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Intrinsic properties of so-called dormant probiotic bacteria, determined by flow cytometric viability assays.

Plate counting and four culture-independent flow cytometric assays were used to determine the viability and intrinsic properties of three probiotic strains during storage. The strains showed reduction in plate counts but were able to maintain esterase activity, intact cytoplasmic membrane, and pH gradient. The apparently uncultivable probiotic cells were active and stress resistant.

Animals↗

Specific response of a novel and abundant Lactobacillus amylovorus-like phylotype to dietary prebiotics in the guts of weaning piglets.

Using 16S rRNA gene-based approaches, we analyzed the responses of ileal and colonic bacterial communities of weaning piglets to dietary addition of four fermentable carbohydrates (inulin, lactulose, wheat starch, and sugar beet pulp). An enriched diet and a control diet lacking these fermentable carbohydrates were fed to piglets for 4 days (n = 48), and 10 days (n = 48), and the lumen-associated microbiota were compared using denaturing gradient gel electrophoresis (DGGE) analysis of amplified 16S rRNA genes. Bacterial diversities in the ileal and colonic samples were measured by assessing the number of DGGE bands and the Shannon index of diversity. A higher number of DGGE bands in the colon (24.2 +/- 5.5) than in the ileum (9.7 +/- 4.2) was observed in all samples. In addition, significantly higher diversity, as measured by DGGE fingerprint analysis, was detected in the colonic microbial community of weaning piglets fed the fermentable-carbohydrate-enriched diet for 10 days than in the control. Selected samples from the ileal and colonic lumens were also investigated using fluorescent in situ hybridization (FISH) and cloning and sequencing of the 16S rRNA gene. This revealed a prevalence of Lactobacillus reuteri in the ileum and Lactobacillus amylovorus-like populations in the ileum and the colon in the piglets fed with fermentable carbohydrates. Newly developed oligonucleotide probes targeting these phylotypes allowed their rapid detection and quantification in the ileum and colon by FISH. The results indicate that addition of fermentable carbohydrates supports the growth of specific lactobacilli in the ilea and colons of weaning piglets.

Animals↗

Simplified gas chromatographic procedure for identification of bacterial metabolic products.

A rapid and simple procedure is described for analysis of fermentation products from anaerobic bacteria grown in glucose broth. A 1-ml sample of the culture is drained through cation-exchange resin in a Pasteur pipette. The effluent fluid is directly analyzed isothermally in a gas chromatograph for volatile fatty acids (C(2) to C(6)) as well as for lactic, pyruvic, and succinic acids. This procedure is considered to be suitable for routine use in clinical bacteriology.

Anaerobiosis↗

Cytological and deoxyribonucleic acid-deoxyribonucleic acid hybridization studies on lactobacillus isolates from San Francisco sourdough.

Molecular taxonomic and electron microscopy studies were performed on four bacterial isolates obtained from different sources of San Francisco sourdough (SD). These bacteria were first isolated by Kline and Sugihara who tentatively described them as a previously unreported species of heterofermentative Lactobacillus; they suggested the name Lactobacillus sanfrancisco. The guanine plus cytosine base composition (%GC) of the deoxyribonucleic acid (DNA) ranged from 38 to 40%. The possible genetic relatedness of these SD isolates to five known species of Lactobacillus with comparable GC contents was assessed in the present work by means of DNA-DNA hybridization competition experiments. Little or no DNA homology was observed between the SD bacteria and the known species. The SD bacteria exhibited a high degree of homology (>88%) among themselves, suggesting that the four isolates were identical taxonomically. Also, the electron photomicrographs revealed structures similar to those of gram-positive bacilli. Accordingly, since these SD isolates have the characteristic phenotypic and morphological properties of the genus Lactobacillus and are not related genetically to any known species, the tentative characterization by the above workers of these isolates as a new species is substantiated.

Bread↗

Analysis of Corynebacterium vaginale by an immunodiffusion technique.

An immunodiffusion technique was employed to study the antigenic relationship of Corynebacterium vaginale isolates, vaginal diphtheroids, and members of the genera Corynebacterium and Lactobacillus. Antisera were prepared against C. vaginale ATCC strain 14018 grown diphasically and on blood agar plates and were tested against extracts of organisms prepared by sonication. Ouchterlony analysis demonstrated that all of the isolates of C. vaginale examined possess a common antigenic determinant. No antigenic relationship was detected between C. vaginale and members of the genera Corynebacterium or Lactobacillus. This study also demonstrated that alterations in the cultural conditions can cause variations in the antigenic composition of C. vaginale.

Agar↗

Effects of feeding a probiotic preparation (SIM) containing inulin on the severity of colitis and on the composition of the intestinal microflora in HLA-B27 transgenic rats.

An overly aggressive immune response to the intestinal microflora in a genetically susceptible host background has been implicated in the pathogenesis of inflammatory bowel diseases. We measured the impact of a probiotic preparation (SIM) containing inulin on the severity of colitis and on intestinal microflora profiles of HLA-B27-beta(2)-microglobulin transgenic (TG) rats. SIM is a mixture of lactobacilli, bifidobacteria, and inulin. Two-month-old TG rats received either SIM or water. Control TG rats received metronidazole, alone or in combination with SIM, for 8 weeks. Nontransgenic rats received SIM or water. The cecal content was removed for analysis of the intestinal microflora by PCR combined with denaturing gradient gel electrophoresis. The colon was scored for histological evidence of inflammation, colonic myeloperoxidase activity and interleukin-1beta RNA levels were measured photometrically or by real-time quantitative PCR. At 4 months, the colonic inflammation of TG rats treated with SIM was histologically diminished compared to that in untreated TG rats (2.2 +/- 0.2 versus 2.9 +/- 0.1; P </= 0.03). The administration of SIM altered the microflora profiles of TG rats by increasing the diversity and stimulating specifically the growth of Bifidobacterium animalis. The probiotic bacteria added to SIM were below the detection level in cecal stool samples at the end of the study period. The administration of SIM resulted in a measurable impact on the cecal microflora profiles of TG rats with attenuation of colitis. The lack of detection of any added probiotic bacteria in the cecal content suggests that prebiotic inulin is the major effective compound.

Animals↗

Probiotics reduce enterohemorrhagic Escherichia coli O157:H7- and enteropathogenic E. coli O127:H6-induced changes in polarized T84 epithelial cell monolayers by reducing bacterial adhesion and cytoskeletal rearrangements.

The aim of this study was to determine if probiotics reduce epithelial injury following exposure to Escherichia coli O157:H7 and E. coli O127:H6. The pretreatment of intestinal (T84) cells with lactic acid-producing bacteria reduced the pathogen-induced drop in transepithelial electrical resistance. These findings demonstrate that probiotics prevent epithelial injury induced by attaching-effacing bacteria.

Actinin↗

Deoxyribonucleic acid base composition of the genus Lactobacillus.

Deoxyribonucleic acids of 45 strains of Lactobacillus and 5 strains of Bifidobacterium which had been analyzed for base composition by chromatographic means were examined at equilibrium in a CsCl density gradient. Regression analysis showed that there have been systematic errors involved in the estimation of guanine plus cytosine (GC) content by the chemical method, and that the relation between buoyant density and base composition is indeed linear and best fitted by the equation GC = 10.309 (rho-1.662), which compares well in slope with the equation of Schildkraut, Marmur, and Doty. With the improved data obtained in this study, the specific groupings of the species of both genera were reevaluated.

Centrifugation, Density Gradient↗

Alteration in bacterial morphology by optochin and quinine hydrochlorides.

Incubation of washed bacterial and ribosomal suspensions with optochin or quinine hydrochloride caused an increase in the turbidity of the suspensions and the appearance of electron-dense cytoplasmic aggregates in the treated cells. These effects were more pronounced with optochin hydrochloride than with quinine hydrochloride, and they did not correlate with the relative sensitivities of different bacteria to growth inhibition by optochin or quinine.

Bacillus subtilis↗

Micronutrients (including zinc) reduce diarrhoea in children: the Pakistan Sprinkles Diarrhoea Study.

AIMS: To examine the effect of the daily use of micronutrients (including zinc) or the same micronutrients plus heat inactivated lactic acid bacteria (LAB), on diarrhoea in children compared to placebo. METHODS: A triple blind randomised clinical trial in an urban slum of Karachi, Pakistan. Micronutrients (including zinc), micronutrients (including zinc and LAB), or placebo, were provided daily for two months to 75 young children (aged 6-12 months) identified at high risk for diarrhoea related mortality on the basis of history of at least one episode of diarrhoea in the preceding two weeks. The longitudinal prevalence of diarrhoea was defined as the percentage of days a child had diarrhoea out of the days the child was observed. RESULTS: Mean longitudinal prevalence of diarrhoea in the micronutrient-zinc group was 15% (SD = 10%) child-days compared to 26% (SD = 20%) child-days in the placebo group and 26% (SD = 19%) child-days in the micronutrient-zinc-LAB group. The difference between the micronutrient-zinc-LAB and placebo groups was not significant. CONCLUSION: The daily provision of micronutrients (including zinc) reduces the longitudinal prevalence of diarrhoea and thus may also reduce diarrhoea related mortality in young children; heat inactivated LAB has negative effects in these children.

Diarrhea, Infantile↗

Survival of lactococci during passage through mouse digestive tract.

One of the important properties of probiotics is the ability to survive in the intestine. There have been few studies on the probiotic property of lactococci, since they are formally not considered to be natural inhabitants of the intestine. To evaluate lactococci as probiotic bacteria, we investigated their ability to survive during gastric transit by in vitro and in vivo tests. When exposed to an in vitro simulated gastrointestinal environment, such as low pH and bile, only Lactococcus lactis subsp. lactis bv. diacetylactis N7 showed a moderate survival rate among the four strains tested. The tested strains were orally administered to mice, and intestinal passage of the ingested strains was monitored by two methods: antibiotics and PCR. Viable cells of strain N7 were recovered from feces within 24-48 h after administration but not at 72 h. Lactococcus lactis subsp. cremoris ATCC 19257, which had a poor survival rate in vitro test, was also detected at 12 h but not at 24 h. These results indicate that lactococci can reach the mouse intestine alive, but not colonize it. If administered daily, viable strain N7 may exist continuously in the intestine. The effect of strain N7 on intestinal microbial balance and on animal health will be the subject of a further study.

Animals↗