Interrelationships between lactobacilli and streptococci in plaque formation on a tooth in an artificial mouth.
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Tests described in this paper show that gelatine membrane filters used in the MD8 microbiological air sampling system collected monodispersed aerosols between 0.7 and 1.0 microns containing viable Bacillus subtilis var. niger spores, with an efficiency of 99.9995%. Gelatine membrane filters linked to the MD8 control pump system were as effective as the well established Casella slit-to-agar device for collecting some viable bacteria, nebulized under controlled experimental conditions and naturally occurring airborne micro-organisms in a pharmaceutical plant. By using a long flexible hose connection to the control pump, the head could be positioned where sampling was required in locations remote from the pump exhaust, making it suitable for microbiological monitoring in critical locations such as laminar flow stations and isolators.
This study evaluated the feasibility of using a colony lift method and DNA probes to enumerate bacterial species cultured on primary isolation plates. Fourteen digoxigenin-labeled whole chromosomal DNA probes representing 12 subgingival species were validated by hybridization with colony lifts prepared from 249 reference strains of 51 species grown on Trypticase soy agar plates supplemented with 5% sheep blood. Colonies of reference strains were lifted onto Nytran filters from plates and treated to lyse cells, remove cellular proteins, denature and fix microbial DNA to the filters. Positive reactions were detected with an anti-digoxigenin antibody conjugated to alkaline phosphatase and revealed by bromo-chloro-indolyl phosphate and nitroblue tetrazolium. Cross-reactions were not observed for 13/14 probes, but 2 strains of Streptococcus mitis reacted with the probe to Streptococcus sanguis II. Subgingival plaque samples were taken by means of a sterile curette from mesiobuccal surfaces of teeth present in each of 26 subjects with differing periodontal disease states. Samples were dispersed, diluted, plated and incubated anaerobically for 7 d at 35 degrees C. Colonies were lifted as described above. Filters were cut into sections and hybridized with the 14 digoxigenin-labeled DNA probes. The probes were used to enumerate the test species and the total number of isolates was determined in 711 plaque samples. The colony lift method and DNA probes provided a sensitive, economical and quantitative method for enumerating cultivable microbial species in subgingival plaque samples. In addition, the amplification provided by growing the organisms on agar plates facilitated determination of numbers of organisms in small plaque samples, such as those from healthy sites.
BACKGROUND/AIMS: Lactobacilli are known to play an important role in the maintenance of health by stimulating natural immunity and contributing to the balance of microflora. However, their role in chronic periodontitis is unclear. We aimed to identify oral lactobacilli in chronic periodontitis and periodontally healthy subjects, and to determine their antimicrobial activity against putative oral pathogens. METHODS: A total of 238 Lactobacillus isolates from the saliva and subgingival sites of 20 chronic periodontitis and 15 healthy subjects were collected. In all, 115 strains were identified using rapid amplified ribosomal DNA restriction analysis. Antimicrobial activity against Streptococcus mutans, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia was assessed. RESULTS: Lactobacilli belonging to 10 species were identified. The most prevalent strains in healthy persons were Lactobacillus gasseri and Lactobacillus fermentum and in chronic periodontitis patients, Lactobacillus plantarum. Obligately homofermentatives, particularly L. gasseri, were less prevalent in chronic periodontitis patients compared with healthy subjects (8% vs. 64% for L. gasseri, P < 0.01). Sixty-nine percent of tested lactobacilli inhibited S. mutans, 88% A. actinomycetemcomitans, 82% P. gingivalis and 65% P. intermedia. The strongest antimicrobial activity was associated with Lactobacillus paracasei, L. plantarum, Lactobacillus rhamnosus, and Lactobacillus salivarius. The strains from periodontally healthy patients showed a lower antimicrobial activity against S. mutans than the strains from chronic periodontitis patients. CONCLUSION: The composition of oral lactoflora in chronic periodontitis and healthy subjects differs, with a higher prevalence of homofermentative lactobacilli, particularly L. gasseri, in the latter group. Both homo- and heterofermentative oral lactobacilli suppress the growth of periodontal pathogens, but the antimicrobial properties are strain, species and origin specific.
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OBJECTIVE: Experimental and clinical studies have shown that a novel symbiotic (known as SCM-III) exerts a beneficial effect on gut translocation and local and systemic inflammatory and microbial metabolic parameters. The present investigation was a preliminary trial on the effectiveness of SCM-III for irritable bowel syndrome (IBS). METHODS: Sixty-eight consecutive adult patients with IBS who were free from lactose malabsorption, abdominal surgery, overt psychiatric disorders and ongoing psychotropic drug therapy or ethanol abuse were studied prospectively and divided into 2 groups that were comparable for age, gender, body size, education and pattern of presenting symptoms. The 2 groups were blindly given for 12 weeks either SCM-III 10 mL t.i.d or the same dosage of heat-inactivated symbiotic. RESULTS: Treatment with SCM-III was 'effective' or 'very effective' in more than 80% of the patients (P < 0.01 vs baseline values and control). Less than 5% reported 'not effective' as the final evaluation compared with over 40% of patients in the control group. After 6 weeks of treatment, a significant improvement of pain and bloating was reported in the treatment group compared with control and baseline values. There was also a benefit for bowel habits, mostly for patients with constipation or alternating bowel habits. No overt clinical or biochemical adverse side-effects were recorded. CONCLUSION: Compared with baseline values and the control group, SCM-III resulted in a significant increase in lactobacilla, eubacteria and bifidobacteria, which suggests that some selected IBS patients could benefit substantially from symbiotics, but the treatment may need to be given on a cyclic schedule because of the temporary modification of the fecal flora.
BACKGROUND: Patients with inflammatory bowel disease have a 10- to 100-fold increased risk of nephrolithiasis, with enteric hyperoxaluria being the major risk factor for these and other patients with fat malabsorptive states. Endogenous components of the intestinal microflora can potentially limit dietary oxalate absorption. METHODS: Ten patients were studied with chronic fat malabsorption, calcium oxalate stones, and hyperoxaluria thought to be caused by jejunoileal bypass (1) and Roux-en-Y gastric bypass surgery for obesity (4), dumping syndrome secondary to gastrectomy (2), celiac sprue (1), chronic pancreatitis (1), and ulcerative colitis in remission (1). For 3 months, patients received increasing doses of a lactic acid bacteria mixture (Oxadrop), VSL Pharmaceuticals), followed by a washout month. Twenty-four-hour urine collections were performed at baseline and after each month. RESULTS: Mean urinary oxalate excretion fell by 19% after 1 month (1 dose per day, P < 0.05), and oxalate excretion remained reduced by 24% during the second month (2 doses per day, P < 0.05). During the third month on 3 doses per day oxalate excretion increased slightly, so that the mean was close to the baseline established off treatment. Urinary oxalate again fell 20% from baseline during the washout period. Calcium oxalate supersaturation was reduced while on Oxadrop, largely due to the decrease in oxalate excretion, although mean changes did not reach statistical significance. CONCLUSION: Manipulation of gastrointestinal (GI) flora can influence urinary oxalate excretion to reduce urinary supersaturation levels. These changes could have a salutary effect on stone formation rates. Further studies will be needed to establish the optimal dosing regimen.
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Proposed herein is a mechanism for virulence by Candida albicans based upon this organism's ability to produce high levels of pyruvate, potentially resulting in localized tissue ketosis and undermining the normal defensive function of neutrophil myeloperoxidase. Neutrophils, a key component of our innate defense against microbial infections, seem to play a particularly important role protecting us against fungal agents such as C. albicans. In this regard, it is myeloperoxidase which is central to many of the antimicrobial properties of neutrophils. We have previously shown that metabolic ketones inactivate myeloperoxidase and impair phagocytosis. Thus, production of pyruvate by C. albicans may indeed be a significant virulence factor.
Oral infections constitute some of the most common and costly forms of infections in humans. The concept of microbial ecological change as a mechanism for preventing dental disease is an important one while altered microbial ecology may lead to dental disease. New methods such as probiotic approaches (i.e. whole bacteria replacement therapy) to eliminate pathogenic members of the microbiota can be investigated. Bacteriotherapy is an alternative and promising way to combat infections by using harmless bacteria to displace pathogenic microorganisms. Probiotics are one of these new agents which are widely used for their therapeutic action. Limited research is available showing that some probiotic cultures may help dental improvement. Present paper focuses on possible oral benefits of probiotics.
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Azide and, to a lesser extent, cyanide inhibit the microbicidal activity of myeloperoxidase and of intact normal leukocytes, but they have little or no effect on peroxidase-negative leukocytes. The contribution of the azide-sensitive (peroxidase-dependent?) systems to the total microbicidal activity of normal leukocytes is considerable. The azide-insensitive antimicrobial systems are more highly developed in peroxidase-negative leukocytes than in normal leukocytes, thus suggesting an adaptation.