermented milk supernatant showing an inhibition of 85.9% and 84.7%, respectively. This soluble fraction was neutralized to eliminate the antimicrobial effect of the organic acids produced; the most sensitive strains were L. innocua and E. coli O157:H7 showing an inhibition of 65.9% and 61.9%, respectively. Finally, the soluble fraction was neutralized and irradiated at 45 kGy using a (60)Co source to eliminate the possible antimicrobial effect of both organic acids and bacteriocin-like substances. Enterococcus faecalis, E. coli O157:H7 and Staph. aureus were the most affected bacteria by this fraction, showing 39.1, 32 and 31.2% inhibition, respectively. CONCLUSIONS: The results obtained in this study suggest the implication of both organic acids and bacteriocin-like inhibitory substances in the antimicrobial activity observed in the soluble fraction of the probiotic preparation. SIGNIFICANCE AND IMPACT OF THE STUDY: This study revealed the antimicrobial mechanisms of action of Lact. acidophilus- and Lact. casei-fermented milk used to prevent antibiotic-associated diarrhoea.')

149 ('>T', 'TI  - The S-layer proteins of Lactobacillus crispatus strain ZJ001 is responsible for competitive exclusion against Escherichia coli O157:H7 and Salmonella typhimurium.', 'AB  - Lactobacillus crispatus ZJ001, isolated from pig intestines and identified by sequencing analysis based on partial 16S rRNA gene, was examined in vitro for probiotic activity exerted by the surface layer proteins (S-layer). The characteristics of L. crispatus ZJ001 were compared to Lactobacillus acidophilus ATCC 4356 from the same genus which also produces the S-layer proteins. The strain ZJ001 was resistant to acidic condition and bile salt. Its antagonistic properties such as adhesion, inhibition of the pathogen growth and competitive exclusion against Escherichia coli O157:H7 and Salmonella typhimurium were apparently advantageous over L. acidophilus ATCC 4356. SDS-PAGE analysis of cell surface proteins revealed the presence of S-layer proteins, approximately at 42 kDa in L. crispatus ZJ001. Removal of the S-layer proteins reduced autoaggregation and adhesion to HeLa cells. The functional role of the S-layer proteins in adhesion was also confirmed by the antibody-mediated inhibition assay using the polyclonal antibody against the S-layer protein. The S-layer proteins from L. crispatus ZJ001 inhibited adhesion of S. typhimurium and E. coli O157:H7 to HeLa cells. These results suggest that L. crispatus ZJ001 possesses probiotic properties and the S-layer proteins are involved in the adhesion and competitive exclusion of pathogens to HeLa cells.')

157 ('>T', 'TI  - Biotic and abiotic factors influencing in vitro growth of Escherichia coli O157:H7 in ruminant digestive contents.', 'AB  - The gastrointestinal tract (GIT) of ruminants is the main reservoir of enterohemorrhagic Escherichia coli, which is responsible for food-borne infections in humans that can lead to severe kidney disease. Characterization of biotic and abiotic factors that influence the carriage of these pathogens by the ruminant would help in the development of ecological strategies to reduce their survival in the GIT and to decrease the risk of contamination of animal products. We found that growth of E. coli O157:H7 in rumen fluid was inhibited by the autochthonous microflora. Growth was also reduced when rumen fluid came from sheep fed a mixed diet composed of 50% wheat and 50% hay, as opposed to a 100% hay diet. In fecal suspensions, E. coli O157:H7 growth was not suppressed by the autochthonous flora. However, a probiotic strain of Lactobacillus acidophilus inhibited E. coli O157:H7 growth in fecal suspensions. The inhibitory effect was dose dependent. These lactic acid bacteria could be a relevant tool for controlling O157:H7 development in the terminal part of the ruminant GIT, which has been shown to be the main site of colonization by these pathogenic bacteria.')

158 ('>T', 'TI  - Antagonistic activity of probiotic lactobacilli and bifidobacteria against entero- and uropathogens.', 'AB  - AIM: To develop in vitro assays for comparing the antagonistic properties and anti-oxidative activity of probiotic Lactobacillus and Bifidobacterium strains against various entero- and urinary pathogens. METHODS AND RESULTS: The antagonistic activity of five probiotic lactobacilli (Lactobacillus rhamnosus GG, Lactobacillus fermentum ME-3, Lactobacillus acidophilus La5, Lactobacillus plantarum 299v and Lactobacillus paracasei 8700:2) and two bifidobacteria (Bifidobacterium lactis Bb12, Bifidobacterium longum 46) against six target pathogens was estimated using different assays (solid and liquid media, anaerobic and microaerobic cultivation) and ranked (low, intermediate and high). Bacterial fermentation products were determined by gas chromatography, and the total anti-oxidative activity of probiotics was measured using linolenic acid test. Pyelonephritic Escherichia coli was highly suppressed by GG and both bifidobacteria strains. Lactobacilli strains 8700:2, 299v and ME-3 were the most effective against Salmonella enterica ssp. enterica in microaerobic while ME-3 and both bifidobacteria expressed high activity against Shigella sonnei in anaerobic milieu. Lact. paracasei, Lact. rhamnosus and Lact. plantarum strains showed intermediate antagonistic activity against Helicobacter pylori under microaerobic conditions on solid media. The highest anti-oxidative activity was characteristic for Lact. fermentum ME-3 (P < 0.05). No efficient antagonist against Clostridium difficile was found. The positive correlations between the pH, lactic acid production and anti-microbial activity for all tested probiotics were assessed. CONCLUSIONS: Developed experimental assays enable to compare the anti-microbial and -oxidative activity of Lactobacillus and/or Bifidobacterium probiotics, which have been claimed to possess the ability of suppressing the growth of various enteric and urinary pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Screening Lactobacillus and Bifidobacterium sp. strains according to their activity in various environmental conditions could precede the clinical efficacy studies for adjunct treatment with probiotics in cure of different gastrointestinal and urinary tract infections.')

161 ('>T', 'TI  - Effect of Lactobacillus isolates on the adhesion of pathogens to chicken intestinal mucus in vitro.', 'AB  - AIMS: The aims of this study were to investigate in vitro the effects of Lactobacillus isolates from a chicken on adhesion of pathogenic Salmonella and Escherichia coli to chicken intestinal mucus obtained from different intestinal regions. METHODS AND RESULTS: Bacteria were labelled by using methyl-1,2-[(3)H]-thymidine. The bacterial adhesion was assessed by measuring the radioactivity of bacteria adhered to the mucus. The results showed that the abilities of Lactobacillus spp. to bind to the same intestinal mucus were higher than those of pathogenic Salmonella and E. coli. Pretreatment of intestinal mucus with Lactobacillus fermentum and Lactobacillus acidophilus, alone or in combination, reduced the adhesion of the tested pathogens, but the reductive extent of pathogenic adhesion by Lactobacillus spp. in combination was relatively high. CONCLUSIONS: The tested bacteria had different adhesions to mucus glycoproteins isolated from different intestinal regions of chicken. Lactobacillus acidophilus and Lact. fermentum in combination revealed a better ability to inhibit attachments of Salmonella and E. coli to chicken intestinal mucus than Lactobacillus sp. alone. SIGNIFICANCE AND IMPACT OF THE STUDY: A mixture of intestinal Lactobacillus spp. from a chicken may play a protective role in excluding pathogenic Salmonella and E. coli from the intestine of chicken.')

162 ('>T', 'TI  - Selection of potentially probiotic Lactobacillus strains towards their inhibitory activity against poultry enteropathogenic bacteria.', 'AB  - Lactobacilli were isolated from chicken gastrointestinal tract and examined for their potentially probiotic properties towards their inhibitory activity against poultry enteropathogenic bacteria. Biochemical tests, ITS-PCR and cell wall protein analysis were used to characterize the Lactobacillus isolates. The identification of isolated Lactobacillus strains based on phenotypic properties was not always satisfactory. ITS-PCR together with protein profile were found to be helpful in strain identification. Lactobacilli were tested for the inhibitory activity against selected strains of poultry enteropathogenic bacteria (Salmonella Enteritidis, Escherichia coli and Clostridium perfringens). Examined supernatants from Lactobacillus broth cultures demonstrated major antimicrobial activity against C. perfringens. Lower antimicrobial activity were observed against E. coli and Salmonella Enteritidis. The strongest inhibition effect were obtained using supernatant of Lactobacillus acidophilus strain 3D. Results received from this study confirmed that identification of Lactobacillus spp. is often tedious. Some isolates, which are in vitro antagonistic against enteropathogenic bacteria may be considered as potential candidates for poultry probiotics, especially in controlling necrotic enteritis caused by C. perfringens.')

164 ('>T', 'TI  - Casein-derived antimicrobial peptides generated by Lactobacillus acidophilus DPC6026.', 'AB  - Three peptides produced by a Lactobacillus acidophilus DPC6026 fermentation of sodium caseinate and showing antibacterial activity against pathogenic strains Enterobacter sakazakii ATCC 12868 and Escherichia coli DPC5063 were characterized. These peptides were all generated from bovine alpha(s1)-casein and identified as IKHQGLPQE, VLNENLLR, and SDIPNPIGSENSEK. These peptides may have bioprotective applicability and potential use in milk-based formula, which has been linked to E. sakazakii infection in neonates.')

168 ('>T', 'TI  - Lactobacillus acidophilus expressing recombinant K99 adhesive fimbriae has an inhibitory effect on adhesion of enterotoxigenic Escherichia coli.', 'AB  - The most common enteric colibacillosis in neonatal and newborns is caused by enterotoxigenic Escherichia coli(ETEC). Colonization of ETEC in the small intestine is associated with adhesions using fimbriae, which is known as a specific adhesion factor and provides highly specific means for anchoring and prerequisite for an infectious agent. In the present study we have engineered Lactobacillus acidophilus to produce recombinant K99 fimbriae, which is used for the colonization to the intestine of pigs. The expression of K99 fimbrial protein was confirmed using SDS-PAGE, immunoblot and agglutination analyses. To evaluate a function of the K99 fimbrial protein, inhibition and competition tests were performed on pre-screened intestinal brush border from pigs. The tests showed that recombinant L. acidophilus, not control L. acidophilus, had a significant inhibitory effect to and competition against K99+ E. coli in a dose dependent manner. In conclusion, we demonstrated that recombinant K99 fimbriae producing L. acidophilus was able to prevent E. coli binding to intestinal brush border.')

178 ('>T', 'TI  - [Evaluation of the effect of probiotic cultures added to commercial yogurt over a known population of Listeria monocytogenes and Escherichia coli O157:H7].', 'AB  - The effect of probiotic cultures over Listeria monocytogenes and Escherichia coli O157:H7 during yogurt storage was evaluated. Two different yogurt brands, one with additional probiotic cultures (Lactobacillus casei and L. acidophilus) were inoculated with known populations (106 UFC/g) of either L. monocytogenes or E. coli O157:H7 in three different times and stored for 32 days at 5 degrees C. Every four days the count of lactic bacteria, the added pathogens and pH was evaluated, according to the methodology described in the Bacteriological Analytical Manual. The pH and lactic bacteria population were constant during the testing period. Yogurt with additional probiotic cultures reduced the population of L. monocytogenes in 8 days, the population of E. coli O157:H7 in 16; yogurt with no additional probiotics took 20 days to reduce L. monocytogenes to non-detectable levels and even after 28 days of storage, E. coli O157:H7 was cultured. In this work, the beneficial effects of additional probiotic cultures in yogurt is confirmed again.')

181 ('>T', 'TI  - Reduction of Escherichia coli O157 in finishing beef cattle by various doses of Lactobacillus acidophilus in direct-fed microbials.', 'AB  - Our objective was to evaluate the effects of three doses of Lactobacillus acidophilus strain NP51 and a combination treatment of strains NP51 and NP45 on prevalence of Escherichia coli O157 in cattle. Three hundred steers were assigned randomly to 60 pens (five steers per pen) and received one of five treatments: (i) control, no added direct-fed microbial; (ii) HNP51, high dose of NP51 at 10(9) CFU per steer daily; (iii) MNP51, NP51 at 10(8) CFU per steer daily; (iv) LNP51, low dose of NP51 at 10(7) CFU per steer daily; and (v) NP51+45, NP51 at 10(9) CFU per steer daily and NP45 at 106 CFU per steer daily. All direct-fed microbial treatments included Propionibacterium freudenreichii at 10(9) CFU per steer. Individual rectal fecal samples were collected on arrival and every 28 days throughout the feeding period. Fecal and hide samples were collected on the day of harvest. Samples were analyzed for presence of E. coli O157 using immunomagnetic separation methods. Cattle receiving HNP51, MNP51, and LNP51 had a lower prevalence (P < 0.01) of E. coli O157 throughout the feeding period compared with the controls, and the dose response for NP51 was a linear decrease in prevalence with increasing dose (P < 0.01). No decrease in prevalence for cattle receiving the combination NP51+45 was detected compared with controls (P = 0.15). E. coli O157 prevalences averaged across collection times were 23.9, 10.5, 9.9, 6.8, and 17.3% for cattle in the control, LNP51, MNP51, HNP51, and NP51 +45 groups, respectively. Least squares mean estimates of fecal prevalence at harvest of E. coli O157 were 31.7, 12.5, 17.4, 8.2, and 41.6% among cattle in the control, LNP51, MNP51, HNP51, and NP51+45 groups, respectively. Least squares mean estimates of the percentage of positive hide samples at harvest were 8.7, 5.9, 4.8, 3.4, and 8.6% among cattle in the control, LNP51, MNP51, HNP51, and NP51+45 groups, respectively. The greatest decrease in E. coli O157 carriage was achieved using NP51 at 10(9) CFU per steer.')

183 ('>T', 'TI  - [Hydrogen peroxide produced by Lactobacillus species as a regulatory molecule for vaginal microflora].', 'AB  - A total of 33 strains of Lactobacillus belonging to 9 species, isolated from vagina, were tested for production of hydrogen peroxide. We observed that the following species: L. delbrueckii, L. acidophilus, L. crispatus, L. johnsonii and L. gasseri dominated over other species in secretion of hydrogen peroxide to the growth medium. Concentration of this substance amounted from 0.05 to 1.06 mM (in case of strong aeration the concentration increased up to 1.8 mM). Moreover, killing properties of the pure hydrogen peroxide exerted toward Escherichia coli and Candida albicans were less prominent than these of the supernatants of cultures of Lactobacillus strains producing H2O2.')

187 ('>T', 'TI  - [Search of promising strains of bifidobacteria and lactobacillus for the development of new biopreparations].', 'AB  - For the first time the species composition of bifidobacteria and lactobacilli in  clinically healthy young children has been studied. As revealed in this study, the dominating species of bifidobacteria are B. longum, B. adolescentis and B. infantis, while the dominating species of lactobacilli are Lactobacillus acidophilus and L. rhamnosus. In 83 isolated cultures of bifidobacteria and 34 isolated species of lactobacilli the activity of acid formation and the antagonistic activity with respect to Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Enterococcus faecalis, Clostridium perfringens have been tested. B. longum strain 58B, B. infantis strain 37B, L. rhamnosus strain 12L and L. acidophilus strain 27L, typical for children of this age group, having good antagonistic activity and pronounced acid-forming properties, have been selected. These strains hold good promise to be used as the basis for the development of a complex probiotic preparation for correcting intestinal microflora in young children.')

188 ('>T', 'TI  - Dietary supplementation with Lactobacillus- and Propionibacterium-based direct-fed microbials and prevalence of Escherichia coli O157 in beef feedlot cattle and on hides at harvest.', 'AB  - The objective of this study was to describe the prevalence of Escherichia coli O157 in the feces and on the hides of finishing beef cattle fed a standard diet and those fed diets supplemented with direct-fed microbials. Two hundred forty steers received one of four treatments throughout the feeding period: (i) control: no added microbials; (ii) HNP51: high dose of Lactohacillius acidophilus strain NP 51 (10(9) CFU per steer daily) and Propionibacterium freudenreichii (10(9) CFU per steer daily); (iii) HNP51+45: high dose of NP 51 (10(9) CFU per steer daily), P. freudenreichii (10(9) CFU per steer daily), and L. acidophilus NP 45 (10(6) CFU per steer daily); or (iv) LNP51+45: low dose of NP 51 (10(6) CFU per steer daily), P. freudenreichii (10(9) CFU per steer daily), and NP 45 (10(6) CFU per steer daily). Samples were collected from each animal and analyzed for the presence of E. coli O157 using immunomagnetic separation methods on day 0 (feces), 7 days before harvest (feces), and at harvest (feces and hide). At the end of the feeding period, cattle receiving HNP51 were 57% less likely to shed detectable E. coli O157 in their feces than were the controls (P < 0.01). For animals receiving HNP51+45 and LNP51+45, fecal prevalence did not differ from that of the controls. The prevalence of positive hide samples was least among cattle receiving HNP51+45 (3.3%); these animals were 79% less likely (P < 0.06) to have a positive hide sample than were the controls (prevalence = 13.8%). There was poor agreement of the culture results between fecal and hide samples collected from the same animal (kappa = 0.08; confidence interval = -0.05 to 0.2). Cattle supplemented with a high dose of NP 51 had reduced E. coli O157 prevalence in both fecal and hide samples, indicating that this treatment may be an efficacious preharvest intervention strategy.')

189 ('>T', 'TI  - Testing two Lactobacillus plantarum and Lactobacillus acidophilus strains for their suitability as a lipoid probiotic.', 'AB  - Two strains of lactobacilli (Lactobacillus acidophilus T-135 and Lactobacillus plantarum 4/97) were selected in order to study their inhibitory properties against frequent udder pathogens (Escherichia coli, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus uberis, Salmonella enteritidis and Bacillus pumilus), their production of organic acids as well as their ability to survive on the teat skin, the teat duct mucosa and in a lipoid emulsion. Both strains inhibited the tested pathogenic microbes and survived on the investigated surfaces and in an emulsion for more than 6 hours and 11 days, respectively.')

193 ('>T', 'TI  - Modulation of anti-pathogenic activity in canine-derived Lactobacillus species by carbohydrate growth substrate.', 'AB  - AIMS: To investigate the effect of various carbon sources on the production of extracellular antagonistic compounds against two Escherichia coli strains and Salmonella enterica serotype Typhimurium by three canine-derived lactobacilli strains. METHODS AND MATERIALS: Cell-free preparations, pH neutralized, were used in antibiotic disc experiments as an initial screening. The bacteria/carbohydrate combinations that showed inhibition of the growth of those pathogens, were further investigated in batch co-culture experiments. The cell-free supernatants of the cultures, that decreased the population number of the pathogens in the co-culture experiments to log CFU ml-1 <or= 4, were tested for inhibition of the pathogens in pure cultures at neutral and acidic pH. CONCLUSIONS: The results showed that the substrate seems to affect the production of antimicrobial compounds and this effect could not just be ascribed to the ability of the bacteria to grow in the various carbon sources. L. mucosae, L. acidophilus and L. reuteri, when grown in sugar mixtures consisting of alpha-glucosides (Degree of Polymerization (DP) 1-4) could produce antimicrobial compounds active against all three pathogens in vitro. This effect could not be attributed to a single ingredient of those sugar mixtures and was synergistic. This inhibition had a dose-response characteristic and was more active at acidic pH. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge of the effect that the carbon source has on the production of antimicrobial compounds by gut-associated lactobacilli allows the rational design of prebiotic/probiotic combinations to combat gastrointestinal pathogens.')

194 ('>T', 'TI  - Growth and lactic acid production by vaginal Lactobacillus acidophilus CRL 1259,  and inhibition of uropathogenic Escherichia coli.', 'AB  - Lactic acid-producing lactobacilli were selected from 134 human vaginal isolates  by testing their capability to inhibit the growth of different pathogenic micro-organisms. Lactobacillus acidophilus CRL 1259 (from the CERELA Culture Collection) was selected to study the effects of temperature, pH and culture medium on growth and lactic acid production. Growth parameters were estimated by using the model of Gompertz. Kinetics of inhibition of uropathogenic Escherichia coli were evaluated in mixed cultures of the pathogen and L. acidophilus. Optimal conditions for growth and lactic acid production by L. acidophilus were pH 6.5 or 8.0 and 37 degrees C. Under these conditions, growth was higher in LAPTg (yeast extract/peptone/tryptone/Tween 80/glucose) broth than in MRS (De Man-Rogosa-Sharpe) broth. However, lactic acid production was more efficient in MRS broth. Under optimal conditions for lactic acid production, L. acidophilus inhibited the growth of E. coli. These results suggest that inclusion of L. acidophilus CRL 1259 in probiotic products for vaginal application would be beneficial.')

203 ('?T', 'TI  - Prevalence of Escherichia coli O157:H7 and performance by beef feedlot cattle given Lactobacillus direct-fed microbials.', 'AB  - Fecal shedding of Escherichia coli O157:H7, the prevalence of Escherichia coli O157:H7 in pens and on carcasses and hides, and cattle performance as a result of daily dietary supplementation with Lactobacillus-based direct-fed microbials (DFMs) were evaluated in a feeding trial involving 180 beef steers. Steers were evaluated for shedding of E. coli O157:H7 by an immunomagnetic separation technique on arrival at the feedlot, just before treatment with the DFMs, and every 14 days thereafter until slaughter. Composite pen fecal samples were collected every 14 days (alternating weeks with animal testing), and prevalence on hides and carcasses at slaughter was also evaluated. Feedlot performance (body weight gain and feed intake) was measured for the period during which the DFMs were fed. Gain efficiency was calculated as the ratio of weight gain to feed intake. Lactobacillus acidophilus NPC 747 decreased (P < 0.01) the shedding of E. coli O157:H7 in the feces of individual cattle during the feeding period. E. coli O157:H7 was approximately twice as likely to be detected in control animal samples as in samples from animals receiving L. acidophilus NPC 747. In addition, DFM supplementation decreased (P < 0.05) the number of E. coli O157:H7-positive hide samples at harvest and the number of pens testing positive for the pathogen. Body weight gains (on a live or carcass basis) and feed intakes during the DFM supplementation period did not differ among treatments. Gain efficiencies on a live-weight basis did not differ among treatments, but carcass-based gain/feed ratios tended (P < 0.06) to be better for animals receiving the two DFM treatments than for control animals. The results of this study suggest that the feeding of a Lactobacillus-based DFM to cattle will decrease, but not eliminate, fecal shedding of E. coli O157:H7, as well as contamination on hides, without detrimental effects on performance.')

205 ('>T', 'TI  - Isolation, selection, and characterization of lactic acid bacteria for a competitive exclusion product to reduce shedding of Escherichia coli O157:H7 in cattle.', 'AB  - Lactic acid bacteria (LAB) were selected on the basis of characteristics indicating that they would be good candidates for a competitive exclusion product (CEP) that would inhibit Escherichia coli O157:H7 in the intestinal tract of live cattle. Fecal samples from cattle that were culture negative for E. coli O157:H7 were collected. LAB were isolated from cattle feces by repeated plating on deMan Rogosa Sharpe agar and lactobacillus selection agar. Six hundred eighty-six pure colonies were isolated, and an agar spot test was used to test each isolate for its inhibition of a four-strain mixture of E. coli O157:H7. Three hundred fifty-five isolates (52%) showed significant inhibition. Seventy-five isolates showing maximum inhibition were screened for acid and bile tolerance. Most isolates were tolerant of acid at pH levels of 2, 4, 5, and 7 and at bile levels of 0.05, 0.15, and 0.3% (oxgall) and were subsequently identified with the API system. Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus delbreukii, Lactobacillus salivarius, Lactobacillus brevis, Lactobacillus cellobiosus, Leuconostoc spp., and Pediococcus acidilactici were the most commonly identified LAB. Nineteen strains were further tested for antibiotic resistance and inhibition of E. coli O157:H7 in manure and rumen fluid. Four of these 19 strains showed susceptibility to all of the antibiotics, 13 significantly reduced E. coli counts in manure, and 15 significantly reduced E. coli counts in rumen fluid (P < 0.05) during at least one of the sampling periods. One of the strains, M35, was selected as the best candidate for a CEP. A 16S rRNA sequence analysis of M35 revealed its close homology to Lactobacillus crispatus. The CEP developed will be used in cattle-feeding trials.')

206 ('>T', 'TI  - Probiotic properties of human lactobacilli strains to be used in the gastrointestinal tract.', 'AB  - AIMS: The study of two human strains of Lactobacillus to be used as probiotics in the gastrointestinal tract. METHODS AND RESULTS: The Lactobacillus acidophilus UO 001 and Lact. gasseri UO 002, were resistant to the gastrointestinal conditions (pH 2 and 3, presence of pepsin, pancreatin or bile salts), the resistance was enhanced in the presence of skimmed milk. Additionally, adhered to Caco-2 cells through glycoproteins in Lact. gasseri and carbohydrates in the case of Lact. acidophilus. These strains are able to inhibit the growth of certain enteropathogens: Salmonella, Listeria and Campylobacter without interfering with the normal microbiota of the gastrointestinal tract, as stated by using the mixed culture and the spot agar test. Finally, strongly adherent Lact. gasseri were found to inhibit the attachment of Escherichia coli O111 to intestinal Caco-2 cells under the condition of exclusion. CONCLUSIONS: These results indicate that the two strains of Lactobacillus from human origin present important properties for survival in, and colonization of, the gastrointestinal tract, that give them potential probiotic. SIGNIFICANCE AND IMPACT OF THE STUDY: Two strains of Lactobacillus isolated from human vagina of healthy premenopausal women could be promising candidates to be used in the preparation of probiotic products and for their use as health-promoting bacteria.')

225 ('>T', 'TI  - Inhibition of in vitro growth of Shiga toxin-producing Escherichia coli O157:H7 by probiotic Lactobacillus strains due to production of lactic acid.', 'AB  - The inhibiting characteristics of lactic acid bacteria on Shiga toxin-producing Escherichia coli (STEC) O157:H7 (three strains, clinically isolated) was investigated by using a batch fermentation system. The species such as Lactobacillus casei strain Shirota or L. acidophilus YIT 0070 exert growth inhibitory and bactericidal activities on STEC. The pH value and undissociated lactic acid (U-LA) concentration of the culture medium of STEC cocultured with L. casei or L. acidophilus dramatically lowered or increased, respectively [corrected], when compared with those of the control culture. The cytotoxic properties of U-LA on STEC strain 89020087 analyzed in vitro was divided into two phases, i.e., the bacteriostatic phase (between 3.2 to 62 mM) and the bactericidal phase (over 62 mM). These data suggest that the bactericidal effect of Lactobacillus on STEC depends on its lactic acid production and pH reductive effect.')

226 ('>T', 'TI  - In vitro adherence properties of Lactobacillus rhamnosus DR20 and Bifidobacterium lactis DR10 strains and their antagonistic activity against an enterotoxigenic Escherichia coli.', 'AB  - Adhesion and colonisation properties of three probiotic strains namely, Lactobacillus rhamnosus DR20, L. acidophilus HN017, and Bifidobacterium lactis DR10, were determined in vitro using the differentiated human intestinal cell-lines including HT-29, Caco-2, and HT29-MTX, and compared with properties of L. acidophilus LA-1 and L. rhamnosus GG (two commercial probiotic strains). Two independent methods were employed to quantitate the "adhesiveness" of each strain. In the first method, the bacteria adhered to human cells were detected by Gram staining and counted in different fields under a microscope. Bacteria were also radio-labelled and extent of adhesion determined by scintillation counting. All three strains showed strong adhesion with the human intestinal cell lines in vitro. Adhesion indices of the three strains to two cell lines, i.e. HT-29, and Caco-2 varied between 99 +/- 17 and 219 +/- 36. With mucus-secreting cell-line HT29-MTX, the adhesion indices of all the strains were 2-3 times higher. The adhesion indices of L. acidophilus LA-1 and L. rhamnosus GG were comparable to the other three probiotic strains. We also investigated the inhibitory effect of adhering strains against the intestinal cell monolayer colonization by a known enterotoxigenic strain of Escherichia coli (strain O157:H7). Pre-treatment of E. coli O157:H7 with 2.5-fold concentrated cell-free culture supernatants from L. acidophilus HN017, L. rhamnosus DR20 and B. lactis DR10 reduced the culturable E. coli numbers on TSB plates and also reduced the invasiveness and cell association characteristics of this toxic strain. The inhibitory molecules secreted into the spent media by these strains were partially affected by treatments with lactate dehydrogenase, trypsin and proteinase K suggesting that overall inhibition may be due to a synergistic action of lactic acid and proteinaceous substances.')

227 ('>T', 'TI  - Lactobacilli in human dental caries and saliva.', 'AB  - Samples (98 plaque and 72 saliva) from 93 patients with dental caries were investigated for Lactobacillus species which comprised 65 (62.5%) of 104 isolates. Yeasts (20.1%), Streptococcus spp. (8.7%), Staphylococcus spp. (2.9%) and a few unidentified species (5.8%), were also found. The Lactobacillus isolates were L. brevis (24.6%) L. fermentum (18.5%) L. casei (16.9%), L. delbrueckii (15.4%), L. plantarum (9.23%), L. acidophilus (7.69%), L. jensenii (4.62%), L. salivarius (1.54%) and L. gasseri (1.54%). The most common species was L. brevis (24.6%). The strains tested for beta-lactamase production showed 75.4% positive. All the Lactobacillus strains were tested for bacteriocin production against Escherichia coli, Salmonella spp., Shigella dysenteriae, S. sonnei, Klebsiella spp. and Campylobacter sp. All the lactobacilli except L. jensenii produced bacteriocin against at least one of the indicator organisms. The involvement of Lactobacillus in dental caries was established, although its role and mechanism is not well understood. The ability of Lactobacillus spp. to protect their host against certain diseases by inhibiting the growth of potential pathogens was evident.')

231 ('>T', 'TI  - Reduction of fecal shedding of enterohemorrhagic Escherichia coli O157:H7 in lambs by feeding microbial feed supplement.', 'AB  - Enterohemorrhagic Escherichia coli O157:H7, an emerging food-borne pathogen, has  been implicated in several outbreaks in the US. Ruminants, including cattle, sheep and deer are reservoirs of E. coli O157:H7 and fecal shedding of the pathogen forms the vehicle of entry into the human food chain. We studied the efficacy of Lactobacillus acidophilus, Streptococcus faecium, a mixture of L. acidophilus and S. faecium and a mixture of L. acidophilus, S. faecium, Lactobacillus casei, Lactobacillus fermentum and Lactobacillus plantarum in reducing fecal shedding of E. coli O157:H7 by sheep experimentally infected with the pathogen prior to administration with the microbials. Following oral inoculation with 10(10)CFU of E. coli O157:H7, 30 Suffolk ram lambs were blocked by body weight (six blocks of five lambs each) and lambs within the block randomly assigned to five groups. The lamb groups were fed daily for 7 weeks a basal diet without microbial supplement (control) or the basal diet with L. acidophilus or with S. faecium or with a mixture of L. acidophilus and S. faecium or with a mixture of L. acidophilus, S. faecium, L. casei, L. fermentum and L. plantarum. The microbial supplements contained stabilized live naturally occurring bacteria and were mixed with the diet at the rate of 6.0x10(6)CFU per kilogram of diet. Fecal samples were collected weekly and analyzed for E. coli O157:H7 using modified tryptic soy broth with novobiocin as a pre-enrichement broth and cefixim-tellurite sorbitol MacConkey agar (CT-SMAC) as a selective media. E. coli O157:H7 was confirmed by its reaction with O157 and H7 antisera. E. coli O157:H7 was shed continuously and in varying numbers in the feces throughout the 7-week experimental period by all five groups. However, lambs administered a mixture of L. acidophilus, S. faecium, L. casei, L. fermentum and L. plantarum shed significantly lower (P=0.0211) average number of E. coli O157:H7 (2.3log(10)CFU per gram of feces per week) than the other lamb groups over the entire experimental period. S. faecium supplemented lambs were comparable (P=0.0884) to lambs fed a mixture of L. acidophulus and S. faecium in fecal shedding of the pathogen (3.5 versus 4.4log(10)CFU per gram of feces) but significantly lower (P=0.0001) than the control lambs (5.6log(10)CFU per gram of feces) and those supplemented with L. acidophilus (5.5log(10)CFU per gram of feces). Average daily gain (ADG) and gain to feed ratio (G:F) were significantly improved (P=0.0145) by the mixed culture microbials (163.0g and 0.33 for the control, 186.4g and 0.37 for L. acidophulus, 168.2g and 0.36 for S. faecium, 213.6g and 0.46 for L. acidophulus and S. faecium, and 219.1g and 0.44, respectively for L. acidophilus, S. faecium, L. casei, L. fermentum and L. plantarum supplemented lambs. The study indicates that supplementing lambs infected with E. coli O157:H7 with S. faecium or a mixture of S. faecium, L. acidophilus, L. casei, L. fermentum and L. plantarum in the diet can reduce total number of E. coli O157:H7 shed in the feces and improve animal meat production performance as well.')

241 ('>T', 'TI  - [The primary screening of bifidobacteria and lactobacilli strains to develop effective probiotic preparations based on them].', 'AB  - 10 Bifidobacterium strains and 10 Lactobacillus strains were studied for their antagonistic activity with respect to Escherichia coli, Klebsiella ozaenae, Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa and for their sensitivity to antibiotics, widely used in clinical practice. L. acidophilus strain 5/4, L. acidophilus strain 18/4, B. adolescentis strain UX, B. longum strain 44 exhibited the highest antagonistic activity and the highest degree of antibiotic resistance. The restriction analysis of the chromosomal DNA of these strains was then made and their plasmid content was studied, making it possible to recognize these strains in future in the course of in vivo experiments.')

242 ('>T', 'TI  - Immediate effect of Lactobacillus acidophilus on the intestinal flora and fecal enzymes of rats and the in vitro inhibition of Escherichia coli in coculture.', 'AB  - The in vitro role of Lactobacillus acidophilus was investigated to explore the potential to inhibit coliforms. A threefold concentrated cell-free extract from L. acidophilus SBT2074 could efficiently inhibit most of the tested Gram-positive and Gram-negative bacteria. Among the three strains of L. acidophilus, SBT2062, SBT2071, and SBT2074, only L. acidophilus SBT2074 showed this inhibitory property. These three strains were also tested in coculture with Escherichia coli 3544 in skim milk medium. The fermentation could result in complete inhibition of E. coli in 36 h. Short-term administration of L. acidophilus SBT2074 in rats with and without E. coli resulted in significant inhibition of coliforms and anaerobes. The E. coli infected rats regained the normal flora in the presence of lactic acid bacteria. The fecal enzyme beta-glucuronidase activity was also decreased significantly when L. acidophilus SBT2074 was administered and was related to the decreased number of bacteria in the intestinal tract. The analysis of the small intestinal contents showed that the concentrations of coliforms in the duodenum, jejunum, and the ileum were significantly reduced by the administration of lactic acid bacteria. The effects are seen in a short period, suggesting that L. acidophilus SBT2074 fermentate may have clinical application for people suffering from gastrointestinal distress caused by coliforms.')

246 ('>T', 'TI  - Purification and characterization of a bacteriocin produced by Lactobacillus acidophilus IBB 801.', 'AB  - Lactobacillus acidophilus IBB 801 produces a small bacteriocin, designated acidophilin 801, with an estimated molecular mass of less than 6.5 kDa. It displays a narrow inhibitory spectrum (only related lactobacilli but including the Gram-negative pathogenic bacteria Escherichia coli Row and Salmonella panama 1467) with a bactericidal activity. The antimicrobial activity of cell-free culture supernatant fluid was insensitive to catalase but sensitive to proteolytic enzymes such as trypsin, proteinase K and pronase, heat-stable (30 min at 121 degrees C), and maintained in a wide pH range. The proteinaceous compound was isolated from cell-free culture supernatant fluid and purified. Crude bacteriocin was isolated as a floating pellicle after ammonium sulphate precipitation (40% saturation) and partially purified by extraction/precipitation with chloroform/methanol (2/1, v/v). Further purification to homogeneity was performed by reversed phase Fast Performance Liquid Chromatography. The amino acid composition was determined. Amino acid sequencing revealed that the N-terminal end was blocked.')

264 ('>T', 'TI  - Adherence of human vaginal lactobacilli to vaginal epithelial cells and interaction with uropathogens.', 'AB  - Three strains of Lactobacillus, identified as Lactobacillus acidophilus, Lactobacillus gasseri, and Lactobacillus jensenii, were selected from among 70 isolates from the vaginas of healthy premenopausal women for properties relevant to mucosal colonization or antagonism. All three self-aggregated and adhered to epithelial vaginal cells, displacing well-known vaginal pathogens, such as G. vaginalis, and inhibiting the growth in vitro of Escherichia coli and Streptococcus agalactiae. The surface components involved in self-aggregation appeared to be proteins for L. gasseri and lipoproteins for L. acidophilus and L. jensenii, as judged by susceptibility to treatment with appropriate degrading enzymes. The factors responsible for adherence to epithelial vaginal cells seemed to be glycoproteins (L. acidophilus and L. gasseri) and carbohydrate (L. jensenii). The receptors of the vaginal cells were glycolipids, which presumably were the targets of the competition observed between the lactobacilli and the pathogenic microbes.')

265 ('>T', 'TI  - Probiotic fermented food mixtures: possible applications in clinical anti-diarrhoea usage.', 'AB  - A probiotic fermented PCMT food mixture was developed by fermentation of an autoclaved and cooled slurry of pearl millet flour, chickpea flour, skim milk powder and fresh tomato pulp (PCMT 2:1:1:1, w/w) with Lactobacillus acidophilus (10(5) cells/ml), a probiotic organism at 37 degrees C for 24 h. Such a fermented mixture inhibited the growth of pathogenic organisms, namely Shigella dysenteriae, Salmonella typhosa and E. coli. A significant decline in pH with a corresponding increase in titratable acidity due to probiotic fermentation occurred in the developed food mixture. Feeding of the freshly developed fermented mixture to mice suffering from E. coli induced diarrhoea, could help to arrest diarrhoea, reduce moisture, protein and ash contents in their faeces. The counts of lactobacilli increased whereas those of E. coli decreased remarkably in the faeces of mice from the 3rd day of the feeding trial till the end of experimental period. The beneficial effect of probiotic feeding may be due to antimicrobial substances produced by L. acidophilus, which might have neutralized the enterotoxins from E. coli. The cost of one 200 ml glass full of this probiotic drink is no more than one rupee.')

270 ('>T', 'TI  - Antibacterial effect of the adhering human Lactobacillus acidophilus strain LB.', 'AB  - The spent culture supernatant of the human Lactobacillus acidophilus strain LB produces an antibacterial activity against a wide range of gram-negative and gram-positive pathogens. It decreased the in vitro viability of Staphylococcus aureus, Listeria monocytogenes, Salmonella typhimurium, Shigella flexneri, Escherichia coli, Klebsiella pneumoniae, Bacillus cereus, Pseudomonas aeruginosa, and Enterobacter spp. In contrast, it did not inhibit lactobacilli and bifidobacteria. The activity was heat stable and relatively sensitive to enzymatic treatments and developed under acidic conditions. The antimicrobialr activity was independent of lactic acid production. Activity against S. typhimurium SL1344 infecting human cultured intestinal Caco-2 cells was observed as it was in the conventional C3H/He/oujco mouse model with S. typhimurium C5 infection and oral treatment with the LB spent culture supernatant.')

276 ('>T', 'TI  - Antagonistic activity of Lactobacillus acidophilus fermented milk against different pathogenic bacteria.', 'AB  - Lactobacillus acidophilus strains tested showed inhibitory activity towards Salmonella typhi, Staphylococcus aureus, Escherichia coli, Proteus vulgaris and Yersinia enterocolitica. However, wide variations in the activity of the strains were observed. Antagonistic activity of the strains exhibited high heat stability (120 min at 92 degrees C, 15 min at 121 degrees C) but it was markedly influenced by changes in pH. Effect of kind of milk on antagonistic activity was variable, whereas skimming had no significant effect.')

284 ('>T', 'TI  - Antagonism of lactic acid bacteria towards Staphylococcus aureus and Escherichia  coli on agar plates and in milk.', 'AB  - The antagonistic effect of lactic acid bacteria (LAB, including Lactobacillus acidophilus, L. bulgaricus, L. casei and Streptococcus thermophilus) on Staphylococcus aureus and Escherichia coli was evaluated on MRS agar with the deferred and cross-streaking techniques, and in milk with the plate counting method. All LAB were repressive to S aureus and E coli on the agar medium. However, their suppressive activity was significantly reduced when the agar medium was buffered to pH 7.2. In normal milk, L acidophilus strains A and B, S thermophilus and its combinations with L acidophilus A and L bulgaricus 6032 were inhibitory to S aureus, while in mastitic milk, only S thermophilus and its combinations showed inhibition. L acidophilus A and L bulgaricus 34104 were repressive to E coli growth in normal milk. S thermophilus and its combinations were inhibitory to E coli in both the normal and mastitic milk samples. These results indicate that the antagonistic activity of LAB on pathogenic bacteria varied with the type of media in which the tests were done, and that testing of in vitro antagonism in milk would be more informative than that in artificial media for in vivo tests concerning the possible roles of competitive microbiological ecology in mastitis control.')

295 ('>T', "TI  - Antimicrobial activity of neutralized extracellular culture filtrates of lactic acid bacteria isolated from a cultured Indian milk product ('dahi').", "AB  - Neutralized extracellular culture filtrate obtained from isolates of Lactobacillus acidophilus, Lactobacillus delbruecki ssp. bulgaricus, Lactobacillus salivarius and Lactococcus lactis ssp. lactis from 'dahi' showed weak to moderate inhibition of Staphylococcus aureus, Bacillus cereus, Escherichia coli, Bacillus brevis, Bacillus circulans, Bacillus coagulans, Bacillus laterosporus, Bacillus subtilis and Pseudomonas aeruginosa when tested by the diffusion agar well assay method. The effective minimum quantity of lactic culture filtrates required to obtain complete inhibition of an inoculum of 10(3) cfu/ml of the bacteria tested was between 20 and 26% (vol/vol), as determined by the agar incorporation method. Neutralized extracellular culture filtrate of these lactic cultures added at a level of 10% in sterile, 10% reconstituted non-fat dry milk was able to either suppress or retard growth of selected bacterial cultures when incubated at 37 degrees C for 24 h. This study indicated the antimicrobial activity of dahi and the potential of using neutralized extracellular culture filtrate of lactic acid bacteria in the biopreservation of foods.")

300 ('>T', 'TI  - Competitive exclusion of diarrheagenic Escherichia coli (ETEC) from human enterocyte-like Caco-2 cells by heat-killed Lactobacillus.', 'AB  - Diarrheagenic Escherichia coli (ETEC) bearing CFA/I or CFA/II adhesive factors specifically adhere onto the brush border of the polarized epithelial human intestinal Caco-2 cells in culture. Heat-killed Lactobacillus acidophilus strain LB, that adheres onto Caco-2 cells, inhibits diarrheagenic Escherichia coli adhesion in a concentration-dependent manner. Since the L. acidophilus does not express ETEC-CFA adhesive factors, it can be postulated that the heat-killed L. acidophilus LB cells inhibit diarrheagenic E. coli attachment by steric hindrance of the human enterocytic ETEC receptors.')

315 ('>T', 'TI  - Detection of a Lactobacillus substance that inhibits Escherichia coli.', 'AB  - Recent studies have shown that certain lactobacilli strains have the ability to interfere with the adherence and growth of uropathogenic bacteria. This interaction is believed to be important in the maintenance of a normal urogenital flora and in the prevention of infection in females. In the present study, Lactobacillus casei ssp. rhamnosus GR-1 and Lactobacillus acidophilus 76 were found to exert an inhibitory effect on pyelonephritogenic mutant Escherichia coli Hu 734 and E. coli ATCC 25922. The bioactivity of the inhibitor produced by strain GR-1 was retained under pH buffered conditions and was bactericidal. The bioactive substance was heat labile, not precipitated by up to 80% ammonium sulphate, and extractable in chloroform. The data indicated that the inhibitor is not lactic acid or hydrogen peroxide and has a molecular weight greater than 12,000-14,000. Human urine supported production of the inhibitor and reduced and delayed outgrowth of the E. coli. The ability of L. casei GR-1 and possibly other lactobacilli strains to produce inhibitors of uropathogenic bacteria may have clinical importance and significance in the microbial ecology of the urogenital tract.')

320 ('>T', 'TI  - Impact of Lactobacillus acidophilus supplements on the human oropharyngeal and intestinal microflora.', 'AB  - The influence of Lactobacillus acidophilus supplements on the oropharyngeal and intestinal microflora was studied before, during and after L. acidophilus administration. 10 healthy volunteers participated in the study. L. acidophilus was given as a fermented milk product containing 5 X 10(8)-2 X 10(9) CFU/ml in a dose of 250 ml twice a day for 7 days. Only minor changes in the number of aerobic and anaerobic microorganisms in the oropharynx were observed, with no increase in the number of lactobacilli. In the aerobic intestinal microflora a decrease in the number of Escherichia coli was observed in 6/10 subjects. Concerning the anaerobic intestinal microflora there was a significant increase in the number of lactobacilli in 9/10 subjects within 7 days of L. acidophilus administration. The increase in the number of lactobacilli remained as long as the subjects were consuming the L. acidophilus preparation. Lactobacilli returned to the same level as before the study 9 days after the L. acidophilus administration was stopped. Anaerobic cocci showed a decrease in 4/10 subjects, while the number of other anaerobic bacteria remained relatively constant throughout the observation time. These studies suggest that L. acidophilus in this type of preparation should be taken continuously in order to maintain high levels of lactobacilli in the intestine.')

332 ('>T', 'TI  - In vivo inhibitory effects of Lactobacillus acidophilus against pathogenic Escherichia coli in gnotobiotic chicks.', 'AB  - Chicks were hatched germfree in gnotobiotic isolators to determine the inhibitory effects of Lactobacillus acidophilus towards pathogeneic Escherichia coli in vivo. Twelve trials were conducted in two flexible film isolators utilizing a total of 221 chicks. One treatment consisted of inoculating 2-day-old chicks with L. acidophilus, then challenging with pathogenic E. coli with subsequent dosing with L. acidophilus. The other treatment consisted of challenging with the E. coli at 2 days of age, then subsequently dosing with L. acidophilus. Statistical analysis of the data showed initial dosing with L. acidophilus prevented excessive mortality when chicks were challenged with E. coli. Also, continued dosing with L. acidophilus lowered the pH in the crop, cecum, and rectum whether chicks were initially given L. acidophilus or E. coli. This strain of L. acidophilus was capable of competing with E. coli in the gut of gnotobiotic chicks.')

334 ('>T', 'TI  - Lactobacillus prophylaxis for diarrhea due to enterotoxigenic Escherichia coli.', 'AB  - In vitro and animal experiments indicated that lactobacilli might prevent Escherichia coli from colonizing the intestine and may produce substances counteracting enterotoxin. Lactinex, a commercial preparation of dried Lactobacillus acidophilus and L. bulgaricus, is marketed for uncomplicated diarrhea. Preliminary experiments in nonfasting volunteers indicated that lactobacilli in this preparation colonized the small intestine for up to 6 h. To evaluate the protective efficacy of Lactinex, a double-blind randomized study was carried out in which 48 volunteers (23 receiving Lactinex and 25 receiving placebos) were challenged with E. coli strains that produced heat-stable or heat-labile enterotoxins or both. No significant differences between the two groups were noted with respect to attack rate, incubation period, duration of diarrhea, volume and number of liquid stools, and coproculture yields. These data suggest that this lactobacillus preparations does not prevent or alter the course of enterotoxigenic E. coli diarrhea in adults. Lack of efficacy occurred despite efforts to maximize small bowel colonization, including administration of Lactinex in milk and in a 6-hour-interval regimen during 36 h before and 96 h after challenge.')

FALSE POSITIVES:
('>F', 'TI  - Determining the role of a probiotic in the restoration of intestinal microbial balance by molecular and cultural techniques.', 'AB  - The human intestine has a vast variety of microorganisms, and their balance is dependent on several factors. Antibiotics affect microfloral balance and allow naturally opportunistic organisms to multiply. Azithromycin is the most widely used macrolide antibiotic, active against a wide number of pathogens including Pseudomonas aeruginosa and Staphylococcus aureus. It is currently used in the treatment of cystic fibrosis patients. The use of probiotics has advantages in gastrointestinal conditions, including infectious diarrhea and imbalance due to antibiotic use. In this research, the effect of azithromycin on the intestinal microbiota of Sprague Dawley rats and the role of Lactobacillus acidophilus in the restoration of the balance by employing molecular and cultural techniques was investigated. PCR with universal primers targeting the V3 region of the 16S rRNA gene followed by DGGE was used to characterize the overall intestinal microbiota composition. Cultivable fecal bacteria count using microbiological media and semi-quantitative PCR with group-specific primers were also utilized to analyze the effects of antibiotic and probiotic on microflora. We found that the total amount of 16S rRNA gene and fecal aerobic bacterial count was reduced following azithromycin administration along with elimination of non-pathogenic Escherichia coli, but it was restored by the use of the probiotic. The results from PCR with group-specific primers showed that Bacteroides sp was present in the control and probiotic groups, but it was nearly eliminated in the antibiotic group. Moreover, semi-quantitative PCR revealed that the numbers of Enterobacteriaceae were nearly the same in the probiotic group and decreased in the antibiotic group, while Bifidobacterium was significantly increased in the probiotic group and decreased in the antibiotic group (P < 0.05) as compared with that in the control group. Azithromycin-induced dysbiosis can result in prolonged deleterious effects on the host. The present study revealed that the use of lactic acid bacteria particularly L. acidophilus helped to restore intestinal microfloral balance.')

('>F', 'TI  - Construction and validation of a mCherry protein vector for promoter analysis in  Lactobacillus acidophilus.', 'AB  - Lactobacilli are widespread in natural environments and are increasingly being investigated as potential health modulators. In this study, we have adapted the broad-host-range vector pNZ8048 to express the mCherry protein (pRCR) to expand the usage of the mCherry protein for analysis of gene expression in Lactobacillus. This vector is also able to replicate in Streptococcus pneumoniae and Escherichia coli. The usage of pRCR as a promoter probe was validated in Lactobacillus acidophilus by characterizing the regulation of lactacin B expression. The results show that the regulation is exerted at the transcriptional level, with lbaB gene expression being specifically induced by co-culture of the L. acidophilus bacteriocin producer and the S. thermophilus STY-31 inducer bacterium.')

('>F', 'TI  - The antibacterial and antifungal activity of essential oils extracted from Guatemalan medicinal plants.', 'AB  - CONTEXT: Essential oils are prevalent in many medicinal plants used for oral hygiene and treatment of diseases. OBJECTIVE: Medicinal plant species were extracted to determine the essential oil content. Those producing sufficient oil were screened for activity against Staphylococcus aureus, Escherichia coli, Streptococcus mutans, Lactobacillus acidophilus, and Candida albicans. MATERIALS AND METHODS: Plant samples were collected, frozen, and essential oils were extracted by steam distillation. Minimum inhibitory concentrations (MIC) were determined using a tube dilution assay for those species yielding sufficient oil. RESULTS: Fifty-nine of the 141 plant species produced sufficient oil for collection and 12 species not previously reported to produce essential oils were identified. Essential oil extracts from 32 species exhibited activity against one or more microbes. Oils from eight species were highly inhibitory to S. mutans, four species were highly inhibitory to C. albicans, and 19 species yielded MIC values less than the reference drugs. DISCUSSION: RESULTS suggest that 11 species were highly inhibitory to the microbes tested and merit further investigation. Oils from Cinnamomum zeylanicum Blume (Lauraceae), Citrus aurantiifolia (Christm.) Swingle (Rutaceae), Lippia graveolens Kunth (Verbenaceae), and Origanum vulgare L. (Lamiaceae) yielded highly significant or moderate activity against all microbes and have potential as antimicrobial agents. CONCLUSION: Teas prepared by decoction or infusion are known methods for extracting essential oils. Oils from 11 species were highly active against the microbes tested and merit investigation as to their potential for addressing health-related issues and in oral hygiene.')

('>F', 'TI  - Effects of Citrus junos by-products fermented with multistrain probiotics on growth performance, immunity, caecal microbiology and meat oxidative stability in broilers.', 'AB  - 1. The present study was conducted to develop Citrus junos probiotics (CJP), using by-products of Citrus junos fermented with multispecies probiotic bacteria including Saccharomyces cerevisiae, Enterococcus faecium, Lactobacillus acidophilus and Bacillus subtilis. The effects of dietary CJP on the growth performance, immune status, caecal microbiology and meat oxidative stability of broiler were investigated. 2. A total of 240 one-day-old Ross broiler chicks were used in a 35-d experiment in which the chicks were randomly allotted to one of the 4 dietary treatments (0, 5, 10 and 20 g CJP/kg diet) in a completely randomised design. 3. Dietary supplementation of 5 g/kg CJP significantly increased body weight, average daily gain and average daily feed intake of broiler during the overall experimental period. 4. Serum immunoglobulin (Ig)M concentration was significantly increased by 10 and 20 g/kg CJP, whereas the IgG and IgA concentration remained unaffected. In addition, 20 g/kg CJP significantly inhibited proliferation of Escherichia coli without affecting the concentration of Lactobacillus or Bacillus spp. 5. A significant reduction in the thiobarbituric acid reactive substances values of breast and thigh meat was observed in response to increasing concentration of dietary CJP. 6. Thus, the results suggest that CJP up to a concentration of 20 g/kg can be used in the diet of broilers to improve immunity and to reduce caecal E. coli and TBARS values of breast and thigh meat without any adverse effects on growth performance.')

('>F', 'TI  - A food additive with prebiotic properties of an alpha-d-glucan from lactobacillus plantarum DM5.', 'AB  - An alpha-d-glucan produced by Lactobacillus plantarum DM5 was explored for in vitro prebiotic activities. Glucan-DM5 demonstrated 21.6% solubility, 316.9% water holding capacity, 86.2% flocculation activity, 71.4% emulsification activity and a degradation temperature (Td) of 292.2 degrees C. Glucan-DM5 exhibited lowest digestibility of 0.54% by artificial gastric juice, 0.21% by intestinal fluid and 0.32% by alpha-amylase whereas the standard prebiotic inulin, showed 25.23%, 5.97% and 19.13%, hydrolysis, respectively. Prebiotic activity assay of glucan-DM5 displayed increased growth of probiotic bacteria such as Bifidobacterium infantis and Lactobacillus acidophilus, but did not support the growth of non-probiotic bacteria such as Escherichia coli and Enterobacter aerogenes. The overall findings indicated that glucan from L. plantarum DM5 can serve as a potential prebiotic additive for food products.')

('?F', 'TI  - Variable efficacy of a vaccine and direct-fed microbial for controlling Escherichia coli O157:H7 in feces and on hides of feedlot cattle.', 'AB  - To evaluate the efficacy of a type-III secreted proteins vaccine and a Lactobacillus-acidophilus-based direct-fed microbial (DFM) for controlling Escherichia coli O157:H7, cattle (n=864) were allocated to the following groups: DFM, finishing diets containing 10(9) colony-forming units (CFU)/animal/day L. acidophilus and Propionibacterium freudenreichii; VAC, finishing diets and 2 mL intramuscular injection of vaccine at allocation and 28 days later; or CON, finishing diets only. Cattle within replicates were stratified by initial levels of E. coli O157:H7 and randomized to experimental groups, with 30 pens allocated on June 15, 2011 (AS1), 18 pens allocated on June 28, 2011 (AS2), and 18 cattle per pen. Rectal fecal samples and perineal swabs were collected at 28-day intervals until shipment to slaughter (103-145 days on trial). Numbers of cattle with enumerable E. coli O157:H7 (>/=1.6 CFU/g feces) were reduced in AS1 and AS2 by VAC (p=0.008), although interventions had no impact on numbers of E. coli O157:H7 shed. For AS1, VAC reduced prevalence of E. coli O157:H7 in feces (p=0.03) and perineal swabs (p=0.04) in the feeding period but not at shipment to slaughter. For AS2, prevalence of E. coli O157:H7 was not reduced in either feces or perineal swabs by VAC at any time. For AS1, DFM reduced prevalence of E. coli O157:H7 in perineal swabs (p=0.01) during the feeding period. For AS2, DFM increased E. coli O157:H7 detection in feces (p=0.03) and perineal swabs (p=0.01) at shipment to slaughter. Seventy-five percent of AS1 E. coli O157:H7 isolates had only stx1, while 87% of AS2 isolates had stx1 and stx2 genes. Of the two interventions, VAC shows the most potential for pre-harvest control of E. coli O157:H7, but due to variable efficacy of both DFM and VAC, additional product development is necessary to ensure more consistent pre-harvest control of E. coli O157:H7.')

('?F <NO EVIDENCE TOWARDS, claims that no inhibition>', 'TI  - Escherichia coli O26 in feedlot cattle: fecal prevalence, isolation, characterization, and effects of an E. coli O157 vaccine and a direct-fed microbial.', 'AB  - Escherichia coli O26 is second only to O157 in causing foodborne, Shiga toxin-producing E. coli (STEC) infections. Our objectives were to determine fecal prevalence and characteristics of E. coli O26 in commercial feedlot cattle (17,148) that were enrolled in a study to evaluate an E. coli O157:H7 siderophore receptor and porin (SRP((R))) vaccine (VAC) and a direct-fed microbial (DFM; 10(6) colony-forming units [CFU]/animal/day of Lactobacillus acidophilus and 10(9) CFU/animal/day of Propionibacterium freudenreichii). Cattle were randomly allocated to 40 pens within 10 complete blocks; pens were randomly assigned to control, VAC, DFM, or VAC+DFM treatments. Vaccine was administered on days 0 and 21, and DFM was fed throughout the study. Pen-floor fecal samples (30/pen) were collected weekly for the last 4 study weeks. Samples were enriched in E. coli broth and subjected to a multiplex polymerase chain reaction (PCR) designed to detect O26-specific wzx gene and four major virulence genes (stx1, stx2, eae, and ehxA) and to a culture-based procedure that involved immunomagnetic separation and plating on MacConkey agar. Ten presumptive E. coli colonies were randomly picked, pooled, and tested by the multiplex PCR. Pooled colonies positive for O26 serogroup were streaked on sorbose MacConkey agar, and 10 randomly picked colonies per sample were tested individually by the multiplex PCR. The overall prevalence of E. coli O26 was higher (p<0.001) by the culture-based method compared to the PCR assay (22.7 versus 10.5%). The interventions (VAC and or DFM) had no impact on fecal shedding of O26. Serogroup O26 was recovered in pure culture from 23.9% (260 of 1089) of O26 PCR-positive pooled colonies. Only 7 of the 260 isolates were positive for the stx gene and 90.1% of the isolates possessed an eaebeta gene that codes for intimin subtype beta, but not the bfpA gene, which codes for bundle-forming pilus. Therefore, the majority of the O26 recovered from feedlot cattle feces was atypical enteropathogenic E. coli, and not STEC.')

('>F', 'TI  - Hydrolysates of glycated and heat-treated peanut 7S globulin (Ara h 1) modulate human gut microbial proliferation, survival and adhesion.', 'AB  - AIMS: Evaluation of an effect of glycation of Ara h 1 on proliferation and survival rate and adhesion of intestinal Enterococcus faecalis, Escherichia coli and Lactobacillus acidophilus. METHODS AND RESULTS: Pure Ara h 1 heated at three different temperature conditions (G37, G60 and C145 degrees C) in the presence or absence of glucose was subjected to enzymatic hydrolysis. Impacts of Ara h 1 hydrolysates on the bacterial proliferation, survival rate and adhesion to Caco-2 cells in mono and heterogeneous cultures were studied with fluorescent techniques: DAPI, LIVE/DEAD staining and FISH. Examined hydrolysates hindered proliferation of E. coli and Ent. faecalis with simultaneous decrease in their survival. Maillard reaction (MR, glycation) of Ara h 1 did not alter the effect of hydrolysates on bacterial proliferation rate. Hydrolysates modified at 60 and 145 degrees C with glucose altered the profile of immobilized bacteria, mostly by lowering the number of adhering E. coli and promoting the adhesion of bacteria from genera Lactobacillus and Enterococcus. CONCLUSIONS: Ara h1 hydrolysates processed in various ways demonstrated their strong modulatory effect on bacterial proliferation, survival rate and adhesion. SIGNIFICANCE AND IMPACT OF THE STUDY: Reducing the adhesion of opportunistic bacteria by hydrolysates of Ara h 1 glycated at 60 and 145 degrees C, together with modulation of immobilization of beneficial lactobacilli and enterococci, may be of relevance in terms of the physiological status of the intestinal barrier.')

('>F', 'TI  - Evaluation of bean and soy tempeh influence on intestinal bacteria and estimation of antibacterial properties of bean tempeh.', 'AB  - In this study the effect of bean tempeh on the growth of Bacillus subtilis, Escherichia coli, Lactobacillus acidophilus and Lactobacillus paracasei bacteria was investigated. Antibacterial activity was observed only in relation to the bacteria Bacillus subtilis. The effect of tempeh products on human intestinal microflora was also assessed. Bean and soy tempeh were culinarily processed and next digested in conditions simulating the human digestive tract (one of the digestive tracts was equipped with a mechanism simulating absorption). Soy tempeh stimulated most the growth of bacteria of the genus Bifidobacterium, while bean tempeh that of Escherichia coli. Using simulation of absorption for the digestion of fried soy tempeh resulted in a higher rise in the bacteria count of the genus Lactobacillus, while after digestion of fried bean tempeh the highest increase was recorded for Bifidobacterium and E. coli.')

('>F', 'TI  - Lactobacillus acidophilus supplementation in human subjects and their resistance  to enterotoxigenic Escherichia coli infection.', 'AB  - To assess the effect of Lactobacillus acidophilus (American Type Culture Collection (ATCC) 700396) on enterotoxigenic Escherichia coli (ETEC) infection, in the present study, a parallel, double-blind, placebo-controlled 4-week intervention was performed in healthy males. The subjects largely consumed their habitual diet, but had to abstain from consuming dairy foods generally high in Ca. The subjects were randomised into the L. acidophilus (dose 10(9) colony-forming units twice daily; n 20) or the placebo (n 19) group. After an adaptation period of 2 weeks, the subjects were orally infected with a live, but attenuated, ETEC vaccine, able to induce mild, short-lived symptoms. Before and after the challenge, the subjects recorded stool consistency, bowel habits, and frequency and severity of gastrointestinal complaints. The ETEC challenge led to a significant increase in faecal output on the 2nd day and a concomitant increase in Bristol stool scale scores. Likewise, abdominal pain, bloating, flatulence, fever, headache and nausea peaked 1 d after the oral challenge. The concentrations of faecal calprotectin and IgA peaked 2 d after and that of serum IgM peaked 9 and 15 d after the oral challenge. The concentrations of serum IgA and IgG were unaffected. The ETEC challenge led to a reduction in the number of Bacteroides-Prevotella, Bifidobacterium, Clostridium cluster XIVab and total faecal bacteria. Probiotic treatment was associated with a larger increase in Bristol stool scale scores and more fever, headache and nausea after the ETEC challenge compared with the placebo treatment. These differences were, however, small and with substantial variation within the groups. Oral application of an attenuated live ETEC vaccine provides a useful model for food-borne infections. Supplementation with L. acidophilus ATCC 700396, however, was ineffective in reducing ETEC infection symptoms in healthy men.')

('>F', 'TI - Secreted aspartic peptidases of Candida albicans liberate bactericidal hemocidins from human hemoglobin.', "AB  - Secreted aspartic peptidases (Saps) are a group of ten acidic hydrolases considered as key virulence factors of Candida albicans. These enzymes supply the fungus with nutrient amino acids as well as are able to degrade the selected host's proteins involved in the immune defense. Our previous studies showed that the human menstrual discharge is exceptionally rich in bactericidal hemoglobin (Hb) fragments - hemocidins. However, to date, the genesis of such peptides is unclear. The presented study demonstrates that the action of C. albicans isozymes Sap1-Sap6, Sap8 and Sap9, but not Sap7 and Sap10, toward human hemoglobin leads to limited proteolysis of this protein and generates a variety of antimicrobial hemocidins. We have identified these peptides and checked their activity against selected microorganisms representative for human vagina. We have also demonstrated that the process of Hb hydrolysis is most effective at pH 4.0, characteristic for vagina, and the liberated peptides showed pronounced killing activity toward Lactobacillus acidophilus, and to a lower degree, Escherichia coli. However, only a very weak activity toward Staphylococcus aureus and C. albicans was noticed. These findings provide interesting new insights into pathophysiology of human vaginal candidiasis and suggest that C. albicans may be able to compete with the other microorganisms of the same physiological niche using the microbicidal peptides generated from the host protein.")

('>F', 'TI  - Impact of 6 different intestinal bacteria on broiler breeder sperm motility in vitro.', 'AB  - Male fertility is often evaluated by measuring sperm parameters, including concentration, viability, and motility. This is important because after copulation occurs, sperm must overcome many barriers in the female reproductive tract to fertilize the ovum. In mammalian species, sperm have been shown to have reduced motility when bacteria are present. In male broiler breeders, bacteria have been associated with spermatozoa, but their effect on motility has not been investigated. The sperm quality index is a modern rapid method of evaluating avian sperm motility. Therefore, the objective of this study was to use the sperm quality index to determine if broiler breeder sperm motility is reduced when semen is exposed to various bacteria. In this experiment, semen was collected from 20 broiler breeders to obtain a pooled neat semen sample. Six different intestinal bacteria, Salmonella enterica, Escherichia coli, Campylobacter jejuni, Clostridium bifermentans, Lactobacillus acidophilus, and Bifidobacterium animalis were cultured overnight. For each bacterium, 50 microL of semen was diluted in 450 microL of saline, sterile broth, or the overnight culture, creating 3 treatments. The experiment was repeated twice. In each treatment, 3 replicates were evaluated at 0 and 10 min postinoculation, creating a completely randomized design with a split plot over time. Also, the pH was measured for each treatment at 0 and 10 min. The results indicated that all broths containing bacteria immediately reduced broiler breeder sperm motility when compared with the controls (P < 0.0001), but broths containing Bifidobacterium or Lactobacillus virtually made sperm immotile. Although broth containing Salmonella, Campylobacter, and Bifidobacterium immediately reduced sperm motility, the reduction did not change over time. Broths containing E. coli, Clostridium, and Lactobacillus reduced sperm motility immediately, but over time motility continued to decrease. However, pH was increased when semen was exposed to the E. coli and Campylobacter treatment, but when semen was exposed to Bifidobacterium and Lactobacillus treatments, pH was reduced. In conclusion, the results indicate that bacteria can reduce broiler breeder sperm motility upon exposure.')

('>F', 'TI  - In vitro assessment of functional properties of lactic acid bacteria isolated from faecal microbiota of healthy dogs for potential use as probiotics.', 'AB  - Lactic acid bacteria were isolated and identified in the faeces of Chinese Crested and Yorkshire terrier pups and their probiotic features were investigated in vitro. Thirty seven isolates were identified as Lactobacillus or Enterococcus. Out of these isolates, 31 were lactic acid bacteria (LAB) and belonged to the species Lactobacillus reuteri (16/37; 43.3%), Lactobacillus animalis (7/37; 18.9%), Lactobacillus acidophilus (3/37; 8.1%), Lactobacillus sanfranciscensis (2/37; 5.4%), Lactobacillus murinus (2/37; 5.4%), and Lactobacillus paraplantarum (1/37; 2.7%), while six other LAB isolates were Enterococcus spp. (6/37; 16.2%). Strains were tested for resistance to gastric acidity (pH 2.5 for 3 h) and bile salts (0.3% ox gall), cell surface hydrophobicity by microbial adhesion to solvents, antagonism against pathogenic bacteria (Staphylococcus aureus, Enterococcus faecalis, Bacillus cereus, Pseudomonas aeruginosa, Escherichia coli, Salmonella enterica serovar Typhimurium and Listeria monocytogenes), production of hydrogen peroxide, and antibiotic susceptibility. Thirty four strains were highly resistant to acidic conditions with slight (18 strains) to moderate (16 strains) growth inhibition by bile salts. Seven isolates had highly hydrophobic cellular surfaces and 28 strains exhibited strong antagonism against the bacterial pathogens tested, although 8 isolates tested against Leptospira interrogans had no effect on pathogen growth. All isolates produced low rates of hydrogen peroxide. Based on these results, two Lactobacillus strains showed promising probiotic-related features and merit investigation as probiotics for dogs.')

('>F', 'TI  - Broad-spectrum antibacterial activity of carbon nanotubes to human gut bacteria.', 'AB  - Carbon nanotubes (CNTs) hold promise in manufacturing, environmental, and biomedical applications, as well as food and agricultural industries. Previous observations have shown that CNTs have antimicrobial activity; however, the impact of CNTs to human gut microbes has not been investigated. Here, the antibacterial activity of CNTs against the microbes commonly encountered in the human digestion system--L. acidophilus, B. adolescentis, E. coli, E. faecalis, and S. aureus--are evaluated. The bacteria studied include pathogenic and non-pathogenic, gram-positive and negative, and both sphere and rod strains. In this study, CNTs, including single-walled CNTs (SWCNTs, 1-3 mum), short and long multi-walled CNTs (s-MWCNTs: 0.5-2 mum; l-MWCNTs: >50 mum), and functionalized multi-walled CNTs (hydroxyl- and carboxyl-modification, 0.5-2 mum), all have broad-spectrum antibacterial effects. Notably, CNTs may selectively lyse the walls and membranes of human gut microbes, depending on not only the length and surface functional groups of CNTs, but also the shapes of the bacteria. The mechanism of antibacterial activity is associated with their diameter-dependent piercing and length-dependent wrapping on the lysis of microbial walls and membranes, inducing release of intracellular components DNA and RNA and allowing a loss of bacterial membrane potential, demonstrating complete destruction of bacteria. Thin and rigid SWCNT show more effective wall/membrane piercing on spherical bacteria than MWCNTs. Long MWCNT may wrap around gut bacteria, increasing the area making contact with the bacterial wall. This work suggests that CNTs may be broad-spectrum and efficient antibacterial agents in the gut, and selective application of CNTs could reduce the potential hazard to probiotic bacteria.')

('>F', 'TI  - Effects of clinoptilolite and modified clinoptilolite on the growth performance,  intestinal microflora, and gut parameters of broilers.', 'AB  - The purpose of this study was to evaluate the effect of natural clinoptilolite (NCLI) and modified clinoptilolite (MCLI) on broiler performance, gut morphology, and its relation to gut circumstances. A total of two hundred forty 1-d-old male chicks were randomly assigned to 3 treatments, each of which comprised 8 pens of 10 chicks per pen. Birds in the control group were fed the basal diet, whereas those in the experimental groups were fed diets supplemented with NCLI at 2% (NCLI group) or MCLI at 2% (MCLI group) for 42 d. The results showed that compared with the control, supplementation with NCLI or MCLI had no significant (P > 0.05) effects on productive parameters from d 1 to 42. Supplementation with MCLI and NCLI was associated with greater (P < 0.05) villus height in the jejunal and ileal mucosa compared with those areas in the controls from d 1 to 42. However, supplementation with NCLI and MCLI had no significant (P > 0.05) influence on the crypt depth in the jejunal and ileal mucosa compared with those in the controls. Total viable counts of Escherichia coli were significantly (P < 0.05) decreased by MCLI and NCLI from d 1 to 21. The NCLI and MCLI significantly increased the total viable counts of Lactobacillus acidophilus from d 22 to 42. Small intestine and cecal pH values in the MCLI group were found to be lower (P < 0.05) than those in other groups. Total volatile fatty acid concentrations were significantly (P < 0.05) decreased in both experimental groups from d 22 to 42. This study showed that NCLI or MCLI, as feed additives for broilers, had a positive effect on gut parameters by acting on microbial populations of the digestive tract.')

('>F', 'TI  - Antimicrobial potential for the combination of bovine lactoferrin or its hydrolysate with lactoferrin-resistant probiotics against foodborne pathogens.', 'AB  - Previous reports have shown that several probiotic strains can resist the antibacterial activity of bovine lactoferrin (bLf), but the results are inconsistent. Moreover, a portion of orally administered apo-bLf is digested in vivo by pepsin to yield bLf hydrolysate, which produces stronger antibacterial activity than that observed with apo-bLf. However, whether bLf hydrolysate affects the growth of probiotic strains is unclear. Therefore, various probiotic strains in Taiwan were collected and evaluated for activity against apo-bLf and bLf hydrolysate in vitro. Thirteen probiotic strains were evaluated, and the growth of Lactobacillus acidophilus ATCC 4356, Lactobacillus salivarius ATCC 11741, Lactobacillus rhamnosus ATCC 53103, Bifidobacterium longum ATCC 15707, and Bifidobacterium lactis BCRC 17394 were inhibited by both apo-bLf and bLf hydrolysate. The growth of 8 strains were not affected by apo-bLf and bLf hydrolysate, including L. rhamnosus ATCC 7469, Lactobacillus reuteri ATCC 23272, Lactobacillus fermentum ATCC 11739, Lactobacillus coryniformis ATCC 25602, L. acidophilus BCRC 14065, Bifidobacterium infantis ATCC 15697, Bifidobacterium bifidum ATCC 29521, and Pediococcus acidilactici ATCC 8081. However, apo-bLf and its hydrolysate inhibited the growth of foodborne pathogens, including Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, and Enterococcus faecalis. Moreover, the supernatants produced by L. fermentum, B. lactis, and B. longum inhibited the growth of most pathogens. Importantly, a combination of apo-bLf or bLf hydrolysate with the supernatants of cultures of the organisms described above showed synergistic or partially synergistic effects against the growth of most of the selected pathogens. In conclusion, several probiotic strains are resistant to apo-bLf and bLf hydrolysate, warranting clinical studies to evaluate the antimicrobial potential for the combination of apo-bLf or its hydrolysate with specific probiotics.')

('>F', 'TI  - Astragalus root and elderberry fruit extracts enhance the IFN-beta stimulatory effects of Lactobacillus acidophilus in murine-derived dendritic cells.', 'AB  - Many foods and food components boost the immune system, but little data are available regarding the mechanisms by which they do. Bacterial strains have disparate effects in stimulating the immune system. In dendritic cells, the gram-negative bacteria Escherichia coli upregulates proinflammatory cytokines, whereas gram-positive Lactobacillus acidophilus induces a robust interferon (IFN)-beta response. The immune-modulating effects of astragalus root and elderberry fruit extracts were examined in bone marrow-derived murine dendritic cells that were stimulated with L. acidophilus or E. coli. IFN-beta and other cytokines were measured by ELISA and RT-PCR. Endocytosis of fluorescence-labeled dextran and L. acidophilus in the presence of elderberry fruit or astragalus root extract was evaluated in dendritic cells. Our results show that both extracts enhanced L. acidophilus-induced IFN-beta production and slightly decreased the proinflammatory response to E. coli. The enhanced IFN-beta production was associated with upregulation of toll-like receptor 3 and to a varying degree, the cytokines IL-12, IL-6, IL-1beta and TNF-alpha. Both extracts increased endocytosis in immature dendritic cells, and only slightly influenced the viability of the cells. In conclusion, astragalus root and elderberry fruit extracts increase the IFN-beta inducing activity of L. acidophilus in dendritic cells, suggesting that they may exert antiviral and immune-enhancing activity.')

('>F#others', 'TI  - [Influence of corynebacteria metabolites on antagonistic activity of H2O2 producing lactobacilli].', 'AB  - AIM: Study combined influence of Corynebacterium genus bacteria metabolites and H2O2 producing lactobacilli on survival rate of Staphylococcus aureus, Escherichia coli and Lactobacillus acidophilus. MATERIALS AND METHODS: The ability to inhibit catalase of the test strains used and to reduce bactericidal effect of hydroxyl radical were determined in corynebacteria. H2O2 containing metabolites were obtained by cultivating lactobacilli in mineral medium, the amount of H2O2 was determined by oxidation of TMB by peroxidase. Bactericidal effect of lactobacilli metabolites for test strains treated by corynebacteria metabolites was evaluated by seeding results. Results. Inhibitio by corynebacteria metabolites of S. aureus catalase activity by 30-40% and E. coli catalase activ ity by 40-70% was shown. A reduction of bactericidal effect of hydroxyl radicals by corynebacteria metabolites by 30-35% for S. aureus, 38-42% for E. coli and 70-73% for L. acidophilus was noted. The enchantment of bactericidal effect of lactobacilli after treatment of the test strain by corynebacteria metabolites against S. aureus and E. coli manifested by reduction of the numbe of viable cells by 2-3 lg CFU. For L. acidophilus the bactericidal effect oflactobacilli metabolite in the same conditions reduced, and that led to the increase ofviability by 2-4 lg PFU. CONCLUSION: A conclusion on the possibility of regulation by associative bacteria the manifestations of antagonistic activity of H2O2 producing dominant microorganisms is made based on the data obtained.')

('>F', 'TI  - Efficacy of a vaccine and a direct-fed microbial against fecal shedding of Escherichia coli O157:H7 in a randomized pen-level field trial of commercial feedlot cattle.', "AB  - Our primary objective was to determine the efficacy of a siderophore receptor and porin proteins-based vaccine (VAC) and a Lactobacillus acidophilus-based direct-fed microbial (DFM) against fecal shedding of Escherichia coli O157:H7 in commercial feedlot cattle fed a corn grain-based diet with 25% distiller's grains. Cattle projected to be on a finishing diet during the summer were randomly allocated into 40 study pens within ten blocks based on allocation dates. Blocks were complete; each of the four pens within a block was randomly assigned one treatment: control, VAC, DFM, or VAC+DFM. The DFM was fed (10(6)CFU/animal/day of Lactobacillus) throughout the study periods (84-88 days) and cattle were vaccinated at enrollment and again three weeks later. Fresh fecal samples (30/pen) from pen floors were collected weekly for four consecutive weeks (study days 52-77). Two concurrent culture procedures were used to enable estimates of E. coli O157:H7 shedding prevalence and prevalence of high shedders. From 4800 total samples, 1522 (31.7%) were positive for E. coli O157:H7 and 169 (3.5%) were considered high shedders. Pen-level linear mixed models were used for data analyses. There were no significant interactions among treatments and time of sampling. However, vaccinated pens had lower (P<0.01) overall prevalence of E. coli O157:H7 (model-adjusted mean +/- SEM=17.4 +/- 3.95%) and lower (P<0.01) prevalence of high shedders (0.95 +/- 0.26%) than unvaccinated pens (37.0 +/- 6.32% and 4.19 +/- 0.81%, respectively). There was no evidence of a DFM effect on either measure of E. coli O157:H7 shedding. Results indicate that a two-dose regimen of the vaccine significantly reduces fecal prevalence of E. coli O157:H7 (vaccine efficacy of 53.0%) and prevalence of E. coli O157:H7 high shedders (vaccine efficacy of 77.3%) in commercial feedlot cattle reared in the summer on a finishing diet with 25% distiller's grains.")

('>F', 'TI  - Antimicrobial activity of Sesbania grandiflora flower polyphenol extracts on some pathogenic bacteria and growth stimulatory effect on the probiotic organism Lactobacillus acidophilus.', 'AB  - Polyphenolic extracts (PE) of edible flower of Sesbania grandiflora were tested to evaluate its antimicrobial effect against some common pathogenic bacteria and growth promoting property against probiotic organism Lactobacillus acidophilus. The antimicrobial activity of S. grandiflora flower PE against selected pathogens was evaluated using both in vitro and in situ methods. In vitro studies suggested that PE has inhibitory effect against Staphylococcus aureus, Shigella flexneri 2a, Salmonella Typhi, Escherichia coli and Vibrio cholerae. The gram-positive organism S. aureus was the most sensitive organism to PE and minimum inhibitory concentration (MIC) was found to be 0.013 mg/mL where as the MIC of PE against V. cholerae was the highest (0.25 mg/mL). On the other hand PE showed growth promoting effect on the common probiotic bacterium L. acidophilus. The major finding was that S. grandiflora PE induced a significant biomass increase of L. acidophilus grown in liquid culture media. PE showed reduction of S. aureus growth in food (fish) during storage at 10 degrees C. High performance liquid chromatography analysis showed that rutin, a major flavonoid of the PE diminished in the culture medium MRS broth with the growth of L. acidophilus.')

('>F', 'TI  - Elevated IgG levels against specific bacterial antigens in obese patients with diabetes and in mice with diet-induced obesity and glucose intolerance.', 'AB  - High fat diets increase the risk for insulin resistance by promoting inflammation. The cause of inflammation is unclear, but germfree mouse studies have implicated commensal gut bacteria. We tested whether diet-induced obesity, diabetes, and inflammation are associated with anti-bacterial IgG. Blood from lean and obese healthy volunteers or obese patients with diabetes were analyzed by ELISA for IgG against extracts of potentially pathogenic and pro-biotic strains of Escherichia coli (LF-82 and Nissle), Bacteroides thetaiotaomicron, and Lactobacillus acidophilus, and for circulating tumor necrosis factor alpha (TNFalpha). C57Bl/6 mice were fed low- or high-fat diets (10% or 60% kcal from fat) for 10 weeks and tested for anti-bacterial IgG, bodyweight, fasting glucose, and inflammation. Obese diabetic patients had significantly more IgG against extracts of E. coli LF-82 compared with lean controls, whereas IgG against extracts of the other bacteria was unchanged. Circulating TNFalpha was elevated and correlated with IgG against the LF-82 extract. Mice fed high-fat diets had increased fasting glucose levels, elevated TNFalpha and neutrophils, and significantly more IgG against the LF-82 extracts. Diabetes in obesity is characterized by increased IgG against specific bacterial antigens. Specific commensal bacteria may mediate inflammatory effects of high-fat diets.')

('>F', 'TI  - Effect of a synbiotic on microbial community structure in a continuous culture model of the gastric microbiota in enteral nutrition patients.', "AB  - Patients with dysphagia require long-term nutritional support. This can be delivered by the enteral route via a percutaneous endoscopic gastrostomy (PEG) tube. Enteral nutrition (EN) bypasses the body's innate defences that prevent the microbial colonization of the proximal gut, which predisposes to microbial overgrowth. A continuous culture model simulating the upper gastrointestinal tract microbiota of EN patients was used to investigate the effects of a synbiotic (Lactobacillus acidophilus DUN-311, Bifidobacterium bifidum BB-02, Bifidobacterium lactis BL-01, Synergy 1) on microbial community structure and metabolism. A PEG tube was inserted into the fermenters to study biofilm formation. The synbiotic delivered in sterile semi-skimmed milk (SSSM) was introduced either 48 h prior to or after PEG tube insertion. The synbiotic reduced biofilm formation on PEG tube surfaces, with suppression of Escherichia coli and Klebsiella pneumoniae when it was added subsequent to PEG insertion. When synbiotic feeding was commenced prior to PEG insertion, colonization by Staphylococcus aureus, Candida albicans and Candida famata was also inhibited. Lactate production increased in response the synbiotic or control (SSSM). These results indicate that the use of a synbiotic has the potential to reduce pathogen colonization on PEG tube surfaces in vivo, thereby reducing the incidence of biofilm-related infectious complications.")

('>F', 'TI  - Heterologous protein display on the cell surface of lactic acid bacteria mediated by the s-layer protein.', 'AB  - BACKGROUND: Previous studies have revealed that the C-terminal region of the S-layer protein from Lactobacillus is responsible for the cell wall anchoring, which provide an approach for targeting heterologous proteins to the cell wall of lactic acid bacteria (LAB). In this study, we developed a new surface display system in lactic acid bacteria with the C-terminal region of S-layer protein SlpB of Lactobacillus crispatus K2-4-3 isolated from chicken intestine. RESULTS: Multiple sequence alignment revealed that the C-terminal region (LcsB) of Lb. crispatus K2-4-3 SlpB had a high similarity with the cell wall binding domains SA and CbsA of Lactobacillus acidophilus and Lb. crispatus. To evaluate the potential application as an anchoring protein, the green fluorescent protein (GFP) or beta-galactosidase (Gal) was fused to the N-terminus of the LcsB region, and the fused proteins were successfully produced in Escherichia coli, respectively. After mixing them with the non-genetically modified lactic acid bacteria cells, the fused GFP-LcsB and Gal-LcsB were functionally associated with the cell surface of various lactic acid bacteria tested. In addition, the binding capacity could be improved by SDS pretreatment. Moreover, both of the fused proteins could simultaneously bind to the surface of a single cell. Furthermore, when the fused DNA fragment of gfp:lcsB was inserted into the Lactococcus lactis expression vector pSec:Leiss:Nuc, the GFP could not be secreted into the medium under the control of the nisA promoter. Western blot, in-gel fluorescence assay, immunofluorescence microscopy and SDS sensitivity analysis confirmed that the GFP was successfully expressed onto the cell surface of L. lactis with the aid of the LcsB anchor. CONCLUSION: The LcsB region can be used as a functional scaffold to target the heterologous proteins to the cell surfaces of lactic acid bacteria in vitro and in vivo, and has also the potential for biotechnological application.')

('>F', 'TI  - D-Fagomine lowers postprandial blood glucose and modulates bacterial adhesion.', 'AB  - D-Fagomine is an iminosugar originally isolated from seeds of buckwheat (Fagopyrum sculentum Moench), present in the human diet and now available as a pure crystalline product. We tested D-fagomine for activities connected to a reduction in the risk of developing insulin resistance, becoming overweight and suffering from an excess of potentially pathogenic bacteria. The activities were: intestinal sucrase inhibition in vitro (rat mucosa and everted intestine sleeves), modulation of postprandial blood glucose in rats, bacterial agglutination and bacterial adhesion to pig intestinal mucosa. When ingested together with sucrose or starch, D-fagomine lowered blood glucose in a dose-dependent manner without stimulating insulin secretion. D-Fagomine reduced the area under the curve (0-120 min) by 20 % (P < 0.01) and shifted the time to maximum blood glucose concentration (Tmax) by 15 min at doses of 1-2 mg/kg body weight when administered together with 1 g sucrose/kg body weight. Moreover, D-fagomine (0.14 mm) agglutinated 60 % of Enterobacteriaceae (Escherichia coli, Salmonella enterica serovar Typhimurium) populations (P < 0.01), while it did not show this effect on Bifidobacterium spp. or Lactobacillus spp. At the same concentration, d-fagomine significantly (P < 0.001) inhibited the adhesion of Enterobacteriaceae (95-99 % cells in the supernatant) and promoted the adhesion of Lactobacillus acidophilus (56 % cells in the supernatant) to intestinal mucosa. D-Fagomine did not show any effect on bacterial cell viability. Based on all this evidence, D-fagomine may be used as a dietary ingredient or functional food component to reduce the health risks associated with an excessive intake of fast-digestible carbohydrates, or an excess of potentially pathogenic bacteria.')

('>F', 'TI  - Regulation of the IL-10/IL-12 axis in human dendritic cells with probiotic bacteria.', 'AB  - In this study, we have used monocyte-derived dendritic cells (DCs) to design a screening model for the selection of microorganisms with the ability to suppress DC-secreted IL-12p70, a critical cytokine for the induction of T-helper cell type 1 immune responses under inflammatory conditions. By the treatment of DCs with cocktails containing TLR agonists and proinflammatory cytokines, the cells increased the secretion of the Th1-promoting cytokine IL-12p70. Clinically used probiotics were tested for their IL-10- and IL-12p70-stimulating properties in immature DCs, and showed a dose-dependent change in the IL-10/IL-12p70 balance. Lactobacillus acidophilus NCFM() and the probiotic mixture VSL#3 showed a strong induction of IL-12p70, whereas Lactobacillus salivarius Ls-33 and Bifidobacterium infantis 35624 preferentially induced IL-10. Escherichia coli Nissle 1917 induced both IL-10 and IL-12p70, whereas the probiotic yeast Saccharomyces boulardii induced low levels of cytokines. When combining these microorganisms with the Th1-promoting cocktails, E. coli Nissle 1917 and B. infantis 35624 were potent suppressors of IL-12p70 secretion in an IL-10-independent manner, indicating a suppressive effect on Th1-inducing antigen-presenting cells. The present model, using cocktail-stimulated DCs with potent IL-12p70-stimulating capacity, may be used as an efficient tool to assess the anti-inflammatory properties of microorganisms for potential clinical use.')

('>F', 'TI  - Effect of supplementation of micronutrients and phytochemicals to fructooligosaccharides on growth response of probiotics and E. coli.', 'AB  - Probiotics and prebiotics, which can change the colonic microenvironment, are the areas of current interest. Unutilizable fractions of the foods and fortificants, which reach the colon can affect the profile of probiotics. Effects of eight such factors viz. zinc sulphate, zinc carbonate, ferrous sulphate, ferric citrate, quercetin, gallic acid, phytic acid, and oxalic acid were, therefore, investigated on 24 H growth of Lactobacillus acidophilus (L1) and Lactobacillus plantarum (L2), two isolates of bifidobacteria (longum (L3) and bifidum (L4)) and a marketed consortium (L5) of eight probiotic cultures. MRS medium with marketed fructooligosaccharide as the only source of carbon was used for study of dose response curves. Quercetin and zinc sulphate showed significant positive effect for L1 and L5 (P < 0.01), whereas there was slight positive effect or no effect on growth of other probiotics. Phytic acid showed a significant inhibitory effect for L2 and a slight inhibitory effect on L3 and L4 whereas L5 were able to tolerate phytic acid. Oxalic acid had slight positive effect for L1 (P < 0.05) and L5 and no effect on growth of other probiotics (P > 0.05). Further, zinc sulphate, ferrous sulphate, quercetin, and oxalic acid significantly inhibited growth of E. coli (P < 0.05)')

('>F', 'TI  - Effects of Lactobacillus acidophilus NCFM on insulin sensitivity and the systemic inflammatory response in human subjects.', 'AB  - According to animal studies, intake of probiotic bacteria may improve glucose homeostasis. We hypothesised that probiotic bacteria improve insulin sensitivity by attenuating systemic inflammation. Therefore, the effects of oral supplementation with the probiotic bacterium Lactobacillus acidophilus NCFM on insulin sensitivity and the inflammatory response were investigated in subjects with normal or impaired insulin sensitivity. In a double-blinded, randomised fashion, forty-five males with type 2 diabetes, impaired or normal glucose tolerance were enrolled and allocated to a 4-week treatment course with either L. acidophilus NCFM or placebo. L. acidophilus was detected in stool samples by denaturating gradient gel electrophoresis and real-time PCR. Separated by the 4-week intervention period, two hyperinsulinaemic-euglycaemic clamps were performed to estimate insulin sensitivity. Furthermore, the systemic inflammatory response was evaluated by subjecting the participants to Escherichia coli lipopolysaccharide injection (0.3 ng/kg) before and after the treatment course. L. acidophilus NCFM was detected in 75 % of the faecal samples after treatment with the probiotic bacterium. Insulin sensitivity was preserved among volunteers in the L. acidophilus NCFM group, whereas it decreased in the placebo group. Both baseline inflammatory markers and the systemic inflammatory response were, however, unaffected by the intervention. In conclusion, intake of L. acidophilus NCFM for 4 weeks preserved insulin sensitivity compared with placebo, but did not affect the systemic inflammatory response.')

('>F', 'TI  - Suppression of human immunodeficiency virus type 1 replication in macrophages by  commensal bacteria preferentially stimulating Toll-like receptor 4.', 'AB  - Protection from primary human immunodeficiency virus type 1 (HIV-1) infection has not yet been accomplished by vaccines inducing HIV-1-specific acquired immunity. Nevertheless, it has been reported that a small subgroup of women remain resistant to HIV-1 infection under natural conditions. If similar conditions can be induced in uninfected individuals, it will contribute the first line of protection against HIV-1 infection, and also improve the effects of anti-HIV-1 vaccines. We reasoned that innate immunity may be involved in the resistance to HIV-1 infection, and investigated the effects of various Toll-like receptor (TLR) ligands and commensal bacteria on HIV-1 replication in macrophages, one of the initial targets of HIV-1 infection and also the main mediators of innate immunity. We established the HIV-1 reporter monocytic cell line, THP-1/NL4-3luc, which could be differentiated into macrophage-like cells in vitro. In these cells, stimulation of TLR3 and TLR4 by their ligands suppressed HIV-1 expression partly through type I interferon (IFN). Among the commensal bacteria tested, Escherichia coli, Veillonella parvula and Neisseria mucosa suppressed HIV-1 expression, whereas Lactobacillus acidophilus, Prevotella melaninogenica, P. bivia and Mycobacterium smegmatis enhanced it. The bacteria with suppressive effects preferentially stimulated TLR4, whereas the ones with enhancing effects stimulated TLR2. Neutralizing antibodies against TLR4 and IFN-alpha/beta receptor abrogated bacterially mediated HIV-1 suppression. Suppressive effects of E. coli, V. parvula and N. mucosa on HIV-1 replication were reproducible in primary monocyte-derived macrophages following acute HIV-1 infection. These findings suggest that certain commensal bacteria preferentially stimulating TLR4 potentially produce local environments resistant to HIV-1 infection.')

('>F', 'TI  - Biodegradable gelatin-chitosan films incorporated with essential oils as antimicrobial agents for fish preservation.', 'AB  - Essential oils of clove (Syzygium aromaticum L.), fennel (Foeniculum vulgare Miller), cypress (Cupressus sempervirens L.), lavender (Lavandula angustifolia), thyme (Thymus vulgaris L.), herb-of-the-cross (Verbena officinalis L.), pine (Pinus sylvestris) and rosemary (Rosmarinus officinalis) were tested for their antimicrobial activity on 18 genera of bacteria, which included some important food pathogen and spoilage bacteria. Clove essential oil showed the highest inhibitory effect, followed by rosemary and lavender. In an attempt to evaluate the usefulness of these essential oils as food preservatives, they were also tested on an extract made of fish, where clove and thyme essential oils were the most effective. Then, gelatin-chitosan-based edible films incorporated with clove essential oil were elaborated and their antimicrobial activity tested against six selected microorganisms: Pseudomonas fluorescens, Shewanella putrefaciens, Photobacterium phosphoreum, Listeria innocua, Escherichia coli and Lactobacillus acidophilus. The clove-containing films inhibited all these microorganisms irrespectively of the film matrix or type of microorganism. In a further experiment, when the complex gelatin-chitosan film incorporating clove essential oil was applied to fish during chilled storage, the growth of microorganisms was drastically reduced in gram-negative bacteria, especially enterobacteria, while lactic acid bacteria remained practically constant for much of the storage period. The effect on the microorganisms during this period was in accordance with biochemical indexes of quality, indicating the viability of these films for fish preservation.')

('>F', 'TI  - Lactobacillus acidophilus induces virus immune defence genes in murine dendritic  cells by a Toll-like receptor-2-dependent mechanism.', 'AB  - Lactobacilli are probiotics that, among other health-promoting effects, have been ascribed immunostimulating and virus-preventive properties. Certain Lactobacillus spp. have been shown to possess strong interleukin-12 (IL-12) -inducing properties. As IL-12 production depends on the up-regulation of type I interferons (IFNs), we hypothesized that the strong IL-12-inducing capacity of Lactobacillus acidophilus NCFM in murine bone-marrow-derived dendritic cells (DCs) is caused by an up-regulation of IFN-beta, which subsequently induces IL-12 and the double-stranded RNA binding Toll-like receptor-3 (TLR-3). The expression of the genes encoding IFN-beta, TLR-3, IL-12 and IL-10 in DCs upon stimulation with L. acidophilus NCFM was determined. Lactobacillus acidophilus NCFM induced a much stronger expression of Ifn-beta, Il-12 and Il-10 compared with the synthetic double-stranded RNA ligand Poly I:C, whereas the levels of expressed Tlr-3 were similar. Whole genome microarray gene expression analysis revealed that other genes related to viral defence were significantly up-regulated and among the strongest induced genes in DCs stimulated with L. acidophilus NCFM. The ability to induce IFN-beta was also detected in another L. acidophilus strain (X37), but was not a property of other probiotic strains tested, i.e. Bifidobacterium bifidum Z9 and Escherichia coli Nissle 1917. The IFN-beta expression was markedly reduced in TLR-2(-/-) DCs, dependent on endocytosis, and the major cause of the induction of Il-12 and Tlr-3 in DCs stimulated with L. acidophilus NCFM. Collectively, our results reveal that certain lactobacilli trigger the expression of viral defence genes in DCs in a TLR-2 manner dependent on IFN-beta.')

('>F', 'TI  - The impact of the stone age diet on gingival conditions in the absence of oral hygiene.', 'AB  - BACKGROUND: The objective of this study was to assess the oral microbiota and clinical data in subjects without access to traditional oral hygiene methods and who ate a diet available in the Stone Age. METHODS: Ten subjects living in an environment replicating the Stone Age for 4 weeks were enrolled in this study. Bleeding on probing (BOP), gingival and plaque indices, and probing depth (PD) were assessed at baseline and at 4 weeks. Microbiologic samples were collected at the mesio-buccal subgingival aspects of all teeth and from the dorsum of the tongue and were processed by checkerboard DNA-DNA hybridization methods. RESULTS: No subject had periodontitis. Mean BOP decreased from 34.8% to 12.6% (P <0.001). Mean gingival index scores changed from 0.38 to 0.43 (not statistically significant) and mean plaque scores increased from 0.68 to 1.47 (P <0.001). PD at sites of subgingival sampling decreased (mean difference: 0.2 mm; P <0.001). At week 4, the total bacterial count was higher (P <0.001) for 24 of 74 species, including Bacteroides ureolyticus, Eikenella corrodens, Lactobacillus acidophilus, Capnocytophaga ochracea, Escherichia coli, Fusobacterium nucleatum naviforme, Haemophilus influenzae, Helicobacter pylori, Porphyromonas endodontalis, Staphylococcus aureus (two strains), Streptococcus agalactiae, Streptococcus anginosis, and Streptococcus mitis. Bacterial counts from tongue samples were higher at baseline (P <0.001) for 20 species, including Tannerella forsythia (previously T. forsythensis), Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans; serotype a), and Streptococcus spp. CONCLUSIONS: The experimental gingivitis protocol is not applicable if the diet (e.g., Stone Age) does not include refined sugars. Although plaque levels increased, BOP and PD decreased. Subgingival bacterial counts increased for several species not linked to periodontitis, whereas tongue bacterial samples decreased during the study period.')

('>F', 'TI  - Characterization of vaginal lactobacilli coaggregation ability with Escherichia coli.', 'AB  - The coaggregation abilities of probiotic strains might enable it to form a barrier that prevents colonization by pathogenic bacteria. In the present study, the characterization of the coaggregation ability of 19 vaginal lactobacilli was studied. Coaggregation ability of all lactobacilli with Escherichia coli ATCC 11229 was positive. Only the highest coaggregation percentage of Lactobacillus acidophilus S1 was obtained with E. coli ATCC 11229 under both aerobic (71%) and anaerobic conditions (62%). The coaggregation abilities of strains occurred higher at acidic pH than at basic pH values. Moreover, the coaggregation abilities of tested strains against E. coli decreased after heat treatment (70 or 85 degrees C). Also, the relationship between hydrophobicity and coaggregation of strains was found to be significant. The effect of sonication, some enzymes (lipase and pepsin) and sodium periodate on coaggregation ability of L. acidophilus S1, which is one of the highest potentials on coaggregation ability, was investigated. Sodium periodate did not have a significant effect on coaggregation ability of L. acidophilus S1. The sonicated cell showed lower coaggregation than the control, the supernatant fluid of this sonicated cells showed similar coaggregation ability to the control. Coaggregation abilities of bacteriotherapeutic lactobacilli with pathogenic bacteria can be used for preliminary screening in order to identify potentially probiotic bacteria suitable for human use against urogenital tract infections.')

('>F', 'TI  - In vitro anti-adhesive activity of green tea extract against pathogen adhesion.', 'AB  - Camellia sinensis polysaccharide has been reported to possess anti-adhesive activity against pathogens. The present study was designed to investigate whether hot water extracts obtained from green tea leaves might inhibit pathogen adhesion to human or mouse cell lines. Green tea extract-4 (CSI-4) with the maximum yield of 4% (w/v) is composed of a major proportion of carbohydrates containing 40% uronic acids, but lack of catechins. It showed strong inhibitory activities against hemagglutination mediated by pathogens Helicobacter pylori, Propionibacterium acnes and Staphylococcus aureus with the minimum inhibitory concentrations of 0.01-0.5 mg/mL. CSI-4 further demonstrated an inhibitory effect on the adhesion of these pathogens to host cell lines with the IC(50) values (50% inhibition of adhesion) of 0.14-2.3 mg/mL. It exhibited the highest activity against P. acnes, but no inhibitory effects were observed against Lactobacillus acidophilus, Bifidobacterium bifidum, Escherichia coli, or Staphylococcus epidermidis. Our results suggest that CSI-4 may exert a selective anti-adhesive effect against certain pathogenic bacteria with no adverse effects against beneficial or commensal bacteria.')

('>F', 'TI  - Oligosaccharide-mediated inhibition of the adhesion of pathogenic Escherichia coli strains to human gut epithelial cells in vitro.', 'AB  - The aim of the study was to investigate the ability of pectic oligosaccharides (POS) to inhibit adhesion of three strains of verotoxigenic Escherichia coli, three strains of enteropathogenic E. coli, and one nonclinical strain of Desulfovibrio desulfuricans to human intestinal epithelial cell cultures. Lactobacillus acidophilus and Lactobacillus gasseri were included for comparison. Attachment was determined in the human HT29 cell line by viable count of adherent bacteria. POS in buffer at pH 7.2 were antiadhesive at a dose of 2.5 mg ml(-1), reducing adhesion of enteropathogenic E. coil and verotoxigenic E. coli strains to less than 30% of control values. Concentrations resulting in 50% inhibition ranged from 0.15 to 0.46 mg ml(-1). L. acidophilus was not significantly affected, but adhesion of L. gasseri was reduced to 29% of the control value. POS reduced the adhesion of D. desulfuricans to 0.33% of the control value. POS also had a protective effect against E. coli verocytotoxins VT1 and VT2 at concentrations of 0.01 and 1 microg ml(-1), respectively.')

('>F', 'TI  - Inhibition of growth of Shiga toxin-producing Escherichia coli by nonpathogenic Escherichia coli.', 'AB  - During routine quality control testing of diagnostic methods for Shiga toxin-producing Escherichia coli (STEC) using stool samples spiked with STEC, it was observed that the Shiga toxin could not be detected in 32 out of 82 samples tested. Strains of E. coli isolated from such stool samples were shown to be responsible for this inhibition. One particular isolate, named E. coli 1307, was intensively studied because of its highly effective inhibitory effect; this strain significantly reduced growth and Shiga toxin levels in coculture of several STEC strains regardless of serovar or Shiga toxin type. The probiotic E. coli Nissle 1917 inhibited growth and reduced Shiga toxin levels in STEC cultures to an extent similar to E. coli 1307, but commensal E. coli strains and several other known probiotic bacteria (enterococci, Bacillus sp., Lactobacillus acidophilus) showed no, or only small, inhibitory effects. Escherichia coli 1307 lacks obvious fitness factors, such as aerobactin, yersiniabactin, microcins and a polysaccharide capsule, that are considered to promote the growth of pathogenic bacteria. We therefore propose strain E. coli 1307 as a candidate probiotic for use in the prevention and treatment of infections caused by STEC.')

('>F', 'TI  - Antimicrobial activity of neuropeptides against a range of micro-organisms from skin, oral, respiratory and gastrointestinal tract sites.', 'AB  - Many neuropeptides are similar in size, amino acid composition and charge to antimicrobial peptides. This study aimed to determine whether the neuropeptides substance P (SP), neurokinin A (NKA), calcitonin gene-related peptide (CGRP), neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP), displayed antimicrobial activity against Streptococcus mutans, Lactobacillus acidophilus, Enterococcus faecalis, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans. SP, NPY, VIP and CGRP displayed variable degrees of antimicrobial activity against all the pathogens tested with the exception of S. aureus. These antimicrobial activities add a further dimension to the immunomodulatory roles for neuropeptides in the inflammatory and immune responses.')

('>F', "TI  - Dendritic cells from Peyer's patches and mesenteric lymph nodes differ from spleen dendritic cells in their response to commensal gut bacteria.", "AB  - Commensal gut bacteria have potent effects on the immune system, which are partially mediated by intestinal dendritic cells (DC). Distinct commensals confer different properties to in vitro-generated DC. The aim of the present study was to reveal strain-dependent maturation patterns in primary DC. To this end, we compared the response of mouse Peyer's patch (PP) DC, mesenteric lymph node (MLN) DC and spleen DC to the commensal bacteria, Bifidobacterium longum Q46, Lactobacillus acidophilus X37 and Escherichia coli Nissle 1917. Bacterial maturation of DC occurred independently of tissue origin. Expression of CCR7 and CD103 on the surface of MLN DC, necessary for the induction of gut-homing regulatory T cells, increased with stimulation by Gram-positive commensals. Bacteria-dependent cytokine production (IL-6, IL-10 and TNF-alpha) was similar in spleen and MLN DC, and contaminant cells in these DC preparations produced IFN-gamma in response to L. acidophilus. In contrast, PP DC produced IL-6 only in response to E. coli, little IL-10 and no TNF-alpha, and this low cytokine production was not due to inhibition by IL-10 or TGF-beta. Bifidobacteria downregulate IL-6, TNF-alpha and IL-12 production induced in in vitro-generated DC by L. acidophilus. Similar inhibition was observed in splenic DC, but not in MLN DC. MLN cells responded to bacterial stimulation with higher IFN-gamma production than spleen cells, possibly due to the presence of more responsive natural killer cells. Commensal bacteria therefore play specific roles in the gut immune system distinguishable from the effect they would have if recognized by the systemic immune system.")

('>F', 'TI  - Electron microscopic analysis of dairy microbes inactivated by ultrasound.', 'AB  - Ultrasonication is a non-thermal method of food preservation that has the advantage of inactivating microbes in food without causing the common side-effects associated with conventional heat treatments, such as nutrient and flavour loss. The aim of this study was to evaluate the use of ultrasound as an alternative to heat pasteurisation and to assess cell damage using transmission electron microscopy (TEM). Three spoilage microbes, previously isolated from pasteurised milk, were used as "test" microbes. Saline solution (SSS) and UHT milk were used as suspension media and were inoculated with exponential growth phase "test" microbes at a microbial concentration of 1 x 10(4) cfu ml(-1). The samples were subjected to power ultrasound (20 kHz, 750 W), at 100%/124 microm wave amplitude for different time intervals. Both Escherichia coli and Saccharomyces cerevisiae were reduced by >99% (for both suspension media) after ultrasonication and Lactobacillus acidophilus was reduced by 72% and 84% in SSS and milk, respectively. Transmission electron microscope micrographs showed that ultrasonication inflicts extensive microbicidal/microbistatic external and internal damage on all three "test" microbes. In E. coli, sonication-induced emulsification caused the formation of unique minute lipopolysaccharide vesicles from the fragmenting cell envelope.')

('>F', 'TI  - Molecular cloning and characterization of a bile salt hydrolase from Lactobacillus acidophilus PF01.', 'AB  - Phenotypic screening for bile salt hydrolase (BSH) activity was performed on Lactobacillus acidophilus PF01 isolated from piglet feces. A gene encoding BSH was identified and cloned from the genomic library of L. acidophilus PF01. The bsh gene and surrounding regions were characterized by nucleotide sequence analysis and were found to contain a single open reading frame (ORF) of 951 nucleotides encoding a 316 amino acid protein. The potential bsh promoter region was located upstream of the start codon. The protein deduced from the complete ORF had high similarity with other BSHs, and four amino acid motifs located around the active site, FGRNXD, AGLNF, VLTNXP, and GXGXGXXGXPGD, were highly conserved. The bsh gene was cloned into the pET21b expression vector and expressed in Escherichia coli BLR(DE3) by induction with 0.1mM of isopropylthiogalactopyranoside. The BSH enzyme was purified with apparent homogeneity using a Ni2+-NTA agarose column and characterized. The overexpressed recombinant BSH enzyme of L. acidophilus PF01 exhibited hydrolase activity against tauroconjugated bile salts, but not glycoconjugated bile salts. It showed the highest activity against taurocholic acid. The maximum BSH activity occurred at approximately 40oC. The enzyme maintained approximately 70% of its maximum activity even at 60 degrees , whereas its activity rapidly decreased at below 37 degrees . The optimum pH was 6, and BSH activity was rapidly inactivated below pH 5 and above pH 7.')

('>F#I::METABOLICINTERACTION', 'TI  - The effect of probiotics and organic acids on Shiga-toxin 2 gene expression in enterohemorrhagic Escherichia coli O157:H7.', 'AB  - Probiotics are known to have an inhibitory effect against the growth of various foodborne pathogens, however, the specific role of probiotics in Shiga-toxin-producing Escherichia coli (STEC) virulence gene expression has not been well defined. Shiga toxins are members of a family of highly potent bacterial toxins and are the main virulence marker for STEC. Shiga toxins inhibit protein synthesis in eukaryotic cells and play a role in hemorrhagic colitis and hemolytic uremic syndrome. STEC possesses Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2), both of which have A and B subunits. Although STEC containing both Stx1 and Stx2 has been isolated from patients with hemorrhagic colitis, Stx2 is more frequently associated with human disease complications. Thus, the effect of Lactobacillus, Pediococcus, and Bifidobacterium strains on stx2A expression levels in STEC was investigated. Lactic acid bacteria and bifidobacteria were isolated from farm animals, dairy, and human sources and included L. rhamnosus GG, L. curvatus, L. plantarum, L. jensenii, L. acidophilus, L. casei, L. reuteri, P. acidilactici, P. cerevisiae, P. pentosaceus, B. thermophilum, B. boum, B. suis and B. animalis. E. coli O157:H7 (EDL 933) was coincubated with sub-lethal concentrations of each probiotic strain. Following RNA extraction and cDNA synthesis, relative stx2A mRNA levels were determined according to a comparative critical threshold (Ct) real-time PCR. Data were normalized to the endogenous control glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and the level of stx2A expression between treated and untreated STEC was compared. Observed for all probiotic strains tested, stx2A was down-regulated, when compared to the control culture. Probiotic production of organic acids, as demonstrated by a decrease in pH, influenced stx2A gene expression.')

('>F', 'TI  - Antimicrobial properties of milkfat globule membrane fractions.', 'AB  - Milkfat globule membranes (MFGMs) were prepared from bovine cream according to standard procedures. These membranes and peptide hydrolysates, which were generated by proteolysis with immobilized digestive enzymes, were screened for antibacterial activity against Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica Typhimurium, Pseudomonas fluorescens, Bacillus cereus, Lactobacillus acidophilus, and Lactobacillus gasseri. Assays were first performed on beef heart infusion (BHI) plates spotted with test protein-peptide fractions and then seeded with lawns of indicator cells to monitor the zone of growth inhibition. Under these experimental conditions, MFGMs were most active against Salmonella Typhimurium and P. fluorescens. However, antibacterial activity was not seen after plating on Luria-Bertani (LB) medium. We determined that the antimicrobial effects observed on BHI plates were due to the generation of H2O2 by xanthine oxidase, a major protein constituent of the MFGMs, as a result of purine catalysis. This substrate is present in BHI but lacking in LB medium. Evaluation of purified xanthine oxidase alone resulted in analogous data trends. The growth of probiotic Lactobacillus strains were affected only marginally when grown on lactobacilli deMan Rogosa Sharpe plates, suggesting the decreased sensitivity of these bacteria to H2O2. In this study, several MFGM hydrolysates exhibited variable antibacterial activity against test food pathogens on agar plates prepared with M9 minimal media, and this variation was not attributable to xanthine oxidase enzymatic activity. The probiotic microorganisms were mostly resilient to these antibacterial fractions. Bovine MFGM fractions may represent an excellent resource material from which to generate native, naturally occurring biodefensive proteins and/or peptides.')

('?F#AMBIGUOUS', 'TI  - Pre-harvest interventions to reduce the shedding of E. coli O157 in the faeces of weaned domestic ruminants: a systematic review.', 'AB  - Our objective was to use formal systematic review methods to evaluate the efficacy of interventions to reduce faecal shedding of Escherichia coli O157 in post-weaned ruminants by increasing animal resistance. The methodology consisted of an extensive search to identify all potentially relevant research, screening of titles and abstracts for relevance to the research question, quality assessment of relevant research, extraction of data from research of sufficient quality, and qualitative summarization of results. The interventions evaluated included probiotics, vaccination, antimicrobials, sodium chlorate, bacteriophages and other feed additives. There was evidence of efficacy for the probiotic combination Lactobacillus acidophilus NP51 (NPC 747) and Propionibacterium freudenreichii and for sodium chlorate in feed or water. The effectiveness of vaccination varied among studies and among vaccine protocols and there was no consistent evidence to suggest that antibiotic use was associated with a decrease in faecal shedding of E. coli O157, or that current industry uses of antimicrobials were associated with increased faecal shedding. There were an insufficient number of studies available to address the effectiveness of bacteriophages and several other feed additives. In general, few of the primary studies evaluated the interventions under commercial housing conditions with a natural disease challenge, there were inconsistencies in the results among study designs and in some cases among studies within study designs, and a relatively large proportion of publications were excluded based on quality assessment criteria. Few studies reported on associations between the proposed intervention and production parameters, such as average daily gain and feed: gain ratio. While the results suggest that some interventions may be efficacious, there are knowledge gaps in our understanding of the efficacy of pre-harvest interventions to increase animal resistance to E. coli O157 that require further targeted research.')

('>F', 'TI  - An experimental study and a randomized, double-blind, placebo-controlled clinical trial to evaluate the antisecretory activity of Lactobacillus acidophilus strain LB against nonrotavirus diarrhea.', 'AB  - OBJECTIVE: Previous studies have shown that selected strains of Lactobacillus have the capacity to antagonize rotavirus-induced diarrhea. However, only a few reports have documented their efficacy against nonrotavirus diarrhea. This study involved an experimental investigation and a clinical trial of the antisecretory activity of Lactobacillus acidophilus strain LB in the context of nonrotavirus diarrhea. METHODS: The activity of a culture of L. acidophilus LB or of the lyophilized, heat-killed L. acidophilus LB bacteria plus their spent culture medium was tested in inhibiting the formation of fluid-formed domes in cultured human intestinal Caco-2/TC7 cell monolayers infected with diarrheagenic, diffusely adhering Afa/Dr Escherichia coli C1845 bacteria. A randomized, double-blind, placebo-controlled clinical trial of male or female children who were 10 months of age and presented with nonrotavirus, well-established diarrhea was conducted to evaluate the therapeutic efficacy of a pharmaceutical preparation that contains 10 billion heat-killed L. acidophilus LB plus 160 mg of spent culture medium. RESULTS: Infection of the cells with C1845 bacteria that were treated with L. acidophilus LB culture or the lyophilized, heat-killed L. acidophilus LB bacteria plus their culture medium produced a dosage-dependent decrease in the number of fluid-formed domes as compared with cells that were infected with untreated C1845 bacteria. The clinical results show that in selected and controlled homogeneous groups of children with well-established, nonrotavirus diarrhea, adding lyophilized, heat-killed L. acidophilus LB bacteria plus their culture medium to a solution of oral rehydration solution shortened by 1 day the recovery time (ie, the time until the first normal stool was passed) as compared with children who received placebo oral rehydration solution. CONCLUSIONS: Heat-killed L. acidophilus LB plus its culture medium antagonizes the C1845-induced increase in paracellular permeability in intestinal Caco-2/TC7 cells and produces a clinically significant benefit in the management of children with nonrotavirus, well-established diarrhea.')

('>F', 'TI  - Are probiotics detectable in human feces after oral uptake by healthy volunteers?', "AB  - GOALS: Assessment of the presence of probiotic bacteria in feces after oral ingestion. BACKGROUND: Probiotic bacteria are said to have beneficial effects on the host. As a precondition for any effect, probiotic strains must survive passage through the gastrointestinal tract. STUDY: The feces of seven volunteers were analyzed for the presence of probiotic strains after one week's oral ingestion of each of six commercially available products: E. coli Nissle 0.5-5 x 10(9) cells (Mutaflor), Enterococcus faecium SF 68 7.5 x 10(7) cells (Bioflorin), Lactobacillus acidophilus and Bifidobacterium infantis both 1 x 10(9) cells (Infloran), Lactobacillus gasseri and Bifidobacterium longum both 1 x 10(8) cells (Omniflora), Lactobacillus casei rhamnosus 1 x 10(9) cells (Antibiophilus), and yoghurt enriched with Lactobacillus casei Immunitas 1 x 10(10) cells (Actimel). Ten colonies were selected from each stool sample, and DNA was extracted and typed using random amplification of polymorphic DNA (RAPD). Typing patterns of the ingested probiotics and the fecal isolates were compared. RESULTS: Fingerprints identical to the ingested probiotic strains were recovered from fecal samples of 4/7 volunteers after one week of Mutaflor, from 4/6 after taking Bioflorin, and from 1/6 after Infloran. Cultivation of strains of the same species from fecal specimens was negative after consumption of Antibiophilus, Omniflora and Actimel. CONCLUSIONS: After oral consumption of probiotics, E. coli and enterococci could be detected in stool samples (57% and 67%, respectively). In contrast, with only one exception, ingested lactobacilli and bifidobacteria could not be detected in human feces.")

('>F', 'TI  - Differential immunomodulating effects of inactivated probiotic bacteria on the allergic immune response.', 'AB  - Bacterial stimulation plays an important role in modulating the allergic immune response. The aim of this study was to investigate the effects of inactivated probiotic Lactobacillus acidophilus and non-pathogenic Escherichia coli strain Nissle on the phenotype and function of T- and B-cells. Peripheral blood mononuclear cells from patients with grass-pollen allergy (n=10) and non-allergic patients (n=19) were co-stimulated with inactivated bacteria and grass-pollen allergen. Expression of CD23, CD80, CD86 and CD69 were analysed, and the intracellular production of interleukin-4 and interferon-gamma was measured by direct ex vivo flow cytometry after stimulation. Both bacteria induced a significant up-regulation of CD69 expression on T-lymphocytes (p=0.001). CD23 expression was significantly increased following stimulation with allergen (p=0.008), but reduced after stimulation with Lactobacillus and significantly reduced with E. coli plus allergen (p=0.029). CD80 expression was reduced after stimulation with Lactobacillus in the allergic group only (p=0.021). By contrast, CD86 expression was significantly increased after stimulation with Lactobacillus (p=0.049) and distinctly increased with E. coli in both groups (p=0.001). The cytokine patterns of CD69-positive T-lymphocytes from allergic patients showed a TH2-dominated response after allergen stimulation (interferon-gamma/interleukin-4-ratio 0.2), directed into a T-helper1-like response by stimulation with both types of bacteria (interferon-gamma/interleukin-4-ratios 1.5-2.0 in both groups). These data show that both types of bacteria modulate the allergic immune response by the alteration of CD23 and co-stimulatory molecule expression. Regarding cytokine production, the data suggest a differential response to both bacteria depending on the atopic state, but a clear promotion of T-helper1-dominated response in allergic donors.')

('>F', 'TI  - Inhibition of pathogenic bacterial adhesion by acidic polysaccharide from green tea (Camellia sinensis).', 'AB  - An acidic polysaccharide CS-F2 from Camellia sinensis was examined to characterize its anti-adhesive effects against pathogenic bacteria, most notably Helicobacter pylori, Propionibacterium acnes, and Staphylococcus aureus. CS-F2 showed marked inhibitory activity against the pathogen-mediated hemagglutination with a minimum inhibitory concentration (MIC) between 0.01 and 0.1 mg/mL, which is lower than the previously reported MIC values for Panax ginseng and Artemisia capillaris. The inhibitory effects of CS-F2 on the adhesion of H. pylori to AGS adenocarcinoma gastric epithelial cells, or P. acnes and S. aureus to NIH 3T3 fibroblast cells, were further assessed resulting in MIC values between 0.063 and 0.13 mg/mL. Importantly, CS-F2 showed no inhibitory effects against Lactobacillus acidophilus, Escherichia coli, or Staphylococcus epidermidis. Our results suggest that CS-F2, which is a pectin-type polysaccharide with a molecular weight of approximately 8.0 x 10(4) Da, may exert a selective anti-adhesive effect against certain pathogenic bacteria, while exerting no effects against beneficial and commensal bacteria.')

('>F', 'TI  - Hydrogen peroxide production and resistance to nonoxinol-9 in Lactobacillus spp.  isolated from the vagina of reproductive-age women.', 'AB  - Lactic acid production is considered to be the major protection mechanism of lactobacilli against vaginal infections due to genital pathogens. However, some species of Lactobacillus are also hydrogen peroxide-producers. Women, who usually use intrauterine dispositive (IUD) and spermicides such as nonoxinol-9 (N-9) as contraceptive methods, increase the risk of acquiring an urinary tract infection and a bacterial vaginosis; some studies have demonstrated that these compounds alter the normal vaginal biota. It is known that they inhibit lactobacilli in vitro at concentrations of 0.1% to 1% and that they do not have an effect on the growth of Escherichia coli. It is probable that the presence of nonoxinol-9 affects the ecological balance of the vagina by inhibiting the protector lactobacilli. In this study, we identified Lactobacillus acidophilus, L. brevis, L. crispatus, L. fermentii and L. jensenii as the species most frequently isolated from women. Seventy-one hydrogen peroxide-producer strains and 48 strains resistant to the inhibitory effect of nonoxinol-9 were detected. L. brevis showed the highest number of resistant strains.')

('>F', 'TI  - Inhibitory effects of Lactobacillus acidophilus lysates on the cytotoxic activity of shiga-like toxin 2 produced from Escherichia coli O157:H7.', 'AB  - AIMS: The purpose of this study was to characterize the degree to which four cell lysates obtained from Lactobacillus acidophilus strains affected the cytotoxic activity of Escherichia coli O157:H7 in vitro and in vivo. METHODS AND RESULTS: In a cytotoxic inhibition test that used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and toxin-binding ELISA assays, the activity of shiga-like toxin 2 (Stx-2) was inhibited profoundly by the cell lysates (10 mg ml(-1)) from two strains of L. acidophilus A4 and 30SC (>85% of survival rates compared with the control) among the five strains tested. In particular, a significant decline in the virulence level of E. coli O157:H7, under the presence of the cell lysates of L. acidophilus A4, was observed by killing assay of Caenorhabditis elegans in vivo model. CONCLUSIONS: According to our results, L. acidophilus strains might be capable of attenuating the virulence of Stx-2 produced from E. coli O157:H7. SIGNIFICANCE AND IMPACT OF THE STUDY: The cell lysates of L. acidophilus can be applied to a variety of foods, and can be used as adjuncts for the inhibition of Stx-2-mediated cytotoxicity.')

('>F', 'TI  - Comparison of the concentrations of phenolic compounds in olive oils and other plant oils: correlation with antimicrobial activity.', 'AB  - The antimicrobial activity of different edible vegetable oils was studied. In vitro results revealed that the oils from olive fruits had a strong bactericidal action against a broad spectrum of microorganisms, this effect being higher in general against Gram-positive than Gram-negative bacteria. Thus, olive oils showed bactericidal activity not only against harmful bacteria of the intestinal microbiota (Clostridium perfringens and Escherichia coli) also against beneficial microorganisms such as Lactobacillus acidophilus and Bifidobacterium bifidum. Otherwise, most of the foodborne pathogens tested (Listeria monocytogenes, Staphylococcus aureus, Salmonella enterica, Yersinia sp., and Shigella sonnei) did not survive after 1 h of contact with olive oils. The dialdehydic form of decarboxymethyl oleuropein and ligstroside aglycons, hydroxytyrosol and tyrosol, were the phenolic compounds that statistically correlated with bacterial survival. These findings were confirmed by testing each individual phenolic compound, isolated by HPLC, against L. monocytogenes. In particular, the dialdehydic form of decarboxymethyl ligstroside aglycon showed a potent antimicrobial activity. These results indicate that not all oils classified as "olive oil" had similar bactericidal effects and that this bioactivity depended on their content of certain phenolic compounds.')

('>F', 'TI  - Post-natal development of the porcine microbiota composition and activities.', 'AB  - The current study describes the development of the porcine microbiota and its metabolic activities during the neonatal and weaning period. Using 16S rRNA-based approaches, we first analysed the ileal and colonic microbiota of neonatal piglets at days 2, 5 and 12 after birth. To further investigate the effect of weaning at 3 weeks of age, 19-day-old piglets (n = 64) were randomly allocated into two groups. Half of the piglets remained with their sows throughout the study, while the remaining piglets were weaned. As revealed by sequence analysis of 16S rRNA gene amplicons, the samples of 2-day-old piglets harboured a consortium of bacteria related to Escherichia coli, Shigella flexneri, Lactobacillus sobrius, Lactobacillus reuteri and Lactobacillus acidophilus. Moreover, species-specific real-time polymerase chain reaction assays unveiled that L. sobrius and L. reuteri predominated in the ileal samples of the neonatal and unweaned piglets with population levels up to 7 x 10(8) cells per gram of lumen content. Following weaning, however, these two lactobacilli were detected at significantly lower levels (< 10(3)) in the ileal samples. Furthermore, a shift in composition and metabolic activities of the predominant microbiota, and emergence of clostridia and E. coli, were encountered in the intestinal samples of the piglets after the early post-weaning period.')

('>F', 'TI  - Pectin-like acidic polysaccharide from Panax ginseng with selective antiadhesive  activity against pathogenic bacteria.', 'AB  - Previous studies have revealed the inhibitory effects of an acidic polysaccharide purified from the root of Panax ginseng against the adhesion of Helicobacter pylori to gastric epithelial cells and the ability of Porphyromonas gingivalis to agglutinate erythrocytes. In this study, this acidic polysaccharide from P. ginseng, PG-F2, was investigated further, in order to characterize its antiadhesive effects against Actinobacillus actinomycetemcomitans, Propionibacterium acnes, and Staphylococcus aureus. The minimum inhibitory concentrations (MIC) were found to be in a range of 0.25-0.5mg/mL. However, results showed no inhibitory effects of PG-F2 against Lactobacillus acidophilus, Escherichia coli, or Staphylococcus epidermidis. PG-F2 is a pectin-type polysaccharide with a mean MW of 1.2 x 10(4) Da, and consists primarily of galacturonic and glucuronic acids along with rhamnose, arabinose, and galactose as minor components. The complete hydrolysis of PG-F2 via chemical or carbohydrolase enzyme treatment resulted in the abrogation of its antiadhesive activity, but limited hydrolysis via treatment with pectinase (EC. 3.2.1.15) yielded an oligosaccharide fraction, with activity comparable to the precursor PG-F2 (the MIC of ca. 0.01 mg/mL against H. pylori and P. gingivalis). Our results suggest that PG-F2 may exert a selective antiadhesive effect against pathogenic bacteria, while having no effects on beneficial and commensal bacteria.')

('>F', 'TI  - Effect of oral bacteria on growth and survival of Candida albicans biofilms.', 'AB  - The purpose of this study was to evaluate the effect of eight aerobic and anaerobic oral commensal bacterial species on in vitro Candida albicans biofilm development. A single isolate of C. albicans 2560 g, and eight different species of oral bacteria comprising, Actinomyces israelii, Lactobacillus acidophilus, Prevotella nigrescens, Porphyromonas gingivalis, Pseudomonas aeruginosa, Escherichia coli, Streptococcus mutans, and Streptococcus intermedius were studied using an in vitro biofilm assay. Biofilm formation was quantified in terms of the ability of Candida to grow on polystyrene plastic surfaces co-cultured with the foregoing bacteria. A viable cell count was used to quantify the sessile yeast growth and scanning electron microscopy was employed to confirm and visualize biofilm formation. Co-culture with differing concentrations of bacteria had variable effects on Candida biofilm formation. Co-culture with the highest concentrations of each of the foregoing bacteria resulted in a consistent reduction in the yeast counts in the candidal biofilm (P<0.05), except for L. acidophilus, S. mutans, and, S. intermedius co-cultures. Further, on regression analysis a significant negative correlation between the co-culture concentration of either P. gingivalis or E. coli and viable yeast counts in the biofilm was noted (P<0.05) although this was not evident for the other bacterial species. Taken together, our data indicate that, quantitative and qualitative nature of the bacteria modulate C. albicans biofilm formation in mixed species environments such as the oral cavity.')

('>F', 'TI  - Preinoculation with the probiotic Lactobacillus acidophilus early in life effectively inhibits murine Citrobacter rodentium colitis.', 'AB  - Enteropathogenic Escherichia coli (EPEC) is a common pathogen in infantile diarrhea, causing a characteristic histopathologic attaching and effacing (A/E) lesion in the intestinal mucosa. The mouse pathogen Citrobacter rodentium causes a similar A/E lesion in the murine intestine. Like EPEC, C. rodentium infection results in colonic crypt hyperplasia, goblet cell depletion, epithelial proliferation, and mucosal disruption. Using this murine model, we tested the hypothesis that preinoculation of murine gut with Lactobacillus acidophilus early in life can enhance host defense against enteric bacterial infection and attenuate bacteria-mediated colitis. Two-week old BALB/c mice were inoculated with L. acidophilus twice per week for 4 weeks before C. rodentium infection or concomitantly with the exposure to C. rodentium at 6-8 weeks of age. The probiotics were administered twice weekly thereafter. We observed that L. acidophilus inoculation in mice inhibits C. rodentium-induced colitis, which is associated with a decrease in C. rodentium colonization and translocation, an increase in its clearance, and a suppression of colonic myeloperoxidase (MPO) activity. Probiotic treatment also stimulates regulatory cytokine expression in the colon [transforming growth factor beta (TGF-beta), interleukin (IL)-10]. Preinoculation with L. acidophilus is more effective than concomitant use of probiotics in the induction of intestinal IgA secretion and in the downregulation of proinflammatory cytokine expression [tumor necrosis factor alpha (TNF-alpha), IL-6, and IL-12]. These observations suggest that inoculation with probiotics can effectively prevent bacteria-induced colitis by limiting enteric bacteria infection and promoting mucosal protective regulatory immune responses. This study may have ramifications for prevention of infectious diarrhea in human infants and children, particularly in developing countries.')

('>F', 'TI  - Caseinoglycomacropeptide inhibits adhesion of pathogenic Escherichia coli strains to human cells in culture.', 'AB  - Caseinoglycomacropeptide (CGMP) derived from kappa-casein was investigated for its ability to inhibit the adhesion of 3 strains of verotoxigenic Escherichia coli (VTEC) and 3 strains of enteropathogenic Escherichia coli (EPEC) to human HT29 tissue cell cultures. Effects on adhesion of Desulfovibrio desulfuricans, Lactobacillus pentosus, Lactobacillus casei, Lactobacillus acidophilus, and Lactobacillus gasseri were also investigated. Generally, CGMP exerted effective anti-adhesive properties at a dose of 2.5 mg/mL, albeit with a high degree of strain specificity. The CGMP reduced adhesion of VTEC strains to <50% of the control and reduced adhesion of EPEC strains to between 80 and 10% of the control. The CGMP also reduced the adhesion of L. pentosus and L. casei to 44 and 42%, respectively. A slight but significant reduction of L. acidophilus, to 81%, was observed, but no significant effects were detected with either Dsv. desulfuricans or L. gasseri. Further investigation of the dose response relationships with the E. coli strains gave IC50 values ranging between 0.12 and 1.06 mg/mL.')

('>F', 'TI  - The effect of probiotics on the genotoxicity of furazolidone.', 'AB  - Antigenotoxic activity of probiotic bacteria against furazolidone was studied using the short-term bacterial assay SOS chromotest, with Escherichia coli PQ37 as the test organism. The supernatants from probiotic and furazolidone co-incubation exhibited rather strong suppression on SOS induction produced by furazolidone on E. coli PQ 37 (sfiA: lacZ). Genotoxicity inhibition was found for all strains of the examined bacteria belonging to three genera. The highest genotoxicity inhibition was detected for Bifidobacterium lactis Bb-12 (92.0%) and for Lactobacillus acidophilus T20 (81.9%).')

('>F', 'TI  - Screening of potential lactobacilli antigenotoxicity by microbial and mammalian cell-based tests.', "AB  - Antigenotoxicity is considered an important property for probiotic lactobacilli.  The ability of non probiotic lactobacilli from dairy products and starters to inhibit two reference genotoxins: 4-nitroquinoline-1-oxide and N-methyl-N'-nitro-N-nitrosoguanidine was evaluated. The study was carried out using short-term assays with different targets, such as procaryotic cells (SOS-Chromotest for genotoxicity in Escherichia coli and Ames test for mutagenicity in Salmonella typhimurium) and eucaryotic cells (Comet assay for genotoxicity in Caco-2 enterocytes). A high proportion of strains inhibiting 4-nitroquinoline-1-oxide activity was found in Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus plantarum. Inhibition of N-methyl-N'-nitro-N-nitrosoguanidine activity occurred in only one L. acidophilus strain. All the strains with antigenotoxic properties also demonstrated antimutagenic activity and produced modifications in genotoxin spectroscopic profiles. Strain viability during and after genotoxin exposure was confirmed. Concordance of the results obtained with microbial and mammalian cell-based tests is underlined.")

('>F#others', 'TI  - Characterization of a bacteriocin produced by Enterococcus faecium GM-1 isolated  from an infant.', 'AB  - AIM: To partially characterize the bacteriocin produced by the GM-1 strain of Enterococcus faecium, isolated from the faeces of a newborn human infant. METHODS AND RESULTS: The bacteriocin produced by E. faecium GM-1 showed a broad spectrum of activity against indicator strains of Escherichia coli, Staphylococcus aureus, Vibrio spp., Salmonella typhimurium, Listeria monocytogenes, Lactobacillus acidophilus, and Streptococcus thermophilus. Treatment of the GM-1 bacteriocin with proteolytic enzymes reduced its inhibitory activities. The bacteriocin was stable at 100 degrees C for 20 min and displayed inhibitory activity at neutral pH. The optimal production of bacteriocin from E. faecium GM-1 was obtained when the culture conditions were pH 6.0-6.5 and 35-40 degrees C. The inhibitory activity of the bacteriocin was not substantially changed by the use of different carbon sources in the media, except when galactose was substituted for glucose. The use of a sole nitrogen source caused a decrease in inhibitory activity. A bacteriocin gene similar to enterocin P was identified from the total DNA of E. faecium GM-1 by PCR and direct sequencing methods. CONCLUSION: E. faecium GM-1, which was isolated from the faeces of a newborn baby, produces an enterocin P-like bacteriocin with inhibitory activity against Gram-positive and Gram-negative bacteria, including food-borne pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: E. faecium GM-1, isolated from infant faeces, produces a new bacteriocin that is similar to enterocin P. This bacteriocin is heat stable and has a broad antibacterial spectrum that includes both Gram-positive and Gram-negative bacteria.')

('>F', 'TI  - Hepatoprotective effect of lactic acid bacteria, inhibitors of beta-glucuronidase production against intestinal microflora.', 'AB  - The hepatoprotective activity of lactic acid bacteria (Lactobacillus brevis HY7401, Lactobacillus acidophilus CSG and Bifidobacterium longum HY8001), which inhibited beta-glucuronidase productivity of intestinal microflora, on t-BHP- or CCl4-induced hepatotoxicity of mice were evaluated. These oral administration of lactic acid bacteria lowered beta-glucuronidase production of intestinal microflora as well as Escherichia coli HGU-3. When lactic acid bacteria at a dose of 0.5 or 2 g (wet weight)/kg was orally administered on CCl4-induced liver injury in mice, these bacteria significantly inhibited the increase of plasma alanine transferase and aspartate transferase activities by 17-57% and 57-66% of the CCl4 control group, respectively. These lactic acid bacteria also showed the potent hepatoprotective effect against t-BHP-induced liver injury in mice. The inhibitory effects of these lactic acid bacteria were more potent than that of dimethyl diphenyl bicarboxylate (DDB), which have been used as a commercial hepatoprotective agent. Among these lactic acid bacteria, L. acidophilus CSG exhibited the most potent hepatoprotective effect. Based on these findings, we insist that an inhibitor of beta-glucuronidase production in intestine, such as lactic acid bacteria, may be hepatoprotective.')

('>F', 'TI  - Selective growth-inhibiting effects of compounds identified in Tabebuia impetiginosa inner bark on human intestinal bacteria.', 'AB  - The growth-inhibiting activity of anthraquinone-2-carboxylic acid and lapachol identified in the inner bark of taheebo, Tabebuia impetiginosa, toward 10 human intestinal bacteria was evaluated by using a paper disk diffusion bioassay and compared to those of seven lapachol congeners (1,4-naphthoquinone, naphthazarin, menadione, lawsone, plumbagin, juglone, and dichlone) as well as two commercially available antibiotics, chloramphenicol and tetracycline. Anthraquinone-2-carboxylic acid exhibited very strong growth inhibition of Clostridium paraputrificum at 1 microg/disk while 100 microg/disk of lapachol was needed for moderate growth inhibition of the same organism. These two isolates exhibited weak inhibition of Clostridium perfringens and Escherichia coli at 100 microg/disk while no adverse effects were observed on the growth of Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium infantis, Lactobacillus acidophilus, and Lactobacillus casei at 1000 microg/disk. Structure-activity relationships indicate that a methyl group in the C-2 position of 1,4-naphthoquinone derivatives might play an important role in antibacterial activity.')

('>F', 'TI  - Growth inhibition of foodborne pathogens and food spoilage organisms by select raw honeys.', 'AB  - Twenty-seven honey samples from different floral sources and geographical locations were evaluated for their ability to inhibit the growth of seven food spoilage organisms (Alcaligenes faecalis, Aspergillus niger, Bacillus stearothermophilus, Geotrichum candidum, Lactobacillus acidophilus, Penicillium expansum, Pseudomonas fluorescens) and five foodborne pathogens (Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica Ser. Typhimurium, and Staphylococcus aureus) using an overlay inhibition assay. They were also tested for specific activity against S. aureus 9144 and B. stearothermophilus using the equivalent percent phenol test--a well diffusion assay corresponding to a dilute phenol standard curve. Honey inhibited bacterial growth due to high sugar concentration (reduced water activity), hydrogen peroxide generation, and proteinaceous compounds present in the honey. Some antibacterial activity was due to other unidentified components. The ability of honey to inhibit the growth of microorganisms varies widely, and could not be attributed to a specific floral source or demographic region produced in this study. Antibacterially active samples in this study included Montana buckwheat, tarweed, manuka, melaleuca, and saw palmetto. Furthermore, the bacteria were not uniformly affected by honey. Varying sensitivities to the antimicrobial properties were observed with four strains of S. aureus thus emphasizing the variability in the antibacterial effect of honey samples. Mold growth was not inhibited by any of the honeys tested. B. stearothermophilus, a heat-resistant spoilage bacteria, was shown to be highly sensitive to honey in both the overlay and well diffusion assays; other sensitive bacteria included A. faecalis and L. acidophilus. Non-peroxide antibacterial activity was observed in both assays; the highest instance was observed in the specific activity assay against B. stearothermophilus. Further research could indicate whether honey has potential as a preservative in minimally processed foods.')

('>F', 'TI  - [In vitro effect of carbohydrates and enteric bacteria on adherence of Candida albicans].', 'AB  - The adherence of Candida albicans to any cell is considered essential in the process that leads to colonization. Our objective in this study was to evaluate the effect of different carbohydrates and the presence of lactobacilli and Escherichia coli on the in vitro adherence of Candida albicans. The adherence to buccal epithelial cells was higher when growing at concentrations of galactose of 50, and 200 mM, as well as 50, 200, and 500 mM of sucrose, and 500 mM of mannose, compared with that obtained when growing in Sabouraud dextrose broth (p < 0.01). The presence of other microorganisms, such as Lactobacillus acidophilus and L. casei, caused a decrease in the in vitro adherence of C. albicans to buccal epithelial cells (p < 0.05), whereas E. coli did not modify this adherence at all.')

('>F', 'TI  - Expression of a heterologous manganese superoxide dismutase gene in intestinal lactobacilli provides protection against hydrogen peroxide toxicity.', 'AB  - In living organisms, exposure to oxygen provokes oxidative stress. A widespread mechanism for protection against oxidative stress is provided by the antioxidant enzymes: superoxide dismutases (SODs) and hydroperoxidases. Generally, these enzymes are not present in Lactobacillus spp. In this study, we examined the potential advantages of providing a heterologous SOD to some of the intestinal lactobacilli. Thus, the gene encoding the manganese-containing SOD (sodA) was cloned from Streptococcus thermophilus AO54 and expressed in four intestinal lactobacilli. A 1.2-kb PCR product containing the sodA gene was cloned into the shuttle vector pTRK563, to yield pSodA, which was functionally expressed and complemented an Escherichia coli strain deficient in Mn and FeSODs. The plasmid, pSodA, was subsequently introduced and expressed in Lactobacillus gasseri NCK334, Lactobacillus johnsonii NCK89, Lactobacillus acidophilus NCK56, and Lactobacillus reuteri NCK932. Molecular and biochemical analyses confirmed the presence of the gene (sodA) and the expression of an active gene product (MnSOD) in these strains of lactobacilli. The specific activities of MnSOD were 6.7, 3.8, 5.8, and 60.7 U/mg of protein for L. gasseri, L. johnsonii, L. acidophilus, and L. reuteri, respectively. The expression of S. thermophilus MnSOD in L. gasseri and L. acidophilus provided protection against hydrogen peroxide stress. The data show that MnSOD protects cells against hydrogen peroxide by removing O(2)(.-) and preventing the redox cycling of iron. To our best knowledge, this is the first report of a sodA from S. thermophilus being expressed in other lactic acid bacteria.')

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

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

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

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

('>F', 'TI  - Comparison of bacterial and tissue cell initial adhesion on hydrophilic/hydrophobic biomaterials.', 'AB  - In this study, interactions of widely-used polymeric biomaterials, i.e. poly(hydroxyethyl methacrylate) (PHEMA) and its copolymer with dimethylaminoethyl methacrylate (PHEMA-20% DMAEMA), polyurethane (PU), polypropylene (PP), poly(vinyl chloride) (PVC), and poly(lactide-glycolide) (PLGA), with three pathogenic bacteria and one nonpathogen were investigated comparatively with the adhesion of two tissue cells in different morphologies, i.e. fibroblast-like baby hamster kidney (BHK 21) cells and epithelial Madine Darby kidney (MDBK) cells. Biomaterials were prepared in the membrane form by bulk polymerization or solvent casting. Surface characterization studies showed that these polymers have different surface free energies in the range of 26.9-63.1 erg cm(-2) and they have smooth surfaces. The bacteria used were; Escherichia coli ATCC 25922, Staphylococcus epidermidis ATCC 12228, Staphylococcus aureus, and Lactobacillus acidophilus B-13. Initial adhesion of bacteria to the polymeric surfaces was examined under static conditions and in a laminar flow cell. The adhesion behaviour of S. aureus and S. epidermidis was found independent of the polymeric surface hydrophobicity. However, the percentage of attached E. coli decreased when increasing the surface free energy of the polymer, while L. acidophilus showed just the opposite behaviour. The comparative results indicated that the adhesion of BHK and MDBK cell was lowest on the most hydrophilic PHEMA surface and highest on the most hydrophobic PP surface. In contrast to the case of bacterial adhesion, no relationship was found between polymer hydrophobicity and mammalian cell adherence.')

('?F', 'TI  - Lactobacillus acidophilus (strain LB) from the resident adult human gastrointestinal microflora exerts activity against brush border damage promoted by a diarrhoeagenic Escherichia coli in human enterocyte-like cells.', 'AB  - BACKGROUND AND AIMS: The normal gastrointestinal microflora exerts a barrier effect against enteropathogens. The aim of this study was to examine whether lactobacilli, a minor genus of the resident gut microflora, exerts a protective effect against the cellular injuries promoted by the diarrhoeagenic Afa/Dr diffusely adhering Escherichia coli (Afa/Dr DAEC) C1845 strain in human intestinal cells. METHODS: Cultured human intestinal fully differentiated enterocyte-like Caco-2/TC7 cells were used. Antibacterial activity was examined by measuring the viability of the adhering C1845 bacteria. The distribution of brush border associated cytoskeleton and functional proteins was examined by immunofluorescence labelling coupled to confocal laser scanning microscopy analysis. RESULTS: The activity of Lactobacillus acidophilus strain LB isolated from the resident human gastrointestinal microflora was examined. A dose dependent decrease in viability of C1845 bacteria was observed after both direct contact in vitro between the spent culture supernatant (LB-SCS) and the bacteria, and when the bacteria were adherent on Caco-2/TC7 cells. Protection against the C1845 induced alterations in expression of F-actin, sucrase-isomaltase, dipeptidylpeptidase IV, alkaline phosphatase, and fructose transporter alterations was observed when the cells were exposed to LB-SCS. CONCLUSION: L acidophilus strain isolated from the resident adult human gastrointestinal microflora, together with its antimicrobial activity, exerts a protective effect against the brush border lesions promoted by the diarrhoeagenic Afa/Dr DAEC strain C1845.')

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

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

('>F', 'TI  - Adsorption effect of activated charcoal on enterohemorrhagic Escherichia coli.', 'AB  - The adsorption property of activated charcoal on verotoxin (VT)-producing Escherichia coli (VTEC) was examined using E. coli O157:H7. In the present study, E. coli O157:H7 strains were effectively adsorbed by activated charcoal. Adsorption was dose-dependent, and the maximum adsorption occurred within 5 min. At 10 mg of activated charcoal, bacteria tested were completely adsorbed. Activated charcoal also had the capacity to adsorb toxin (verotoxin 2) activity from the bacterial extract. Furthermore, the adsorption efficiency of activated charcoal for the normal bacterial flora in the intestine was assessed using Enterococcus faecium, Bifidobacterium thermophilum, and Lactobacillus acidophilus. Activated charcoal showed lower binding capacity to the normal bacterial flora tested than that to E. coli O157:H7 strains. These results suggest that activated charcoal could be a good adsorbent system for the removal of VTEC and verotoxin.')

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

('>F', 'TI  - Molecular analysis of mutated Lactobacillus acidophilus promoter-like sequence P15.', 'AB  - The promoter-like sequence P15 that was previously cloned from the chromosome of  Lactobacillus acidophilus ATCC 4356 is active in Lactobacillus reuteri, Lactobacillus plantarum, Lactobacillus acidophilus, and Escherichia coli, but not in Lactococcus lactis. N-methyl-N-nitroso-N-guanidine (MNNG) mutagenesis of P15 was used to select for a promoter active in L. lactis MG1363. Molecular analysis of the mutated promoter (designated P16) revealed a 90 bp deletion and a T-->A transversion. This deletion, in combination with the addition to the transversion, created a promoter with putative -35 and -10 hexamers identical to the consensus promoter sequence found in E. coli and Bacillus subtilis vegetative promoters. The activity of P16 was measured by its ability to promote chloramphenicol resistance in different bacteria when inserted in the promoter-probe plasmid pBV5030 (designated pLA16). The MIC of chloramphenicol in L. lactis, L. reuteri, L. plantarum, E. coli, and L. acidophilus harbouring pLA16 were 30, 170, 180, > 500, and 3 micrograms/mL, respectively. This represents an increase in promoter activity compared to P15 in L. reuteri of 3-fold, in L. plantarum of 9-fold, and in E. coli of at least 2.5-fold, but a decrease in L. acidophilus of 7-fold.')

('>F', 'TI  - Effect of supplements with lactic acid bacteria and oligofructose on the intestinal microflora during administration of cefpodoxime proxetil.', 'AB  - Thirty healthy volunteers in three groups participated in a study of the effect on the intestinal microflora of oral supplementation with Bifidobacterium longum, Lactobacillus acidophilus and oligofructose, an indigestible oligosaccharide, during oral administration of cefpodoxime proxetil bd for 7 days. Those in group A also received an oral supplement with c.1011 cfu of B. longum BB 536 and L. acidophilus NCFB 1748 and 15 g oligofructose daily, those in group B received a supplement with oligofructose only and those in group C received placebo, for 21 days. In all three groups there was a marked decrease in aerobic microorganisms, involving mainly a rapid and almost complete disappearance of Escherichia coli (P: < 0.05) during antimicrobial administration and, thereafter, an overgrowth of enterococci (P: < 0.05). The number of intestinal yeasts also increased significantly (P: < 0.05) in groups A and B over the same period. There was a dramatic decrease in anaerobic microorganisms on day 4 of administration, mainly caused by loss of bifidobacteria (P: < 0.05) in all groups. The number of lactobacilli also decreased but was significantly higher in group A than in group C at the end of cefpodoxime proxetil administration. Clostridium difficile was found in only one person from group A, but six persons each in groups B and C. Of the bifidobacterial strains isolated from the faecal samples in group A, one was similar to the strain of B. longum administered, but most volunteers were colonized by several different strains of B. longum during the investigation period. The administered strain of L. acidophilus was recovered from six patients in group A.')

('>F', 'TI  - Cordycepin: selective growth inhibitor derived from liquid culture of Cordyceps militaris against Clostridium spp.', "AB  - 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.")

('>F', 'TI  - Analysis of promoter sequences from Lactobacillus and Lactococcus and their activity in several Lactobacillus species.', 'AB  - Promoter-active fragments were isolated from the genome of the probiotic organism Lactobacillus rhamnosus strain GG using the promoter-probe vector pNZ272. These promoter elements, together with a promoter fragment isolated from the vaginal strain Lactobacillus fermentum BR11 and two previously defined promoters (Lactococcus lactis and Lactobacillus acidophilus ATCC 4356 slpA), were introduced into three strains of Lactobacillus. Primer-extension analysis was used to map the transcriptional start site for each promoter. All promoter fragments tested were functional in each of the three lactobacilli and a purine residue was used to initiate transcription in most cases. The promoter elements encompassed a 52- to 1,140-fold range in promoter activity depending on the host strain. Lactobacillus promoters were further examined by surveying previously mapped sequences for conserved base positions. The Lactobacillus hexamer regions (-35: TTgaca and -10: TAtAAT) closely resembled those of Escherichia coli and Bacillus subtilis, with the highest degree of agreement at the -10 hexamer. The TG dinucleotide upstream of the -10 hexamer was conserved in 26% of Lactobacillus promoters studied, but conservation rates differed between species. The region upstream of the -35 hexamer of Lactobacillus promoters showed conservation with the bacterial UP element.')

('>F', 'TI  - Antagonistic effect of Lactobacillus acidophilus, Saccharomyces boulardii and Escherichia coli combinations against experimental infections with Shigella flexneri and Salmonella enteritidis subsp. typhimurium in gnotobiotic mice.', 'AB  - Lactobacillus acidophilus, Saccharomyces boulardii and Escherichia coli are probiotic strains used individually to protect against enteropathogenic agents. In order to determine if a synergistic effect of the individual protective mechanisms ordinarily attributed to each of these biotherapeutic agents is possible, we orally administered Lact. acidophilus H2B20, S. boulardii and E. coli EMO (LSE) to germfree mice. Ten days after colonization of the digestive tract, groups of animals associated (experimental) or not (control) with LSE were challenged orally with streptomycin resistant (Sfr) or streptomycin sensitive (Sfs) Shigella flexneri strains or Salmonella enteritidis subsp. typhimurium. Bacterial counts in faeces from experimental mice showed that the Sfr strain was eliminated 11 d after challenge while Sfs and S. enteritidis subsp. typhimurium colonized the digestive tract and continued to be present at high population levels (108 CFU g-1 of faeces), which is similar to that observed in control animals. All possible di- and monoassociations of the three probiotics with gnotobiotic mice were also performed before experimental oral infection with Sfr. The data showed that antagonism was obtained only when E. coli EMO was present. Different sensitivity of Sh. flexneri Sfr and Sfs to E. coli EMO antagonism could be explained by the different generation times between Sfr and Sfs, as shown by colonization kinetic experiments in the digestive tract of gnotobiotic mice.')

('>F', 'TI  - Enterocin 012, a bacteriocin produced by Enterococcus gallinarum isolated from the intestinal tract of ostrich.', 'AB  - Enterococcus gallinarum strain 012, isolated from the duodenum of ostrich, produced enterocin 012 which is active against Ent. faecalis, Lactobacillus acidophilus, Lact. sake, Listeria innocua, Propionibacterium acidipropionici, Propionibacterium sp., Clostridium perfringens, Pseudomonas aeruginosa and Salmonella typhimurium. One of the four pathogenic strains of Escherichia coli isolated from the intestinal tract of ostrich was inhibited by enterocin 012. No antimicrobial activity was recorded against Bacillus cereus, Cl. sporogenes, Cl. tyrobutyricum, Leuconostoc cremoris, Pediococcus pentosaceus, Staphylococcus carnosus and Streptococcus thermophilus. Enterocin 012 was resistant to treatment with lysozyme, catalase, lipase and papain, but sensitive to Proteinase K, alpha-chymotrypsin, trypsin and pepsin. Treatment of enterocin 012 with gastric juice from the duodenum resulted in a 50% loss of antibacterial activity. Half of the activity was lost when incubated at 80 degrees C for 30 min, or when kept overnight at a pH of 1.0-5.0 and pH 11.0 and 12.0, respectively. Enterocin 012 production started in mid-logarithmic growth and reached a maximum of 800 AU ml-1, but increased further to 1600 AU ml-1 in the stationary growth phase. The peptide is approximately 3.4 kDa in size, as determined after partial purification with Amberlite XAD-1180 and ammonium sulphate precipitation, followed by tricine-sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The mechanism of antimicrobial activity against Lact. sake LMG 13558 is bactericidal and caused cell lysis of active growing cells.')

('>F', 'TI  - Effects of intestinal bacteria on the development of colonic neoplasm II. Changes in the immunological environment.', 'AB  - To study the effects of intestinal bacteria on the development of colonic neoplasm, we have established gnotobiotic mice with a single species of intestinal bacteria. In the previous study, the incidence of colonic adenoma induced with 1,2-dimethylhydrazine (DMH) in the gnotobiotic mice with Lactobacillus acidophilus, gnotobiotic mice with Escherichia coli and germ-free mice were 30, 50 and 74%, respectively. In this study, 7-week-old mice in each group were sacrificed without the administration of DMH to examine the constituents of immuno-competent cells in various mouse organs using flow cytometry. In the gnotobiotic mice, CD3 intermediate interleukin (IL)-2Rbeta positive cells were observed predominantly in the liver. In the gnotobiotic mice with L. acidophilus, Mac-1 positive Gr-1 positive cells were observed predominantly in the colonic lamina propria. The activation of extrathymic T cells in the liver and granulocytes in the colonic mucosa may be related to anti-neoplastic effects of L. acidophilus in this experimental model.')

('>F', 'TI  - Growth-inhibiting effects of Coptis japonica root-derived isoquinoline alkaloids  on human intestinal bacteria.', 'AB  - 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.')

('>F#OPPOSITE', 'TI  - Antibacterial activity of Lactobacillus species against Vibrio species.', 'AB  - Forty-one Lactobacillus strains were tested for antagonistic activity against nine strains of Vibrio. L. plantarum and L. casei were the most effective, and L. brevis was the least effective in inhibiting the growth of Vibrio species. L. gasseri and L. helveticus strains showed higher activity, while L. reuteri and L. fermentum showed lower inhibitory activity against Vibrio species. L. acidophilus strains exhibited various degrees of antagonistic activities against Vibrio species. However, none of the Lactobacillus species were able to inhibit the growth of Salmonella enteritidis, S. typhimurium, Escherichia coli, and Staphylococcus aureus. Inhibition of the Vibrio species was probably due to the production of organic acids by the Lactobacillus species.')

('>F', 'TI  - Mechanism of inhibition of tannic acid and related compounds on the growth of intestinal bacteria.', 'AB  - Tannic acid, propyl gallate and methyl gallate, but not gallic acid, were found to be inhibitory to the growth of intestinal bacteria Bacteroides fragilis ATCC 25285, Clostridium clostridiiforme ATCC 25537, C. perfringens ATCC 13124, C. paraputrificum ATCC 25780, Escherichia coli ATCC 25922, Enterobacter cloacae ATCC 13047, Salmonella typhimurium TA98 and S. typhimurium YG1041 at 100-1000 microg/ml in culture broth. Neither Bifidobacterium infantis ATCC 15697 nor Lactobacillus acidophilus ATCC 4356 was inhibited by any of the above compounds up to 500 microg/ml. Tannic acid has a much greater relative binding efficiency to iron than propyl gallate, methyl gallate or gallic acid. The inhibitory effect of tannic acid to the growth of intestinal bacteria may be due to the strong iron binding capacity of tannic acid; whereas the effect of propyl gallate and methyl gallate probably occurs by a different mechanism. The growth of E. coli was restored by the addition of iron to the medium after the precipitate caused by tannic acid was removed. Neither B. infantis nor L. acidophilus require iron for growth. This probably contributes to their resistance to tannic acid. Because tannins are abundant in the human diet, tannins may affect the growth of some intestinal bacteria and thus may have an impact on human health.')

('>F', 'TI  - Interference in initial adhesion of uropathogenic bacteria and yeasts to silicone rubber by a Lactobacillus acidophilus biosurfactant.', "AB  - The ability of the Lactobacillus acidophilus RC14 biosurfactant 'surlactin' to inhibit the initial adhesion of various uropathogenic bacteria and two yeast strains to silicone rubber was investigated in a parallel-plate flow chamber in filter-sterilised pooled human urine. A parallel-plate flow chamber with a silicone rubber bottom plate was filled with a 1.0 mg/ml biosurfactant solution for adsorption overnight (18 h). Subsequently, the adhesion of the bacterial or yeast cells from a urine suspension under low flow (shear rate 15 s(-1)) was followed in situ by automated image analysis. Control tests were with untreated silicone rubber. Initial deposition rates and numbers of adhering cells after 4 h of flow were determined. Surlactin layers caused a marked inhibition of the initial deposition rates and adhesion numbers after 4 h for the majority of the bacteria (11 of 15 strains tested) and this inhibition was particularly effective against Enterococcus faecalis, Escherichia coli and Staphylococcus epidermidis. Although the initial deposition rates of the two Candida albicans strains were reduced by c. 50% in comparison with the controls, the numbers of yeast cells adhering after 4 h were similar.")

('>F', 'TI  - Increased antibody production against gut-colonizing Escherichia coli in the presence of the anaerobic bacterium Peptostreptococcus.', 'AB  - Germ-free rats were colonized with E. coli alone, or with E. coli plus Lactobacillus acidophilus and a strain of the obligate anaerobic gram-positive species, Peptostreptococcus. The presence of Peptostreptococcus reduced translocation of E. coli, but increased the serum antibody response to E. coli antigen. Whereas the immunoglobulin G (IgG) anti-E. coli antibodies largely represented cross-reactive antibodies, those of the immunoglobulin M (IgM) isotype represented true anti-E. coli antibodies because they could not be absorbed by L. acidophilus or Peptostreptococcus but could with E. coli. We suggest that peptostreptococci prime the gut immune system to other bacterial antigens and that this could be a mechanism behind the reduced translocation of facultative anaerobes in the presence of obligate anaerobes.')

('>F', 'TI  - Growth-inhibitory effects of Galla Rhois-derived tannins on intestinal bacteria.', 'AB  - The growth-inhibitory activity of Galla Rhois-derived materials towards 17 intestinal bacteria was evaluated using an impregnated paper disc method. The biologically active components of Galla Rhois were characterized as the tannins methyl gallate (MG) and gallic acid (GA) by spectral analysis. The growth responses varied with bacterial strain tested. In the test using 10 mg disc-1, MG and GA produced a clear inhibitory effect on harmful bacteria such as Clostridium perfringens, Cl. paraputrificum, Eubacterium limosum, Bacteroides fragilis, Staphylococcus aureus and Escherichia coli. Methyl gallate showed no growth-inhibitory activity towards Bifidobacterium adolescentis or B. longum whereas the growth of B. bifidum, B. breve, B. infantis, B. animalis, B. thermophilum, Lactobacillus acidophilus, Lact. plantarum and Streptococcus faecalis was slightly affected. However, GA did not adversely affect the growth of the bifidobacteria and lactobacilli. At 5 mg disc-1, MG significantly inhibited the growth of Cl. perfringens and Cl. paraputrificum but did not affect the growth of the bifidobacteria and lactobacilli. At 1 mg disc-1, MG greatly inhibited the growth of Cl. perfringens alone. These results may be an indication of at least one of the pharmacological actions of Galla Rhois.')

('>F#FAILURE', 'TI  - Note: lack of influence of adherent Lactobacillus isolates on the attachment of Escherichia coli to the intestinal epithelial cells of chicken in vitro.', 'AB  - Two Lactobacillus isolates, Lact. acidophilus I 26 and Lact. fermentum I 25, were selected, based on their poor aggregation with Escherichia coli and strong ability to adhere to ileal epithelial cells (IEC), to study in vitro interactions with E. coli O1:K1, O2:K1 and O78:K80 in an IEC radioactive-assay under the conditions of exclusion (lactobacilli and IEC, followed by the addition of E. coli), competition (lactobacilli, IEC and E. coli together) and displacement (E. coli and IEC, followed by the addition of lactobacilli). The results indicated that Lact. acidophilus I 26 and Lact. fermentum I 25 could not significantly reduce the attachment of E. coli O1:K1, O2:K1 and O78:K80 to IEC under the three conditions tested in vitro, except that the attachment of E. coli O1:K1 was slightly reduced by Lact. fermentum I 25 in the test for competition.')

('>F', 'TI  - Adhesion of some probiotic and dairy Lactobacillus strains to Caco-2 cell cultures.', 'AB  - The adhesion of 12 different Lactobacillus strains was studied using Caco-2 cell  line as an in vitro model for intestinal epithelium. Some of the strains tested have been used as probiotics, and most of them are used in the dairy and food industry. Human and bovine enterotoxigenic Escherichia coli strains were used as positive and negative control, respectively. Bacterial adhesion to Caco-2 cell cultures was quantitated using radiolabelled bacteria. The adherence of bacteria was also observed microscopically after Gram staining. Viability of bacteria prior to adhesion was verified using flow cytometry. Among the tested strains, L. casei (Fyos) was the most adhesive strain and L. casei var. rhamnosus (Lactophilus) was the least adhesive strain, approximately 14 and 3% of the added bacteria adhered to Caco-2 cell cultures, respectively. The corresponding values for positive and negative control E. coli strains were 14 and 4%, respectively. The Lactobacillus strains tested could not be divided into distinctly adhesive or non-adhesive strains, since there was a continuation of adhesion rates. The four most adhesive strains were L. casei (Fyos), L. acidophilus 1 (LC1), L. rhamnosus LC-705 and Lactobacillus GG (ATCC 53103). No significant differences in the percentage adhesion were observed between these strains. Adhesion of all the strains was dependent on the number of bacteria used, since an approximately constant number of Caco-2 cells was used, indicating that the Caco-2 cell binding sites were not saturated. Viability of bacteria was high since approximately 90% of the bacteria were viable with the exception of L. acidophilus 1 which was 74% viable. Microscopic evaluations agreed with the radiolabelled binding as evidenced by observing more bacteria in Gram-stained preparations of good adhering strains compared to poorly adhering strains.')

('>F', 'TI  - Nonlipopolysaccharide component(s) of Lactobacillus acidophilus stimulate(s) the  production of interleukin-1 alpha and tumor necrosis factor-alpha by murine macrophages.', 'AB  - Previous studies in our laboratory suggested that Lactobacillus acidophilus strain DDS-1 (LA1) has a suppressive effect on chemically induced tumors in experimental animals. In an effort to understand the possible mechanisms underlying this effect, we investigated the ability of LA1 to induce the production of interleukin-1 alpha (IL-1 alpha) and tumor necrosis factor-alpha (TNF-alpha), which have potent cytocidal and cytostatic effects on tumor cells. The mouse macrophage cell line RAW264.7 was incubated with live or heat-killed cells of four strains of L. acidophilus or Bifidobacterium bifidum. Escherichia coli was used as a source of lipopolysaccharide that is known to induce the above cytokines. The amount of the cytokines present in the culture fluid was quantitated by an enzyme-linked immunosorbent assay. LA1 induced the production of higher levels of IL-1 alpha and TNF-alpha than other lactobacilli and bifidobacteria. Stimulation of the production of the cytokines was not due to the lipopolysaccharide (LPS) component, since LPS at concentrations equivalent to, or 100-fold greater than, that of LA1 induced only negligible amounts of IL-1 alpha and TNF-alpha. These results reveal that non-LPS component(s) of LA1 stimulate(s) the production of IL-1 alpha and TNF-alpha by macrophages, indicating that this organism stimulates the production of immunologic factors.')

('>F', 'TI  - Deoxycytidine kinase and deoxyguanosine kinase of Lactobacillus acidophilus R-26  are colinear products of a single gene.', 'AB  - Three of the four deoxynucleoside kinases required for growth of Lactobacillus acidophilus R-26 exist as heterodimeric pairs specific for deoxyadenosine (dAK) and deoxycytidine (dCK) or dAK and deoxyguanosine (dGK). However, only two tandem genes, dak/dgk, are found, and are expressed only as dAK/dGK in transformed Escherichia coli. Sequencing peptides spanning 63% of the native dCK subunit revealed a sequence identical to that deduced from dgk (beginning MTVIVL...), except that dCK lacks residues 2 and 3 (dCK is M..IVL; dGK is .TVIVL). Also, mass spectrometry indicates that native dCK and dGK subunits are identical in mass adjusted for the first three residues. Furthermore, the native enzymes have identical isoelectric pH values, indicating an equal number of charged residues. To enable E. coli to express peptide having the native dCK sequence, codons 2 and 3 were deleted from the dgk portion of the tandem genes, resulting in expression of protein having the specificities and regulatory properties of native dAK/dCK, including heterotropic stimulation of dAK activity by deoxycytidine or dCTP (not deoxyguanosine or dGTP) and end-product inhibition of the respective activities by dATP and dCTP. Subcloning normal and mutant dgk yielded homodimeric dGK and dCK, respectively. The dCK homodimer strongly resembles human dCK, with a low K(m) for deoxycytidine, the ability to phosphorylate deoxyadenosine and deoxyguanosine at much higher K(m) values, and end-product inhibition by dCTP. Thus two distinct and specific enzymes evidently are derived from a single Lactobacillus gene. The mechanism by which this occurs in vivo has yet to be elucidated.')

('>F', 'TI  - The potential of Lactobacillus as a carrier for oral immunization: development and preliminary characterization of vector systems for targeted delivery of antigens.', 'AB  - Oral administration of lactobacilli evokes mucosal and systemic immune responses  against epitopes associated with these organisms (Gerritse et al., 1990, 1991). The adjuvant function of different Lactobacillus species was investigated under the conditions of intraperitoneal (i.p.) injection or oral administration. After i.p. injection of trinitrophenylated chicken gamma-globulin, high DTH responses were observed with Lactobacillus casei and Lactobacillus plantarum, but low responses with Lactobacillus fermentum and Lactobacillus delbrueckii subsp. bulgaricus. In different experimental model systems L. casei and L. plantarum consistently showed significant adjuvanticity. A series of expression and expression-secretion vectors containing the strong constitutive promoter of the L. casei L-ldh gene or the regulatable promoter of the Lactobacillus amylovorus amy gene (Pouwels and Leer, 1995) was used for the intracellular, extracellular and surface-bound expression of an influenza virus antigenic determinant fused to Escherichia coli beta-glucuronidase. Intracellular expression of the fusion protein amounted to 1-2% of total soluble protein. Lactobacilli synthesizing the fusion protein intracellularly evoked an oral immune response after subcutaneous priming.')

('>F', 'TI  - Role of intestinal bacteria in ileal ulcer formation in rats treated with a nonsteroidal antiinflammatory drug.', 'AB  - The role of intestinal bacteria in induction and repression of ulcer formation in the ileum of rats treated with one of the nonsteroidal antiinflammatory drugs (NSAIDs), 5-bromo-2-(4-fluorophenyl)-3-(4-methylsulfonylphenyl) thiophene (BFMeT), was examined in this study. BFMeT was administered by intragastric gavage once at doses of 500-1,500 mg/kg of body weight to Wistar rats treated with and without antibiotics (bacitracin, neomycin, streptomycin), germ-free rats and gnotobiotic rats, and 72 hr later their gastrointestinal tracts were examined for ulcer formation. A single oral administration of BFMeT induced ileal ulcers in specific pathogen-free rats. However, the rats given antibiotics to reduce the intestinal bacteria had no ulcers. BFMeT-treated germ-free rats and gnotobiotic rats mono-associated with Bifidobacterium adolescentis or Lactobacillus acidophilus also had no intestinal ulcers. However, the drug induced ileal ulcers in gnotobiotic rats mono-associated with Eubacterium limosum or Escherichia coli. An overnight culture of B. adolescentis or L. acidophilus or yogurt containing Bifidobacterium breve and Streptococcus thermophilus, when given as drinking water, inhibited ulcer formation in the ileum of rats treated with BFMeT. Gram staining of the ileal contents of normal rats revealed that 97.4% of the stained microorganisms were Gram-positive rods and only 1.2% were Gram-negative rods. In the group of rats with ulcers induced by BFMeT, the Gram-positive rods decreased by 56.4% and the Gram-negative rods including Escherichia coli, Klebsiella, Proteus and Bacteroides increased by 37.3%. However, in the group of rats administered the Bifidobacterium culture, the Lactobacillus culture or yogurt, the percentages of the Gram-negative rods were decreased. Although Lactobacillus was a major bacterium in the ileum of normal rats, the Gram-negative facultatively anaerobic rods E.coli, Klebsiella and Proteus were increased in the ulcerated ileum of rats treated with BFMeT, suggesting that these bacteria are associated with ulcer formation in rats treated with NSAIDs, and that Lactobacillus and Bifidobacterium inhibit it by repressing the growth of ulcer-inducing bacteria.')

('>F', 'TI  - Positive selection, cloning vectors for gram-positive bacteria based on a restriction endonuclease cassette.', 'AB  - Lactococcus lactis contains numerous restriction and modification (R/M) systems of different specificities. A novel IIS type R/M system encoded by the LlaI operon has previously been characterized from the L. lactis conjugative plasmid pTR2030. The LlaI operon is composed of six genes: First, a small regulatory gene llaIC precedes the methylase gene llaIM. The following three genes, llaI.1, llaI.2, llaI.3, are all essential for restriction endonuclease activity and are designed as the restriction cassette llaIR. The forth open reading frame of unknown function follows the llaIR gene cassette. We have successfully subcloned the three llaIR genes, llaI.1, llaI.2, and llaI.3, without llaIM, as a suicide cassette into the three shuttle vectors pTRKL2, pTRKH2, and pBV5030. A promoter (P6) from Lactobacillus acidophilus ATCC4356, which is functional in E. coli, lactococci, and lactobacilli (Djordjevic and Topisirovic, unpublished) was cloned upstream of the three gene cassette. Restriction activity was evaluated in Escherichia coli and several gram-positive bacteria. The llaIR restriction cassette was not functional in E. coli, but its presence was lethal to L. lactis, Lactobacillus gasseri, Lactobacillus plantarum, Lactobacillus johnsonii, Lactobacillus acidophilus, Carnobacterium pisicola, Enterococcus faecalis, Bacillus subtilis, and Leuconostoc gelidum. Several novel, positive selection cloning vectors were developed that can exploit unique cloning sites within the llaIR cassette. Insertions in llaI.1 resulted in complete inactivation of restriction activity and provided unconditional selection for recombinant plasmids in surviving transformants. These positive selection cloning vectors are the first for gram-positive bacteria that are based on a restriction endonuclease cassette. Functional activity of the llaIR genes in various gram-positive bacteria would also enable use of these cloning vectors for positive selection of promoters, terminators, and regulatory sequences across these genera.')

('>F', 'TI  - Role of Escherichia coli P fimbriae in intestinal colonization in gnotobiotic rats.', 'AB  - Adherence via P fimbriae is associated with long-term persistence of Escherichia  coli in the human large intestine, but a causal relationship has not been proven. In the present study, germfree rats were colonized with a mixture of two isogenic E. coli strains, one P fimbriated and the other type 1 fimbriated. Both types of fimbriae conferred adherence to rat colonic epithelial cells. With two mutant strains from a pyelonephritogenic isolate of serotype O75:K5:H-, the P-fimbriated strain 824 attained much higher numbers than its type 1-fimbriated counterpart when colonized in vivo for 2 weeks (10(10) versus 10(6) bacteria per g, respectively; P < 0.0001). The expression of P fimbriae by 824 was also retained during colonization. With transformant isogenic strains obtained from a normal fecal isolate incapable of phase variation, no benefit of P fimbriae was seen and most bacteria lost their plasmids during in vivo colonization. When the pyelonephritogenic mutant and fecal transformant strains were combined, the former colonized at high levels while the latter were suppressed. In contrast, no suppression was seen when the transformant E. coli strains colonized in combination with Lactobacillus acidophilus or Peptostreptococcus sp. The results indicate that P fimbriae, but also other bacterial traits linked to uropathogeneicity, could play an important role for persistence in the gut normal microbiota. Neither P nor type 1 fimbriae seemed to contribute to the ability to translocate to the mesenteric lymph nodes.')

('>F', 'TI  - Genetic and biochemical characterization of the Lactobacillus delbrueckii subsp.  lactis bacteriophage LL-H lysin.', 'AB  - LL-H, a virulent phage of Lactobacillus delbrueckii subsp. lactis, produces a peptidoglycan-degrading enzyme, Mur, that is effective on L. delbrueckii, Lactobacillus acidophilus, Lactobacillus helveticus, and Pediococcus damnosus cell walls. In this study, the LL-H gene mur was cloned into Escherichia coli, its nucleotide sequence was determined, and the enzyme produced in E. coli was purified and biochemically characterized. Mur was purified 112-fold by means of ammonium sulfate precipitation and cation-exchange chromatography. The cell wall-hydrolyzing activity was found to be associated with a 34-kDa protein. The C-terminal domain of Mur is not essential for catalytic activity since it can be removed without destroying the lytic activity. The N-terminal sequence of the purified lysin was identical to that deduced from the nucleotide sequence, but the first methionine is absent from the mature protein. The N-terminal part of this 297-amino-acid protein had homology with several Chalaropsis-type lysozymes. Reduction of purified and Mur-digested L. delbrueckii cell wall material with labeled NaB3H4 indicated that the enzyme is a muramidase. The temperature optimum of purified Mur is between 30 and 40 degrees C, and the pH optimum is around 5.0. The LL-H lysin Mur is stable at temperatures below 60 degrees C.')

('>F', 'TI  - Cloning and expression of the heterodimeric deoxyguanosine kinase/deoxyadenosine  kinase of Lactobacillus acidophilus R-26.', 'AB  - Two uniquely paired deoxynucleoside kinases, deoxycytidine kinase/deoxyadenosine  kinase (dCK/dAK) and deoxyguanosine kinase/deoxyadenosine kinase (dGK/dAK) are required, together with thymidine kinase (TK), for deoxynucleotide synthesis in Lactobacillus acidophilus R-26. Using polymerase chain reaction-generated probes based on N-terminal amino acid sequences, we have cloned tandem genes for 25- and 26-kDa polypeptides, whose derived amino acid sequences and size correspond to wild-type Lactobacillus enzyme subunits. Expression in Escherichia coli uses a single endogenous promoter and yields active dGK/dAK (approximately 3% of extracted protein) closely resembling wild-type dGK/dAK in specificity, kinetics, heterotropic activation, and end product inhibition. Alignment of cloned genes reveals 65% identity in their DNA sequences and 61% identity in derived amino acid sequences. Comparison with herpes-viral TKs reveals three conserved regions: glycine- and arginine-rich ATP-binding motifs and a D/E-R-S/H motif at the putative TK deoxynucleoside site. Greater homology, however, is seen upon multiple alignment of dGK with mammalian deoxycytidine kinases, yielding the consensus sequence-D/E-R-S-I/V-Y-x-D-.dGK also shares a sequence (-Y-D-P-T-I/L-E-D-S/Y-Y-) required for GTP hydrolysis by p21ras.')

('>F#AGG', 'TI  - The effect of Lactobacillus spp. on the attachment of enterotoxigenic Escherichia coli to isolated porcine enterocytes.', 'AB  - A total of 43 strains of lactobacilli were isolated from the gastrointestinal tract of piglets at the time of weaning. Isolates, grown on solid media, were allocated to strongly adherent or non/weakly adherent groups on the basis of numbers attaching to isolated porcine enterocytes. Strains of Lactobacillus fermentum were disproportionally represented amongst the strongly-adherent strains and Lact. acidophilus and Lact. salivarius amongst the non/weakly-adherent group. Lactobacilli showed significantly better attachment ability when grown on agar than when grown in broth culture. Strongly adherent strains were not found to effect the attachment of enterotoxigenic Escherichia coli to porcine enterocytes, tested under the conditions of exclusion (lactobacilli added to the enterocytes before E. coli), competition (lactobacilli and E. coli added simultaneously) and displacement (E. coli added before lactobacilli). Tests were made with [14C]-labelled E. coli. Suspensions of bacteria and enterocytes were passed through a filter selected to retain enterocytes but pass free bacterial cells. Counts (dpm) obtained from filters after solubilization were taken as a measure of E. coli attachment. Some strains of lactobacilli coaggregated with enterotoxigenic E. coli with K88 fimbriae, but not with a K88-negative mutants strain. These were excluded from the competitive exclusion experiments. In the apparent absence of a direct effect on the association of E. coli with host tissue, removal of potential gut pathogens by aggregation could contribute to the probiotic properties ascribed to lactic acid bacteria.')

('>F#AGG', 'TI  - Lactobacillus acidophilus LA 1 binds to cultured human intestinal cell lines and  inhibits cell attachment and cell invasion by enterovirulent bacteria.', 'AB  - Four human Lactobacillus acidophilus strains were tested for their ability to adhere onto human enterocyte like Caco-2 cells in culture. The LA 1 strain exhibited a high calcium independent adhesive property. This adhesion onto Caco-2 cells required a proteinaceous adhesion promoting factor, which was present in the spent bacterial broth culture supernatant. LA 1 strain also strongly bound to the mucus secreted by the homogeneous cultured human goblet cell line HT29-MTX. The inhibitory effect of LA 1 organisms against Caco-2 cell adhesion and cell invasion by a large variety of diarrhoeagenic bacteria was investigated. As a result, the following dose dependent inhibitions were obtained: (a) against the cell association of enterotoxigenic, diffusely adhering and enteropathogenic Escherichia coli, and Salmonella typhimurium; (b) against the cell invasion by enteropathogenic Escherichia coli, Yersinia pseudotuberculosis, and Salmonella typhimurium. Incubations of L acidophilus LA 1 before and together with enterovirulent E coli were more effective than incubation after infection by E coli.')

('>f', 'TI  - Adhering heat-killed human Lactobacillus acidophilus, strain LB, inhibits the process of pathogenicity of diarrhoeagenic bacteria in cultured human intestinal cells.', 'AB  - Heat-killed L. acidophilus, strain LB, was tested for its ability to adhere in vitro onto human enterocyte-like Caco-2 and muco-secreting HT29-MTX cells in culture. The heat-killed LB bacteria exhibited a high adhesive property. A diffuse pattern of adhesion was observed to the undifferentiated cells, the apical brush border of the enterocytic cells, and to the mucus layer that covered the surface of the mucus-secreting cells. The inhibitory effect of heat-killed LB organisms against the human intestinal Caco-2 cell-adhesion and cell-invasion by a large variety of diarrhoeagenic bacteria was investigated. The following dose-dependent inhibitions were obtained: (i) against the cell-association of enterotoxigenic, diffusely-adhering and enteropathogenic Escherichia coli, Listeria monocytogenes, Yersinia pseudotuberculosis, and Salmonella typhimurium; (ii) against the cell-invasion by enteropathogenic Escherichia coli, Yersinia pseudotuberculosis, Listeria monocytogenes and Salmonella typhimurium.')

('>F', 'TI  - S-layer protein of Lactobacillus acidophilus ATCC 4356: purification, expression  in Escherichia coli, and nucleotide sequence of the corresponding gene.', 'AB  - The cell surfaces of several Lactobacillus species are covered by a regular layer composed of a single species of protein, the S-protein. The 43-kDa S-protein of the neotype strain Lactobacillus acidophilus ATCC 4356, which originated from the pharynx of a human, was purified. Antibodies generated against purified S-protein were used to screen a lambda library containing chromosomal L. acidophilus ATCC 4356 DNA. Several phages showing expression of this S-protein in Escherichia coli were isolated. A 4.0-kb DNA fragment of one of those phages hybridized to a probe derived from an internal tryptic fragment of the S-protein. The slpA gene, coding for the surface layer protein, was located entirely on the 4.0-kb fragment as shown by deletion analysis. The nucleotide sequence of the slpA gene was determined and appeared to encode a protein of 444 amino acids. The first 24 amino acids resembled a putative secretion signal, giving rise to a mature S-protein of 420 amino acids (44.2 kDa). The predicted isoelectric point of 9.4 is remarkably high for an S-protein but is in agreement with the data obtained during purification. The expression of the entire S-protein or of large, C-terminally truncated S-proteins is unstable in E. coli.')

('>F', 'TI  - Growth of lactobacilli, Staphylococcus aureus and Escherichia coli in normal and  mastitic milk and whey.', 'AB  - The growth of three lactobacilli (Lactobacillus acidophilus, L. bulgaricus and L. casei), Staphylococcus aureus and Escherichia coli was followed in normal and mastitic milk and whey using the standard plate count method. L. acidophilus, L. bulgaricus 6032 and L. casei 6028 grew well in normal milk, but had decreased growth in mastitic milk if not pre-adapted in mastitic whey. S. aureus 26003 and E. coli 44102 showed enhanced growth in mastitic milk as compared with their growth in normal milk. These mastitis pathogens grew faster than the lactobacilli in both the normal milk and the mastitic milk. Among the lactobacilli, L. acidophilus and L. bulgaricus grew faster than L. casei in both types of milk samples. All the bacteria tested grew well in the normal and mastitic whey samples. However, they seemed to have enhanced growth in mastitic whey with the exception of L. casei. Pre-adaptation of the bacteria in mastitic whey in subculture markedly improved the growth of both pathogenic and non-pathogenic bacteria in mastitic milk. All the bacteria showed decreased replication in mastitic milk as compared with mastitic whey.')

('>F#AGG', 'TI  - Inhibition of adhesion of enteroinvasive pathogens to human intestinal Caco-2 cells by Lactobacillus acidophilus strain LB decreases bacterial invasion.', 'AB  - Salmonella typhimurium and enteropathogenic Escherichia coli (EPEC) were found to adhere to the brush border of differentiated human intestinal epithelial Caco-2 cells in culture, whereas Yersinia pseudotuberculosis and Listeria monocytogenes adhered to the periphery of undifferentiated Caco-2 cells. All these enterovirulent strains invaded the Caco-2 cells. Using a heat-killed human Lactobacillus acidophilus (strain LB) which strongly adheres both to undifferentiated and differentiated Caco-2 cells, we have studied inhibition of cell association with and invasion within Caco-2 cells by enterovirulent bacteria. Living and heat-killed Lactobacillus acidophilus strain LB inhibited both cell association and invasion of Caco-2 cells by enterovirulent bacteria in a concentration-dependent manner. The mechanism of inhibition of both adhesion and invasion appears to be due to steric hindrance of human enterocytic pathogen receptors by whole-cell lactobacilli rather than to a specific blockade of receptors.')

('>F', 'TI  - A quantitative assessment of the antimicrobial activity of garlic (Allium sativum).', 'AB  - An aqueous extract of freeze-dried garlic (Allium sativum), when incorporated into growth media, inhibited many representative bacteria, yeasts, fungi and a virus. All microorganisms tested were susceptible to garlic. Quantitative assessment of the minimum inhibitory concentrations for bacteria and yeasts showed values ranging from 0.8 to 40.0 mg garlic ml(-1). Fungal radial colony growth was inhibited by at least 25% at concentrations as low as 2.0 mg garlic ml(-1). The 50% endpoint neutralization titre for rotavirus was 2.4 to 2.8 mug ml(-1). Lactic acid bacteria were the least sensitive microorganisms to the inhibitory effects of garlic. In mixed culture studies of Lactobacillus acidophilus and Escherichia coli, garlic prevented the establishment of E. coli, although the final outcome of competition was not affected.')

('>F', 'TI  - Heat-killed Lactobacillus acidophilus inhibits adhesion of Escherichia coli B41 to HeLa cells.', 'AB  - Escherichia coli B41 (O101: K99: F41: ST+) adheres to HeLa 229 cells in a diffuse pattern. Heat-killed (100-105 degrees C) Lactobacillus acidophilus (Lacteol strain) was found to inhibit this adhesion in a dose-dependent manner. This inhibitory action was lost after lysis of the L acidophilus, suggesting steric hindrance of E coli adhesion sites rather than competition for a common binding site. A thermostable factor (100-105 degrees C) excreted by L acidophilus into the medium may be required for the adhesion of L acidophilus to HeLa cells, and for the inhibition of adhesion of E coli to these cells.')

('>F', 'TI  - Incompatibility of Lactobacillus Vectors with Replicons Derived from Small Cryptic Lactobacillus Plasmids and Segregational Instability of the Introduced Vectors.', 'AB  - Three new Lactobacillus vectors based on cryptic Lactobacillus plasmids were constructed. The shuttle vector pLP3537 consists of a 2.3-kb plasmid from Lactobacillus pentosus MD353, an erythromycin resistance gene from Staphylococcus aureus plasmid pE194, and pUC19 as a replicon for Escherichia coli. The vectors pLPE317 and pLPE323, which do not contain E. coli sequences, were generated by introducing the erythromycin resistance gene of pE194 into a 1.7- and a 2.3-kb plasmid from L. pentosus MD353, respectively. These vectors and the shuttle vector pLP825 (M. Posno, R. J. Leer, J. M. M. van Rijn, B. C. Lokman, and P. H. Pouwels, p. 397-401, in A. T. Ganesan and J. A. Hoch, ed., Genetics and biotechnology of bacilli, vol. 2, 1988) could be introduced by electroporation into Lactobacillus casei, L. pentosus, L. plantarum, L. acidophilus, L. fermentum, and L. brevis strains with similar efficiencies. Transformation efficiencies were strain dependent and varied from 10 to 10 transformants per mug of DNA. Plasmid DNA analysis of L. pentosus MD353 transformants revealed that the introduction of pLP3537 or pLPE323 was invariably accompanied by loss of the endogenous 2.3-kb plasmid. Remarkably, pLPE317 could only be introduced into an L. pentosus MD353 strain that had been previously cured of its endogenous 1.7-kb plasmid. The curing phenomena are most likely to be explained by the incompatibility of the vectors and resident plasmids. Lactobacillus vectors are generally rapidly lost when cells are cultivated in the absence of selective pressure. However, pLPE323 is stable in three of four Lactobacillus strains tested so far.')

('>F', 'TI  - Population of Salmonella serovar typhimurium in the cecum of gnotobiotic chickens with Escherichia coli and intestinal bacteria.', 'AB  - To test the interaction between various species of bacteria and Salmonella serovar typhimurium (S. typhimurium), the population of S. typhimurium was measured in the cecum of gnotobiotic chickens in the presence of Escherichia coli (E. coli) and one of the four intestinal bacteria; Lactobacillus acidophilus. Clostridium perfringens, Bifidobacterium thermophilum and Bacteroides vulgatus. Competitive exclusion of S. typhimurium by di-flora chicken was not demonstrated. But the population of S. typhimurium was temporarily suppressed in di-flora chickens with E. coli and L. acidophilus. In penta-flora chickens with E. coli and these four intestinal bacteria, the population of S. typhimurium was suppressed for only 2 days. In normalized chickens, the population of S. typhimurium was markedly suppressed.')

('?F#see if bacteriocin', 'TI  - Cloning, phenotypic expression, and DNA sequence of the gene for lactacin F, an antimicrobial peptide produced by Lactobacillus spp.', 'AB  - Lactacin F is a heat-stable bacteriocin produced by Lactobacillus acidophilus 11088. A 63-mer oligonucleotide probe deduced from the N-terminal lactacin F amino acid sequence was used to clone the putative laf structural gene from plasmid DNA of a lactacin F-producing transconjugant, L. acidophilus T143. One clone, NCK360, harbored a recombinant plasmid, pTRK160, which contained a 2.2-kb EcoRI fragment of the size expected from hybridization experiments. An Escherichia coli-L. acidophilus shuttle vector was constructed, and a subclone (pTRK162) containing the 2.2-kb EcoRI fragment was introduced by electroporation into two lactacin F-negative strains, L. acidophilus 89 and 88-C. Lactobacillus transformants containing pTRK162 expressed lactacin F activity and immunity. Bacteriocin produced by the transformants exhibited an inhibitory spectrum and heat stability identical to those of the wild-type bacteriocin. An 873-bp region of the 2.2-kb fragment was sequenced by using a 20-mer degenerate lactacin F-specific primer to initiate sequencing from within the lactacin F structural gene. Analysis of the resulting sequence identified an open reading frame which could encode a protein of 75 amino acids. The 25 N-terminal amino acids for lactacin F were identified within the open reading frame along with an N-terminal extension, possibly a signal sequence. The lactacin F N-terminal sequence, through the remainder of the open reading frame (57 amino acids; 6.3 kDa), correlated extremely well with composition analyses of purified lactacin F which also predicted a size of 51 to 56 amino acid residues. Molecular characterization of lactacin F identified a small hydrophobic peptide that may be representative of a common bacteriocin class in lactic acid bacteria.')

('>F', 'TI  - Effect of lactic acid producing bacteria on the human intestinal microflora during ampicillin treatment.', 'AB  - 20 healthy volunteers participated in a double blind study concerning the effect  of lactic acid producing bacteria on the intestinal microflora during ampicillin treatment. 10 volunteers received 500 mg ampicillin tablets t.i.d. together with capsules containing lactic acid producing bacteria (Lactobacillus acidophilus and Bifidobacterium bifidum) for 7 days, and the other 10 volunteers were given 500 mg ampicillin tablets together with placebo capsules t.i.d. for 7 days. Both groups of volunteers continued the intake of the capsules t.i.d. for another 14 days after the ampicillin administration had been completed. The number of enterococci, streptococci and corynebacteria decreased during ampicillin administration but returned to normal levels after 14 days. Yeasts increased during the antibiotic treatment but returned to the same levels as before treatment within 14 days. Escherichia coli strains were suppressed in most volunteers during ampicillin administration. The numbers of anaerobic gram-positive cocci and rods decreased in most subjects during ampicillin treatment but were normalized within 2 weeks. Bacteroides strains were recovered in higher numbers in the lactic acid producing bacteria group compared to the placebo group. The volunteers receiving lactic acid producing bacteria were recolonized slightly faster than those having placebo. There were adverse effects observed in 3 subjects receiving ampicillin plus placebo. In the lactic acid producing bacteria group, one subject had diarrhoea on day 3 to on day 3 to day 7.')

('?F#ATT', 'TI  - Exclusion of uropathogen adhesion to polymer surfaces by Lactobacillus acidophilus.', 'AB  - The ability of bacteria to adhere to surfaces is a major cause of concern in the  use of biomaterial substrates. The adhesion of Staphylococcus epidermidis strain 1938 was examined using image analysis and was found not to correlate with polymer surface tension, unlike that of Lactobacillus acidophilus, which adhered to more hydrophobic polymers. A fimbriated uropathogenic E. coli strain showed very low levels of adherence to the biomaterials. Precoating the polymers with lactobacilli significantly reduced the staphylococcal and E. coli adhesion, a result which could have clinical significance. An additional finding was that the interaction of staphylococci and E. coli with lactobacilli coated polymers altered the adhesion profile of the latter. Lactobacilli appeared to detach from polymers of low surface tension and reattach to polymers with high surface tensions. This resulted in the highest levels of exclusion of uropathogens being found for lactobacilli-coated glass and sulfonated polystyrene, both of which are hydrophilic (with high surface tensions). These results demonstrate that lactobacilli can be used to coat biomaterial surfaces leading to a reduced adhesion of uropathogens.')

('>F', 'TI  - Population of Salmonella typhimurium in the cecum of gnotobiotic chickens.', 'AB  - To test the interaction of various species of bacteria with Salmonella typhimurium, levels of S. typhimurium were measured in the cecum of gnotobiotic chickens inoculated with various test bacteria and subsequently with S. typhimurium. The population of S. typhimurium was suppressed in the cecum of chickens previously inoculated with Escherichia coli. Populations of Bacteroides vulgatus, Clostridium perfringens, Bifidobacterium thermophilum, or Lactobacillus acidophilus were transiently, though not significantly, suppressed by S. typhimurium. Inoculation of chickens with C. perfringens caused a terminal increase in the population of S. typhimurium. Thus the effect of cecal contents on reducing the levels of S. typhimurium in the cecum may be due to the presence of E. coli in the cecal contents.')

('>F', 'TI  - Characterization and promoter selectivity of Lactobacillus acidophilus RNA polymerase.', "AB  - DNA-dependent RNA polymerase has been purified from gram-positive Lactobacillus acidophilus and found to be composed of 4 protein subunits, alpha, beta, beta', and sigma, with molecular weights of 40,000, 150,000, 135,000, and 45,000 kD, respectively, estimated on the basis of SDS-polyacrylamide gel electrophoresis. The purified enzyme exhibits optimal activity in the presence of Mn2+, while Mg2+ shows only a slight effect. The L. acidophilus enzyme transcribes several Escherichia coli promoters examined so far, such as promoters of trp operon, lacUV5, and bla P3 from pBR322, whereas it lacks the ability to recognize bla P1 and tet P2 promoters from pBR322. Thus, the specificity of L. acidophilus RNA polymerase in recognizing the promoters is somehow different from that of the E. coli enzyme. By means of an in vitro transcription assay system for L. acidophilus RNA polymerase, 2 promoters have been identified in the DNA of an L. acidophilus cryptic plasmid (pRNL5). These promoters possess nucleotide sequences in the -10 region similar to the consensus sequence for the E. coli promoters.")

('>F#OPP', 'TI  - Impact of Lactobacillus acidophilus on the normal intestinal microflora after administration of two antimicrobial agents.', 'AB  - Twenty healthy volunteers participated in a comparative study concerning the influence of Lactobacillus acidophilus supplements on the normal intestinal microflora after the administration of two antimicrobial agents, enoxacin and clindamycin, respectively. L. acidophilus NCFB 1748 was given as a fermented milk product containing 5 x 10(8)-2 x 10(9) CFU/ml to ten of the volunteers immediately after the administration of the antimicrobial agents. On the seventh day of enoxacin administration enterobacteria were eliminated in nine of ten subjects. Enterococci disappeared or decreased significantly in five subjects. During the L. acidophilus supplementation, there was a significant increase in the number of Escherichia coli in one subject, while enterococci returned to the same level as before enoxacin administration in all subjects.')

('>F', 'TI  - Effect of infection with Eimeria tenella upon the cecal bacterial population in monoflora chickens.', 'AB  - Monoflora chickens were established at 2 days of age by an oral inoculation with  one of six species of bacteria (Lactobacillus acidophilus, Bifidobacterium thermophilum, Bacteroides vulgatus, Clostridium perfringens, Escherichia coli, or Streptococcus faecalis) and were infected with Eimeria tenella (5 X 10(4) oocysts per bird) 2 days later. There were two experimental groups for each bacterium: birds infected with bacteria alone and birds infected with a combination of bacteria and E. tenella. Seven days after E. tenella infection, counts of B. thermophilum in the cecal contents were significantly lower for E. tenella-infected birds than for those infected by B. thermophilum alone, whereas 10 days after E. tenella infection, counts were higher for E. tennella-infected birds. The population of L. acidophilus in the cecal contents of the E. tenella-infected chickens 10 days after inoculation was significantly greater than that in uninfected chickens. No significant differences were observed between the numbers of B. vulgatus, C. perfringens, and S. faecalis in cecal contents of groups with and without E. tenella infections.')

('>F', 'TI  - [Effect of the administration of killed Lactobacillus acidophilus on the survival of suckling mice infected with a strain of enterotoxigenic Escherichia coli].', 'AB  - After oral administration of 10(2) to 10(5) CFU of Escherichia coli B41 (0101:K 99+:ST+) to 24-48 h old suckling mice (Swiss OF1), a 80 to 100% mortality rate is observed within three days. We compared the effect of the oral treatment with a lyophilized preparation of heat-killed Lactobacillus acidophilus and with sterile water on the mortality rate of newborn mice. In six out of seven assays, the heat-killed L. acidophilus administration extended survival of infected suckling mice (P ranging from 0.019 to less than 0.001). In another test, result was contradictory. A global analysis indicated that the two treatments were statistically different (P less than 0.001). Lactic acid was unable to induce protection.')

('>F', 'TI  - Further studies on thymidine kinase: distribution pattern of the enzyme in bacteria.', "AB  - Various micro-organisms (131 strains of 73 species) were studied for their ability to produce thymidine kinase (TK; EC 2.7.1.21). Taking the specific TK activity of Escherichia coli K12 [specific activity of sonicated cell extracts 95-194 pmol min-1 (mg protein)-1] as 100%, the test organisms had the following relative specific TK activities. In the Gram-positive cocci, Staphylococcus aureus (21-84%) showed higher activity than Staph. epidermidis (1-20%) and Streptococcus (1-7%) except for one strain of Strep. pyogenes (29%). Neisseria sicca, a Gram-negative coccus, lacked TK. Gram-positive endospore-forming rods showed significant activity (Bacillus, 13-51%; Clostridium perfringens, 9-18%) except for one strain of B. megaterium (2%) and C. difficile (1-3%). Among the Gram-positive asporogenous rods, Listeria monocytogenes and six species of Lactobacillus (especially L. brevis, L. buchneri and L. casei) had moderate to high activity (23-348%) but L. acidophilus, L. bulgaricus, L. lactis and L. cellobiosus had low activity (0-8%). Of the species of Pseudomonas studied, most lacked TK but Ps. fluorescens and Ps. maltophilia had significant TK activity (15-53%). Of the Gram-negative facultative anaerobes, Vibrio lacked TK, while Enterobacteriaceae, including Salmonella (148-1120%), Escherichia (59-141%), Klebsiella (78-299%) and Serratia (61-110%), had a high activity. Proteus had a somewhat lower activity (0-34%) except for 'Pr. rettgerella' (307%). Propionibacterium and Bifidobacterium and related organisms other than Streptomyces, Nocardia, Rhodococcus, Corynebacterium and Mycobacterium lacked TK. The seven species of Candida tested, and Cryptococcus neoformans, essentially lacked TK.(ABSTRACT TRUNCATED AT 250 WORDS)")

('>F', 'TI  - Reverse effect of gram-positive bacteria vs. gram-negative bacteria on adjuvant-induced arthritis in germfree rats.', 'AB  - Germfree (GF) F344 rats developed severe adjuvant-induced arthritis with a 100% incidence after a single intradermal injection of heat-killed Mycobacterium bovis (BCG). Specific pathogene-free (SPF) rats developed less severe arthritis with a lower incidence. The rats colonized with Escherichia coli or Bacteroides developed mild disease comparable to that in SPF rats. The rats colonized with Bifidobacterium, Propionibacterium acnes, Lactobacillus casei, L. fermentum, L. murini, and L. acidophilus developed more severe disease than that in GF rats. Furthermore, the rats colonized with a mixture of E. coli and the above lactobacilli developed very mild disease similar to that in SPF rats. These results suggest that gram-negative bacteria, such as E. coli and Bacteroides, may suppress the disease, possibly through their lipopolysaccharides, and may be responsible for the lower susceptibility of SPF rats; gram-positive bacteria, such as Bifidobacterium, P. acnes, and lactobacilli, may enhance the disease, possibly through their peptidoglycans; and E. coli may play a dominant role in modulating the development of adjuvant-induced arthritis.')

('>F', 'TI  - Importance of bile tolerance of Lactobacillus acidophilus used as a dietary adjunct.', 'AB  - Cultures of lactobacilli identified as Lactobacillus acidophilus from the intestinal contents of young calves varied in their ability to grow in broth containing .3% oxgall compared with control broth. Frozen concentrated cultures were prepared from a strain exhibiting low tolerance to bile and from a strain exhibiting high tolerance to bile. Plate counts were comparable from the concentrated cultures before and after frozen storage on lactobacilli MRS agar with and without .15% oxgall. In a feeding trial involving newborn dairy calves supplementation of the diet with the more bile resistant strain of Lactobacillus acidophilus caused greater increases of numbers of facultative lactobabilli in the upper small intestines than did the strain exhibiting lower resistance to bile. It was not possible to determine whether the lactobacilli would prevent intestinal infections in the calves challenged with enteropathogenic Escherichia coli. This portion of the study failed as the challenge with Escherichia coli did not cause infections even in control animals.')

('>F', 'TI  - Exogenous lactobacilli fed to man - their fate and ability to prevent diarrheal disease.', 'AB  - Clinical and microbiologic studies were conducted to evaluate the capability of exogenous lactobacilli to implant in the upper small intestine, the ability of exogenous lactobacilli to prevent enterotoxigenic Escherichia coli (ETEC) diarrhea, and the effect of lactobacillus therapy in reduction of diarrhea due to neomycin. Microbiologic studies of jejunal aspirates indicated that orally ingested Lactobacillus acidophilus and L. bulgaricus in LactinexR and L. acidophilus in Infloran BernaR survived passage through the stomach and remained viable in the proximal small bowel for three to six hours in most, not all, individuals. Challenge studies with ETEC revealed that despite the ability of lactobacilli in LactinexR to persist in the upper small intestine for several hours, the lactobacillus preparation did not prevent or alter the course of ETEC diarrhea in adults. Lactobacillus therapy with one lot of LactinexR reduced the volume and duration of neomycin-associated diarrhea, whereas another lot of LactinexR had no therapeutic effect. Thus, lot to lot variations in lactobacillus preparations may account for some conflicting results observed with lactobacillus therapy for intestinal disorders.')

('>F', 'TI  - Influence of dietary lactose on the gut flora of chicks.', 'AB  - 1. The gut microflora of chicks fed on a purified diet containing 300 g lactose plus 300 g starch/kg was compared with that of control birds receiving a diet containing 600 g starch/kg. 2. In 14-d-old conventional chicks, lactose in the diet decreased the incidence of lactobacilli and clostridia in the caecal contents, although when presenting lactose-fed chicks the counts of lactobacilli exceeded those of control chicks. 3. High counts of Proteus sp. were present in the caeca of control birds but they were completely suppressed in conventional birds fed on the lactose diet. In vitro tests showed that this inhibition was partially due to Escherichia coli and Streptococcus faecalis. 4. The growth of Lactobacillus acidophilus was inhibited by lactose when gnotobiotic chicks were monoassociated but not when polyassociated. The protective effect was shown in vitro to be due to L. salivarius. 5. The pH was markedly lowered in the caecum of conventional and polyassociated chicks receiving dietary lactose. Of the strains used in gnotobiotic experiments E. coli, S. faecalis and L. salivarius produced the lowest pH values in the caeca.')

('>F', 'TI  - Inhibitory effect of some intestinal bacteria on liver tumorigenesis in gnotobiotic C3H/He male mice.', 'AB  - Liver tumorigenesis in gnotobiotic C3H/He male mice was markedly promoted by association with a bacterial combination of Escherichia coli, Streptococcus faecalis, and Clostridium paraputrificum. This study demonstrated that this promoting effect was suppressed by addition of certain intestinal bacteria, such as Bifidobacterium longum, Lactobacillus acidophilus, and Eubacterium rectale.')

('>F', 'TI  - The formation of germtubes by Candida albicans, when grown with Staphylococcus pyogene, Escherichia coli, Klebsiella pneumoniae, Lactobacilius acidophilus and Proteus vulgaris.', 'AB  - The formation of germtubes by twelve clinical isolates of C. albicans was studied in human serum containing per millilitre 10(3) to 10(9) organisms as: Staphylococcus pyegene, Escherichia coli, Klebsiella pneumoniae, Lactobacillus acidophilus and Proteus vulgaris. All the five bacteria inhibited formation of germtubes by C. albicans at all concentrations and the percent germtube formed diminished with increasing concentration of the bacteria. Lactobacillus acidophilus inhibited the formation of germtubes maximally followed by Staphylococcus pyogene, Escherichia coli and Klebsiella pneumoniae. Proteus vulgaris in the concentrations of 10(3) to 10(7) bacteria per millilitre produced only insignificant inhibition of formation of germtubes by C. albicans. Since germtubes of C. albicans are invasive, it is suggested that inhibition of "blastospore-germtube transformation" may be significantly responsible for prevention of infection by C. albicans by coexisting bacterial flora.')

('>F', 'TI  - Changes in the microflora and physiology of the anterior intestinal tract of pigs weaned at 2 days, with special reference to the pathogenesis of diarrhea.', 'AB  - The gastrointestinal microflora and gastric physiology of piglets weaned at 2 days was compared with that of piglets allowed to continue sucking the sow. Although there was a significantly higher count of Escherichia coli in the stomach, duodenum, and jejunum of the early-weaned compared with sow-reared pigs, these differences were not detectable in samples from the ileum. There were no quantitative differences in lactobacilli and in streptococci between the two treatments. Lactobacillus fermentum, L. acidophilus, Streptococcus salivarius, S. bovis, and related biotypes were isolated from both groups of pigs. L. fermentum and S. salivarius were isolated more frequently from sow-reared piglets. The weight of digesta in the stomach was greater in weaned than in sucking pigs and was even greater in scouring weaned pigs, suggesting that in scouring pigs there may be gastric stasis. The gastric pH was higher in the weaned pigs at 4 days of age, but gradually decreased up to 10 days, during which time the lactic acid concentration rose. In weaned pigs there was a highly significant negative correlation between pH and lactic acid concentration in the stomach digesta, and also a positive correlation between pH and number of E. coli. These correlations suggest that lactic acid, from bacterial fermentation, is the major component in the regulation of gastric pH in weaned pigs. Three of twenty sucking pigs, but none of the weaned pigs, were secreting HCl (chloride concentration > 3 mg/g, pH < 3.5). In sucking pigs there was an inverse relationship between the chloride and lactic acid concentrations in the digesta. In weaned scouring pigs there was a nonsignificant increase in pepsin concentration in the stomach tissue. There was a threefold increase in the total proteolytic activity of the stomach tissue.')

('>F', 'TI  - Deconjugation of bile acids by human intestinal bacteria.', 'AB  - The purpose of this report is to present the deconjugation of bile acids by numbers of strains of bacteria in the small intestine and feces. The small intestinal juice was aseptically aspirated by a double lumen tube with a rubber cover on the tip devised by us ("Fukushima Type 1"). Bile acids were analyzed with thin layer chromatography. THE RESULTS: 1) Among aerobic bacteria, species of which all of the strains split conjugated bile acids was enterococcus, and most of the strains split were Staphylococcus (S.) epidermidis and Lactobacillus (L.) bifidus. Species of which none of the strains split were Escherichia (E.) coli, E. communior, E. freundii, L. plantarum, L. acidophilus, L. buchneri, L. cellobiosus, L. bulgaricus, S. aureus, Aerobacter aerogenes, Pseudomonas aeruginosa, candida, proteus, serratia, and almost none of the species split was Intermediate coliform bacilli. 2) Among anaerobic bacteria, species of which all of the strains split were Bacteroides (B.) vulgatus, B. thetaiotaomicron, B. uniformis, Corynebacterium (C.) granulosum, C. avidum, Peptostreptococcus (Peptostrept.) putridus, Eubacterium (Eubact.) lentum, Peptococcus (Pept.) grigoroffii, Pept. anaerobius, Veillonella (V.) orbiculus, and most of the strains split were Coryne. diphtheroides, Eubact. parvum, Peptostrept. intermedius. Species of which none of the strains split were Coryne, parvum, Peptostrept. micros, V. alcalescens, V. parvula, Catenabacterium (Catena.) catenaforme, and Catena. filamentosum. 3) All or none, or almost all or none, of the strains of each species tested split conjugated bile acids, and it seems probably that the presence or absence of this ability would be a proper character of eachspecies.')

('@F', 'TI  - [Study of the antagonism between Lactobacillus, E. coli and Bac. Anthracoides].', '##PADDING##')

('>F', 'TI  - Nicotinamide deamidation by microorganisms in rat stomach.', 'AB  - We have extended our investigation of nicotinamide deamidation in the stomach of  conventional rats. The bacterial species in the pars preventricularis were identified as Flavobacterium peregrinum, Escherichia coli, Streptococcus faecalis, and Lactobacillus acidophilus, listed in order of decreasing deamidase activity. Nicotinamide-7-(14)C ingested into rat stomach was rapidly deamidated to nicotinic acid. These results contribute to the accumulated evidence that microorganisms present in the pars preventricularis of rat stomach are responsible for the deamidation of nicotinamide to nicotinic acid, a known precursor of mammalian pyridine nucleotides.')

('>F', 'TI  - Myeloperoxidase-halide-hydrogen peroxide antibacterial system.', 'AB  - An antibacterial effect of myeloperoxidase, a halide, such as iodide, bromide, or chloride ion, and H(2)O(2) on Escherichia coli or Lactobacillus acidophilus is described. When L. acidophilus was employed, the addition of H(2)O(2) was not required; however, the protective effect of catalase suggested that, in this instance, H(2)O(2) was generated by the organisms. The antibacterial effect was largely prevented by preheating the myeloperoxidase at 80 C or greater for 10 min or by the addition of a number of inhibitors; it was most active at the most acid pH employed (5.0). Lactoperoxidase was considerably less effective than was myeloperoxidase when chloride was the halide employed. Myeloperoxidase, at high concentrations, exerted an antibacterial effect on L. acidophilus in the absence of added halide, which also was temperature- and catalase-sensitive. Peroxidase was extracted from intact guinea pig leukocytes by weak acid, and the extract with peroxidase activity had antibacterial properties which were similar, in many respects, to those of the purified preparation of myeloperoxidase. Under appropriate conditions, the antibacterial effect was increased by halides and by H(2)O(2) and was decreased by catalase, as well as by cyanide, azide, Tapazole, and thiosulfate. This suggests that, under the conditions employed, the antibacterial properties of a weak acid extract of guinea pig leukocytes is due, in part, to its peroxidase content, particularly if a halide is present in the reaction mixture. A heat-stable antibacterial agent or agents also appear to be present in the extract.')

FALSE NEGATIVES:
('?T', 'TI  - Pre-treatment with probiotics prolongs survival after experimental infection by multidrug-resistant Pseudomonas aeruginosa in rodents: an effect on sepsis-induced immunosuppression.', 'AB  - Based on several randomised clinical studies indicating benefit from oral probiotic intake for the prevention of hospital-acquired infections in critically ill patients, this study aimed to explain the mechanism of action of probiotics for the prevention of lethal experimental infection by multidrug-resistant (MDR) Pseudomonas aeruginosa. Experiments using an Escherichia coli strain susceptible to all antimicrobials were also conducted. C57BL/6 mice were pre-treated intraperitoneally with sterile water for injection or Lactobacillus plantarum. Survival was recorded and mice were sacrificed for measurement of apoptosis and tissue bacterial overgrowth and for isolation and culture of splenocytes for cytokine production. Experiments were repeated after pre-treatment with a commercial preparation of four probiotics (L. plantarum, Lactobacillus acidophilus, Saccharomyces boulardii and Bifidobacterium lactis; LactoLevure((R))). Peripheral blood mononuclear cells (PBMCs) of healthy volunteers were stimulated by heat-killed P. aeruginosa following pre-treatment with medium or probiotics. Pre-treatment with L. plantarum significantly prolonged survival after challenge by either MDR P. aeruginosa (66.7% vs. 31.3%; P=0.026) or E. coli (56.0% vs. 12.0%, P=0.003). Survival benefit was even more pronounced when mice were pre-treated with LactoLevure((R)). Tissue bacterial outgrowth and apoptosis of white blood cells and splenocytes were not altered. TNFalpha and IL-10 production by splenocytes of mice pre-treated with probiotic was increased and IFNgamma production was decreased. Pre-treatment with LactoLevure((R)) restored production of IL-17. Stimulation of human PBMCs after probiotic pre-treatment was accompanied by reduced gene expression of SOCS3. The results suggest that the protective effect of probiotics is mediated through prevention of sepsis-induced immunosuppression.')

('>T', 'TI  - Effect of Lactobacillus acidophilus strain NP51 on Escherichia coil O157:H7 fecal shedding and finishing performance in beef feedlot cattle.', 'AB  - A 2-year study was conducted during the summer months (May to September) to test  the effectiveness of feeding Lactobacillus acidophilus strain NP51 on the proportion of cattle shedding Escherichia coli O157:H7 in the feces and evaluate the effect of the treatment on finishing performance. Steers (n = 448) were assigned randomly to pens, and pens of cattle were assigned randomly to NP51 supplementation or no supplementation (control). NP51 products were mixed with water and applied as the feed was mixed daily in treatment-designated trucks at the rate of 10(9) CFU per steer. Fecal samples were collected (n = 3,360) from the rectum from each animal every 3 weeks, and E. coli O157:H7 was isolated by standard procedures, using selective enrichment, immunomagnetic separation, and PCR confirmation. The outcome variable was the recovery of E. coli O157:H7 from feces, and was modeled using logistic regression accounting for year, repeated measures of pens of cattle, and block. No significant differences were detected for gain, intakes, or feed efficiency of control or NP51-fed steers. The probability for cattle to shed E. coli O157:H7 varied significantly between 2002 and 2003 (P = 0.004). In 2002 and 2003, the probability for NP51-treated steers to shed E. coli O157:H7 over the test periods was 13 and 21%, respectively, compared with 21 and 28% among controls. Over the 2 years, NP51-treated steers were 35% less likely to shed E. coli O157: H7 than were steers in untreated pens (odds ratio = 0.58, P = 0.008). This study is consistent with previous reports that feeding NP51 is effective in reducing E. coli O157:H7 fecal shedding in feedlot cattle.')

('>T', 'TI  - Lipoteichoic acids from Lactobacillus johnsonii strain La1 and Lactobacillus acidophilus strain La10 antagonize the responsiveness of human intestinal epithelial HT29 cells to lipopolysaccharide and gram-negative bacteria.', 'AB  - Intestinal epithelial cells (IECs) respond to lipopolysaccharide (LPS) from gram-negative bacteria in the presence of the soluble form of CD14 (sCD14), a major endotoxin receptor. Since sCD14 is also known to interact with gram-positive bacteria and their components, we looked at whether sCD14 could mediate their effects on human IECs. To this end, we examined the production of proinflammatory cytokines following exposure of the IECs to specific gram-positive bacteria or their lipoteichoic acids (LTAs) in the absence and presence of human milk as a source of sCD14. In contrast to LPS from Escherichia coli or Salmonella enteritidis, neither the gram-positive bacteria Lactobacillus johnsonii strain La1 and Lactobacillus acidophilus strain La10 nor their LTAs stimulated IECs, even in the presence of sCD14. However, both LTAs inhibited the sCD14-mediated LPS responsiveness of IECs. We have previously hypothesized that sCD14 in human milk is a means by which the neonate gauges the bacterial load in the intestinal lumen and liberates protective proinflammatory cytokines from IECs. The present observations suggest that gram-positive organisms, via their LTAs, temper this response and prevent an exaggerated inflammatory response.')