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PubMed2002

One repeat of the cell wall binding domain is sufficient for anchoring the Lactobacillus acidophilus surface layer protein.

The N-terminal repeat (SAC1) of the S-protein of Lactobacillus acidophilus bound efficiently and specifically to cell wall fragments (CWFs) when fused to green fluorescent protein, whereas the C-terminal repeat (SAC2) did not. Treatment of CWFs with hydrofluoric acid, but not phenol, prevented binding. Apparently, SAC1 is necessary and sufficient for cell wall binding. Our data suggest that SAC anchors the S-protein to a cell wall teichoic acid.

Bacterial Proteins↗

PubMed2004

Effects of probiotic Lactobacillus acidophilus strain DSM13241 in healthy adult dogs.

OBJECTIVE: To evaluate viability of a probiotic strain of Lactobacillus acidophilus in a dry dog food, determine its ability to survive transit through the gastrointestinal tract and populate the colon, and assess its effects on intestinal and systemic parameters. ANIMALS: 15 adult dogs. PROCEDURE: Dogs were sequentially fed a dry control food for 2 weeks, the same food supplemented with > 10(9) L. acidophilus for 4 weeks, and the control food again for 2 weeks. Fecal score was assessed daily, and fecal and blood samples were collected for enumeration of bacterial populations and measurement of hematologic variables. RESULTS: Recovery of L. acidophilus from the supplemented food was 71% and 63% at the start and end of the study, respectively, indicating that the bacteria were able to survive manufacture and storage. The probiotic bacterium was detected in feces via ribotyping and RNA gene sequencing during the probiotic administration phase but not 2 weeks after cessation of administration. Administration of the probiotic-supplemented food was associated with increased numbers of fecal lactobacilli and decreased numbers of clostridial organisms. There were significant increases in RBCs, Hct, hemoglobin concentration, neutrophils, monocytes, and serum immunoglobin G concentration and reductions in RBC fragility and serum NO concentration. CONCLUSIONS AND CLINICAL RELEVANCE: These data indicate that L. acidophilus can be successfully incorporated into a dry dog food, survive transit through the canine gastrointestinal tract, and populate the colon and are associated with local and systemic changes. This probiotic bacterium may have the potential to enhance intestinal health and improve immune function in dogs.

Analysis of Variance↗

PubMed1976

Death of Lactobacillus acidophilus caused by incorporated 3H-thymine during incubation without amino acids.

Of the cells of Lactobacillus acidophilus R-26 incorporated 3H-thymine (specific radioactivity 1.57 Ci/mmol or 3.15 Ci/mmol), their transfer to a medium without essential amino acids resulted in their death. This death may be interpreted in such a way that cell damage caused by disintegration of tritium cannot be effectively repaired under conditions of amino acid deprivation. The experimental eonditions make it possible to explain this death either as a result of inhibition of protein or RNA synthesis or as a result of the absence of amino acids. These possibilities were tested in experiments, in which the synthesis of proteins and RNA was inhibited by specific inhibitors in the presence of amino acids. Under these conditions no death of cells was detected, thus indicating that free amino acids play a role in the repair of radiation damage.

Amino Acids↗

PubMed2002

The survival and persistence of Lactobacillus acidophilus LF221 in different ecosystems.

Being a human isolate and bacterocin producer with wide antimicrobial spectrum, Lactobacillus acidophilus LF221 fulfills preliminary conditions as a potential probiotic strain. To investigate the LF221 strain with respect to its persistence in complex ecosystems, viable LF221 cells were orally administered to laboratory animals (mice and piglets) and added to cheese milk. During the 10-day feeding of mice with LF221 viable cells, 0.9% of lactobacilli isolated from faeces were identical with LF221 strain. Within the piglets' feeding trial, 920 lactobacilli colonies randomly selected from faecal samples were screened for the presence of LF221-like colonies. During the six times of sampling of piglets' faeces, LF221 colonies were always detected. Even the final sampling 10 days after the last LF221 cell suspension was dosed into piglets revealed the presence of LF221 strain at a concentration of about 3 x 10(8) cells/g of faeces. LF221 intake did not significantly influence the total count of lactobacilli in faecal samples and it had no negative health effects on animals. Analyses of cheese demonstrated that the LF221 strain maintained a high viability during ripening and that it could be detected for at least 6 weeks at a level of about 6.8 x 10(6) cells/g of cheese. According to these results, semihard cheese represents a suitable medium for introducing a potential probiotic LF221 into a diet.

Administration, Oral↗

PubMed2005

Effect of regular ingestion of Saccharomyces boulardii plus inulin or Lactobacillus acidophilus LB in children colonized by Helicobacter pylori.

AIM: To evaluate the effect of a probiotic, Lactobacillus acidophilus LB (LB), or a synbiotic, Saccharomyces boulardii plus inulin (SbI), on Helicobacter pylori (Hp) colonization in children. SUBJECTS AND METHODS: A clinical trial was carried out in a school from a low socio-economic area of Santiago. Two hundred and fifty-four asymptomatic children (8.40+/-1.62 y) were screened for Hp by the (13)C-Urea Breath Test ((13)C-UBT). Hp-positive children were randomly distributed into three groups to receive either antibiotic treatment (lanzoprazole, clarythromycin and amoxicillin) for 8 d, or SbI or LB daily for 8 wk. A second (13)C-UBT was carried out at this time. Spontaneous clearance was evaluated in the same way in 81 infected, untreated children. The differences in the delta(13)CO(2) over baseline values before and after treatments (?DOB) were evaluated. RESULTS: 182 subjects (71.7%) were colonized by Hp, and 141 of them completed their treatment (22.5% dropout). Hp was eradicated in 66%, 12% and 6.5% of the children from the Ab, SbI and LB groups, respectively, while no spontaneous clearance was observed in the children without treatment. A moderate but significant difference in ?DOB was detected in children receiving living SbI (-6.31; 95% CI: -11.84 to -0.79), but not in those receiving LB (+0.70; 95% CI: -5.84 to +7.24). CONCLUSION: S. boulardii seems promising as an agent that interferes with Hp in colonized individuals. More studies are needed to confirm these results and to elucidate the mechanisms by which Sb inhibits Hp.

Child↗

PubMed1996

Evaluation of media for selective enumeration of Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus acidophilus, and bifidobacteria.

Fifteen media were evaluated to determine their suitability for selective enumeration of Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus acidophilus, and bifidobacteria using 5 to 6 strains of each of the four groups of organisms. Streptococcus thermophilus agar was found to be suitable for selective enumeration of S. thermophilus under aerobic incubation at 37 degrees C for 24 h. The MRS agar at pH 5.2 or reinforced clostridial agar at pH 5.3 could be used for the selective enumeration of L. delbrueckii ssp. bulgaricus when the incubation was carried out at 45 degrees C for > or = 72 h. However, the recovery of this organism was lower on MRS agar at pH 5.2 or reinforced clostridial agar at pH 5.3 than that obtained on MRS agar. The recovery of L. acidophilus and bifidobacteria on MRS agar and MRS-maltose agar was similar; MRS-maltose agar could be used to enumerate total counts of L. acidophilus and bifidobacteria. For selective enumeration of L. acidophilus, MRS-salicin agar or MRS-sorbitol agar could be used. For selective enumeration of bifidobacteria, MRS NNLP (nalidixic acid, neomycin sulfate, lithium chloride, and paromomycin sulfate) agar was suitable; however, determination of bifidobacteria by differential counts between L. acidophilus enumerated on MRS-salicin agar or MRS-sorbitol agar and the total counts of L. acidophilus and bifidobacteria obtained from MRS-maltose agar resulted in higher recovery of some strains of bifidobacteria. Other media that were evaluated in this study were not suitable for selective enumeration.

Bifidobacterium↗

PubMed1998

Survival Rate and Enzyme Activities of Lactobacillus acidophilus Following Frozen Storage.

The ability of two strains of Lactobacillus acidophilus, CRL 640 and CRL 800, to survive and retain their biological activities under frozen storage was determined. Freezing and thawing, as well as frozen storage, damaged the cell membrane, rendering the microorganisms sensitive to sodium chloride and bile salts. Both lactic acid production and proteolytic activity were depressed after 21 days at -20 degreesC, whereas beta-galactosidase activity per cell unit was increased. Cell injury was partially overcome after repair in a salt-rich medium. Copyright 1998 Academic Press.

Journal Article↗

PubMed1981

Calcium-Induced Alteration of Cellular Morphology Affecting the Resistance of Lactobacillus acidophilus to Freezing.

Examination of factors affecting the resistance of Lactobacillus acidophilus NCFM culture concentrates to freeze injury induced during frozen storage at -20 degrees C revealed that calcium supplementation of the growth medium contributed to the storage stability of cells prepared in static culture. Culture concentrates of L. acidophilus NCFM were prepared from cells propagated in MRS broth or MRS broth supplemented with 0.1% calcium carbonate, calcium chloride, or calcium phosphate. After 28 days of frozen storage at -20 degrees C, concentrated cells (3.2 x 10 colony-forming units per ml) prepared from MRS broth cultures showed an 84% reduction in viable cells. Of the remaining viable cells, 88% were sublethally injured and unable to form colonies on MRS agar supplemented with 0.15% bile. Cells prepared in calcium-supplemented MRS broths demonstrated more resistance to frozen storage. Viability and injury losses in the frozen concentrates were limited to 10 to 39% and 3 to 23%, respectively. It was observed that calcium supplementation of MRS medium resulted in a morphological transition of L. acidophilus NCFM from filamentous to bacilloid rods, and the bacilloid cells were more resistant to freezing and storage at conventional freezer temperatures. The results suggest that the morphology of the L. acidophilus cell may be an important consideration in the preparation of freeze-stable culture concentrates.

Journal Article↗

PubMed2006

Effects of Lactobacillus acidophilus DSM13241 as a probiotic in healthy adult cats.

OBJECTIVE: To evaluate the effect of dietary supplementation with the probiotic strain Lactobacillus acidophilus DSM13241 in healthy adult cats. ANIMALS: 15 adult cats. PROCEDURES: Cats were fed a nutritionally complete dry food for 5 weeks. Fecal character was assessed daily, and a single fecal sample and 3-mL blood sample were collected for bacterial enumeration and hematologic analysis, respectively. Cats were then fed the same diet supplemented with L acidophilus DSM13241 (2 x 10(8) CFU/d) for 4.5 weeks. Repeat fecal and hematologic measurements were taken prior to the return to control diet for a 4-week period. RESULTS: The probiotic species was recovered from feces, demonstrating survival through the feline gastrointestinal tract. Probiotic supplementation was associated with increased numbers of beneficial Lactobacillus and L acidophilus groups in feces and decreased numbers of Clostridium spp and Enterococcus faecalis, indicating an altered bacterial balance in the gastrointestinal tract microflora. Fecal pH was also decreased suggesting a colonic environment selective for the beneficial lactic acid bacterial population. Systemic and immunomodulatory effects were associated with administration of L acidophilus DSM13241 including altered cell numbers within WBC subsets and enhanced phagocytic capacity in the peripheral granulocyte population. In addition, plasma endotoxin concentrations were decreased during probiotic feeding, and RBCs had a decreased susceptibility to osmotic pressure. CONCLUSIONS AND CLINICAL RELEVANCE: Probiotic strain L acidophilus DSM13241 fed at 2 x 10(8) CFU/d can alter the balance of gastrointestinal microflora in healthy cats. Furthermore, administration of this probiotic results in beneficial systemic and immunomodulatory effects in cats.

Animal Feed↗

PubMed2000

Effect of some nutritional and environmental parameters on the production of diacetyl and on starch consumption by Pediococcus pentosaceus and Lactobacillus acidophilus in submerged cultures.

Three series of 5-day submerged cultures with Pediococcus pentosaceus MITJ-10 and Lactobacillus acidophilus Hansen 1748 were carried out in starch-based media, and the effect of cultural factors on the changes of starch, diacetyl and amylase activity determined. In axenic cultures, Ped. pentosaceus MITJ-10 produced more diacetyl (63.27 mg l(-1)) by adding glucose, yeast extract and CaCO3 (P < 0.01), at 28 degrees C (P < 0.05); but more starch was consumed (18.4 g l(-1)) in the absence of glucose (P < 0.01). Lact. acidophilus Hansen 1748 consumed more starch (26.56 g l(-1)) at 28 degrees C, with CaCO3, glucose (P < 0.01) and yeast extract (P < 0.05); however, the amylolytic activity (10077U l(-1)) was favoured at 35 degrees C (P < 0.01). Little starch was consumed in mixed cultures due to the low pH; nevertheless, diacetyl content rose to 135.76 mg l(-1) at 32 degrees C (P < 0.01). Therefore, both studied strains might be useful to produce aromatic extensors from starchy substrates. These natural aromatic extensors are of interest to the food industry.

Amylases↗

PubMed1998

Antagonistic activity against Helicobacter infection in vitro and in vivo by the human Lactobacillus acidophilus strain LB.

The purpose of the present study was to examine the activity of the human Lactobacillus acidophilus strain LB, which secretes an antibacterial substance(s) against Helicobacter pylori in vitro and in vivo. The spent culture supernatant (SCS) of the strain LB (LB-SCS) dramatically decreased the viability of H. pylori in vitro independent of pH and lactic acid levels. Adhesion of H. pylori to the cultured human mucosecreting HT29-MTX cells decreased in parallel with the viability of H. pylori. In conventional mice, oral treatment with the LB-SCS protected against infection with Helicobacter felis. Indeed, at both 8 and 49 days post-LB-SCS treatment (29 and 70 days postinfection), inhibition of stomach colonization by H. felis was observed, and no evidence of gastric histopathological lesions was found. LB-SCS treatment inhibits the H. pylori urease activity in vitro and in H. pylori that remained associated with the cultured human mucosecreting HT29-MTX cells. Moreover, a decrease in urease activity was detected in the stomach of the mice infected with H. felis and treated with LB-SCS.

Animals↗

PubMed2005

Reduction of Escherichia coli O157 in finishing beef cattle by various doses of Lactobacillus acidophilus in direct-fed microbials.

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.

Animals↗

PubMed2006

Similarity and differences in the Lactobacillus acidophilus group identified by polyphasic analysis and comparative genomics.

A set of lactobacilli were investigated by polyphasic analysis. Multilocus sequence analysis, DNA typing, microarray analysis, and in silico whole-genome alignments provided a remarkably consistent pattern of similarity within the Lactobacillus acidophilus complex. On microarray analysis, 17 and 5% of the genes from Lactobacillus johnsonii strain NCC533 represented variable and strain-specific genes, respectively, when tested against four independent isolates of L. johnsonii. When projected on the NCC533 genome map, about 10 large clusters of variable genes were identified, and they were enriched around the terminus of replication. A quarter of the variable genes and two-thirds of the strain-specific genes were associated with mobile DNA. Signatures for horizontal gene transfer and modular evolution were found in prophages and in DNA from the exopolysaccharide biosynthesis cluster. On microarray hybridizations, Lactobacillus gasseri strains showed a shift to significantly lower fluorescence intensities than the L. johnsonii test strains, and only genes encoding very conserved cellular functions from L. acidophilus hybridized to the L. johnsonii array. In-silico comparative genomics showed extensive protein sequence similarity and genome synteny of L. johnsonii with L. gasseri, L. acidophilus, and Lactobacillus delbrueckii; moderate synteny with Lactobacillus casei; and scattered X-type sharing of protein sequence identity with the other sequenced lactobacilli. The observation of a stepwise decrease in similarity between the members of the L. acidophilus group suggests a strong element of vertical evolution in a natural phylogenetic group. Modern whole-genome-based techniques are thus a useful adjunct to the clarification of taxonomical relationships in problematic bacterial groups.

Genetic Variation↗

PubMed2006

Improvement of protein production in lactic acid bacteria using 5'-untranslated leader sequence of slpA from Lactobacillus acidophilus. Improvement in protein production using UTLS.

The 5'-untranslated leader sequence (UTLS) of the slpA gene from Lactobacillus acidophilus contributes to mRNA stabilization by producing a 5' stem and loop structure, and a high-level expression system for the lactic acid bacteria was developed using the UTLS in this study. A plasmid, which expresses alpha-amylase under the control of the ldh promoter, was constructed by integrating the core promoter sequence with the UTLS. The role of the UTLS in increasing the copies of the alpha-amylase mRNA was proved by measuring alpha-amylase activity in the culture supernatant and the relative expression of alpha-amylase mRNA was determined by the quantitative real-time PCR analysis. Moreover, several expression systems were constructed by combining the core promoter sequence with the UTLS or with the partially deleted UTLS and the expression level was evaluated. The use of the UTLS led to the success in improving alpha-amylase expression in the two strains of Lactobacillus casei and Lactococcus lactis. The current study showed that the improvement in protein production using the UTLS could be applied to the expression system in the lactic acid bacteria.

5' Untranslated Regions↗

PubMed1980

Genetic heterogeneity among Lactobacillus acidophilus strains.

Physiological characteristics, DNA base composition (% GC) and DNA-DNA reassociation values were determined for 138 Lactobacillus acidophilus strains. Twenty seven strains were received from various culture collections and 111 strains were freshly isolated during a study on the composition of the intestinal lactic microflora of piglets and suckling calves. All strains had physiological characteristics which were substantially similar. The strains isolated from pigs were unable to ferment trehalose. The % GC ranged from 35.8 to 43.4. On the basis of the results of DNA-DNA hybridization the strains were divided into four genetic groups.

Animals↗

PubMed1999

Identification of the pH-inducible, proton-translocating F1F0-ATPase (atpBEFHAGDC) operon of Lactobacillus acidophilus by differential display: gene structure, cloning and characterization.

The influence of low pH on inducible gene expression in Lactobacillus acidophilus was investigated by the use of differential display. Logarithmic phase cultures were exposed to pH 3.5 for various intervals, and RNA was isolated and reverse transcribed. The resultant cDNAs were subjected to PCR and the products were resolved by electrophoresis. Several cDNA products were induced after exposure to pH 3.5. One of these products, a 0.7 kb fragment, showed sequence similarity to bacterial atpBEF genes of the atp operon, whose genes encode the various subunits of the F1F0-ATPase. With the 0.7 kb differential display product as a probe, hybridizations with total RNA from untreated and acid-treated L. acidophilus verified the acid inducibility of this operon. The increase in atp mRNA induced by low pH was accompanied by an increase in the activity of the enzyme in membrane extracts. The full-length atp operon was sequenced, and its genes were in the order of atpBEFHAGDC, coding for the a, c, b, delta, alpha, gamma, beta and epsilon subunits respectively. The operon contained no i gene, but was preceded by a 122 bp intergenic space, which contained putative extended -10 and -35 promoter regions. Primer extension analysis of RNA from cultures that were shifted from pH 5.6 to pH 3. 5, and held for 0, 30 or 45 min, revealed that the transcriptional start site did not change position as a function of culture pH or time after exposure to pH 3.5. The primary structure and genetic organization indicated that the H+-ATPase of L. acidophilus is a typical F1F0-type ATPase. The similarity to streptococcal ATPases and the acid inducibility of this operon suggest that it may function in the ATP-dependent extrusion of protons and maintenance of cytoplasmic pH. Finally, the use of differential display RT-PCR was an effective approach to identify genes in L. acidophilus induced by an environmental stimulus.

Amino Acid Sequence↗

PubMed1980

Production of Hydrogen peroxide by Lactobacillus acidophilus.

Hydrogen peroxide production in reconstituted skim milk (10%) and low-fat milk by four strains lf Lactobacillus acidophilus was studied at 37 and 4 C. Strains A and B produced little, but strains C and D produced larger amounts, especially if agitated continuously during growth at 37 C or storage at 4 C. Continuous shaking was required at 4 C for C or D (4.0 X 10(8)/ml) to produce sufficient hydrogen peroxide to retard growth of Pseudomonas fragi. Flavin adenine dinucleotide stimulated the oxidation of reduced nicotinamide adenine dinucleotide by dialyzed cell-free extracts of C and D, which indicated that the reduced nicotinamide adenine dinucleotide oxidases of these strains produce hydrogen peroxide as an end product.

Animals↗

PubMed1975

Participation of exogenous thymine and thymidine in deoxyribonucleic acid synthesis in Lactobacillus acidophilus.

The degree of participation (DP) of exogenous thymidine and thymine in overall DNA synthesis was studied in Lactobacillus acidophilus R-26. The DP of thymidine remains constant under a variety of conditions (except at low thymidine concentrations, when the DP is influenced by deoxyribonucleosides and folic acid). A 5-bromodeoxyuridine-resistant mutant was isolated, which displayed cross-resistance to 5-bromouracil and a significantly lower DP of thymidine than the parental strain. Thymine was poorly incorporated in the parental strain even in the presence of deoxyribosides. The results of this investigation would be compatible with the possibility of an alternative pathway for thymidylate synthesis other than the known thymidylate synthase pathway.

Bromodeoxyuridine↗