Interactions between black-pigmented gram-negative anaerobes and other species which may be important in disease development.
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Bacterial Adhesion↗
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Black-pigmented gram-negative anaerobes in periodontitis.
Black-pigmented Gram-negative anaerobes have been associated with periodontal disease and tooth loss since they were first isolated by Burdon in 1928. Porphyromonas gingivalis, which is usually not isolated from children, adolescents or adults with no periodontal breakdown, has been recognized as one of the most important periodontopathogens. Its presence is strongly correlated with deep periodontal pockets, which are assumed to be its main habitat. Correlations have been shown also with attachment loss, clinical inflammation and serum antibody levels, indicating an aetiological role in the periodontal disease. Their pathogenicity in animal models resembling periodontal disease is documented. They are frequently isolated from periodontal abscesses. The relationship between Prevotella intermedia and periodontal disease is not clear. It is frequently isolated from advanced periodontitis, often as the only black-pigmented Gram-negative anaerobic species; however, the prevalence in adults with no periodontal breakdown is high. It is found frequently in periodontal abscesses and in acute necrotizing and ulcerative gingivitis. Serogroup I is found predominantly in deep periodontal pockets, whereas all serogroups (I-III) are found in shallow pockets and gingivitis. No conclusive difference in pathogenicity between serogroups has been found. Pr. melaninogenica, Pr. denticola and Pr. loescheii are frequently found in the gingival crevice in preschool children and other age groups with gingivitis, but are seldom found in deep periodontal pockets.
Adolescent↗
Black-pigmented gram-negative anaerobes in endodontic infections.
Necrotic dental root canal infections are polymicrobial infections dominated by anaerobic bacteria. The number of different species in one canal is usually low, approx. 4-7 species. The species isolated most frequently belong to the genera Prevotella, Porphyromonas, Fusobacterium, Peptostreptococcus, Eubacterium and Streptococcus. The frequency of isolation of black-pigmented Gram-negative anaerobes in endodontic infections varies from 25% to > 50%. Pr. intermedia is the most commonly found pigmented species, followed by Pr. denticola and two Porphyromonas species, P. gingivalis and P. endodontalis. Several studies have shown that P. gingivalis and P. endodontalis are closely related to the presence of acute symptoms in endodontic infections, whereas other black-pigmented Gram-negative anaerobes are not. However, several other species may also be involved in acute infections. Moreover, Porphyromonas species have occasionally been isolated from cases with no symptoms. Although Porphyromonas spp. are clearly related to symptoms at the beginning of therapy, they are not important for the prognosis of the treatment.
Bacteroidaceae↗
Black-pigmented gram-negative anaerobes in genito-urinary tract and pelvic infections.
Black-pigmented Gram-negative anaerobes are part of the normal vaginal flora and contribute to a range of superficial and deep genital infections. Prevotella melaninogenica is found in moderate numbers (10(4-6) cfu/g) in healthy women; the numbers and detection rates increase in anaerobic vaginosis (where it may be a significant contributor to changed microbial metabolic activity that gives the signs and symptoms of this condition) and in other vaginal infective conditions. Pr. melaninogenica is also part of the mixed flora in deep pelvic infections: endometritis, post-partum and post-abortal uterine infections; salpingitis and tubo-ovarian abscesses; PID and pelvic abscesses. Porphyromonas asaccharolytica is probably not a vaginal commensal, but may be isolated from patients with vaginal or pelvic disease. It is more specifically associated with superficial abscesses (e.g. Bartholin's abscess) and ulcers of the genitalia and perineum. P. asaccharolytica was the commonest species isolated from men with genital ulcers of various primary causes and ranging in severity from superficial balanitis/balanoposthitis to synergic gangrene.
Abscess↗
Antibiotic susceptibilities of black-pigmented gram-negative anaerobes.
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Anti-Bacterial Agents↗
Detection and identification of Mobiluncus species by direct filter hybridization with an oligonucleotide probe complementary to rRNA.
A hybridization assay for direct detection and identification of Mobiluncus species has been developed and tested. A [32P]-labelled synthetic oligonucleotide probe, complementary to a nucleotide sequence in the variable region V8 of Mobiluncus 16S ribosomal RNA, was utilized. One of the advantages of using rRNA as target molecule for the hybridization assays is the copy number of rRNA, which can be as high as 10(4), and that additionally three to six sites on the minus strand of the DNA gene can be utilized. This probe was found to be sensitive and to react with 62 of 68 tested typical or atypical Mobiluncus isolates. It was also specific, and was shown not to react with 96 tested unrelated bacterial species and isolates, including taxonomically closely related species like Actinomyces or Bifidobacterium spp., or with bacteria isolated from the vagina of both healthy persons with an undisturbed flora, as well as from patients suffering from the bacterial vaginosis syndrome (BV).
Bacteroidaceae↗
Microbiologic response to treatment of bacterial vaginosis with topical clindamycin or metronidazole.
To compare the frequencies, concentrations, and antimicrobial susceptibilities of vaginal microbes isolated from women with bacterial vaginosis (BV) before and after therapy, 119 nonpregnant women aged 18 to 45 with clinical and Gram stain evidence of BV were randomized to receive intravaginal clindamycin or metronidazole. Vaginal swabs were collected at baseline and 7 to 12 days, 35 to 45 days, and 70 to 90 days following therapy for quantitative vaginal culture. For the 99 women completing all four visits, statistical analyses were performed comparing differences in vaginal microflora between the two treatment arms and between visits in the same treatment group. Antimicrobial susceptibility testing using the agar dilution method was performed for anaerobic gram-negative rods. Although both therapies resulted in decreased colonization by Gardnerella vaginalis and Mycoplasma hominis, only metronidazole treatment resulted in a significant decrease in the frequency and concentration of Prevotella bivia and black-pigmented Prevotella species. Of the 865 anaerobic gram-negative rods evaluated for susceptibility, only 3 (0.3%) were resistant to metronidazole, whereas clindamycin resistance increased significantly for P. bivia and black-pigmented anaerobic gram-negative rods persisting following clindamycin therapy. Clindamycin-resistant subpopulations of P. bivia and black-pigmented Prevotella species emerged 7 to 12 days after therapy even among women colonized initially by clindamycin-susceptible strains. These resistant subpopulations persisted at high frequencies (42 to 50%) 70 to 90 days following therapy. The two topical agents for treatment of BV have differing microbiologic effects on the vaginal microflora. The emergence of clindamycin-resistant anaerobic gram-negative rods following therapy is of concern.
Administration, Topical↗
Porphyromonas gingivalis prevalence related to other micro-organisms in adult refractory periodontitis.
Forty-six adult periodontal patients, selected on the basis of clinical examination, and 46 adult healthy subjects were examined. The subgingival plaque samples from one inflammatory and one non-inflammatory site of each periodontal patient were studied to determine Porphyromonas gingivalis prevalence related to other periodontal micro-organisms and to periodontal tissue destruction. The results showed Porphyromonas gingivalis as the main pathogenic micro-organism isolated in the inflammatory sites together with Bacteroides forsythus. Peptostreptococcus sp., Actinomyces sp. and Prevotella sp. were found as a normal oral flora in the healthy subjects. Fusobacterium nucleatum, Prevotella intermedia, Campylobacter rectus and Eikenella corrodens were detected both in inflammatory and in non-inflammatory sites of periodontal patients as well as in the healthy subjects.
Adult↗
Impaired vocal cord mobility in the setting of acute suppurative thyroiditis.
BACKGROUND: Acute suppurative thyroiditis is an increasingly rare entity, accounting for less than 0.1% of thyroid surgery. This is the first report in the literature of acute thyroiditis causing a unilateral reversible vocal cord paralysis. METHODS: A 41-year-old woman was initially seen with a unilateral vocal cord paralysis resulting from acute suppurative thyroiditis. RESULTS: The vocal cord paralysis resolved following surgical drainage of the thyroid abscess and treatment with intravenous antibiotics. CONCLUSIONS: We present this case to emphasize the need for a complete and thorough head and neck exam including preoperative documentation of vocal cord mobility.
Acute Disease↗
The relationship between the presence of periodontopathogenic bacteria in saliva and halitosis.
OBJECTIVE: To evaluate the association between the presence of periodontal pathogenic bacteria in saliva and halitosis in mouth air. DESIGN: Cross-sectional microbiological and clinical oral examination of adult patients. SUBJECTS: 101 adult patients (25 males, 76 females) who attended the Preventive Dentistry and Breath Odour Clinic of Kyushu Dental College. Their average age was 50.0+/-13.5 years old (mean +/- SD). SETTING: The subjects were classified into three groups: halitosis subjects with a probing depth (PD) > or = 4mm (P group), halitosis subjects without PD > or = 4mm (H group), and non-halitosis subjects without PD > or = 4mm (C group). METHODS: All subjects received a periodontal examination. Volatile sulphur compounds (VSC: hydrogen sulphide and methyl mercaptan) were measured using gas chromatography. The presence of Bacteroides forsythus, Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Prevotella intermedia in the saliva was detected by polymerase chain reaction. RESULTS AND CONCLUSION: The presence of B. forsythus, P. gingivalis and P. intermedia influenced the production of VSC. Specifically, the presence of B. forsythus in subjects with periodontitis was strongly correlated to the concentration of VSC in mouth air.
Actinobacillus Infections↗
Methyl mercaptan production by periodontal bacteria.
Oral malodour is principally caused by volatile sulphur compounds (VSC) such as hydrogen sulphide, methyl mercaptan and dimethyl sulphide. Methyl mercaptan is highly toxic, and its presence within a periodontal pocket suggests involvement in the induction and/or progression of periodontal disease. Methyl mercaptan is produced from L-methionine by L-methionine- alpha -deamino- gamma -mercaptomethane-lyase (METase). METase catalyses the alpha,gamma-eliminating reaction of L-methionine, which results in the release of alpha-ketobutyrate, methyl mercaptan and ammonia. Although methyl mercaptan is produced by a variety of microorganisms, Porphyromonas gingivalis is considered to be the most potent producer. METases of P. gingivalis have been characterised and the genes responsible for their production, the mg/genes, have been sequenced. To ascertain the role of METase in P. gingivalis pathogenicity, a METase-deficient mutant strain (M1217) from P. gingivalis strain W83 was engineered. Only 7.7% of the mice infected with W83 survived 4 days after subcutaneous injection, whereas 36% of the mice infected with M1217 survived over the same time period. Many papers have reported the periodontal pathogenesis of VSC. It has been argued that methyl mercaptan may play a significant role in the pathogenicity of P. gingivalis.
Ammonia↗
Porphyromonas gingivalis lipopolysaccharide induces shedding of syndecan-1 expressed by gingival epithelial cells.
Syndecans are constitutively shed from growing epithelial cells as the part of normal cell surface turnover. However, increased serum levels of the soluble syndecan ectodomain have been reported to occur during bacterial infections. The aim of this study was to evaluate the potential of lipopolysaccharide (LPS) from the periodontopathogen Porphyromonas gingivalis to induce the shedding of syndecan-1 expressed by human gingival epithelial cells. We showed that the syndecan-1 ectodomain is constitutively shed from the cell surface of human gingival epithelial cells. This constitutive shedding corresponding to the basal level of soluble syndecan-1 ectodomain was significantly increased when cells were stimulated with P. gingivalis LPS and reached a level comparable to that caused by phorbol myristic acid (PMA), an activator of protein kinase C (PKC) which is well known as a shedding agonist. The syndecan-1 shedding was paralleled by pro-inflammatory cytokine interleukin-1 beta (IL-1beta), IL-6, IL-8, and tumor necrosis factor alpha (TNF-alpha) release. Indeed, secretion of IL-1beta and TNF-alpha increased following stimulation by P. gingivalis LPS and PMA, respectively. When recombinant forms of these proteins were added to the cell culture, they induced a concentration-dependent increase in syndecan-1 ectodomain shedding. A treatment with IL-1beta converting enzyme (ICE) specific inhibitor prevented IL-1beta secretion by epithelial cells stimulated by P. gingivalis LPS and decreased the levels of shed syndecan-1 ectodomain. We also observed that PMA and TNF-alpha stimulated matrix metalloproteinase-9 secretion, whereas IL-1beta and P. gingivalis LPS did not. Our results demonstrated that P. gingivalis LPS stimulated syndecan-1 shedding, a phenomenon that may be mediated in part by IL-1beta, leading to an activation of intracellular signaling pathways different from those involved in PMA stimulation.
Adult↗
A clinical study: Melaleuca, Manuka, Calendula and green tea mouth rinse.
A novel mouthrinse (IND 61,164) containing essential oils and extracts from four plant species (Melaleuca alternifolia, Leptospermum scoparium, Calendula officinalis and Camellia sinensis) were tested. This study aimed to evaluate the safety, palatability and preliminary efficacy of the rinse. Fifteen subjects completed the Phase I safety study. Seventeen subjects completed the Phase II randomized placebo-controlled study. Plaque was collected, gingival and plaque indices were recorded (baseline, 6 weeks, and 12 weeks). The relative abundance of two periodontal pathogens (Actinobacillus actinomycetemcomitans, Tanerella forsythensis) was determined utilizing digoxigenin-labeled DNA probes. ANCOVA was used at the p = 0.05 level of significance. Two subjects reported a minor adverse event. One subject withdrew from the study. Several subjects objected to the taste of the test rinse but continued treatment. Differences between gingival index, plaque index or relative abundance of either bacterial species did not reach statistical significance when comparing nine placebo subjects with eight test rinse subjects. Subjects exposed to the test rinse experienced no abnormal oral lesions, altered vital signs, changes in liver, kidney, or bone marrow function. Larger scale studies would be necessary to determine the efficacy and oral health benefits of the test rinse.
Adult↗
L-Methionine-gamma-lyase, as a target to inhibit malodorous bacterial growth by trifluoromethionine.
Methyl mercaptan is a major component responsible for oral malodor. This compound arises from the bacterial metabolism of methionine. Here we show that the growth of Porphyromonas gingivalis, a periodontal microorganism that produces large amounts of methyl mercaptan, was strongly inhibited by l-trifluoromethionine (TFM), a fluorinated derivative of methionine. In contrast, TFM had no effect on the growth of bacteria which do not produce methyl mercaptan. In addition, the survival rate of P. gingivalis-infected mice was remarkably increased by the co-injection of TFM. These results suggest that TFM is a promising antibacterial agent specific to the malodorous oral bacteria.
Animals↗
Trypsin-like protease activity of Porphyromonas gingivalis as a potential virulence factor in a murine lesion model.
Porphyromonas gingivalis possesses a large number of enzymatic activities which might be important in the virulence of this putative periodontopathogen. The purpose of this study was to examine these enzymatic activities in vivo in a murine model to assess their role in soft tissue destruction. Whole cells of P. gingivalis strains whether grown on blood agar plates or in broth exhibited high levels of alkaline phosphatase (ALPase), a trypsin-like protease (TLPase), acid phosphatase (ACPase), N-acetyl beta-glucosaminidase (Na beta-Gase) enzymes and collagenolytic activities. P. gingivalis W50 treated with 2 mM Na-P-tosyl-L-lysine chloromethyl ketone (TLCK)/phenylmethylsulfonyl fluoride (PMSF) prior to subcutaneous infection of mice failed to induce a phlegmonous abscess and lethality characteristic of animals challenged with untreated P. gingivalis. Comparison of wild type P. gingivalis strain 3079.03 with its protease-deficient (TLPase-negative) mutant NG4B19 revealed the mutant to be avirulent (no lesion and no death) in this model. P. gingivalis BEI and SW5 mutants (parent W50), which partially lacked TLPase enzyme activity produced only localized lesions, and no death. Thus, the TLPase enzyme appears to be correlated with the lesion type (spreading or localized), lesion size, and death in this mouse abscess model. Therefore, the enzymatic activities of P. gingivalis and specifically the TLPase enzyme could play an important role in periodontal disease by enhancing bacterial spread and degrading gingival tissues.
Acetylglucosaminidase↗
Host modulation of tissue destruction caused by periodontopathogens: effects on a mixed microbial infection composed of Porphyromonas gingivalis and Fusobacterium nucleatum.
These studies determined the ability of selected periodontopathogens to synergistically initiate soft tissue destruction in a murine abscess model. The development of immunity following recovery from infection or by active immunization was also examined. Mice were infected with P. gingivalis W50, F. nucleatum T18, or a combination of the two microorganisms. F. nucleatum caused only a localized lesion in contrast to P. gingivalis which caused a spreading suppurative inflammatory lesion of the skin and subcutaneous tissues, which, depending upon the dose, could result in death. Infection of mice with a combination of P. gingivalis and F. nucleatum elicited a significantly greater lesion size (P<0.001) and lethality compared with P. gingivalis alone. Mice infected with a subclinical dose (no visible lesion) of P. gingivalis failed to develop protective immunity to a secondary P. gingivalis challenge. Mice that had recovered from P. gingivalis lesions demonstrated partial protection against subsequent P. gingivalis challenge; however, the immunity was less protective against the mixed F. nucleatum + P. gingivalis infection. Active immunization with P. gingivalis protected against both the P. gingivalis and F. nucleatum + P. gingivalis challenges and this protection was correlated with the levels of specific serum immunoglobulin G (IgG) antibody. The results indicated that the murine model is ideally suited to examine bacterially-mediated mixed infections that result in soft tissue destruction. This destruction can be minimized, but not abrogated, with development of immunity. Challenge with sufficient numbers of the pathogens can overwhelm the acquired immunity.
Animals↗
Porphyromonas gingivalis virulence in a murine lesion model: effects of immune alterations.
This study utilized various mouse strains with documented alterations in immune system components to assess their contribution to modify the virulence of Porphyromonas gingivalis. P. gingivalis W50 was cultivated on blood agar plates, harvested and used to challenge mice by subcutaneous injection on the dorsolateral surface of the back. Soft tissue lesion development was estimated by measuring the area of the spreading lesion formed by this microorganism over a period of 15 days. Challenge of various normal inbred and outbred mouse strains including: BALB/cN, BALB/cJ, BALB/c nu/+, ICR, B10.A(4R), B10.MBR, A/J, C57BL/6J, CBA/CaH, C.B-17/Icv Tacf DF and C3H/HeN with 2 x 10(10) bacteria showed similar lesion size among these strains (approximately 400 mm2). Genetically deficient mouse strains [C.B-17/Icr Tac (SCID); DBA/2 (C5 deficient); BALB/c nu/nu (T cell deficient); CBA/CaHN-XID/J (B cell deficient) and C3H/HeJ (LPS hyporesponsive)] demonstrated a lesion size which was similar to normal animals. C57BL/6J-BgJ (NK cell deficient) mice exhibited a significantly more severe lesion than the other strains tested. Following healing of the lesions, we initiated a secondary infection of the surviving animals to estimate the acquisition of protective immunity following recovery from the primary infection. Normal mice demonstrated a delayed onset and decrease in lesion size of 15 to 30% compared with the primary infection. In contrast, each of the immunodeficient strains appeared unable to develop immune protection to the secondary challenge. The findings suggest that protection against primary infections with P. gingivalis are mediated by innate immune mechanisms (PMN. NK cells). Additionally, it appears that T-cell-dependent humoral responses are critical to developing immunity to subsequent P. gingivalis infection.
Animals↗
Comparison of pathogenic properties between two types of arginine-specific cysteine proteinases (gingipains-R) from Porphyromonas gingivalis.
Two major arginine-specific cysteine proteinases (gingipains R) from Porphyromonas gingivalis have been compared with regard to their potential participation in the pathology of periodontal disease. Both the high and low molecular mass forms, HRgpA and RgpB, cleaved oligopeptide fluorogenic substrates at the P1-arginine residue with essentially identical specificity but different efficiencies, with HRgpA being about 1.5 to seven-fold less potent than RgpB. In contrast HRgpA, which occurs as a non-covalent complex of catalytic and hemagglutinin/adhesion domains, was about two-fold more active than RgpB in degrading fibrinogen and fibrin, while both enzymes activated prekallikrein with similar efficiency. These data indicate the likelihood that both activities could be involved in both the bleeding tendency and production of gingival crevicular fluid, which occur at infected periodontitis sites. Significantly, however, is the fact that HRgpA, but not RgpB, was able to bind phospholipids in the presence of calcium ions, the effect dramatically enhancing the activation of clotting factors by this proteinase. This suggests that HRgpA may play a more important role in the virulence of Porphyromonas gingivalis, relative to RgpB, almost certainly because of the presence of the hemagglutinin/adhesion domain which can bind phospholipid and apparently modulate enzyme activity.
Adhesins, Bacterial↗