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A lyophilized and inactivated culture of Lactobacillus acidophilus increases Helicobacter pylori eradication rates.

BACKGROUND: Acid suppression plus two antibiotics is considered the reference anti-Helicobacter pylori treatment. Reported eradication rates are around 65-80%. Human Lactobacillus acidophilus shows an in vitro inhibitory effect on the attachment of H. pylori to gastric epithelial cell lines. Culture supernatant of this bacillus seems to decrease the in vitro viability of H. pylori. AIM: To evaluate whether the supplementation with an inactivated preparation of L. acidophilus could improve the efficacy of a standard anti-H. pylori therapy. METHODS: One-hundred and twenty H. pylori-positive patients were randomly assigned to a 7-day triple therapy based on rabeprazole (20 mg b.d.), clarithromycin (250 mg t.d.s.) and amoxicillin (500 mg t.d.s.) (RCA group: 60 subjects), or to the same regimen supplemented with a lyophilized and inactivated culture of Lactobacillus acidophilus (t.d.s.) (RCAL group: 60 subjects). RESULTS: In the RCA group, eradication was successful in 72% (42 out of 58 patients) from a per protocol (PP) analysis, or 70% (42 out of 60 patients) using an intention-to treat (ITT) analysis. In the RCAL group a significant increase in the eradication rate was observed: 88% (52 out of 59 patients) from PP analysis (P=0.03), 87% (52 out of 60 patients) from ITT analysis (P=0.02). CONCLUSIONS: These results seem to confirm the in vitro anti-H. pylori effect of L. acidophilus, suggesting that the inactivated L. acidophilus could be effective in increasing eradication rates of a standard anti-H. pylori therapy.

2-Pyridinylmethylsulfinylbenzimidazoles↗

Lectin-like activity of Lactobacillus acidophilus strain JCM 1026.

The lectin-like activity of the Lactobacillus acidophilus strain JCM 1026 was studied by hemagglutination and hemagglutination-inhibition assays. L. acidophilus strain JCM 1026 was found to hemagglutinate human and animal erythrocytes. Neuraminidase-treatment of human-type O erythrocytes enhanced the activity. Treatment of the bacterial cells with proteinase K reduced hemagglutinating activity significantly. Although several mono- and disaccharides did not inhibit hemagglutination, several different glycoproteins did. These data indicate that a proteinaceous lectin-like component(s) recognizing carbohydrate-containing molecules is located on the cell surface of L. acidophilus.

Animals↗

Effects of a milk product, fermented by Lactobacillus acidophilus and with fructo-oligosaccharides added, on blood lipids in male volunteers.

OBJECTIVE: To investigate in adult male volunteers the effect of a new fermented milk product, fermented by Lactobacillus acidophilus and with fructo-oligosaccharides added, on blood lipids. DESIGN: Randomized placebo-controlled double-blind two-way cross over trial with two treatment periods of three weeks, separated by a wash-out period of one week. SETTING: the study was performed at the TNO Nutrition and Food Research Institute in Zeist, The Netherlands. SUBECTS: Thirty normal healthy men, aged 33-64y (mean serum total cholesterol level: 5.23 +/- 1.03 (s.d.)), were selected for this study. Normal health was assessed by pre study screening. All subjects were used to an average Dutch food pattern. INTERVENTIONS: During the treatment periods subjects consumed three times daily a 125 ml of either test or reference product as a part of their habitual diet. The test product was a milk, fermented by yogurt starters and Lactobacillus acidophilus, and contained 2.5% fructo-oligosaccharides, 0.5% vegetable oil and 0.5% milk fat. The reference product was a traditional yogurt (milk fermented only by yogurt strains), containing 1% milk fat. Blood samples for serum lipid analysis and blood glucose measurements were taken before the start of the experiment and at the end of both treatment periods. RESULTS: As compared to the reference product, consumption of the test product resulted in significantly lower values for serum total cholesterol (P < 0.001), LDL-cholesterol (P < 0.005), and the LDL/HDL-ratio (P < 0.05) by 4.4, 5.4 and 5.3% respectively. Levels of serum HDL-cholesterol, triglycerides and blood glucose remained essentially unchanged. The beneficial effects of the test product on serum cholesterol were largely related to an increase of this parameter during the consumption of the reference product. CONCLUSIONS: As compared to traditional yogurt, daily consumption of three times 125 ml of test product specifically lowered serum LDL-cholesterol levels in normal healthy male adult subjects with borderline elevated levels of serum total cholesterol within three weeks.

Adult↗

Desmutagenicity of milk cultured with Lactobacillus acidophilus strains against mutagenic heated tauco.

Desmutagenicity of milk cultured with Lactobacillus acidophilus strains on the mutagenicity of heated salty and sweet tauco were examined using streptomycin dependent (SD) 510 strain of Salmonella typhimurium TA98 as tester culture. Cultured milk samples widely exhibited desmutagenic effects against mutagenic heated salty and sweet tauco. Mutagenicity of heated salty tauco was inhibited by acidophilus cultured milks stronger than that of heated sweet tauco. Milk cultured with strains SBT2054, SBT0299, SBT0274, SBT10238. SBT1702, SBT10240, SBT10241 and SBT10239 showed high inhibition against the mutagenicity of both heated salty and sweet taucos. Maximum inhibition was reached after 24 hr of incubation which corresponded to stationary growth phase. Desmutagenic activities of the acidophilus cultured milks against mutagenic heated tauco were mainly attributed to the bacterial cells and also to casein of milk.

Animals↗

Production of organic acids from fermentation of mannitol, fructooligosaccharide and inulin by a cholesterol removing Lactobacillus acidophilus strain.

AIMS: Assessment of individual production of organic acids by Lactobacillus acidophilus ATCC 4962 in the presence of mannitol, fructooligosaccharide (FOS) and inulin. METHODS AND RESULTS: The production patterns of individual organic acids by L. acidophilus ATCC 4962 were assessed using the experimental region for optimum cholesterol removal from the interaction between L. acidophilus ATCC 4962 and prebiotics selected in our previous study. The production of acetic and formic acids was growth associated and was greatly influenced by the inoculum size of the organism and the concentration of mannitol. The growth of the organism was repressed with the fermentation end products of FOS and inulin, which subsequently exhibited repressed production of acetic and formic acids as well. The inoculum size, mannitol and FOS linearly affected the formation of butyric acid and the response surface generated showed a correlation between butyric acid and acetic acid. The experimental regions with increased production of lactic acid showed cessation of growth of the organism, indicating inhibition of growth at high concentration of lactic acid. CONCLUSIONS: The production of individual organic acids was dependent on growth and the fermentability of prebiotics. Mannitol, FOS and inulin favoured the production of formic, lactic and butyric acids respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: The fermentability of prebiotics to produce metabolites has been a controversial issue. Information gathered in this study provides a better understanding on the production of organic acids from fermentation of mannitol, FOS and inulin by L. acidophilus ATCC 4962, and on changes in their production as a response from interaction of factors.

Acetates↗

Systemic augmentation of the immune response in mice by feeding fermented milks with Lactobacillus casei and Lactobacillus acidophilus.

This study investigates the effect of feeding fermented milks with Lactobacillus casei, Lactobacillus acidophilus and a mixture of both micro-organisms on the specific and non-specific host defence mechanisms in Swiss mice. Animals fed with fermented milk for 8 days (100 micrograms/day) showed an increase in both phagocytic and lymphocytic activity. This activation of the immune system began on the 3rd day, reached a maximum on the 5th, and decreased slightly on the 8th day of feeding. In the 8-day treated mice, boosted with a single dose (100 micrograms) on the 11th day, the immune response increased further. The feeding with fermented milk produced neither hepatomegaly nor splenomegaly. These results suggest that L. casei and L. acidophilus, associated with intestinal mucosae, can influence the level of activation of the immune system. The possible clinical application of fermented milks as immunopotentiators is also discussed.

Animals↗

Prevention of colonic preneoplastic lesions by the probiotic Lactobacillus acidophilus NCFMTM in F344 rats.

The experiments described here were aimed at developing novel probiotic strains that may aid in the reduction of colon cancer risk. We assessed the potential anticancer properties of Lactobacillus acidophilus NCFMTM in male F344 rats using inhibition of the formation of azoxymethane (AOM)-induced aberrant crypt foci (ACF) in the colon as the measure of preventive efficacy. At 6 weeks of age, groups of rats were fed the experimental diets containing 0, 2% or 4% lyophilized cultures of L. acidophilus NCFMTM. At 7 weeks of age, all animals in each dietary group, except the vehicle-treated rats, were s.c. injected with AOM (15 mg/kg body weight) once weekly for two weeks. The vehicle-treated groups were given s.c. injections of normal saline. All rats were necropsied 10 weeks after the last AOM injection and ACF in formalin-fixed, methylene blue-stained colonic tissues were counted under the light microscope. The contents of the cecum were analyzed for bacterial beta-glucuronidase activity. Diet supplementation with the probiotic strain NCFMTM significantly suppressed AOM-induction of colonic ACF, in terms of total number, as well as crypt multiplicity and number of ACF/cm2 colon (P<0.01 - 0.001). NCFMTM inhibited AOM-induced colonic ACF formation in a dose-dependent manner (P<0.01). A significant dose-dependent reduction of cecal beta-glucuronidase activities was observed in the rats fed 2% (P<0.04) and 4% (P<0.0001) NCFMTM. These results suggest that Lactobacillus acidophilus NCFMTM may potentially prevent colon cancer development. Further studies are warranted to determine the full potential of this probiotic strain in preclinical efficacy studies.

Animals↗

Inhibitory effect of Lactobacillus acidophilus on Helicobacter pylori in peptic ulcer patients: in vitro study.

Lactobacillus acidophilus (L. acidophilus) have been introduced into many fermented dairy products. The presence of L. acidophilus appears to decrease Helicobacter pylori (H. pylori) density in the human stomach and could enhance antibiotic therapy for H. pylori eradication. This study was designed to determine the optimal density of L. acidophilus that has a maximum inhibitory effect on H. pylori in peptic ulcer patients. To determine whether L. acidophilus has an inhibitory effect on H. pylori isolated from peptic ulcer patients and to determine whether the optimal density of L. acidophilus has a maximum inhibitory effect on H. pylori isolated from peptic ulcer patients. H. pylori was isolated from gastric biopsy specimens of peptic ulcer patients. The suspension of pure H. pylori colonies were inoculated into the broth and adjusted to match the density of No.3 MacFarland standard (approximately 9x10(8) cells/ml). Forty microliters of the suspension were equally spread onto each quadrant of the plate and left to dry. L. acidophilus was prepared from LC-1 (Nestle Research Center, Switzerland). They were grown on blood agar and incubated overnight at 37 degrees C. The suspension of L. acidophilus was inoculated into the broth and adjusted to match the density of No.1, No.2, No.3 and No.4 MacFarland standard (approximately 3x10(8), 6x10(8), 9x10(8) and 12x10(8) cells/ml respectively). Ten microliters of each density of L. acidophilus was dropped onto each quadrant of a previously inoculated H. pylori plate and then the plate was kept under microaerophilic conditions for 72 hours. Inhibition clear halo zone of H. pylori around the colonies of L. acidophilus was interpreted as the inhibitory effect. H. pylori were isolated from gastric biopsy specimens of fifteen peptic ulcer patients (eleven patients with gastric ulcer and four patients with duodenal ulcer). A total of sixty tests of inhibitory effect of L. acidophilus on H. pylori were evaluated in the present study. L. acidophilus had inhibitory effect on H. pylori in 13/15 patients (86.67%) and L. acidophilus in the density of No.3 MacFarland standard had a significantly higher inhibitory effect on H. pylori in the density of No.3 MacFarland than that of No.1 (60% vs 20%; p<0.05), No.2 (60% vs 20%; p<0.05) and No.4 (60% vs 20%; p<0.05) MacFarland standard. In conclusion, L. acidophilus has an inhibitory effect on H. pylori isolated from peptic ulcer patients. Approximately an equal density of L. acidophilus on H. pylori has the most favorable effect. This optimal density of L. acidophilus should have maximum effect on H. pylori clearance and could enhance antibiotic therapy for H. pylori eradication in humans.

Adult↗

Serum cholesterol levels in axenic mice colonized with Enterococcus faecium and Lactobacillus acidophilus.

Hypocholesterolemic effect was shown in axenic, mono, bicolonized and conventional mice: the effect was different depending on probiotic properties of intestinal microorganisms. Contamination by Enterococcus faecium CX determined the highest effect: haematic cholesterol level decrease was 16.9% in females and 7.8% in males. In mice contaminated by Lactobacillus acidophilus N5 the decrease of haematic cholesterol levels was less and not relevant in mice contaminated by conventional microflora. Enterococcus faecium CX and Lactobacillus acidophilus N5 strains were able to grow in presence of bile salts, to colonize intestinal tract, to survive at gastric conditions and to assimilate cholesterol (E. faecium more than L. acidophilus). The authors consider the possibility to associate probiotic strains with these characteristics for the health of consumers.

Animal Feed↗

Genetic analysis of acidocin B, a novel bacteriocin produced by Lactobacillus acidophilus.

The genes encoding the production of acidocin B, a bacteriocin produced by Lactobacillus acidophilus strain M46 which is active against Listeria monocytogenes, Clostridium sporogenes, Brochothrix thermosphacta, Lactobacillus fermentum and Lactobacillus delbrueckii subsp. bulgaricus, but inactive against most other Lactobacillus species, were previously localized on a 4 kb XbaI-HindIII fragment of plasmid pCV461. In the present work, DNA sequence analysis revealed the presence of three consecutive ORFs, which potentially code for hydrophobic peptides composed of 60, 91 and 114 amino acids, respectively, and a fourth ORF of opposite polarity which could potentially encode a peptide of 59 amino acids. The middle ORF (ORF-2; acdB) was identified as the gene encoding acidocin B by comparing the amino acid composition of highly purified acidocin B with the deduced amino acid sequence of ORF-2. Our results suggest that acidocin B is synthesized as a precursor which is processed at a site which conforms to the ' -3, -1' rules of von Heijne to yield active acidocin B (59 amino acids). The presence of an immunity-protein-encoding gene on the 4 kb XbaI-BamHI fragment was deduced from the capacity of a plasmid vector harbouring this fragment to confer immunity upon transformation of L. fermentum NCK127. One of the three non-assigned ORFs may encode this immunity protein.

Amino Acid Sequence↗

Preservation of intestinal integrity during radiotherapy using live Lactobacillus acidophilus cultures.

Twenty-four female patients suffering from gynaecological malignancies and scheduled for internal and external irradiation of the pelvic area (pelvic dose 5000 cGy) were selected for a study on prevention of intestinal side-effects by live Lactobacillus acidophilus cultures. The patients were randomised into two groups. Both groups received dietary counselling recommending a low-fat and low-residue diet during radiotherapy. The control group received dietary counselling only. The test group received 150 ml of a fermented milk test product supplying them with at least 2 X 10(9) live Lactobacillus acidophilus bacteria daily and 6.5% lactulose as substrate for the bacteria. The results indicated that the test product appeared to prevent radiotherapy-associated diarrhoea. However, flatulence was increased probably due to lactulose ingestion in the test group.

Adult↗

Antimicrobial activity of lactobacilli: preliminary characterization and optimization of production of acidocin B, a novel bacteriocin produced by Lactobacillus acidophilus M46.

Approximately 1000 lactobacillus strains were isolated and screened for the production of antimicrobial activity, using a target panel of spoilage organisms and pathogens. Only eight positive strains were found; two of these were studied in more detail. Lactobacillus salivarius M7 produces the new broad spectrum bacteriocin salivaricin B which inhibits the growth of Listeria monocytogenes, Bacillus cereus, Brochothrix thermosphacta, Enterococcus faecalis and many lactobacilli. A new atypical bacteriocin produced by Lact. acidophilus M46, acidocin B, combines the inhibition of Clostridium sporogenes with a very narrow activity spectrum within the genus Lactobacillus and was selected for further characterization. Acidocin B is sensitive to trypsin, heat-stable (80 degrees C for 20 min) and can be extracted from the culture supernatant fluid with butanol. Native acidocin B occurs as a large molecular weight complex (100 kDa), while with SDS-PAGE the partly purified activity migrates as a peptide of 2.4 kDa. Optimization of the cultivation conditions resulted in an eightfold increase of the amount of acidocin B produced during growth. Growth is not necessary for acidocin B production; washed producer cells can synthesize the bacteriocin in a chemically defined production medium. The application potential of acidocin B is discussed.

Bacteriocins↗

Identification and purification of a protein that induces production of the Lactobacillus acidophilus bacteriocin lactacin B.

Lactacin B is a heat-stable bacteriocin produced by Lactobacillus acidophilus N2 that is active against closely related lactobacilli, including Lactobacillus delbrueckii subsp. lactis (formerly Lactobacillus leichmannii) ATCC 4797. Pure producer cultures propagated in MRS broth (initial pH 6.5) contain no lactacin B; it is detected only in cultures maintained at pH 5.0 to 6.0 and produced optimally at pH 6.0 S. F. Barefoot and T. R. Klaenhammer, Antimicrob. Agents Chemother. 26:328-334, 1984). Associative growth of producer and indicator, L. delbrueckii subsp. lactis ATCC 4797, resulted in production of an inhibitor identical to lactacin B. Associative growth increased lactacin B production from nondetectable levels (< 100 activity units [AU]/ml) to between 3,200 and 6,400 AU/ml in MRS broth (initial pH 6.5) and resulted in early but equal production of lactacin B (approximately 25,600 AU/ml) in broth maintained at pH 6.0. Indicator cells, but not spent culture filtrates, induced lactacin B production. Indicator cells disrupted by a French pressure cell yielded cell-free filtrates containing inducing activity. Chromatofocusing and gel filtration high-performance liquid chromatography of cell-free filtrates yielded a protein with a pI of 4.1 and a molecular size of approximately 58 kDa that induced lactacin B production. Analytical isoelectric focusing yielded a single protein band. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels contained a 28-kDa protein suggesting a two-subunit structure. Protein sequencing identified an N-terminal serine and 18 additional amino acids. To our knowledge, there are not previous descriptions of proteins that induce bacteriocin production in lactic acid bacteria.

Amino Acid Sequence↗

Characterization and purification of acidocin CH5, a bacteriocin produced by Lactobacillus acidophilus CH5.

AIMS: To characterize and to purify a bacteriocin produced by Lactobacillus acidophilus strain with its activity restricted to Gram-positive bacteria. METHODS AND RESULTS: Native acidocin CH5, a bacteriocin produced by L. acidophilus CH5 an isolate from a dairy starter culture forms in MRS (Oxoid, Basingstoke, UK) broth high-molecular weight aggregates which can dissociate into smaller units (retained by 5 kDa membrane) with higher activity. Acidocin CH5 was purified using combinations of chromatographic methods based on hydrophobic and cation exchange principles and the N-terminal region was sequenced. CONCLUSIONS: Based on our results it is evident that acidocin CH5 belongs, according to bacteriocin classification, to the class II bacteriocins with identical N-terminal amino acid sequence described in the literature previously. SIGNIFICANCE AND IMPACT OF THE STUDY: The study has provided further data on bacteriocin acidocin CH5 from class II with wide spectrum of antimicrobial activity atypical for bacteriocins produced by L. acidophilus sharing the same homology.

Amino Acid Sequence↗

Transmission of Lactobacillus jensenii and Lactobacillus acidophilus from mother to child at time of delivery.

The presence of Lactobacillus jensenii and Lactobacillus acidophilus has been studied in specimens from the rectum and vagina of the mother, from the mouth of the infant at the time of delivery, and from the mouth and rectum of infants six days of age. L. jensenii could be differentiated from other species of lactobacilli by the following combination of characteristics: production of only D-lactate, hydrolysis of arginine, and fermentation of cellobiose, galactose, and ribose, but not of lactose. L. jensenii and L. acidophilus were common inhabitants of the vagina. In spite of a contamination of the infant's mouth by L. jensenii and L. acidophilus during delivery, neither of these organisms became established in the mouth of the newborn infants.

Adult↗

Impact of Lactobacillus acidophilus supplements on the human oropharyngeal and intestinal microflora.

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.

Adult↗