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Effect of SnF2, administered as mouthrinses or topically applied, on Streptococcus mutans, Streptococcus sanguis and lactobacilli in dental plaque and saliva.

Mouthrinsing with SnF2 reduced the Streptococcus mutans population in plaque and saliva and the proportion of Streptococcus sanguis in plaque. The effect was of short duration: 2 weeks after treatment the values of S. mutans in plaque and saliva were even higher than the pretreatment values. Topical SnF2 applications reduced the S. mutants population in plaque and saliva but did not reduce the proportion of S. sanguis in plaque. The effect was more prolonged : 4 weeks after treatment the S. mutans population in interproximal plaque remained significantly reduced and the salivary levels of the organism had not fully returned to pretreatment levels. Both SnF2 treatments significantly increased the salivary levels of lactobacilli. The values of lactobacilli in saliva remained significantly increased 4 weeks after the SnF2 mouthrinsing but had almost returned to pretreatment levels within 2 weeks after the topical SnF2 applications. The findings suggest that the cariogenic potential of dental plaque is differently affected depending on whether a drug is administered as a mouthrinse or is applied topically.

Bacteria↗

Oral glucose clearance in subjects with high or low salivary levels of Streptococcus mutans and lactobacilli.

The ability to eliminate sugar from the oral cavity was studied in subjects with high or low numbers of salivary Streptococcus mutans and lactobacilli. The glucose concentration in saliva was determined after consumption of biscuits using the Gloxtest, and the logarithms of the glucose values were plotted against time. The sugar clearance time was measured where the slope of the plot intersected with a glucose level of 1 mg/ml. Subjects with a high level of S. mutans had a significantly longer clearance time than subjects with a low level. More salivary lactobacilli were found in a subject group with a long sugar clearance time than in a group with a fast clearance rate.

Adolescent↗

Proportions of Streptococcus mutans, lactobacilli and Actinomyces spp in root surface plaque.

The incidence of Streptococcus mutans, lactobacilli and Actinomyces was examined in root surface dental plaque from 24 subjects with root surface caries and in 24 subjects with exposed root surfaces as a consequence of periodontitis but without root surface caries. Plaque was collected from the most posterior teeth in each quadrant: in the caries group from carious lesions and in the periodontitis group from buccal root surfaces. There were significantly higher proportions of S. mutans and lactobacilli in dental plaque samples taken from subjects suffering from root surface caries than in samples from subjects without root surface caries. The level of the group A. viscosus/naeslundii in the periodontitis group was higher than that found in the root surface caries group, but the difference was not statistically significant.

Actinomyces↗

Effects of Weight Watchers' diet on salivary secretion rate, buffer effect and numbers of mutans streptococci and lactobacilli.

The aim of this investigation was to study how a change to the dietary regimen of the Weight Watchers' (WW) affected salivary secretion rate, buffer effect and number of cariogenic bacteria. Thirty-three women joining the WW were saliva sampled at the evening prior the day of the start of the dietary regimen and after 4 and 8 wk on the dietary regimen. No significant changes were found regarding saliva secretion rate and buffer effect. A significant decrease of salivary numbers of mutans streptococci and lactobacilli occurred. Between the 4th and 8th wk of dietary regimen an increase of cariogenic bacteria occurred but not back to the baseline values. At baseline 15 of the WW carried Streptococcus sobrinus in addition to Streptococcus mutans. The total number of salivary mutans streptococci and lactobacilli was significantly higher in those subjects. In conclusion the dietary regimen of the WW, based on food choice and distributions of meals in agreement with a non-cariogenic diet, reduced the number of cariogenic bacteria in most subjects, but not in all, while the secretion rate and buffer effect remained the same.

Adult↗

Mutans streptococci and lactobacilli in the elderly.

The prevalence of salivary mutans streptococci and lactobacilli, salivary flow rate, and the type of dentition were studied in connection with a medical survey of 76-, 81-, and 86-yr-old inhabitants of Helsinki living at home. High bacterial counts were associated with low salivary flow rates. High counts of mutans streptococci (> 10(6) CFU/ml) were found in 68% of wearers of full dentures, as compared with 53% of subjects having natural teeth. High counts of lactobacilli (< 10(6) CFU/ml) were found in 44% of subjects having removable partial dentures and in 39% of subjects having natural teeth. The bacterial counts correlated neither with medicines taken daily nor with diseases among this population.

Aged↗

Inconsistent levels of mutans streptococci and lactobacilli measured in stimulated whole saliva.

The counts of mutans streptococci (MS) and lactobacilli (LB) in saliva have been used for prediction of dental caries. The results have not justified putting the methods into clinical practice as the explanatory values of the bacteria for caries are low. There is therefore reason to question how representative the methods for sampling and estimation of those bacteria are. The aims of this study were to investigate if the variation in caries is better explained by the bacteria, estimated as means of numbers in double saliva samples, than by numbers in single samples, and to determine whether chewing on either the preferred or the non-preferred side influences the bacterial counts. Twenty-five healthy subjects were included in the study. Two saliva samples were collected with a 1-wk interval by chewing on a piece of sterile paraffin wax on the preferred side only. One month later this was repeated on the opposite side. In order to test if there were variations in the numbers of bacteria due to the laboratory methods, a paired cultivation of 20 samples was carried out. The results showed that the coefficients of variation (c.v.) for the differences between the individual double samples varied between 110 and 276% of the total mean of the double samples. The paired cultivation gave r values for MS of 0.93 and for LB of 0.99. Using double samples did not increase the explanatory values for caries. There were higher explanatory values for caries on the preferred side than on the non-preferred side, this being much more pronounced for lactobacilli than for mutans streptococci.

Adult↗

Metabolism of some drugs by intestinal lactobacilli and their toxicological considerations.

The bacterial metabolism of three drugs (sulphasalazine, phthalylsulphathiazole and chloramphenicol palmitate) and two dyes (tartrazine and methyl red) has been studied using a resting culture technique. The strains used were isolated intestinal lactobacilli, E. coli and mixed cultures of faeces of mice. As high as 94.3% degradation of sulphasalazine was found with a strain of Lactobacillus acidophilus. The highest degradation of phthalylsulphathiazole and chloramphenicol palmitate was found to be 17.6% and 8%, respectively. A kinetic study was conducted on the rate of degradation of sulphasalazine, phthalylsulphathiazole and methyl red. The toxicological aspects of degradation products in relation to the use of lactobacilli as dietary supplement or therapeutic aid are discussed.

Animals↗

Transient colonization of the gut of newborn infants by orally administered bifidobacteria and lactobacilli.

We investigated if orally administered bifidobacteria and/or lactobacilli could be cultured from faeces of infants after antibiotic treatment, when these bacterial species are usually absent. Lyophilized Bifidobacterium longum, strain BB-536, B. breve, strain BB-576, or Lactobacillus acidophilus, strain LAC-343, were used. Doses of 3 x 10(9) cells of one strain, or a mixture of all three strains 3 x 10(9) cells each were fed three times daily at mealtimes to 11 infants aged 0-8 weeks. Treatment was started the first day after antibiotic treatment and was continued for 5 days. The bacterial species were isolated in 9 of 11, 7 of 10 and 2 of 9 specimens obtained on the last day of bifidobacteria or lactobacilli administration, 5 and 15 days thereafter, respectively. No side effects were noted.

Administration, Oral↗

Lactobacilli isolated from the stomach of conventional mice.

Twenty strains of lactobacilli isolated from the stomach of conventional mice were tested for their ability to ferment or hydrolyze substrates that may be present in the stomach habitat. The lactobacilli could be placed in four groups (A to D) depending on their ability to ferment N-acetylglucosamine, dextrin, cellobiose, gum arabic, and xylan. The majority of the isolates belonged to groups A and D. Group A strains did not resemble previously described Lactobacillus species, but group D strains were identified as L. leichmannii. A representative group A isolate colonized the surface of the nonsecretory epithelium of the stomach of gnotobiotic mice; a group D isolate did not.

Agglutination↗

Gas chromatography analysis of cellular fatty acids and neutral monosaccharides in the identification of lactobacilli.

Cellular fatty acids and monosaccharides in a group of 14 lactobacilli were analyzed by gas chromatography and the identity of the components was confirmed by gas chromatography-mass spectrometry. From the same bacterial sample, both monosaccharides and fatty acids were liberated by methanolysis, and in certain experiments, fatty acids alone were released by basic hydrolysis. The results indicate that basic hydrolysis gave more comprehensive information about the fatty acids, but the analysis of monosaccharides was found to be much more useful in distinguishing between different species of lactobacilli. The method described allowed differentiation of 11 of 14 Lactobacillus species, and even single colonies isolated from agar plates could be used for analysis without subculturing.

Chromatography, Gas↗

Lactobacilli and azoreductase activity in the murine cecum.

Azoreductase activity in the ceca of mice lacking lactobacilli as members of the normal microflora (reconstituted-lactobacillus-free [RLF] mice) was compared with that of RLF mice whose gastrointestinal tracts were colonized by strains of Lactobacillus delbrueckii and Lactobacillus fermentum. Azoreductase activity was 31% lower in the ceca of mice colonized by lactobacilli.

Animals↗

Deconjugation of bile acids by lactobacilli in the mouse small bowel.

Conjugated and unconjugated bile acid concentrations in the small bowel contents and portal serum samples collected from mice with and without lactobacilli (RLFL and RLF, respectively) as gastrointestinal inhabitants were measured. The major portion of bile acids in the small bowel contents of RLFL mice was unconjugated (67.9%) in contrast to that of RFL animals in which a smaller portion of bile acids was unconjugated (23.5%). This study demonstrated that bile salt hydrolase produced by lactobacilli is active under the conditions prevailing in the proximal small bowels of mice.

Amidohydrolases↗

Effects of lactobacilli on yeast-catalyzed ethanol fermentations.

Normal-gravity (22 to 24 degrees Plato) wheat mashes were inoculated with five industrially important strains of lactobacilli at approximately 10(5), approximately 10(6), approximately 10(7), approximately 10(8), and approximately 10(9) CFU/ml in order to study the effects of the lactobacilli on yeast growth and ethanol productivity. Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus #3, Lactobacillus rhamnosus, and Lactobacillus fermentum were used. Controls with yeast cells but no bacterial inoculation and additional treatments with bacteria alone inoculated at approximately 10(7) CFU/ml of mash were included. Decreased ethanol yields were due to the diversion of carbohydrates for bacterial growth and the production of lactic acid. As higher numbers of the bacteria were produced (depending on the strain), 1 to 1.5% (wt/vol) lactic acid resulted in the case of homofermentative organisms. L. fermentum, a heterofermentative organism, produced only 0.5% (wt/vol) lactic acid. When L. plantarum, L. rhamnosus, and L. fermentum were inoculated at approximately 10(6) CFU/ml, an approximately 2% decrease in the final ethanol concentration was observed. Smaller initial numbers (only 10(5) CFU/ml) of L. paracasei or Lactobacillus #3 were sufficient to cause more than 2% decreases in the final ethanol concentrations measured compared to the control. Such effects after an inoculation of only 10(5) CFU/ml may have been due to the higher tolerance to ethanol of the latter two bacteria, to the more rapid adaptation (shorter lag phase) of these two industrial organisms to fermentation conditions, and/or to their more rapid growth and metabolism. When up to 10(9) CFU of bacteria/ml was present in mash, approximately 3.8 to 7.6% reductions in ethanol concentration occurred depending on the strain. Production of lactic acid and a suspected competition with yeast cells for essential growth factors in the fermenting medium were the major reasons for reductions in yeast growth and final ethanol yield when lactic acid bacteria were present.

Ethanol↗

Analysis of fecal populations of bifidobacteria and lactobacilli and investigation of the immunological responses of their human hosts to the predominant strains.

The bifidobacterial and lactobacillus populations of fecal samples collected from 10 human subjects were studied. The numbers of bifidobacteria were similar in the fecal samples of all of the subjects, but lactobacillus numbers varied, even between samples collected from the same individual. Analysis of the composition of the bacterial populations by ribotyping and pulsed-field gel electrophoresis to differentiate between strains showed that, at least for the numerically predominant strains, each subject harbored a unique collection of bifidobacteria and lactobacilli. Predominant bifidobacterial and lactobacillus strains detected in the feces of each subject were used in immunological assays (lymphocyte transformation, serum antibody titers) to determine the influence of the bacteria on the immune system of their host. Immunoglobulin G antibodies reactive with lactobacilli were detected at high concentrations; antibodies reactive with bifidobacteria were present at lower concentrations. The antibodies appeared to be genus specific rather than strain specific. The results of the study emphasized the complexity of the relationship that exists between the intestinal microflora and the human host.

Adult↗

Urea hydrogen peroxide reduces the numbers of lactobacilli, nourishes yeast, and leaves no residues in the ethanol fermentation.

Urea hydrogen peroxide (UHP) at a concentration of 30 to 32 mmol/liter reduced the numbers of five Lactobacillus spp. (Lactobacillus plantarum, L. paracasei, Lactobacillus sp. strain 3, L. rhamnosus, and L. fermentum) from approximately 10(7) to approximately 10(2) CFU/ml in a 2-h preincubation at 30 degrees C of normal-gravity wheat mash at approximately 21 g of dissolved solids per ml containing normal levels of suspended grain particles. Fermentation was completed 36 h after inoculation of Saccharomyces cerevisiae in the presence of UHP, even when wheat mash was deliberately contaminated (infected) with L. paracasei at approximately 10(7) CFU/ml. There were no significant differences in the maximum ethanol produced between treatments when urea hydrogen peroxide was used to kill the bacteria and controls (in which no bacteria were added). However, the presence of L. paracasei at approximately 10(7) CFU/ml without added agent resulted in a 5.84% reduction in the maximum ethanol produced compared to the control. The bactericidal activity of UHP is greatly affected by the presence of particulate matter. In fact, only 2 mmol of urea hydrogen peroxide per liter was required for disinfection when mashes had little or no particulate matter present. No significant differences were observed in the decomposition of hydrogen peroxide in normal-gravity wheat mash at 30 degrees C whether the bactericidal agent was added as H(2)O(2) or as urea hydrogen peroxide. NADH peroxidase activity (involved in degrading H(2)O(2)) increased significantly (P = 0.05) in the presence of 0.75 mM hydrogen peroxide (sublethal level) in all five strains of lactobacilli tested but did not persist in cells regrown in the absence of H(2)O(2). H(2)O(2)-resistant mutants were not expected or found when lethal levels of H(2)O(2) or UHP were used. Contaminating lactobacilli can be effectively managed by UHP, a compound which when used at ca. 30 mmol/liter happens to provide near-optimum levels of assimilable nitrogen and oxygen that aid in vigorous fermentation performance by yeast.

Carbamide Peroxide↗

Colonization of the stratified squamous epithelium of the nonsecreting area of horse stomach by lactobacilli.

Selective adhesion to only certain epithelia is particularly common among the bacterial members of the indigenous microflora of mammals. We have found that the stratified squamous epithelium of the nonsecreting area of horse stomach is colonized by gram-positive rods. The microscopic features of a dense layer of these bacteria on the epithelium were found to be similar to those reported in mice, rats, and swine. Adhering microorganisms were isolated and identified as Lactobacillus salivarius, L. crispatus, L. reuteri, and L. agilis by DNA-DNA hybridization and 16S rRNA gene sequencing techniques. These lactobacilli associated with the horse, except for L. reuteri, were found to adhere to horse epithelial cells in vitro but not to those of rats. A symbiotic relationship of these lactobacilli with the horse is suggested.

Animals↗

Inhibition of Giardia intestinalis by extracellular factors from Lactobacilli: an in vitro study.

The aim of the present work was to evaluate the effect of spent culture supernatants of different strains of lactobacilli on giardia trophozoites. The growth of Giardia intestinalis strain WB, as well as the attachment to the human intestinal epithelial cell line Caco-2, was evaluated by using proliferation and adhesion assays with radiolabeled parasites. In addition, scanning electron microscopy and flow cytometric analysis were performed. The effect of spent culture supernatants from lactobacilli was strain dependent. Lactobacillus johnsonii La1 significantly inhibited the proliferation of G. intestinalis trophozoites. Although the effect was strongly pH dependent, it was not simply due to lactic acid. According to flow cytometric analysis, trophozoites were arrested in G(1) phase but neither significant necrosis nor apoptosis could be detected. Bacterial cells or their spent culture supernatants were unable to modify trophozoite attachment to Caco-2 cells. However, trophozoites treated with spent culture supernatants had little, if any, proliferative capacity. These results suggest that La1 produces some substance(s) able to inhibit proliferation of Giardia trophozoites. Partial characterization of the factors involved in the antigiardiasic action showed that they have a low molecular mass and are inactivated by heating. On this basis, it seems worthwhile to explore how colonization of the proximal small bowel with these lactic acid bacteria could interfere with giardiasis in vivo.

Animals↗

Microbiological characterization of wet wheat distillers' grain, with focus on isolation of lactobacilli with potential as probiotics.

Wet wheat distillers' grain (WWDG), a residue from ethanol fermentation, was examined from a microbiological perspective. After storage, WWDG was characterized by a high content of lactobacilli, nondetectable levels of other bacteria, occasional occurrence of yeasts, and a pH of about 3.6 and contained a mixture of lactic acid, acetic acid, and ethanol. The composition of lactobacilli in WWDG was simple, including primarily the species Lactobacillus amylolyticus, Lactobacillus panis, and Lactobacillus pontis, as determined by 16S rRNA gene sequencing. Since the use of WWDG as pig feed has indicated a health-promoting function, some relevant characteristics of three strains of each of these species were examined together with basal physiological parameters, such as carbohydrate utilization and growth temperature. Seven of the strains were isolated from WWDG, and two strains from pig feces were included for comparison. It was clear that all three species could grow at temperatures of 45 to 50 degrees C, with L. amylolyticus being able to grow at temperatures as high as 54 degrees C. This finding could be the explanation for the simple microflora of WWDG, where a low pH together with a high temperature during storage would select for these organisms. Some strains of L. panis and L. pontis showed prolonged survival at pH 2.5 in synthetic stomach juice and good growth in the presence of porcine bile salt. In addition, members of all three species were able to bind to immobilized mucus material in vitro. Especially the isolates from pig feces but, interestingly, some isolates from WWDG as well possessed properties that might be of importance for colonization of the gastrointestinal tracts of pigs.

Animal Feed↗