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PubMed1976

Intestinal pH and propulsion: an explanation of diarrhoea in lactase deficiency and laxation by lactulose.

Subjects deficient in lactase may experience bloating, cramps and diarrhoea after ingesting milk, due to the unhydrolysed and poorly-absorbed lactose. The diarrhoea may result from an osmotic effect of the lactose itself or its poorly-absorbed acidic products of fermentation (Weijers, van de Kamer & others, 1961; Christopher & Bayless, 1971), possibly together with an alteration of sodium and water absorption due to the lowered colonic pH (Rousseau & Sladen, 1971). Laxation by lactulose (1-4-beta-galactosidofructose) may operate through an analogous mechanism. The drug is a synthetic dissaccharide which, in oral doses of 10-20 g, relieves chronic constipation (Wesselius-de Casparis, Braadbaart & others, 1968). It is neither hydrolysed by intestinal dissaccharidase (Dahlqvist & Gryboski, 1965) nor absorbed in the gut, but it is converted in the colon mainly to lactic and acetic acids by various bacteria including Lactobacillus acidophilus. Apart from the increased osmotic effect, the pH in the proximal colon falls markedly (Bown, Gibson & others, 1974), and larger doses may reduce stool pH. Weijers & others (1961) inferred that the acidic products formed from lactose in the colon stimulate propulsion, and K.S. Liem (Philips-Duphar) suggested to us that lactulose may relieve constipation partly by stimulation of propulsion due to the lowered pH. The experiments described below support this view.

Animals↗
PubMed2006

Biotic and abiotic factors influencing in vitro growth of Escherichia coli O157:H7 in ruminant digestive contents.

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.

Animals↗
PubMed1991

Bacteriocin-mediated inhibition of Clostridium botulinum spores by lactic acid bacteria at refrigeration and abuse temperatures.

The bacteriocinogenicity of Lactococcus lactis ATCC 11454, Pediococcus pentosaceus ATCC 43200, P. pentosaceus ATCC 43201, Lactobacillus plantarum BN, L. plantarum LB592, L. plantarum LB75, and Lactobacillus acidophilus N2 against Clostridium botulinum spores at 4, 10, 15, and 35 degrees C was investigated by modified deferred and simultaneous antagonism methods. All the strains, except L. acidophilus N2, produced inhibition zones on lawns of C. botulinum spores at 30 degrees C. L. plantarum BN, L. lactis ATCC 11454, and P. pentosaceus ATCC 43200 and 43201 were bacteriocinogenic at 4, 10, and 15 degrees C. Supplementation of brain heart infusion agar with 0 to 5% NaCl increased the radii of inhibition zones during simultaneous antagonism assays. Detectable bacteriocin activities were extracted from freeze-thawed agar cultures of L. plantarum BN and L. lactis ATCC 11454 which had been grown at 4 and 10 degrees C. These results suggest that low levels of L. plantarum BN or L. lactis ATCC 11454, in the presence of 3 or 4% NaCl, could be formulated into minimally processed refrigerated food products for protection against possible botulism hazards.

Bacteriocins↗
PubMed1993

Rapid Mini-Prep Isolation of High-Quality Plasmid DNA from Lactococcus and Lactobacillus spp.

A simple, rapid plasmid mini-prep procedure for lactococci and lactobacilli which gives high yields and can be performed on overnight broth cultures is presented. Large plasmids were isolated from both lactococci and lactobacilli, including a 70-kb plasmid from Lactobacillus acidophilus C7. The purity of the resulting plasmid DNA makes it suitable for subsequent molecular manipulations. The convenience of the technique makes this rapid mini-prep procedure suitable for routine plasmid isolation from lactic acid bacteria.

Journal Article↗
PubMed1995

A Collagen-Binding S-Layer Protein in Lactobacillus crispatus.

Two S-layer-expressing strains, Lactobacillus crispatus JCM 5810 and Lactobacillus acidophilus JCM 1132, were assessed for adherence to proteins of the mammalian extracellular matrix. L. crispatus JCM 5810 adhered efficiently to immobilized type IV and I collagens, laminin, and, with a lower affinity, to type V collagen and fibronectin. Strain JCM 1132 did not exhibit detectable adhesiveness. Within the fibronectin molecule, JCM 5810 recognized the 120-kDa cell-binding fragment of the protein, while no bacterial adhesion to the amino-terminal 30-kDa or the gelatin-binding 40-kDa fragment was detected. JCM 5810 but not JCM 1132 also bound (sup125)I-labelled soluble type IV collagen, and this binding was efficiently inhibited by unlabelled type IV and I collagens and less efficiently by type V collagen, but not by laminin or fibronectin. L. crispatus JCM 5810 but not L. acidophilus JCM 1132 also adhered to Matrigel, a reconstituted basement membrane preparation from mouse sarcoma cells, as well as to the extracellular matrix prepared from human Intestine 407 cells. S-layers from both strains were extracted with 2 M guanidine hydrochloride, separated by electrophoresis, and transferred to nitrocellulose sheets. The S-layer protein from JCM 5810 bound (sup125)I-labelled type IV collagen, whereas no binding was seen with the S-layer protein from JCM 1132. Binding of (sup125)I-collagen IV to the JCM 5810 S-layer protein was effectively inhibited by unlabelled type I and IV collagens but not by type V collagen, laminin, or fibronectin. It was concluded that L. crispatus JCM 5810 has the capacity to adhere to human subintestinal extracellular matrix via a collagen-binding S-layer.

Journal Article↗
PubMed1996

PCR detection and quantitation of predominant anaerobic bacteria in human and animal fecal samples.

PCR procedures based on 16S rRNA gene sequences specific for 12 anaerobic bacteria that predominate in the human intestinal tract were developed and used for quantitative detection of these species in human (adult and baby) feces and animal (rat, mouse, cat, dog, monkey, and rabbit) feces. Fusobacterium prausnitzii, Peptostreptococcus productus, and Clostridium clostridiiforme had high PCR titers (the maximum dilutions for positive PCR results ranged from 10(-3) to 10(-8)) in all of the human and animal fecal samples tested. Bacteroides thetaiotaomicron, Bacteroides vulgatus, and Eubacterium limosum also showed higher PCR titers (10(-2) to 10(-6)) in adult human feces. The other bacteria tested, including Escherichia coli, Bifidobacterium adolescentis, Bifidobacterium longum, Lactobacillus acidophilus, Eubacterium biforme, and Bacteroides distasonis, were either at low PCR titers (less than 10(-2)) or not detected by PCR. The reported PCR procedure including the fecal sample preparation method is simplified and rapid and eliminates the DNA isolation steps.

Adult↗
PubMed2001

Surface expression of the conserved C repeat region of streptococcal M6 protein within the Pip bacteriophage receptor of Lactococcus lactis.

The C repeat region of the M6 protein (M6c) from Streptococcus pyogenes was expressed within the Pip bacteriophage receptor on the surface of Lactococcus lactis. M6c was also detected in the culture medium. The pip-emm6c allele was integrated into the chromosome and stably expressed without antibiotic selection. The level of cell-associated surface expression of PipM6c was 0.015% of total cellular protein. The amount of PipM6c on the cell surface was increased about 17-fold by expressing pip-emm6c from a high-copy-number plasmid. Replacing the native pip promoter with stronger promoters isolated previously from Lactobacillus acidophilus increased surface expression of PipM6c from the high-copy-number plasmid up to 27-fold. Concomitantly, the amount of PipM6c in the medium increased 113-fold. The amount of PipM6c did not vary greatly between exponential- and stationary-phase cultures. Western blots indicated that the full-length PipM6c protein and most of the numerous proteolytic products were found only on the cell surface, whereas only one proteolytic fragment was found in the culture medium.

Antigens, Bacterial↗
PubMed1995

Role of Escherichia coli P fimbriae in intestinal colonization in gnotobiotic rats.

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.

Animals↗
PubMed2004

[Efficient prevention of prematurity - the Thuringian model].

In two prospective projects, the effectiveness of the self-care programme for prematurity prevention developed by Saling was prevented. Pregnant women in Erfurt were shown how to perform self-measurements of their vaginal pH by means of test gloves twice a week in order to screen for any disturbances in the vaginal millieu. The women were instructed to see their physician immediately, if abnormal values (ph > or = 4.7) or other risk factors were present, in order to start Lactobacillus acidophilus therapy or, in cases of bacterial vaginosis, treatment with intravaginal clindamycin cream. Patients who were not interested in the programme, served as a control group. 73 out of 381 women in the intervention group were identified as risk cases. In this study, the prematurity rate was 8.1% in the self-measurement/intervention group versus 12.3% in the control group (p < 0.05, n = 2,341); 0.3 versus 3.3% of the neonates belonged to the group of very early prematures with a gestational age of <32 + 0 weeks (p < 0.01). Starting on March 1, 2000, a similar statewide pH screening programme was initiated in order to reduce prematurity in the State of Thuringia. According to the study design, a significant decrease in prematurity was hypothetically expected for the second half of 2000. Data from 16,276 women are available. On this basis, a significant reduction of early prematurity from 1.58 to 0.99% was seen (p < 0.001). Comparing low birth weights, a significant reduction of cases was also achieved in all groups. On the basis of data obtained, we recommend that the campaign should be extended to the whole of Germany. This recommendation is also supported by the observation that after the campaign had been discontinued the prematurity rates monitored in 2002 were as high as they had been prior to the introduction of the statewide programme.

Anti-Bacterial Agents↗
PubMed2002

Comparison of bacterial and tissue cell initial adhesion on hydrophilic/hydrophobic biomaterials.

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.

Animals↗
PubMed2006

Lactobacillus plantarum gene clusters encoding putative cell-surface protein complexes for carbohydrate utilization are conserved in specific gram-positive bacteria.

BACKGROUND: Genomes of gram-positive bacteria encode many putative cell-surface proteins, of which the majority has no known function. From the rapidly increasing number of available genome sequences it has become apparent that many cell-surface proteins are conserved, and frequently encoded in gene clusters or operons, suggesting common functions, and interactions of multiple components. RESULTS: A novel gene cluster encoding exclusively cell-surface proteins was identified, which is conserved in a subgroup of gram-positive bacteria. Each gene cluster generally has one copy of four new gene families called cscA, cscB, cscC and cscD. Clusters encoding these cell-surface proteins were found only in complete genomes of Lactobacillus plantarum, Lactobacillus sakei, Enterococcus faecalis, Listeria innocua, Listeria monocytogenes, Lactococcus lactis ssp lactis and Bacillus cereus and in incomplete genomes of L. lactis ssp cremoris, Lactobacillus casei, Enterococcus faecium, Pediococcus pentosaceus, Lactobacillius brevis, Oenococcus oeni, Leuconostoc mesenteroides, and Bacillus thuringiensis. These genes are neither present in the genomes of streptococci, staphylococci and clostridia, nor in the Lactobacillus acidophilus group, suggesting a niche-specific distribution, possibly relating to association with plants. All encoded proteins have a signal peptide for secretion by the Sec-dependent pathway, while some have cell-surface anchors, novel WxL domains, and putative domains for sugar binding and degradation. Transcriptome analysis in L. plantarum shows that the cscA-D genes are co-expressed, supporting their operon organization. Many gene clusters are significantly up-regulated in a glucose-grown, ccpA-mutant derivative of L. plantarum, suggesting catabolite control. This is supported by the presence of predicted CRE-sites upstream or inside the up-regulated cscA-D gene clusters. CONCLUSION: We propose that the CscA, CscB, CscC and CscD proteins form cell-surface protein complexes and play a role in carbon source acquisition. Primary occurrence in plant-associated gram-positive bacteria suggests a possible role in degradation and utilization of plant oligo- or poly-saccharides.

Antigens, Surface↗
PubMed2005

Bifidobacterium Lactis sp. 420 up-regulates cyclooxygenase (Cox)-1 and down-regulates Cox-2 gene expression in a Caco-2 cell culture model.

Cyclooxygenases (Cox) -1 and -2 play important roles in gastrointestinal health; chronic overexpression of Cox-2 is associated with inflammatory and cancerous disease, whereas Cox-1 is expressed constitutively. We studied the effects of two probiotic (Bifidobacterium lactis sp. 420 and Lactobacillus acidophilus) and two control microorganisms (Escherichia coli and Salmonella enteritidis) and four microbial metabolites (acetate, butyrate, lactate and propionate) on the expression levels of the Cox isoforms in the enterocyte-like cell line Caco-2. Butyrate, which is anticarcinogenic, resulted in an 85% down-regulation of Cox-2 and a 37-fold increase in Cox-1 transcription. Propionate gave similar results (72% reduction of Cox-2, 23-fold induction of Cox-1), but lactate and acetate had no effect on Cox expression profile. Bifidobacterium sp. 420, which produces acetate and lactate but no butyrate or propionate, shared the Cox-1-increasing and Cox-2-silencing properties of butyrate and propionate, whereas L. acidophilus was similar to E. coli and S. enteritidis in having no effect on the Cox-1/Cox-2 ratio. For the first time, we therefore demonstrate evidence for a direct relationship between a probiotic bacterial strain and host Cox expression profile, suggesting that modulation of Cox expression may be an important factor in the potential anti-inflammatory and anticarcinogenic properties of some probiotics.

Acetates↗
PubMed2002

New rapid enzyme-linked immunosorbent assay to detect antibodies against bacterial surface antigens using filtration plates.

An easy and rapid ELISA system, Filtration ELISA, to detect antibodies against bacterial cell surface antigens was developed using a 96-well filtration plate fitted with a 0.22microm membrane (MultiScreen-GV, Millipore). Bacterial whole cells were used as antigens without fixing the cells with formalin etc. The whole cell antigens were washed by vacuum filtration through a filter and resuspended in washing buffer. Assay reactions could be done in the wells without losing the solution. The technique was established using antisera of mice immunized with Escherichia coli, and then evaluated by assaying antibodies to Shiga-toxin producing E. coli O157:H7 (STEC), Staphylococcus aureus and Lactobacillus acidophilus in fecal extracts of 157 children who had eaten school lunches contaminated with STEC in comparison with 25 age-matched control children. The lunch group showed significantly higher IgA antibody titers against STEC than the control group (p<0.0005), but not against L. acidphilus. The results indicate that Filtration ELISA is a quantitative and specific technique for measuring antibodies against antigens on the surface of bacteria without extracting antigens from the bacteria. This technique is widely applicable to the assay of antibodies in various samples including serum and fecal extract against various kinds of bacteria.

Animals↗
PubMed1999

Evaluation of yeast extracts as growth media supplements for lactococci and lactobacilli by using automated spectrophotometry.

An automated spectrophotometric (AS) method was used to evaluate the growth-promoting ability of yeast extracts (YE) on cultures of Lactobacillus acidophilus and Lactococcus lactis subsp. cremoris. The AS data were compared to that obtained from classical shake flask fermentations and from 250 ml bioreactors equipped with pH control. In assays involving the evaluation of 26 different commercial YE, maximum growth rate (&mgr;(max)) values determined with the AS unit ranged from 0.25 to 0.45 h(-1) for Lb. acidophilus and from 0.10 to 0.40 h(-1) for Lc. cremoris. Good correlations were obtained between AS data and manual sampling from the shake flasks or the bioreactors for mmax, as well as maximum optical density (OD(max)). The AS method is thus useful as a screening tool for the selection of YE lots in media formulation. Species reacted differently to the 26 YE, but less variation was observed between strains of the same species. This suggests that a producer of various lactococci or lactobacilli can expect a relatively constant response to a given YE lot between strains of the same species. However, it should not be assumed that the YE having the best growth-promoting properties for Lb. acidophilus will also be the best media supplements for the growth of Lc. cremoris.

Journal Article↗
PubMed1985

Influence of realimentation diet on recovery of rumen activity and feed intake in beef steers.

Two trials were conducted to determine the influence of realimentation diet energy, protein, B-vitamin (BV) and Lactobacillus acidophilus (LAC) content on recovery of rumen activity and feed consumption in beef steers. In trial 1, ruminal-fistulated steers were fasted and refed 1) prairie hay, 2) 10% protein (LCP), 3) 12.5% protein (MCP), 4) LCP + BV or 5) LCP + LAC. In trial 2, calves were fasted and refed 1) 60% cottonseed hulls-40% alfalfa dehy (high roughage), 2) LCP, 3) 15% protein (HCP), 4) LCP + BV or 5) LCP + LAC. Rumen fermentative capacity declined 74% (P less than .05) during feed and water deprivation, but returned to control levels by d 7 of realimentation. On d 3 of realimentation, steers fed the LCP and MCP diets had molar proportions of ruminal butyrate in excess of 35%. Steers fed the hay, LAC and BV diets did not have a high butyrate fermentation. In trial 2, calves lost about 15% of their body weight during feed and water deprivation. Calves fed the high roughage diet appeared to return to prefast feed and energy intakes more slowly than steers fed the medium roughage diets. Results of this study indicate that rumen fermentative capacity is a factor limiting feed intake in fasted calves for 7 to 14 after the reintroduction of feed and water.

Animal Feed↗
PubMed2004

Suppressive effect of functional drinking yogurt containing specific egg yolk immunoglobulin on Helicobacter pylori in humans.

Helicobacter pylori is a human pathogen that infects over 50% of the population worldwide. It is the most important etiologic agent of gastroduodenal ulcers and malignancies. Helicobacter pylori urease enzyme is considered the main factor for the organism's colonization in the gastroduodenal mucosa. Hens immunized with the purified urease produce a highly specific anti-H. pylori urease immunoglobulin (IgY-urease) in their egg yolks. Immunoglobulin Y-urease was stable at 60 to 65 degrees C for 30 min and at pH 4.0 for 7 h. Its activity was lost at 80 degrees C for 20 min and at pH 2 for 4 h. Specially designed functional drinking yogurt containing Lactobacillus acidophilus and Bifidobacterium spp. with 1% egg yolk IgY-urease was produced commercially. Immunoglobulin Y-urease activity showed stability in the product up to 7 d, and then decreased to 85% after 3 wk of storage. A clinical study was conducted to determine the effectiveness of IgY-urease yogurt to suppress infection in humans. Forty-two volunteers who tested positive for H. pylori using a 13C-urea breath test were recruited. A total of 450 mL of IgY-urease (test group) or IgY-urease-free yogurt (control group) was consumed in 150-mL portions 3 times daily for 4 wk. Volunteers were tested after 2 and 4 wk; urea breath test values significantly decreased in the test group compared with the control group. The results indicate that suppression of H. pylori infection in humans could be achieved by consumption of drinking yogurt fortified with IgY-urease.

Adult↗
PubMed1991

In vitro binding of mutagenic pyrolyzates to lactic acid bacterial cells in human gastric juice.

The binding of mutagenic pyrolyzates to freeze-dried cells of Streptococcus cremoris Z-25 was investigated in human gastric juice and compared with the ability of these pyrolyzates to bind in distilled water (pH 6.30). The pH of donated gastric juice ranged from 1.2 to 7.8. Binding that occurred in gastric juice was pH dependent, and binding was affected by NaCl, CaCl2, and MgCl2. Les sof Trp-P-2 was bound in gastric juice at low pH (1.2, 1.48, and 2.1) than at pH 4 to 8. The strain bound simultaneously Trp-P-1, Trp-P-2, and Glu-P-1. The maximum amounts of Trp-P-2 bound to S. cremoris Z-25 were approximately 47.50 micrograms/mg of cells. Freeze-dried cells treated at 120 degrees C for 15 min or 80 degrees C for 3 h showed increased binding of Trp-P-2. However, binding decreased 10% after heating at 120 degrees C for 15 min. Cells nonthermally killed by gastric juice (pH 1.5) had the same Trp-P-2 binding capacity as viable cells. Lactobacillus acidophilus IFO 13951 and Bifidobacterium bifidum IFO 14252 representation of the intestinal bacteria nonthermally killed by gastric juice also showed larger binding of Trp-P-2.

Calcium Chloride↗