agar and Periodontitis

The chewing stick (miswak) is used for oral hygiene in many parts of the world. In addition to the mechanical removal of plaque, an antibacterial effect has been postulated; however, tests of miswak extract from Salvadora persica (Arak) disclosed only low to moderate antibacterial effects. This may be attributable to the extraction process. Our aim was to test in vitro the antibacterial effect of miswak pieces, without extraction, on bacteria implicated in the etiology of periodontitis and caries.. Miswak pieces were standardized by size and weight (0.07 and 0.14 g) and tested against Streptococcus mutans, Lactobacillus acidophilus, Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), Porphyromonas gingivalis, and, as a reference, Haemophilus influenzae. The miswak pieces were tested in two ways: embedded in the agar plate or suspended above the agar plate.. The inhibitory effect was most pronounced on P. gingivalis, A. actinomycetemcomitans, and H. influenzae, less on S. mutans, and least on L. acidophilus. Suspended miswak had comparable or stronger effects than miswak embedded in agar. The 0.14-g suspended miswak exhibited significantly greater inhibition on A. actinomycetemcomitans and H. influenzae than the 0.14-g miswak embedded in agar (P<0.01 and P<0.001, respectively).. Miswak embedded in agar or suspended above the agar plate had strong antibacterial effects against all bacteria tested. The antibacterial effect of suspended miswak pieces suggests the presence of volatile active antibacterial compounds.

Topics: Agar; Aggregatibacter actinomycetemcomitans; Anti-Bacterial Agents; Bacteria; Culture Media; Dental Caries; Haemophilus influenzae; Humans; Lactobacillus acidophilus; Materials Testing; Periodontitis; Plant Preparations; Plant Roots; Porphyromonas gingivalis; Salvadoraceae; Streptococcus mutans

Actinobacillus actinomycetemcomitans is considered to be one of the major oral putative pathogens, especially in cases of juvenile periodontitis. This microorganism requires nutritionally complex media for growth, and therefore the media for its primary isolation usually include blood agar or serum in their base. In this study we present a new medium, Dentaid-1, which improves the detection of A. actinomycetemcomitans in periodontal samples. In its composition, blood and serum have been omitted, hence reducing its cost and making it a more restrictive medium against the growth of other microorganisms with high nutritional requirements. The growth yields of pure cultures of the bacteria on Dentaid-1 were comparable to those on nonselective blood agar. Moreover, clinical efficacy was evaluated in subgingival samples from 77 subjects with adult periodontitis. Dentaid-1 detected A. actinomycetemcomitans in 24 subjects, while a previously described tryptic soy-serum-bacitracin-vancomycin agar detected the microorganism in only 19 subjects (79.1%). Dentaid-1 is a low-cost, noninhibitory formula for the improved diagnosis and monitoring of patients subgingivally infected by this important oral putative pathogen.

Topics: Actinobacillus Infections; Adult; Agar; Aggregatibacter actinomycetemcomitans; Bacteriology; Blood; Costs and Cost Analysis; Culture Media; Dental Plaque; Gingiva; Humans; Periodontitis; Polymerase Chain Reaction; Reproducibility of Results; Spain

Peptostreptococcus micros, an anaerobic gram-positive coccus, has been associated with periodontal and endodontic lesions, including those refractory to treatment, as well as many human polymicrobial infections in other body locations. A selective and differential medium for the primary isolation of P. micros was developed and evaluated. Columbia CNA agar, a selective medium for gram-positive cocci, was supplemented with glutathione and lead acetate (P. micros medium: PMM). P. micros has a characteristic of rapidly utilizing the reduced form of glutathione to form hydrogen sulfide, which reacts with lead acetate producing a black precipitate in the medium. When grown on PMM, P. micros can be easily identified by its typical colonial morphology and the presence of a black precipitate directly under the colony. PMM was compared for the growth of P. micros with phenylethyl alcohol agar (PEA) and Columbia base medium (CBM) with 80 strains of P. micros and 30 strains of other gram-positive cocci. All P. micros isolates tested grew and showed the typical morphology of P. micros on PMM. Using colony counts on CBM as controls, there was an average 81.8% recovery in the number of P. micros colonies on PMM, in contrast to an average 6.1% recovery on PEA. Subgingival plaque and tongue samples from 12 adult periodontitis and 6 early-onset periodontitis patients were cultured onto PMM for the isolation of P. micros. P. micros was isolated on PMM and identified biochemically and enzymatically from both adult periodontitis and early-onset periodontitis patients with higher percentages isolated from the diseased periodontal pockets of adult periodontitis patients; furthermore, this is the first isolation of P. micros from tongue samples taken from periodontally diseased patients. This medium in cultural studies will further our understanding and assist future investigations of P. micros involved in disease processes.

Topics: Agar; Analysis of Variance; Bacteriological Techniques; Colony Count, Microbial; Culture Media; Dental Plaque; Evaluation Studies as Topic; Glutathione; Humans; Hydrogen Sulfide; Organometallic Compounds; Peptostreptococcus; Periodontitis; Sensitivity and Specificity; Staphylococcus; Statistics, Nonparametric; Streptococcus; Tongue

In four young adult monkeys, periodontitis was experimentally induced by inserting elastic rubber bands around the necks of two teeth on each side of the mandible. On one side the teeth were not cleaned during the experimental period. On the control side, the teeth were cleaned by scaling and polishing weekly. After 8 weeks there were 6 mm deep pockets and great bone loss on the experimental side. Subgingival plaque was collected from experimental and control sides under anaerobic conditions and transported in a reduced transport fluid. The microorganisms were incubated anaerobically for 7 days on four different media: 1) Horse blood agar, 2) Rabbit blood agar, 3) Kanamycin-vancomycin blood agar, 4) Rabbit blood agar with Kanamycin and supernatant of filtrated Propionibacterium acnes culture (BPBSM). All media contained 0.5 microgram/ml menadione and 5.0 micrograms/ml hemin. Colony forming units of black-pigmented Bacteroides were found on all four media from all animals and were increased in numbers in the experimental side. One hundred and fifteen isolates of black-pigmented colonies were identified biochemically. Recovery of black-pigmented Bacteroides on BPBSM was significantly higher than on the other three media.

Topics: Agar; Alveolar Bone Loss; Animals; Bacteroides; Colony Count, Microbial; Dental Plaque; Dental Scaling; Female; Macaca fascicularis; Periodontitis

On the basis of in vitro susceptibility testing of antibiotics, dyes, and other antimicrobial agents, we developed and evaluated a medium, TBBP, for the selective isolation of oral Capnocytophaga spp. TBBP medium consists of 4% Trypticase soy agar (BBL Microbiology Systems, Cockeysville, Md.), 5% sheep blood, 0.1% yeast extract, 50 micrograms of bacitracin per ml, and 100 micrograms of polymyxin B per ml. A total of 34 Capnocytophaga stock cultures grew well on TBBP medium. Except for some streptococcal strains, TBBP medium inhibited growth of all test stock culture isolates of common oral gram-positive and gram-negative species. In a clinical study of 15 deep periodontal pockets, TBBP medium demonstrated Capnocytophaga recoverability that was similar to or higher than that shown by a nonselective blood agar medium. Typical Capnocytophaga colonial morphology enabled us to readily distinguish this organism from the few other bacteria which could grow on TBBP medium.

Topics: Adolescent; Adult; Agar; Animals; Anti-Bacterial Agents; Bacitracin; Bacteria; Capnocytophaga; Coloring Agents; Culture Media; Cytophagaceae; Humans; Microbial Sensitivity Tests; Periodontitis; Polymyxin B; Sheep

A selective medium, malachite green bacitracin agar, was developed for the isolation of Actinobacillus actinomycetemcomitans from subgingival plaque of periodontally diseased patients. The medium consisted of Trypticase soy agar 40 gm/liter, bacitracin 128 micrograms/ml, malachite green 8 micrograms/ml and 5% defibrinated sheep blood. The medium, when incubated in an atmosphere of air plus 10% CO2 for 5 days, permitted greater than 80% recovery of pure cultures of A. actinomycetemcomitans when compared with a nonselective medium. The most frequent contaminant in plaque samples from different clinical conditions was Haemophilus aphrophilus. Decomposition of H2O2 was useful in differentiating these two species. Clinical studies employing the malachite green bacitracin medium revealed a significant association between the presence of the organism, A. actinomycetemcomitans and juvenile periodontitis.

Topics: Actinobacillus; Agar; Bacitracin; Bacteriological Techniques; Culture Media; Dental Plaque; Humans; Periodontitis; Rosaniline Dyes

Both prereduced molten agar and broth and aerobic molten agar and broth were inoculated with blood samples collected from patients with periodontitis, but in otherwise good health, both before and after extraction of two or more teeth. Postoperative blood samples from 23 of 25 patients sampled yielded anaerobic and facultative species. Colony counts from nine samples yielded from less than 1 to over 100 colonies per ml of blood. Organisms detected were species belonging to the genera Bacteroides, Fusobacterium, Peptostreptococcus, Leptotrichia, Propionibacterium, Peptococcus, Veillonella, plus Streptococcus mitis, S. salivarius, vibrio forms, and strains resembling S. mutans. The data indicate that prereduced anaerobically sterilized culture medium with polyanethol sulfonate is effective for detecting anaerobic species in bacteremia and that anaerobic species can be prevalent in bacteremias immediately after tooth extraction in patients with periodontitis.

Topics: Aerobiosis; Agar; Anaerobiosis; Animals; Anticoagulants; Bacteria; Bacteriological Techniques; Culture Media; Gastric Juice; Humans; Periodontitis; Postoperative Complications; Rumen; Sepsis; Sterilization; Sulfonic Acids; Tooth Extraction