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PubMed2000

Cordycepin: selective growth inhibitor derived from liquid culture of Cordyceps militaris against Clostridium spp.

The growth responses of nine human intestinal bacteria to liquid culture of Cordyceps militaris Link. Pt. (Ascomycotina: Clavicipitaceae) collected from a pupa of Bombyx mori L. (Lepidoptera: Bombycidae) were examined using spectrophotometric and impregnated paper disk methods and compared to those of tetracycline and chloramphenicol, as well as those of Coptis japonica root-derived berberine chloride. The biologically active constituent of the cultures was characterized as cordycepin (3'-deoxyadenosine) by spectroscopic analysis. This compound revealed potent growth-inhibiting activity toward Clostridium paraputrificum and Clostridium perfringens at 10 microgram/disk without adverse effects on the growth of Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium adolescentis, Lactobacillus acidophilus, and Lactobacillus casei, whereas tetracycline and chloramphenicol inhibited the growth of these lactic acid-producing bacteria, clostridia and Escherichia coli. However, C. militaris-derived materials revealed no growth stimulation on the bifidobacteria and lactobacilli. These results may be an indication of at least one of the pharmacological actions of C. militaris. As a naturally occurring antibacterial agent, cordycepin could be useful as a new preventive agent against various diseases caused by clostridia.

Animals↗
PubMed2000

Stimulatory effect of honey on multiplication of lactic acid bacteria under in vitro and in vivo conditions.

The effect of honey and sucrose on lactic acid bacteria in vitro and in rat gut was studied to determine whether these organisms were affected differently by honey compared with sucrose. Under in vitro conditions, the number of Lactobacillus acidophilus and Lactobacillus plantarum counts increased 10-100 fold in the presence of honey compared with sucrose. Feeding of honey to rats also resulted in significant increase in counts of lactic acid bacteria. Although there was no significant difference in the counts of lactic acid bacteria in the small and large intestines of different groups, the honey-fed group showed a significant increase (P<0.05) in counts over the control and sucrose-fed animals. The results support the fact that consumption of honey has a beneficial effect on the physiological constitution of animals fed with it.

Animals↗
PubMed2001

Probiotics in foods not containing milk or milk constituents, with special reference to Lactobacillus plantarum 299v.

Lactic acid fermentation is the simplest and safest way of preserving food and has probably always been used by humans. Species such as Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus acidophilus, and Lactobacillus salivarius are common in the human mucosa, from the mouth to the rectum. In food, L. paracasei and L. rhamnosus are usually associated with dairy products whereas L. plantarum is found in fermented foods of plant origin. A probiotic food product containing no milk constituent was launched in Sweden in 1994. The product is a lactic acid fermented oatmeal gruel that is mixed in a fruit drink. It contains approximately 5 x 10(10) colony-forming units of L. plantarum 299v/L. The strain L. plantarum 299v originates from the human intestinal mucosa and has been shown in rats to decrease translocation, improve mucosal status, improve liver status, improve the immunologic status of the mucosa, and reduce mucosal inflammation. In humans, L. plantarum 299v can increase the concentration of carboxylic acids in feces and decrease abdominal bloating in patients with irritable bowel disease. It can also decrease fibrinogen concentrations in blood. Should probiotics be administrated through foods, the probiotic organism must remain vigorous in the food until consumption and the food must remain palatable, ie, the food carrier and the organism must suit each other. L. plantarum 299v not only affects the bacterial flora of the intestinal mucosa but may also regulate the host's immunologic defense. The mechanisms involved need to be clarified.

Animals↗
PubMed1994

Effects of H2O2-producing lactobacilli on Neisseria gonorrhoeae growth and catalase activity.

In the vagina and endocervix, Neisseria gonorrhoeae must interact with complex microflora. Among these are lactobacilli, which may inhibit the growth of gonococci. Lactobacillus acidophilus, which produce H2O2 (LB+), and L. acidophilus and Lactobacillus casei, which do not produce H2O2 (LB-), were coincubated with catalase-positive and -deficient strains of N. gonorrhoeae. When the incubation medium was maintained at pH 7.3, neither LB+ nor LB- affected gonococcal growth. However, LB+ caused a significant increase in expression of gonococcal catalase, which could be offset by exposure of the bacteria to exogenous catalase. When coincubation medium was at lower pH (4.8-5.0), there was a significant decrease in gonococcal survival and catalase activity, which was only partly reversed by exogenous catalase. Lysates of LB+ also effectively inhibited gonococcal catalase. This inhibition was retained upon heating of the lysate to 100 degrees C for 15 min but was lost with proteinase K treatment. Thus, LB+ may inhibit growth of gonococci by acidification of the environment, secretion of H2O2, and production of protein inhibitors.

Catalase↗
PubMed2005

Lactobacillus suntoryeus sp. nov., isolated from malt whisky distilleries.

Eight strains of Lactobacillus with identical partial 16S rRNA gene sequences and similar randomly amplified polymorphic DNA patterns were isolated from fermentation samples from Japanese and Scottish malt whisky distilleries. Phylogenetic analysis of almost complete 16S rRNA gene sequences from three representative strains (two from Japan, one from Scotland) placed them in the genus Lactobacillus as members of the Lactobacillus acidophilus group. Lactobacillus helveticus and Lactobacillus gallinarum were the most closely related species, with 16S rRNA gene similarities of 99.3 and 98.1 %, respectively. A similar phylogeny was derived from partial sequences of elongation factor Tu (tuf) genes in which the alleles from the three distillery isolates were identical and shared 99.0 % similarity with L. helveticus and L. gallinarum tuf genes. S-layer (slp) gene sequences suggested different relationships among the strains and the distillery isolates no longer formed a monophyletic group. The alleles from the Japanese and Scottish strains shared only 54 % similarity. Chromosomal DNA from the distillery strains gave DNA-DNA hybridization values between 79 and 100 % but showed less than 43 and 22 % reassociation with L. helveticus and L. gallinarum DNA, respectively. The name Lactobacillus suntoryeus sp. nov. is proposed for this novel taxon; the type strain is strain SAT (=LMG 22464T=NCIMB 14005T).

Alcoholic Beverages↗
PubMed2006

Antagonistic activity of probiotic lactobacilli and bifidobacteria against entero- and uropathogens.

AIM: To develop in vitro assays for comparing the antagonistic properties and anti-oxidative activity of probiotic Lactobacillus and Bifidobacterium strains against various entero- and urinary pathogens. METHODS AND RESULTS: The antagonistic activity of five probiotic lactobacilli (Lactobacillus rhamnosus GG, Lactobacillus fermentum ME-3, Lactobacillus acidophilus La5, Lactobacillus plantarum 299v and Lactobacillus paracasei 8700:2) and two bifidobacteria (Bifidobacterium lactis Bb12, Bifidobacterium longum 46) against six target pathogens was estimated using different assays (solid and liquid media, anaerobic and microaerobic cultivation) and ranked (low, intermediate and high). Bacterial fermentation products were determined by gas chromatography, and the total anti-oxidative activity of probiotics was measured using linolenic acid test. Pyelonephritic Escherichia coli was highly suppressed by GG and both bifidobacteria strains. Lactobacilli strains 8700:2, 299v and ME-3 were the most effective against Salmonella enterica ssp. enterica in microaerobic while ME-3 and both bifidobacteria expressed high activity against Shigella sonnei in anaerobic milieu. Lact. paracasei, Lact. rhamnosus and Lact. plantarum strains showed intermediate antagonistic activity against Helicobacter pylori under microaerobic conditions on solid media. The highest anti-oxidative activity was characteristic for Lact. fermentum ME-3 (P < 0.05). No efficient antagonist against Clostridium difficile was found. The positive correlations between the pH, lactic acid production and anti-microbial activity for all tested probiotics were assessed. CONCLUSIONS: Developed experimental assays enable to compare the anti-microbial and -oxidative activity of Lactobacillus and/or Bifidobacterium probiotics, which have been claimed to possess the ability of suppressing the growth of various enteric and urinary pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Screening Lactobacillus and Bifidobacterium sp. strains according to their activity in various environmental conditions could precede the clinical efficacy studies for adjunct treatment with probiotics in cure of different gastrointestinal and urinary tract infections.

Antibiosis↗
PubMed1988

Effects of Lactic Acid Bacteria and Organic Acids on Growth and Germination of Bacillus cereus.

Growth and germination of vegetative cells and endospores of Bacillus cereus were affected by Streptococcus lactis, Streptococcus thermophilus, Lactobacillus acidophilus, and Lactobacillus bulgaricus in nonfat milk medium and by salts of organic acids in broth medium. Growth of the lactic acid bacteria was not affected by B. cereus. B. cereus increased rapidly to about 10 CFU/ml when cells were added at the beginning of growth of lactic acid bacteria; it was inactivated slowly when added after 24 h and rapidly when added after 72 h of lactic acid bacterial growth. Streptococci were more inhibitory to the growth of B. cereus than lactobacilli were at 24 h. Spore germination was not affected after 24 h, but it was inhibited after 48 and 72 h of lactic acid bacterial growth. Acetate was more inhibitory to the growth of vegetative cells, while formate was more inhibitory to spore germination. Acetate, formate, and lactate (all at 0.1 M) completely inactivated multiplication of B. cereus at pH 6.1, 6.0, and 5.6, respectively. Spores of B. cereus were more resistant to these organic acids compared with the resistance of vegetative cells. Formate, lactate, and acetate (all at 0.1 M) caused 50% inhibition of spore germination at pH 4.4, 4.3, and 4.2, respectively.

Journal Article↗
PubMed1999

Molecular analysis of the microbial diversity present in the colonic wall, colonic lumen, and cecal lumen of a pig.

Random clones of 16S ribosomal DNA gene sequences were isolated after PCR amplification with eubacterial primers from total genomic DNA recovered from samples of the colonic lumen, colonic wall, and cecal lumen from a pig. Sequences were also obtained for cultures isolated anaerobically from the same colonic-wall sample. Phylogenetic analysis showed that many sequences were related to those of Lactobacillus or Streptococcus spp. or fell into clusters IX, XIVa, and XI of gram-positive bacteria. In addition, 59% of randomly cloned sequences showed less than 95% similarity to database entries or sequences from cultivated organisms. Cultivation bias is also suggested by the fact that the majority of isolates (54%) recovered from the colon wall by culturing were related to Lactobacillus and Streptococcus, whereas this group accounted for only one-third of the sequence variation for the same sample from random cloning. The remaining cultured isolates were mainly Selenomonas related. A higher proportion of Lactobacillus reuteri-related sequences than of Lactobacillus acidophilus- and Lactobacillus amylovorus-related sequences were present in the colonic-wall sample. Since the majority of bacterial ribosomal sequences recovered from the colon wall are less than 95% related to known organisms, the roles of many of the predominant wall-associated bacteria remain to be defined.

Anaerobiosis↗
PubMed2002

Evolution of the lactic acid bacterial community during malt whisky fermentation: a polyphasic study.

The development of the lactic acid bacterial community in a commercial malt whisky fermentation occurred in three broad phases. Initially, bacteria were inhibited by strong yeast growth. Fluorescence microscopy and environmental scanning electron microscopy revealed, in this early stage, both cocci and rods that were at least partly derived from the wort and yeast but also stemmed from the distillery plant. Denaturing gradient gel electrophoresis (DGGE) of partial 16S rRNA genes and sequence analysis revealed cocci related to Streptococcus thermophilus or Saccharococcus thermophilus, Lactobacillus brevis, and Lactobacillus fermentum. The middle phase began 35 to 40 h after yeast inoculation and was characterized by exponential growth of lactobacilli and residual yeast metabolism. Lactobacillus casei or Lactobacillus paracasei, L. fermentum, and Lactobacillus ferintoshensis were detected in samples of fermenting wort examined by DGGE during this stage. Bacterial growth was accompanied by the accumulation of acetic and lactic acids and the metabolism of residual maltooligosaccharides. By 70 h, two new PCR bands were detected on DGGE gels, and the associated bacteria were largely responsible for the final phase of the fermentation. The bacteria were phylogenetically related to Lactobacillus acidophilus and Lactobacillus delbrueckii, and strains similar to the former had previously been recovered from malt whisky fermentations in Japan. These were probably obligately homofermentative bacteria, required malt wort for growth, and could not be cultured on normal laboratory media, such as MRS. Their metabolism during the last 20 to 30 h of fermentation was associated with yeast death and autolysis and further accumulation of lactate but no additional acetate.

Alcoholic Beverages↗
PubMed2005

Antioxidative activity of lactobacilli measured by oxygen radical absorbance capacity.

The reducing ability and antioxidative activity of some species of Lactobacillus were compared under in vitro conditions. Cultures of Lactobacillus delbrueckii ssp. lactis, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus acidophilus, and Lactobacillus casei were grown at 37 degrees C in de Man, Rogosa, Sharpe (MRS) broth supplemented with 0.5% 2,3,5 triphenyl tetrazolium chloride (TTC) to evaluate reducing activity. Reduced TTC was extracted from the cultures with acetone, and the intensity of the red color measured colorimetrically at 485 nm was an indication of reducing activity. The lactobacilli varied significantly in relative ability to reduce TTC when grown in MRS broth for 15 h. The relative amounts of growth as indicated by pH values at 18 h appeared to influence the amount of reduction. Antioxidative activity was evaluated by the ability of the whole cells or the cell-free extracts from cultures to protect a protein from being attacked by free radicals. These analyses were performed using the oxygen radical absorbance capacity method. All cultures tested exhibited some degree of antioxidative activity. Among the treatments, the cell-free extracts from cells grown in MRS broth exhibited significantly higher values than did whole cells. There was no apparent relationship between the reducing and antioxidative activities of the cultures evaluated. The results from this study show that these cultures can provide a source of dietary antioxidants. Furthermore, selection of cultures that produce antioxidants as starters could provide yet another health or nutritional benefit from cultured or culture-containing dairy products.

Antioxidants↗
PubMed2005

Microbiological quality of sous and tamarind, traditional drinks consumed in Jordan.

This study was conducted to evaluate the microbiological quality of sous (a drink prepared by extracting dried roots of Glycyrrhiza glabra) and tamarind (a drink prepared by infusing Tamarindus indica dried pulp), traditional drinks consumed in Jordan. Twenty-one samples of sous and 44 samples of tamarind were collected from the local market in Amman, Jordan. Water is the major component of the drinks. Sous drink is characterized by having an alkaline pH (range, 6.6 to 9.9; mean, 8.6), whereas tamarind drink has an acidic pH (range, 1.8 to 3.7; mean, 2.8). The drinks are not processed for safety before serving, and at some vendors drinks are not properly refrigerated. The mean counts for aerobic bacteria, lactic acid bacteria, and yeasts in sous drink samples were 5.9, 5.0, and 3.8 log CFU/ml, respectively; those in tamarind drink samples were 4.0, <1, and 5.8 log CFU/ml, respectively. The lactic acid bacteria isolated were Enterococcus raffinosus, Enterococcus hirae, Enterococcus durans, Lactobacillus acidophilus, and Lactobacillus buchneri. The yeast isolates in sous drink were from the genera Candida, Filobasidium, Hanseniaspora, Lodderomyces, Pichia, and Williopsis, and those in tamarind drink were from Arthroascus, Brettanomyces, Candida, Debaromyces, Filobasidiella, Hanseniaspora, Klavispora, Lodderomyces, Pichia, Saccharomycodes, Trichosporon, and Zygosaccharomyces. Enterobacteriaceae were detected in two sous samples and were identified as Enterobacter sakazakii and Erwinia sp., and in two tamarind samples and were identified as Citrobacter freundii and Klebsiella pneumoniae. Salmonella was detected in one sous and one tamarind sample. Pseudomonas aeruginosa was detected in only one sous sample. These findings highlight the importance of application of hygienic practices throughout preparation and vending of drinks, starting with raw ingredients and continuing through preparation, storage, display, and serving.

Bacteria↗
PubMed2002

[The nutritional status change the effectiveness of a dietary supplement of lactic bacteria on the emerging of respiratory tract diseases in children].

One hundred children 6 to 24 month old, normal or undernourished according to weight for height index, received during three months--autumn to winter--a dietary supplement of live Lactobacillus acidophilus and Lactobacillus Casei, 10(7)-10(8)/ml in fermented milk (LB) or an equivalent amount of fluid milk (L) as control. Children's follow-up was performed as outpatients in the Hospital Posadas (Great Buenos Aires). Episodes of respiratory tract infections were recorded and classified according to severity as: Pneumonia (N); Bronchitis (B), Recurrent Obstructive Bronchitis (BOR) and upper respiratory tract infections (CVAS). 58% of children fitted the study protocol, 22 in the LB and 36 in the L group; 21 were undernourished and 37 presented normal weight/height. No deaths were recorded. Total episodes were 103: 34 in LB and 69 in L, that means a frequency of 1.55 and 1.92 episodes/children respectively. In LB a maximum of 3 episodes/children was recorded, meanwhile the number reached 7 in L (p = 0.0373). Severity was higher in L than LB: 0.06 vs. 0 for N; 0.69 vs. 0.45 for B + BOR and 1.17 vs. 1.09 for CVAS. In the control group frequency of severe pathologies was about twice in undernourished than in normal: 0.08 vs. 0.04 for N; 1.08 vs. 0.50 for B + BOR; no difference was found for CVAS. Live lactobacillus supplement suppressed pneumonia and decreased bronchitis in undernourished as well as in normal. In this study undernutrition not only increased the chance of suffering severe acute respiratory tract infections but also impaired the effectiveness of the supplement to decrease severity. The effect is explained on the basis of the immunocompetence depression linked to an inadequate nutritional status.

Animals↗
PubMedDate not supplied

In vitro antibiotic susceptibility of lactobacilli isolated from commercial products containing active lactobacilli.

To survey the antibiotic susceptibility patterns of some commercially available Lactobacillus, we collected four commercial products that contain active Lactobacillus. We incubated individual product and identified these colonies by the methods of API50 CH test kit and RAPID ID 32A kit. Strains of Streptococcus thermophilus, Bifidobacterium infantis, Lactobacillus acidophilus and Lactobacillus casei were collected. By agar dilution method, each identified strain was inoculated to Brucella blood agar-MIC plates. Each plate contained one of the following antibiotics with different concentrations: amoxicillin, cephalothin, gentamicin, vancomycin, erythromycin, rifampin, tetracyclin and penicillin G, clindamycin, chloramphenicol, cefmetazole, metronidazole, ampicillin/sulbactum, cefoxtin, etc. After incubation, the growth condition of each Brucella blood agar-MIC plate was observed and the breakpoint of each antibiotic to different Lactobacillus products determined. The MICs of amoxicillin, ampicillin/sulbactum and penicillin-G to all identified strains were < or =2 microg/ml and those of vancomycin, clindamycin, erythromycin, metronidazole, cefmetazole and cefoxtin for L. casei were >32 microg/ml. L. casei was more resistant to all the testing antibiotics than the other strains. According to the MICs of the above antibiotics, proper active lactobacillus products could be chosen to prevent antibiotic-associated diarrhea in the pediatric field.

Amoxicillin↗
PubMedDate not supplied

[Applyed aspects of molecular biology of Bifidobacteria and Lactobacillus].

The data obtained in the molecular genetic studies of the representatives of the genera Bifidobacterium and Lactobacillus are given. At present the genomes of Bifidobacterium longum, Lactobacillus acidophilus and Lactobacillus plantarum sized 2,256.640, 1,993.564 and 3,308.274 base pairs respectively, were completely sequenced. The use of different methods of gene indication and gene typing essentially widened the existing notions of the species variety of these bacteria, made it possible to change the classification of known species and to find out new ones, vegetating in different biotopes of the human body. Molecular genetic approaches also ascertained the taxonomic position of production strains of lactic-acid bacteria used in food industry, helped determine the kinetics of the passage of probiotic strains of bifidobacteria and lactobacilli along the gastrointestinal tract, as well as determine the duration of their preservation in the intestine after oral administration. In addition, the data on the evaluation of the therapeutic effect of some preparations, probiotics and synbiotics, with determination of the population level of endogenic and orally introduced exogenic bifidobacteria and lactobacilli are presented.

Administration, Oral↗
PubMedDate not supplied

[The electron microscopic study of cell-to-cell interactions between antagonistic microorganisms].

The electron microscopic study of thin sections and positively stained specimens of cells taken from particular cocultures of Lactobacillus acidophilus D75, Lactobacillus casei YIT 9018, Shigella flexnery 2a, Bacillus subtilis ATCC 6633, and Staphylococcus aureus ATCC 25923 (some of these bacteria are antagonistic to others) showed the presence of specific ultrastructural elements indicating cell specialization and cooperation. The responses of antagonistic bacteria manifested themselves at the cellular and population levels.

Antibiosis↗
PubMed2006

Identification and antibiotic susceptibility of bacterial isolates from probiotic products available in Italy.

This study was carried out to assay the bacterial viability and the probable contamination of a range of probiotic products available in Italy and to test the susceptibility of the isolates. Eleven dried food supplements and five fermented functional foods were examined using different isolation media under standardized cultivation conditions. The identification was made by conventional phenotypic characteristics and biochemical tests. Among isolates from the probiotic products antibiotic susceptibility was detected using the E-test (ABBiodisk). Our results demonstrate that nine food supplements and two fermented foods claimed species which could not be isolated, whereas potential pathogens (i.e. Micromonas micros) were isolated. Lactobacilli displayed species-dependent antibiotic resistance. Atypical resistance occurred for penicillin in Lactobacillus acidophilus and Lactobacillus bulgaricus and for erythromycin in Lactobacillus lactis and Lactobacillus salivarius. A broad range of MICs was observed for cephalosporins and fluroquinolones. Aminoglycosides had poor activity against Lactobacillus isolates. Two of the four isolates of Bifidobacterium exhibited high resistance to trimethroprim/sulfametoxazole and to fluoroquinolones. Our results suggest that some probiotic products claim species that cannot always be isolated, and are sometimes contaminated by potential pathogens. Moreover, the probable transferable erythromycin or penicillin resistance among the lactobacilli isolated should be taken into account.

Anti-Bacterial Agents↗
PubMed1999

The effect of vaginal microbes on in vivo and in vitro expression of human papillomavirus 16 E6-E7 genes.

Since other viruses can influence expression of human papillomavirus (HPV) 16 E6-E7 genes in vitro, this study addressed whether specific vaginal bacteria do so as well. The vaginal microflora of 18 women with cervical intraepithelial neoplasia (CIN) or normal histology and HPV 16 infection, was evaluated by quantitative culture. Expression of HPV 16 E6-E7 oncogenes was assessed in exfoliated cervical cells by quantitative polymerase chain reaction. HPV 16 expression was also quantitated in CaSki carcinoma cell line cocultured with Bacteroides fragilis or Lactobacillus acidophilus. Isolation of Lactobacillus sp. (p = 0.05) and expression of the E6*II transcript (p = 0.03) were associated with low-grade CIN or normal histology. However, changes in E6-E7 expression were not associated independently with isolation of a specific microorganism. Similarly, expression of HPV 16 E6-E7 oncogenes in vitro was unaltered in the presence of bacteria. These results suggest that vaginal microorganisms are unlikely to alter the natural history of HPV-associated CIN by influencing HPV oncogene expression.

Bacteroides fragilis↗