Research Abstracts 

  1. Chemokine Interleukin-8 Production as a Host Pro-Inflammatory Response Induced by Gastrointestinal Pathogens

  2. Effect of Lactobacillus on Enterohemorrhagic E. coli -Induced Alterations in Intestinal Epithelial Cell Permeability

  3. Production of exopolysaccharide by Lb. rhamnosus R and analysis of its enzymatic degradation during prolonged fermentation

  4. Effects of Lactic Acid Bacteria on cytokine production by a human intestinal epithelial cell line

  5. Induction of Interleukin-6 and Tumour Necrosis Factor Production by Lactic Acid Bacteria

  6. The Immunomodulatory Effects of Selenium , as a Component of Mineral Enriched Yeast, in a Murine System

  7. Lactobacillus species inhibit adherence of enteropathogenic E. coli and enterohemorrhagic E. coli to host epithelial cells

  8. Protection against an infectious disease by enterohaemorrhagic E. coli 0-157

  9. Immunomodulatory Effects of Different Lactobacillus spp . on Proliferation and Cytokine Production by Murine Splenocyte Fractions

  10. Casein-Derived Opioid Peptides and Lactobacilli : Effects on Cell Proliferation and Antibody Production

  11. Characterization of bifidobacteria by random DNA amplification

  12. Varied immunomodulatory effects of different Lactobacillus spp. on cytokine production by murine splenocytes, murine macrophages and macrophage cell lines

  13. Effect of yeast-supplemented feed on Salmonella and Campylobacter populations in broilers

  14. Possible mitogenic effects of Lactobacillus ssp. on murine B, T and macrophage cell lines

  15. Lb. acidophilus in the Treatment of Children with Gastrointestinal Tract Disease

  16. Differentiation of bifidobacteria by use of pulsed-field gel electrophoresis and polymerase chain reaction

  17. Lactobacilli in the Treatment of Dyspepsia in Dysmicrobia in Different Aetiology

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Research Abstracts

 

  1. Chemokine Interleukin-8 Production as a Host Pro-Inflammatory Response Induced by Gastrointestinal Pathogens. Yeung HP, Johnson-Henry K and Sherman PM (2001). Presented at the 2001 Paediatric.Academic Societies meeting, Baltimore, MD, April 30, 2001.

    Infection by enteropathogens Helicobactor pylori, enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic Escherichia coli (EHEC) involves attachment by the bacteria to host epithelial cells and induction of host cell secretion of pro-inflammatory cytokines, including interleukin-8 (IL-8). Previous studies suggest that Lactobacillus, which composes part of the commensal gut flora, may exert a protective effect against infection by preemptively binding the epithelial cells, thereby decreasing the available surface area for further attachment by harmful organisms. However, it is not yet known whether exposure of exogenous Lactobacilli in host epithelial cells provokes a pro-inflammatory response as observed with pathogenic bacteria. Our study aimed to assess the response by epithelial cells infected with pathogenic bacteria (H. pylori, EPEC, and EHEC) versus non-pathogenic bacteria (L. acidophilus R0052 and L. rhamnosus R0011).

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  2. Effect of Lactobacillus onEnterohemorrhagic Escherichia coli-Induced Alterations in Intestinal Epithelial Cell Permeability. Johnson-Henry KC, Tompkins TA, Sherman PM (2001). Presented at the Annual Meeting of the American Society for Microbiology in Orlando, Florida, May 20-24, 2001.

    Enteropathogenic Escherichia coli (EPEC) E2348/69 and enterohemorrhagic Escherichia coli (EHEC) O157:H7 infection result in watery diarrhea, hemorrhagic colitis and the hemolytic-uremic syndrome in children. EPEC and EHEC infection decrease the transepithelial electrical resistance of polarized human T84 cell monolayers thereby inducing an increase in permeability. The aims of this study were to define the effects of probiotic bacteria on transepithelial electrical resistance (TER) of T84 epithelial cells and to determine if Lactobacillus species (L. rhamnosus R0011 and L. acidophilus R0052) were able to prevent E. coli strains E2348/69(O127:H6) and CL-56 (O157:H7) induced changes in TER. Alterations of the TER were measured after an 18 hour incubation. Both EPEC and EHEC decreased resistance to approximately 20% of control values, whereas L. rhamnosus R0011 and L. acidophilus R0052 did not drop TER. There was a time dependent attenuation of the drop in resistance produced by EPEC and EHEC when T84 cells were pre-treated with L. rhamnosus R0011 and L. acidophilus R0052. At 6 hours, there was a significant increase of the TER towards control values. Further studies defining the mechanisms of action may lead to a novel approach for treating and preventing EPEC and EHEC-induced diarrhea.

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  3. Production of exopolysaccharide by Lactobacillus rhamnosus R and analysis of its enzymatic degradation during prolonged fermentation. Pham PL, Dupont I, Roy D, Lapointe G, Cerning J (2000). Appl Environ Microbiol 66:2302-2310

    The potential of Lactobacillus rhamnosus R for producing exopolysaccharide (EPS) when grown on basal minimum medium supplemented with glucose or lactose was investigated. EPS production by L. rhamnosus R is partially growth associated and about 500 mg of EPS per liter was synthesized with both sugars. The product yield coefficient (Y(EPS/S)) was 3.15 (0.0315 g of EPS [g of lactose](-1)) and 2.88 (0.0288 g of EPS [g of glucose](-1)). It was clearly shown that the amount of EPS produced declined upon prolonged fermentation. Degradation of EPS in fermentation processes was also assessed by measuring its molecular weights and viscosities. As these reductions might have a negative effect on the yield and viscosifying properties of EPS, it was essential to examine possible causes related to this breakdown. The decrease in viscosities and molecular weights of EPS withdrawn at different cultivation times permitted us to suspect the presence of a depolymerizing enzyme in the fermentation medium. Our study on enzymatic production profiles showed a large spectrum of glycohydrolases (alpha-D-glucosidase, beta-D-glucosidase, alpha-D-galactosidase, beta-D-galactosidase, beta-D-glucuronidase, and some traces of alpha-L-rhamnosidase). These enzymes were localized, two of them (alpha-D-glucosidase and beta-D-glucuronidase) were partially purified and characterized. When incubated with EPS, these enzymes were capable of lowering the viscosity of the polymer as well as liberating some reducing sugars. Upon prolonged incubation (27 h), the loss of viscosity was increased up to 33%.

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  4. Effects of Lactic Acid Bacteria on cytokine production by a human intestinal epithelial cell line. Wallace TD, Tompkins TA and Green-Johnson JM (2000). Presented at the Danone Symposium on Fermented Food, Fermentation and Intestinal Flora, New York, NY, May 2000.

    Orally ingested lactic acid bacteria (LAB) have been shown to have immunostimulatory effects in several different systems. One route through which they may be able to exert their effects is through interactions with intestinal epithelial cells. This cell type has recently been shown to be capable of c ytokine production, and thus to play an active role in the immune response. We have examined the effects of lactic acid bacteria on the production of three cytokines (Interleukin-6, Interleukin-8 and RANTES) by the human intestinal epithelial cell line HT-29. Two of the five strains of LAB tested, Lactobacillus delbreuckii ssp.lactis (R0187), and L. rhamnosus (R0011), were found to exert effects on cytokine production, resulting in up-regulation of Interleukin-6 production and down-regulation of IL-8 production, with no significant effects on RANTES production. These effects were only observed with industrial preparations of these bacteria, and not with strains grown in Mann Rogosa Sharpe (MRS) broth, suggesting that growth conditions may be a critical factor in determining the immunostimulatory potential of certain LAB.

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  5. Induction of Interleukin-6 and Tumour Necrosis Factor Production by Lactic Acid Bacteria. Wallace TD, Measham JD, Tompkins TA and Green-Johnson JM (2000). Presented at the Annual Meeting of the American Association of Immunologists in Seattle, WA, May 2000.

    Interest in the immunostimulatory effects of probiotic lactic acid bacteria (LAB) has raised many questions about their potential mode of action. We examined the effects on cytokine production by intestinal epithelial cells and by splenic macrophages, comparing different species and strains of LAB, as well as strains grown under different conditions (lab-grown (LG) versus industrial preparations (IND)). Effects on cytokine production by epithelial cells were measured using the human intestinal cell line HT-29. IL-6 production was enhanced by IND strains of Lactobacillus delbreuckii ssp. lactis (R0187) and L. rhamnosus (strains R0011 and R0049), but not by LG preparations of the same strains. The same trend was observed for Bifidobacterium longum (R0175), with less marked effects on IL-6 production. Initial results suggest strain and growth-condition-related variation in the ability to up-regulate production of IL-6 and TNF by murine splenocytes and macrophages. These findings support the potential for the lactic acid bacteria to be used as immunostimulants, and suggest that their relative efficacy may vary not only with strain, but also with growth conditions. Effects on the production of these cytokines may reflect an important route through which probiotic lactic acid bacteria can influence the immune response.

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  6. The Immunomodulatory Effects of Selenium, as a Component of Mineral Enriched Yeast, in a Murine System. Measham, JD and Green-Johnson, JM (2000). Presented at the International Symposium on Probiotics in Montreal, QC, October 2000.

    Introduction: Micronutrients, such as selenium (Se) and Zinc (Zn) are essential components of mammalian diets. Se is a known, potent anti-oxidant. However, observations of increased susceptibility to infection associated with Se deficiency suggest an additional role as an immunomodulator. The data presented here aims to elucidate the action of Se and Zn, as components of mineral enriched yeast (MEY), on both humoral and cell mediated immunity.Methods: The immunomodulatory actions of MEY were explored by stimulating murine splenocytes with MEY, preparations provided by the Institute Rosell (IR), Montreal, QC. Effects on proliferation were measured using an MTT Colorimetric assay. Antibody (Ab) production was measured using by ELISA. Cytokine induction was measured by ELISA (for Interleukin 6) or by bioassay (for Tumor necrosis factor alpha).Results: All of the yeast preparations tested, Zn-enriched MEY (Zn-50; IR), Se-enriched MEY (Selena; IR) and non-mineral enriched yeast controls (LBI 2133; IR) significantly affected proliferation of murine spleen cells (ANOVA P<0.000, N=4). Initial results suggest that MEY are not effective inducers of Ab production. A dose-dependent trend for TNF-? and IL-6 induction was observed for both Zn and Se MEY, relative to control yeast preparations. Se MEY was a more potent inducer of both cytokines, and cytokine production increased with increasing Se concentration. In contrast more cytokine was detected when lower Zn MEY concentrations were used. Interestingly, Zn MEY at 3000 ng/mL completely suppressed production of both TNF-? and IL-6, yet induced significant levels of proliferation.Conclusion: The most dramatic immunostimulatory effects of MEY were on cytokine production. Dose dependent cytokine down-regulation induced by Zn MEY does not appear to be a result of direct toxicity, as proliferation was enhanced. Together, these results suggest that MEY can affect the immune system.

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  7. Lactobacillus species inhibit adherence of enteropathogenic Escherichia coli and enterohemorrhagic E. coli to host epithelial cells. Johnson-Henry, K., T.A. Tompkins and P.M. Sherman (2000a). Presented at the Danone Symposium on Fermented Food, Fermentation and Intestinal Flora, New York, NY May 2000.

    The role of probiotics as therapeutic agents to prevent diarrheal disease caused by enteropathogenic E. coli and enterohemorrhagic E. coli that cause protracted diarrhea in infants and hemorrhagic colitis, respectively has not been evaluated in detail. The purpose of this study was to determine if Lactobacillus species bind to human epithelial cells in tissue culture and inhibit binding of diarrheagenic E. coli to host cell surfaces. Tissue culture cells (HEp-2) were incubated with Lactobacillus acidophilus (R0052) and L. rhamnosus (R0011) grown industrially or in MRS medium either alone or in combination with enteropathogenic E. coli strain E2348/69 (serotype O127:H6) or Shiga-toxin producing E. coli strain CL-56 (O157:H7) for 3 hrs at 37oC. Brightfield microscopy and quantitative binding assays were employed as complementary measures of bacterial adhesion to eukaryotic cell surfaces. Adherence of probiotics to the surface of tissue culture cells varied depending on the growth conditions employed (maximum adhesion was observed when grown in MRS medium). Both lactobacillus strains inhibited binding, in a dose dependent manner, of enteropathogenic E. coli and enterohemorrhagic E. coli to host cells thereby preventing rearrangements of host cytoskeleton proteins. Lactobacillus species inhibit initial adhesion of diarrheagenic E. coli strains that potentially may be mediated by their ability to adhere to host cell surfaces. Future studies will evaluate the effects of the probiotic strains on E. coli-induced changes in ion fluxes and transepithelial electrical resistance in polarized intestinal cell monolayers.

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  8. Lactobacillus species inhibit adherence of diarrheagenic Escherichia coli to host epithelial cells. Johnson-Henry, K., T. A. Tompkins and P. M. Sherman (2000b). Presented at the International Symposium on Probiotics in Montreal, QC, October 2000.

    Probiotics could prevent diarrheal disease caused by enteropathogenic E. coli and enterohemorrhagic E. coli that cause protracted diarrhea in infants and hemorrhagic colitis, respectively. The aims of this study were to determine if Lactobacillus species adhere to human epithelial cells in tissue culture and inhibit binding of diarrheagenic E. coli to host cell surfaces. Tissue culture cells (HEp-2 and T84) were infected with Lactobacillus acidophilus (R0011) and L. rhamnosus (R0052) grown industrially or in Mann Rogosa Sharpe (MRS) broth either alone or in combination with enteropathogenic E. coli strain E2348/69 (serotype O127:H6) or Shiga-toxin producing E. coli, strain CL-56 (O157:H7) for 3 hrs at 37oC. Quantitative binding assays and Giemsa staining were employed as complementary measures of bacterial adhesion to eukaryotic cell surfaces. Adherence of probiotics to the surface of tissue culture cells varied depending on the growth conditions, bacterial species, and cell type employed. Both lactobacillus strains inhibited binding, in a dose dependent manner, of enteropathogenic E. coli and enterohemorrhagic E. coli to host cells. Future studies will evaluate the effects of the probiotic strains on E. coli-induced changes in ion fluxes and transepithelial electrical resistance in polarized intestinal cell monolayers.

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  9. Protection against an infectious disease by enterohaemorrhagic E. coli 0-157. Ota A (1999). Med Hypotheses 53:87-88.

    Preventive measures against infection by enterohaemorrhagic E. coli 0157 are described. Eating yoghurt and Kefir induces more bifidobacteria and lactic acid bacteria to colonize in the intestines, thereby protecting humans from infection by E. coli 0157. Some foods, such as plum extract, act as a mild antibiotic and produce an acidic environment within the intestine, thus interfering with growth of the E. coli 0157. The natural colonization of harmless E. coli or other bacteria that are more powerful than E. coli 0157 can possibly protect against infection. A vaccination against E. coli 0157 H7 may also be effective. In addition, it has been suggested that the correct levels of nitric oxide and calcium in the blood may activate immunity and protect against infection by E. coli 0157.

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  10. Immunomodulatory Effects of Different Lactobacillus spp. On Proliferation and Cytokine Production by Murine Splenocyte Fractions. Measham, J., A. A. Babcock, and J. M. Green-Johnson (1999). Presented at the 49th Annual Meeting of the Canadian Society of Microbiologists, Montreal, June 1999.

    The lactic acid bacteria (LAB) have been plenocyte as active components of the gastrointestinal microflora. Recently LAB have been the focus of investigations seeking to determine their potential as immunomodulatory agents. We investigated cell proliferation and cytokine production as a function of co-incubation of murineplenocytes fractions and cell lines, with various Lactobacillus spp. And strains of heat-killed LAB isolated from yoghurt starter packs (Institut Rosell Inc.) and purchased from  the ATCC. In most cases, co-incubation induced a dose dependent proliferative response. Based on considerable strain difference noted previously in the ability to induce antibody production, we anticipated strain-dependent variations in the ability to induce cytokine production. Cytokine production by murine splenocytes was seen following co-incubation with certain of the LAB tested. LAB also induced cytokine production following co-incubation with some of the cell lines tested. The immunomodulatory potential of these ecologically important probiotic flora warrants future study.

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  11. Casein-Derived Opioid Peptides and Lactobacilli: Effects on Cell Proliferation and Antibody Production. Babcock, A. A., J. Measham, and J. M. Green-Johnson (1999). Presented at the 49th Annual Meeting of the Canadian Society of Microbiologists, Montreal, QC, June 1999.

    Purified peptide regions from bovine casein have been shown to display bioactivity similar to that of the endogenous opioids, neuropeptides now known to play an immunomodulatory role. Released by the lactic acid bacteria (LAB) during the fermentation of milk, casein-derived opioid peptides use endorphin receptors to modify immune function. We investigated the effects of the casein-derived peptides bovine beta-casomorphin and morphiceptin on the proliferative response of the HT-29 human intestinal epithelial cell line, and on proliferation and antibody (Ab) production by murine splenocytes. Both peptides were shown to modify HT-29 proliferation in a time- and dose-dependent manner, when stimulated with Lipopolysaccharide (LPS), Escherichia coli, or Lactobacilli. The casein peptides did not activate murine splenocytes, but did modify Ab production by splenocytes stimulated with LPS or LAB, and significantly suppressed the proliferation of LAB-stimulated splenocytes. These investigations have provided further insight into the effects of milk products on the immune system, and may elucidate the mechanism through which they influence immune function.

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  12. Characterization of bifidobacteria by random DNA amplification. Vincent D, Roy D, Mondou F, and Dery C (1998). Int J Food Microbiol 43:185-193.

    RAPD conditions were optimized to generate reproducible banding patterns by testing primers, thermocyclers and overall reproducibility in repeat DNA analysis and separate DNA extractions. Five primers were chosen on the basis of band intensity and distribution (between 2 and 10 bands) which clearly distinguished among strains of Bifidobacterium adolescentis, B. animalis, B. bifidum, B. breve, B. infantis and B. longum. The use of five single-primer reactions under optimized conditions improved the resolution and accuracy of the RAPD method for the characterization of dairy-related bifidobacteria. The results indicated that this method was highly reproducible in repeated analysis. Similarity between bifidobacteria strains was evaluated based on their RAPD profiles. Using a set of five primers, it was demonstrated that it may be possible to distinguish three different species of Bifidobacterium (B. breve, B. bifidum and B. adolescentis), based on similarity of the RAPD profiles to known reference strains. Furthermore, application of the RAPD technique may also be useful and faster, than traditional systematics for placement of industrial strains into specific clusters (either B. longum/infantis or B. animalis/lactis).

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  13. Varied immunomodulatory effects of different Lactobacillus spp. on cytokine production by murine splenocytes, murine macrophages and macrophage cell lines. Measham JD and Green-Johnson JM (1998). International Symposium for Microbial Ecology-8, Halifax, NS, 1998.

    The lactic acid bacteria (LAB) have been recognized as active components of the gastrointestinal (GI) microflora. Recently, LAB have been the focus of investigations seeking to determine their potential as immunomodulatory agents. We investigated cell proliferation and cytokine production as a function of coincubation of murine splenocytes, murine macrophages and macrophage cell lines with heat-killed Lactobacillus acidophilus and L. bulgaricus isolated from yoghurt starter packs. Similar studies were conducted with different strains purchased from the ATCC. In most cases, co-incubation induced a dose dependent proliferative response. Based on considerable strain differences noted previously in the ability to induce antibody production, we anticipated strain-dependent variations in the ability to induce cytokine production. Significant IL-6 production by murine splenocytes was seen following co-incubation with both strains of L. bulgaricus. Similarly, induction of IL-6 production by murine splenocytes was noted as a function of coincubation with L. acidophilus strains. LAB also induced IL-6 production following co-incubation with the macrophage cell line PU51-8. We are currently examining the ability of these strains of Lactobacilli to induce IL-6 production by purified murine splenic macrophages and are also investigating the effects of LAB on other members of the Interleukin family. The immunomodulatory potential of these ecologically important probiotic flora warrants future study.

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  14. Effect of yeast-supplemented feed on Salmonella and Campylobacter populations in broilers. Line JE, Bailey JS, Cox NA, Stern NJ, Tompkins TA (1998). Poult Sci. 77:405-410.

    The effect of the yeast, Saccharomyces boulardii, on experimental cecal colonization of broilers with Salmonella typhimurium and Campylobacter jejuni was investigated. Duplicate pens of broiler chicks were given ad libitum access to a standard feed supplemented with no yeast (control), or 1 g (1x), or 100 g (100x) dried S. boulardii/kg feed. All chicks except negative controls were challenged on Day 4 with 3.2 x 10(8) cfu S. typhimurium and 6.5 x 10(8) cfu C. jejuni by oral gavage. After 3 wk, the broilers were euthanatized and ceca were aseptically removed and analyzed for Salmonella and Campylobacter. Frequency of Salmonella colonization was significantly (P < 0.05) reduced due to yeast treatment. Of the positive control birds, 70% were colonized with Salmonella; whereas only 20 and 5% of the 1x and 100x yeast-treated birds were colonized. Mean number of Salmonella per gram of ceca and contents were log 1.64, 0.35, and 0.15, respectively, for the control, 1x, and 100x yeast-treated birds. Campylobacter colonization was not significantly affected by yeast treatment. Similar results were obtained from a second trial conducted in larger isolation floor pens.

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  15. Bifidobacteria and their role in yoghurt-related products. Roy D, Mainville I, and Mondou F (1997). Microecology and Therapy 26: 167-180.

    Bifidobacterium bifidum R0071, B. breve R0070, B. infantis R0033, and B. longum R0175 were used in combination with yoghurt culture organisms to produce yoghurt-related products with pH values of 4.2 (Yoghurt 4.2) and 4.6 (Yoghurt 4.6) obtained after fermentation and cooling. In Yoghurt 4.2 stored at 4oC, the number of viable cells of B. breve was >106 CFU/ml until the 10th day of storage whereas viable cells of B. bifidum, B. infantis and B. longum were <106 CFU/ml after 3, 3 and 7 days of storage at 4oC, respectively. In Yoghurt 4.2 stored at 12oC, B. breve also exhibited higher survival rates than all other species although the decline was more rapid at storage temperature of 12oC as compared to 4oC. In Yoghurts 4.6, the survival of bifidobacteria was better than in Yoghurts 4.2 because the extent of acidification was lower; B. breve survived very well during 16 days of storage at both temperatures and the number of viable cells of B. bifidum, B. infantis and B. longum was >106 CFU/ml during 10 days of storage at 4oC. In this study, the number of lactobacilli was high, except in yoghurt-related products made by a combination of streptococci, lactobacilli and B. breve because this latter strain was metabolically active and could modify the ratio of streptococci and lactobacilli during the preparation of the products which resulted in a reduction of postacidification. A slight loss of beta-galactosidase activity was observed during the preparation of yoghurt-related products with bifidobacteria although the lactose content in these products was lower than in the control.

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  16. Possible mitogenic effects of Lactobacillus ssp. on murine B, T and macrophage cell lines. Easo JG, Measham JD and Green-Johnson J (1997). Presented at the 47th Annual Meeting of the Canadian Society of Microbiology, Quebec City, QC, June 1997.

    Summary: Heat-killed L. bulgaricus (Rosell Yogurt Start Culture, YS) and L. acidophilus (R0052) both induced significant IgM production by murine splenocytes. L. acidophilus (R0052) also induced significant IgG production by murine splenocytes, indicative of a mitogenic effect. High Ab levels were still observed after removal of L. acidophilus-specific Ab from spleen cell supernatants providing evidence for polyclonal B cell activation. L. bulgaricus (ATCC) and L. acidophilus (ATCC) failed to induce significant levels of Ab production by murine splenocytes, indicating a marked strain difference within species of lactobacilli. All lactobacilli tested failed to induce Ab production by murine B cell lines A20 and X16C8.5. L. bulgaricus (ATCC) induces IL-6 production in murine splenocytes. IL-6 production has been noted in both the macrophage cell line PU51-8 and in murine splenocytes stimulated by L. bulgaricus (Y.S.) and L. acidophilus (YS) respectively.

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  17. Lactobacillus acidophilus in the Treatment of Children with Gastrointestinal Tract Disease. Tláskal P, Michková E, Klazarová H, Jerábková L, Nevoral J, Balácková J, Tejnická J, Valtrová V, Simandlová M, and Kejvalová L (1996). Cesko-Slovenská, 51: 615-619.

    The authors evaluated two groups of child patients with diseases of the gastrointestinal tract. The first group was formed by 33 children (12 infants, 13 toddlers and 8 older children) who were given lyophilysate of active Lactobacillus acidophilus bacteria (LA; Institut Rosell Inc., Canada). The second group of children was formed by 42 patients (23 infants, 13 toddlers, 6 older children) who did not receive LA and were treated by preparations of mixtures of adsorption clay, smectite (Smecta pulvis) and a concentrate of metabolic products of common intestinal symbiotic bacteria (preparation Hylak forte). Otherwise treatment in the two groups was the same. Comparison of the results of the two groups revealed that LA contributed in a significant way to the regression of diarrhoea (quantity and quality of bowel movements) in patients with acute gastroenteritis (p<0.01). The incidence of Citrobacter freundii decreased, in particular in infants. The presence of bacteria Salmonella enteritidis persisted however after treatment of children of both groups. LA contributed also significantly (p<0.01) to more rapid recovery from diarrhoea in children treated with antibiotics for other than gastrointestinal disease. When evaluating other clinical symptoms of gastrointestinal disease (temperature, vomiting, abdominal pain, meteorism), the two groups did not differ significantly. Complete recovery from clinical complaints was recorded sooner in children (p<0.01) given LA. When lyophilysate of LA bacteria was administered, complaints associated also with other diseases of the digestive tract (chronic enterocolitis, short bowel syndrome, irritable bowel syndrome) improved.

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  18. Differentiation of bifidobacteria by use of pulsed-field gel electrophoresis and polymerase chain reaction. Roy D, Ward P, Champagne G (1996). Int J Food Microbiol 29:11-29.

    Several different genomic fingerprints can be obtained from various commercially-important species of Bifidobacterium using pulsed-field gel electrophoresis (PFGE) following digestion of DNA with XbaI and SpeI. Four different genomic finger printings were discernible for reference strains of Bifidobacterium animalis, five for B. bifidum, three for B. breve, five for B. infantis and three for B. longum. Standard commercially-available industrial strains of B. animalis are identical to the reference strain ATCC 27536, previously isolated from chicken feces. There was more genomic heterogeneity among industrial strains of B. longum, in that only one gave profiles similar to the type strain of this species (ATCC 15707). The other 14 commercially-available strains of B. longum (mainly isolated from Japanese commercial preparations) were divided into four new molecular types based on their PFGE patterns. The PFGE method indicated that only five distinct strains of B. longum and one strain of B. animalis are used in commercial preparations. Additionally, the use of polymerase chain reaction amplification of portions of 16S rDNA provides a highly specific technique to discriminate between the species B. breve, B. infantis and B. longum.

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  19. Irritable Bowel Syndrome. Kocián J (1994a). Vnitrni Lékarstvi, 40: 246-249.

    Irritable bowel is a functional gastrointestinal disorder with chronic or relapsing symptoms of abdominal pain and impaired frequency and consistency of the faeces caused by obscure structural or biochemical deviations. The frequency of the condition in civilized countries is estimated to amount to 15-20% of the population and it accounts for 25-50% of all patients in gastroenterological ambulatory departments. From the clinical aspect the type with dominant diarrhoea, typically in the morning and very compelling, and the type with pain and constipation are known but even combinations of the two types are encountered. A psychosomatic disorder of the motility of the large bowel and its tonus is involved associated with enhanced pain perception. Despite great efforts to find aetiopathogenetic factors, knowledge still is at the level of obscure theories. The diagnosis is still established per exclusionem after all organic causes are ruled out, i.e. we always have to differentiate between an irritable bowel from an irritated one. In therapy the patient's confidence in his doctor is most important and it is essential to gain the patient's active cooperation. In case of diarrhoea a low-residue diet is used, calcium carbonate, codeine, loperamide, conversely in constipation adequate dietary fibre, intake metoclopramide or cisapride. Pain is relieved by spasmolytics or Ca channel blockers in the smooth musculature of the large bowel. The associated dysbiosis is transformed into eubiosis by Lactobacillus or other related bacterial products.

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  20. Further Possibilities in Treatment of Lactose Intolerance-Lactobacilli. Kocián J (1994b). Prakticky Lékar, 74: 212-214.

    After administration of 2 billion lactobacilli per day from two weeks the lactose tolerance improved in 19 patients with lactose intolerance from milk on average by 1.62 g, cream cheese by 0.96 g and cheese by 0.27 g. The calcium intake increased from milk on average by 50 mg, cream cheese only by 20 mg but from cheese by 103 mg. The favourable effect of lactobacilli is probably due to the content of beta-galactosidase in their cells, which breaks down lactose into simple and readily absorbed hexoses - glucose and galactose. In the small intestine bile must be present which increases the permeability of the cellular wall for lactobacilli and lactose thus enters their cells more easily and it makes easier penetration of the enzyme into the intestinal contents possible and thus promotes further breakdown of lactose in the intestine.


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  21. Lactobacilli in the Treatment of Dyspepsia in Dysmicrobia in Different Aetiology. Kocián J (1994c). Vnitrni Lékarstvi, 40: 79-83.

    In 30 patients with dyspepsia caused by dysbacteriosis of the gastrointestinal tract the authors administered the preparation Lactobacillus acidophilus (Institut Rosell Inc., Canada) - 1 Capsule with 2 billion live bacteria, in the morning after breakfast. The patients were divided into four groups: maldigestion, malabsorption, radiation enterocolitis and administration of antibiotics. The patients recorded themselves their subjective symptoms: pain, pressure, bloating, flatulence and appetite, and as to objective symptoms, the number and consistency of bowel movements, changes of body weight. The most rapid effect was achieved in dysbioses after antibiotics - within 3-4 days normalization occurred which persisted even after discontinuation of the drug. In maldigestion after one week bloating, flatulence, abdominal pain and pressure in the epigastrium was milder, and within two weeks the condition improved further. An excellent effect was achieved in radiation enterocolitis. In patients with lactose intolerance the tolerance of dairy products improved. No side-effects were observed, the preparation was very well tolerated; the mean body weight increment was 0.75 kg in three weeks. The preparation proved a new useful probiotic which is highly effective in dyspepsias caused by dysbiosis of the intestinal microflora.

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