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Enhancement of lysozyme trypsin-mediated decay of intestinal bifidobacteria and lactobacilli.

Lysozyme-mediated lysis of Bifidobacteria and Lactobacilli was studied in in vitro tests using the agar gel plate and turbidometric Micrococcus luteus (lysodeikticus) procedure as a standard. Suspensions of the strains Bifidobacterium infantis, B. infantis liberorum, B. breve, B. longum, B. ssp, and Lactobacillus acidophilus proved to be resistant to egg white lysozyme and human milk lysozyme when incubated at 37 degrees C in concentrations of 5, 50, and 500 mg lysozyme/L, respectively, through 30 and 60 min. Heat treatment at 100 degrees C for 1 h and pretreatment with ether, acetone, ascorbic acid, and hydrogen peroxide failed to incline the bacteria to the lytic effects of lysozyme. Consecutive incubation of the lysozyme-pretreated bacteria with trypsin resulted in a significantly enhanced bacteriolysis in all strains of bacteria, with the exception of B. longum. The mode of action of lysozyme and proteolytic enzymes on Bifidobacteria and Lactobacilli offers an explanation for the release of microbial building blocks and their colonic absorption and retention in the breast-fed baby.

Acetone↗

Segregation of deoxyribonucleic acid in bacteria: association of the segregating unit with the cell envelope.

Cells of the gram-positive organism Lactobacillus acidophilus R-26 were labeled with (3)H-thymine to measure the segregation of radioactive deoxyribonucleic acid (DNA) into daugher cells. Such cells were found to contain 8 conserved units of DNA which would correspond to two replicating chromosomes per cell. Fluorescent antibody (FA) against this organism was used to demonstrate that portions of the cell surface (2 to 4 units per cell) were conserved during growth and division. The permanent association of DNA with these conserved cell surface units was measured by combining autoradiography with FA techniques. DNA synthesized immediately before FA labeling was not associated with the fluorescent cell surface, whereas DNA synthesized a generation previously was. The results are consistent with a model in which DNA becomes permanently fixed to the cell surface when it is first used as a template.

Animals↗

Incipient caries lesions on cementum by mono- and co-culture oral bacteria.

OBJECTIVES: There is increasing prevalence of root caries. We hypothesized different biofilms will cause varying demineralization in cementum. This study investigated the extent of demineralization of cementum by oral biofilm formed from three major cariogenic microorganisms: Streptococcus mutans, Lactobacillus acidophilus, and Actinomyces israelii. Sound cementum tooth blocks were incubated with mono-, bi-, and tri-species combinations of the bacteria under investigation. MATERIALS AND METHODS: The matrix (amide I) and phosphate content of the lesions was analyzed by Fourier-transform infrared spectroscopy (FTIR), and calcium and phosphorus levels were analyzed by scanning electron microscopy and energy-dispersive spectroscopy (SEM-EDX). RESULTS: The log[amide I:HPO(4)(2-) absorbance] values showed that A. israelii mono-culture caused significantly more demineralization than the other bacterial cultures. log[Ca:P] showed that all carious lesions were confined to the cementum. CONCLUSIONS: Oral biofilm arising from bacterial species A. israelii alone was the most cariogenic of those tested and produced the most demineralization in incipient carious lesions in cementum.

Actinomyces↗

Myeloperoxidase-halide-hydrogen peroxide antibacterial system.

An antibacterial effect of myeloperoxidase, a halide, such as iodide, bromide, or chloride ion, and H(2)O(2) on Escherichia coli or Lactobacillus acidophilus is described. When L. acidophilus was employed, the addition of H(2)O(2) was not required; however, the protective effect of catalase suggested that, in this instance, H(2)O(2) was generated by the organisms. The antibacterial effect was largely prevented by preheating the myeloperoxidase at 80 C or greater for 10 min or by the addition of a number of inhibitors; it was most active at the most acid pH employed (5.0). Lactoperoxidase was considerably less effective than was myeloperoxidase when chloride was the halide employed. Myeloperoxidase, at high concentrations, exerted an antibacterial effect on L. acidophilus in the absence of added halide, which also was temperature- and catalase-sensitive. Peroxidase was extracted from intact guinea pig leukocytes by weak acid, and the extract with peroxidase activity had antibacterial properties which were similar, in many respects, to those of the purified preparation of myeloperoxidase. Under appropriate conditions, the antibacterial effect was increased by halides and by H(2)O(2) and was decreased by catalase, as well as by cyanide, azide, Tapazole, and thiosulfate. This suggests that, under the conditions employed, the antibacterial properties of a weak acid extract of guinea pig leukocytes is due, in part, to its peroxidase content, particularly if a halide is present in the reaction mixture. A heat-stable antibacterial agent or agents also appear to be present in the extract.

Animals↗

[Effectiveness of acipole in prevention of enteric dysbacteriosis due to antibacterial therapy].

Acipole (Lactobacillus acidophilus + Kefir greins) was used to manage antibiotic dysbacteriosis as an adverse reaction of antibacterial therapy. 120 patients treated with antibacterial drugs for acute pneumonia and exacerbations of chronic bronchitis were observed. 54 of them were treated under the routine regimen with antibiotics and 66 were additionally treated with the eubiotic acipole: 1 tablet (5 doses) 3 times a day 30 minutes before meal. Routine bacteriological examination of the feces was applied to all the patients. High frequency of bacteriologically revealed dysbacteriosis was stated. The therapy under the antibiotic + acipole regimen lowered the frequency of dysbacteriosis events and their severity. The fact that the use of acipole simultaneously with the routine antibacterial therapy prevented the development of dysbacteriosis clinical signs is of practical importance.

Adult↗

Survival of viruses in fermented edible waste material.

The survival of selected viruses in fermented edible waste material was studied to determine the feasibility of using this material as a livestock feed ingredient. Seven viruses, including pseudorabies, Newcastle disease, infectious canine hepatitis, avian infectious bronchitis, measles, vesicular stomatitis, and a porcine picornavirus were inoculated into a mixture of ground food waste (collected from a school lung program) containing Lactobacillus acidophilus. Mixtures were incubated at 5 C, 10 C, 20 C, and 30 C for 96 hours. Temperature, pH, and redox potential were monitored. Samples for virus isolation were obtained daily. Newcastle disease virus and infectious canine hepatitis virus survived the entire test period. The porcine picornavirus was inactivated at 30 C after 74 hours, but survived for the entire test period at the other temperatures. Pseudorabies virus was inactivated at 20 C and 30 C within 24 hours, but survived for 48 hours at 10 C and 96 hours at 5 C. Avian infectious bronchitis virus was inactivated at 20 C and 30 C within 24 hours, but survived 72 hours at 5 C and 10 C. Measles and vesicular stomatitis viruses were rapidly inactivated at all 4 temperatures.

Adenoviruses, Canine↗

Probiotic bacteria stimulate virus-specific neutralizing antibodies following a booster polio vaccination.

BACKGROUND: Orally ingested probiotic bacteria may modulate the immune response and increase antibody titers against enteric infections by bacteria or viruses. Even though positive effects of probiotics on respiratory tract infections have been reported, overall only few studies have examined effects on virus infections concerning organs other than the gastrointestinal tract. AIM OF THE STUDY: It was the aim of the study to investigate whether and how probiotics affect the immune response to a standardized enterovirus challenge (polio) and infections not limited to the gastrointestinal tract in healthy adults. METHODS: In a randomized, controlled and double-blind study 64 volunteers consumed for 5 weeks chemically acidified clotted milk without bacteria or with 10(10)/serving (Lactobacillus rhamnosus ) GG or Lactobacillus acidophilus CRL431 added. In the second week subjects were vaccinated orally against polio 1, 2 and 3. Polio virus neutralizing serum activity, the primary parameter, was determined by the standard neutralization test (WHO) before and three times after vaccination. Polio-specific IgA, IgG and IgM were detected by ELISAs. RESULTS: Probiotics increased poliovirus neutralizing antibody titers (NT) and affected the formation of poliovirus-specific IgA and IgG in serum. The maximum increase after immunization was about 2, 2.2, or 4-fold higher, respectively, for NT, IgG or, IgA, in volunteers consuming probiotics instead of placebo. No consistent difference was noted between bacterial strains. CONCLUSIONS: Probiotics induce an immunologic response that may provide enhanced systemic protection of cells from virus infections by increasing production of virus neutralizing antibodies.

Adult↗

In vitro growth responses of bifidobacteria and enteropathogens to bovine and human lactoferrin.

A series of in vitro experiments was performed to test the ability of bovine and human lactoferrin to influence the growth of the gram-positive probiotic bacteria, Bifidobacterium bifidum, Bifidobacterium infantis, and Lactobacillus acidophilus, as well as the gram-negative enteric bacteria, E. coli O157:H7 and Salmonella typhimurium. None of the lactoferrin preparations stimulated the growth of the tested strains. However, iron-free apo-lactoferrin (bovine and human) and 66% iron-saturated bovine lactoferrin dramatically slowed the growth of E. coli O157:H7 in single culture experiments, while 98% iron-saturated preparations had no effect. In coculture experiments of B. infantis and E. coli, the iron-limited preparations of lactoferrin also slowed the growth of the latter without inhibiting the bifidobacteria. These results suggest that lactoferrin in iron-limited forms may have the potential to be combined with probiotic bacteria in biotherapeutic products, which could help balance human gut microflora and limit the overgrowth of certain enteric microorganisms.

Animals↗

[Long-term experience in the therapy and prevention of unspecific vaginal discharge with a Lactobacillus vaccine].

This report presents a study of 28 women with chronic non-specific vaginitis treated by Solco Trichovac, a vaccine containing at least 7 X 10(9) inactivated microorganisms of various strains of Lactobacillus acidophilus in each 0.5 ml dose. 3 injections were administered at 2-week intervals, and the results of therapy investigated with regard to quantitative increase in lactobacilli in the vaginal secretion, change in grade according to Schröder and reduction of the vaginal fluor. The vaccination caused a significant (p less than 0.01) quantitative increase in physiological lactobacilli. Accordingly, there was a significant improvement (p less than 0.01) in grade already 4 weeks after completion of the primary course of treatment. Re-examination after one year showed that nearly 80% of the patients were grade I to II; the vaginal fluor was clinically cured or at least markedly improved in 85.6% of cases. In our experience the vaccination presents a promising new concept of therapy and shows a special effect in patients with chronic non-specific vaginitis and with reduced Döderlein's vaginal bacillus due to restoration of the biological protective mechanisms.

Adolescent↗

Lactobacilli express cell surface proteins which mediate binding of immobilized collagen and fibronectin.

Binding of immobilized collagen-I (Cn-I) and fibronectin (Fn) by Lactobacillus acidophilus CRL 639 depends on cell-surface proteins. Capsule formation during the stationary growth phase has a negative effect on adherence of Cn-I and Fn. However, cells from the exponential growth phase, which produce no capsule, exhibit maximal binding. Binding is sensitive to trypsin, proteinase K, pronase E, and heat. Gelatin and soluble Cn-I partially inhibit binding of Cn-I although various proteins, sugars and amino acids do not affect binding to Fn. These results indicate that protein-protein interactions mediate adhesion to extracellular matrix proteins. SDS-PAGE and Western blot analyses of surface proteins revealed that several proteins including the major 43-kDa protein of the S-layer are expressed. Monoclonal antibodies showed that Fn binds to a 15-kDa protein, while Cn-I binds to proteins of 45 and 58 kDa.

Animals↗

Immunomodulation of human blood cells following the ingestion of lactic acid bacteria.

Because of the lack of data that convincingly show immunomodulatory properties of lactic acid bacteria in humans, a study was performed in which healthy volunteers were divided into two groups and given a fermented milk product supplemented with Lactobacillus acidophilus strain La1 or Bifidobacterium bifidum strain Bb 12 for 3 wk. Blood was sampled throughout the study to assess changes in lymphocyte subsets or leukocyte phagocytic activity following consumption of the fermented products. No modifications of lymphocyte subpopulations were detected. In contrast, phagocytosis of Escherichia coli sp. in vitro was enhanced after the administration of both fermented products. The increment in phagocytosis was coincident with fecal colonization by the lactic acid bacteria and persisted for 6 wk after ingestion of the fermented products. By this time, the fecal lactobacilli and bifidobacteria had returned to concentrations prior to consumption. Nonspecific, anti-infective mechanisms of defense can be enhanced by the ingestion of specific lactic acid bacteria strains. These strains can be used as nutritional supplements to improve the immune function of particular age groups, i.e., the neonate or the elderly, for which these functions are diminished.

Adult↗

Studies of the subgingival microflora in patients with acquired immunodeficiency syndrome.

Two unique forms of periodontal disease, HIV-gingivitis and HIV-periodontitis, have been described in patients with Acquired Immunodeficiency Syndrome (AIDS). In order to determine the bacterial species associated with periodontitis in AIDS patients, the predominant cultivable microflora was examined in 21 subgingival plaque samples from 11 AIDS patients with periodontitis. The presence of putative periodontal pathogens including Actinobacillus actinomycetemcomitans, Bacteroides intermedius, Porphyromonas gingivalis (formerly B. gingivalis), and Wolinella recta was examined by immunofluorescence in 128 subgingival dental plaque samples from 50 AIDS patients including 32 patients with periodontitis. Of 666 bacterial strains isolated from the 21 subgingival plaque samples, Streptococcus sanguis II was the most frequently recovered species comprising 18.5% of the total number of isolates followed by Lactobacillus acidophilus (12.2%), Porphyromonas gingivalis (12%), Fusobacterium nucleatum (11.4%), Staphylococcus epidermidis (8.7%), Actinomyces naeslundii (7.5%), and Actinomyces viscosus (4.7%). Fusobacterium nucleatum was the most prevalent species and was found in 76% of the sites and 91% of the patients. Enteric species including Enterococcus avium and Enterococcus faecalis, Clostridium clostridiiforme and Clostridium difficle as well as Klebsiella pneumoniae also were recovered. Immunofluorescence assays detected similar carriage rates of A. actinomycetemcomitans, B. intermedius, and P. gingivalis in both gingivitis patients and periodontitis patients, while four times more periodontitis patients demonstrated W. recta. Subgingival yeast was a frequent finding in these AIDS patients, present in 62% of the subjects and 55% of the sites. This study indicates that subgingival plaque in AIDS patients with periodontitis can harbor high proportions of the same periodontal pathogens as are associated with periodontitis in non-HIV infected subjects as well as high proportions of opportunistic pathogens.

Acquired Immunodeficiency Syndrome↗

Alterations of the intestinal microflora by diet, oral antibiotics, and Lactobacillus: decreased production of free amines from aromatic nitro compounds, azo dyes, and glucuronides.

F344 rats were fed compounds containing a nitro, azo, or amine-glucuronide group-2-nitronaphthalene (CAS: 581-89-5), 2-nitrofluorene (CAS: 607-57-8; 2-nitro-9H-fluorene), 4-phenylazo-2-naphthol (CAS: 6410-10-2; 1-[(4-nitrophenyl)azo]-2-naphthol), and 2-naphthylamine-N-D-glucuronide--and the formation of free amines was measured in the feces. The effects of diet (mean vs. grain), administration of antibiotic, and the feeding of Lactobacillus acidophilus were evaluated. Meat-fed rats given 2-nitronaphthalene had approximately 4 times higher concentration of free amines in their feces as compared to the concentration in grain-fed rats. In a similar experiment with 2-naphthylamine-N-D-glucuronide, the meat-fed animals had approximately 1.5 times more of the free amines and 50% less of the conjugates in their feces. When meat-fed animals were given L. acidophilus with 2-nitrofluorene or 2-naphthylamine-N-D-glucuronide, they had significantly lower free amines and higher concentrations of conjugates in their feces than did the meat-fed controls. Similarly, erythromycin or tetracycline decreased the fecal amine production in meat-fed animals that had received 2-nitrofluorene or 4-phenylazo-2-naphthol. In another experiment, erythromycin inhibited the production of free amines from 2-naphthylamine-N-D-glucuronide in rats fed a high-fat diet. Studies of fecal enzymes revealed lower levels of beta-D-glucuronidase, nitroreductase, and azoreductase in grain-fed rats and in meat-fed animals also given antibiotics or lactobacilli. These experiments showed that intestinal flora have the ability to convert exogenously administered aromatic nitro and azo compounds and an amine-glucuronide compound to free amines. The rate of these conversions was affected by diet and by oral administration of antibiotics and lactobacilli.

Amines↗

[Lactobacilli in the treatment of dyspepsia due to dysmicrobia of various causes].

In 30 patients with dyspepsia caused by dysbacteriosis of the gastrointestinal tract the authors administered the preparation Lactobacillus acidophilus (Rossel Co. Canada)--1. capsule with 2 billion live bacteria, in the morning after breakfast. The patients were divided into four groups: maldigestion, malabsorption, radiation enterocolitis and administration of antibiotics. The patients recorded themselves their subjective symptoms: pain, pressure, bloating, flatulence and appetite, and as to objective symptoms, the number and consistency of bowel movements, changes of body weight. The most rapid effect was achieved in dysbioses after antibiotics--within 3-4 days normalization occurred which persisted even after discontinuation of the drug. In maldigestion after one week bloating, flatulence, abdominal pain and pressure in the epigastrium was milder, and within two weeks the condition improved further. An excellent effect was achieved in radiation enterocolitis. In patients with lactose intolerance the tolerance of dairy products improved. No side-effects were observed, the preparation was very well tolerated; the mean body weight increment was 0.75 kg in three weeks. The preparation proved a new useful probiotic which is highly effective in dyspepsias caused by dysbiosis of the intestinal microflora.

Antibiosis↗

Bacterial and fungal microleakage of AH26 and AH Plus root canal sealers.

AIM: To evaluate the penetration of Candida albicans alone and a combination of bacteria through root canals filled with gutta-percha and one or other root canal sealers, AH26 and AH Plus. METHODOLOGY: Eighty teeth were randomly divided into two groups of 40 teeth each and obturated with gutta-percha using either AH26 or AH Plus sealer. A further 10 teeth served as negative controls and 10 as positive controls. The external surface of each root, except the apical 2 mm, was covered with two layers of nail varnish. The teeth were inserted into Eppendorf plastic tubes and suspended in glass bottles containing sterile Schaedler broth. Streptococcus mutans, Streptococcus mitis, Prevotella melaninogenica and Lactobacillus acidophilus were placed in the access cavities of 20 teeth filled with AH26 and 20 with AH Plus. Candida albicans was placed in the access cavities of the other teeth. The culture medium with microorganisms was changed every 7 days. Every 72 h bacterial or fungal growth in the broth was tested up to a period of 90 days. RESULTS: Leakage in the experimental teeth occurred between 14 and 87 days. Leakage was present in 47% of all samples. From the samples with AH26, 45% leaked bacteria and 60% leaked fungi; whilst from the samples with AH Plus, 50% leaked bacteria and 55% fungi. There was no statistically significant difference in penetration of bacteria and fungi between the sealers. CONCLUSION: In this in vitro study, gutta-percha and the sealers AH26 and AH Plus allowed leakage of bacteria and fungi.

Bacteria↗

The effect of soy consumption on the urinary 2:16-hydroxyestrone ratio in postmenopausal women depends on equol production status but is not influenced by probiotic consumption.

Some epidemiologic studies reported an association between a low ratio of urinary 2-hydroxyestrogens (2-hydroxyestradiol + 2-hydroxyestrone) to 16alpha-hydroxyestrone (2:16OHE(1)) and increased breast cancer risk. Some studies show that soy consumption increases this ratio, and it is suggested that this effect may reduce breast cancer risk. We hypothesized that consumption of probiotic bacteria would alter fecal bacteria and enzymes involved in soy isoflavone metabolism, thereby increasing isoflavone bioavailability and enhancing the beneficial effects of soy on estrogen metabolism. Breast cancer survivors (n = 20) and controls (n = 20) were given 4 treatments for 6 wk each, separated by 2-wk washout periods, in a randomized, crossover design: soy protein (26.6 +/- 4.5 g protein/d containing 44.4 +/- 7.5 mg isoflavones/d); soy protein + probiotics (10(9) colony-forming units Lactobacillus acidophilus DDS(R)+1 & Bifidobacterium longum, 15-30 mg fructooligosaccharide/d); milk protein (26.6 +/- 4.5 g protein/d); and milk protein + probiotics. Survivors tended to have a lower baseline urine 2:16OHE(1) ratio than controls (P = 0.10). In the group as a whole, soy consumption tended to increase urinary 2-hydroxyestrogens (P = 0.07) and 16alpha-hydroxyestrone (P = 0.11) but had no effect on the urinary 2:16OHE(1) ratio. When subjects were divided into groups by plasma concentrations and urinary levels of the daidzein metabolite equol, soy increased urinary 2-hydroxyestrogens (P = 0.01) and the 2:16OHE(1) ratio (P = 0.04) only in subjects with high plasma equol concentrations. None of these results were influenced by probiotic consumption. These results are consistent with studies that found lower urine 2:16OHE(1) ratios in women with breast cancer and suggest that soy consumption increases this ratio only in women who are equol producers.

Bifidobacterium↗

Effect of ginseng polysaccharide on the stability of lactic acid bacteria during freeze-drying process and storage.

Lactic acid bacteria (LAB) quickly attenuate or are killed during the freeze-drying process and storage. The effect of some natural polysaccharides, which are known as potent antitumor and immunomodulating substances, on the viability of the LAB, Lactobacillus acidophilus and Bifidobacterium breve, on freeze-drying and storage were investigated. Among the polysaccharides tested, red ginseng polysaccharide (RGP) and chitosan significantly inhibited the cell death of the LAB during freeze-drying, and fucoidan and RGP most potently protected the cell death of the LAB during storage. The stabilities of the LAB on the addition of RGP and fucoidan were comparable to that of skimmed milk. However, white ginseng polysaccharide (WGP) did not promote storage stability. When 5% skimmed milk/5% RGP treated LAB were freeze-dried and stored, their viabilities were found to be significantly higher those treated with 5% or 10% RGP. The stabilizing effect of 5% RGP/5% skimmed milk during LAB freeze-drying and storage stability was comparable to that of treatment with 10% skimmed milk. Based on these findings, we believe that RGP beneficially improves the stability of LAB during the freeze-dry process and storage.

Animals↗

Effect of spray-dried yogurt and lactic acid bacteria on the initiation and promotion stages of chemically induced skin carcinogenesis in mice.

The effects of dietary spray-dried yogurt powder product (YPP) and two strains of lactic acid bacteria on the initiation and promotion stages of carcinogenesis were investigated using the 7,12-dimethylbenz[a]anthracene (DMBA)-12-O-tetradecanoylphorbol-13-acetate (TPA) mouse skin carcinogenesis model. In two independent studies, Sencar mice were fed a diet containing 86%, 43%, or 0% YPP or the 0% YPP diet supplemented with viable cultures of Lactobacillus acidophilus or bifidobacteria. Animals were initiated with a single topical application of DMBA (10 nmol). Promotion began three weeks later with twice weekly treatment of TPA (1 microgram/200 microliters acetone). During the initiation study (Study 1) the experimental diets were fed for four weeks before and one week after DMBA treatment. All mice were then switched to the AIN-76 diet. For the promotion study (Study 2) the experimental diets were begun one week after initiation and fed during the remainder of the study. Gross appearance of tumors was assessed weekly. No statistically significant differences in body weight or food disappearance were observed among the diet groups during the studies. For Studies 1 and 2, final histologically verified papilloma incidence and multiplicity and carcinoma incidence were not statistically different. These data suggest that different levels of YPP or lactic acid bacteria fed during the initiation or promotion stage of carcinogenesis do not significantly affect chemically induced skin tumor development.

9,10-Dimethyl-1,2-benzanthracene↗