Bacteremia after plate removal and tooth extraction.

Our aim was to investigate the occurrence of bacteremia associated with removal of a semirigid osteosynthesis plate and an adjacent third molar. Ten patients with fixed mandibular angle fracture were bacteriologically sampled from the second molar's distal gingival pocket, from the third molar's extraction socket and from the osteosynthesis plate. Blood samples from the ante-cubital vein were taken 10 times until 30 min postoperatively. Established culture, isolation and identification methods for the bacterial species were used. Bacteremia was detected in 60% of the subjects, most frequently 1.5 min after removal of the plate (20%) and 1.5 and 5 min after extraction of the tooth (20%), but also 10 min (10%) and 30 min (10%) postoperatively. 13 different bacterial species or groups were isolated, mean 2.5 +/- 1.9 per bacteremia-positive subject. The majority (85%) were anaerobes with Actinomyces, Campylobacter and Lactobacillus species predominating. In all the blood culture-positive cases the corresponding species was also recovered from one or more of the oral samples. These results show that oral surgical procedures are associated with a high frequency of longstanding anaerobic bacteremia, which could be harmful in patients at risk.

Characterisation of aerobic gram-negative bacteria from subgingival sites of dogs--potential bite wound pathogens.

Ninety-eight aerobic, gram-negative bacterial isolates from subgingival samples from family-owned dogs with naturally occurring periodontitis were characterised phenotypically by conventional biochemical testing, by cellular fatty acid profiling and by the use of commercial identification systems. The majority (48, 81%) of the fermentative isolates but only 18% of the non-fermenters were identified by conventional biochemical testing alone. With additional cellular fatty acid profiling, another 7 (12%) fermentative and 23 (59%) non-fermentative isolates were identified to genus or group level. Cellular fatty acid analysis was essential for the identification of most non-fermenters, many of which are difficult to identify due to a paucity of positive reactions in routine biochemical tests. Commercial identification systems were less useful and did not contribute to further identification of these problematic isolates. This study underlines the difficulties encountered in the identification of canine oral bacteria--a group of potential bite wound pathogens--and presents schemes for microbiology laboratories to characterise such isolates.

Oral colonization with Actinomyces species in infants by two years of age.

In early childhood, the human mouth is already colonized by actinomycetes. Due to recent taxonomic changes within the genus Actinomyces, up-to-date data are warranted on the time and succession of different Actinomyces species in the oral cavity. By using a longitudinal study design and culture techniques, we examined the age-related occurrence of Actinomyces species in saliva from 39 healthy infants at 2, 6, 12, 18, and 24 months of age. Altogether 428 Actinomyces isolates were available for this study. Identification was based on biochemical tests and gas chromatographic demonstration of metabolic end-products, and when needed, cellular fatty acid profiles were determined. The frequency of the total actinomycetal flora increased from 31% to 97% within 2 years. A. odontolyticus was the most prominent Actinomyces colonizer at all five sampling occasions. A. naeslundii was the second most common Actinomyces sp. but was not detected before the age of 1 year. As a novel observation, we found A. graevenitzii in the oral cavity. The number of A. graevenitzii isolates indicates that this species is not just occasionally present in infants' mouths. We also found A. viscosus, A. gerencseriae, A. israelii, and A. georgiae. Based on the present results, we suggest that A. odontolyticus is the main primary Actinomyces species on oral mucosal surfaces in infants up to 2 years of age.

Establishment of oral anaerobes during the first year of life.

Anaerobic species constitute a significant part of the bacterial community of the mouth. Although the time and species involved in the primary colonization of infants are of great importance by forming the basis for further colonization, the development of the oral anaerobic microflora with age is still inadequately understood. In the present study, time and succession of colonization of oral anaerobes were longitudinally examined in 44 healthy Caucasian infants at 2, 6, and 12 months of age. Unstimulated saliva samples were quantitatively cultured on non-selective Brucella blood agar and several selective media for the isolation of anaerobic micro-organisms. The most frequent anaerobic finding in two-month-old infants was Veillonella spp. The Prevotella melaninogenica group also represented early colonizing species, and the frequency increased remarkably during the first year of life, whereas the Prevotella intermedia group organisms seemed to be late colonizers. Fusobacterium nucleatum, non-pigmented Prevotella spp., and Porphyromonas catoniae were occasional findings in subjects at 2 months but frequent findings in those at one year of age. F. nucleatum was the most frequent strictly anaerobic species in one-year-old infants; other fusobacteria were also occasionally found. The frequency of facultative/micro-aerophilic corroding rods and Capnocytophaga spp. started to increase toward the end of the first year. Except for the common presence of facultative/micro-aerophilic Actinomyces spp., other anaerobic gram-positive species were only occasionally present in these infants. Once established, early-colonizing species tended to persist in the mouth. Our longitudinal study demonstrated the establishment of several anaerobic species with steadily increasing frequencies during the first year of life.

beta-lactamase production and antimicrobial susceptibility of oral heterogeneous Fusobacterium nucleatum populations in young children.

Oral Fusobacterium nucleatum populations from 20 young, healthy children were examined for beta-lactamase production. Ten children (50%) harbored, altogether, 25 beta-lactamase-positive F. nucleatum isolates that were identified as F. nucleatum subsp. polymorphum, F. nucleatum subsp. nucleatum, and F. nucleatum subsp. vincentii (J. L. Dzink, M. T. Sheenan, and S. S. Socransky, Int. J. Syst. Bacteriol. 40:74-78, 1990). In vitro susceptibility of these beta-lactamase-producing and 26 non-beta-lactamase-producing F. nucleatum isolates was tested with penicillin G, amoxicillin-clavulanic acid, tetracycline hydrochloride, metronidazole, trovafloxacin, and azithromycin. Except for penicillin G, the antimicrobials exhibited good activity against all F. nucleatum isolates.

Susceptibilities of bovine summer mastitis bacteria to antimicrobial agents.

The susceptibility to 9 antimicrobial agents of 32 aerobic bacterial isolates and to 10 antimicrobial agents of 37 anaerobic bacterial isolates from 23 cases of bovine summer mastitis (16 Actinomyces pyogenes isolates, 8 Streptococcus dysgalactiae isolates, 3 S. uberis isolates, 3 S. acidominimus isolates, 2 Streptococcus spp., 15 Peptostreptococcus indolicus isolates, 10 Fusobacterium necrophorum isolates, and 12 isolates of anaerobic gram-negative rods) was determined by the agar dilution method. All isolates except one Bacteroides fragilis isolate (beta-lactamase producer) were susceptible to penicillin G, amoxicillin, amoxicillin-clavulanate, cefoxitin, clindamycin, and chloramphenicol (the B. fragilis strain was susceptible to the last four), which had MICs at which 90% of isolates were inhibited (MIC90s) of < or = 0.06, < or = 0.06, < or = 0.06 0.25, < or = 0.06, and 4.0 micrograms/ml, respectively. Spiramycin was active against the gram-positive aerobes (MIC90, 1.0 microgram/ml) but not against the anaerobes (MIC90, 16.0 micrograms/ml). Similar trends were noted for susceptibilities of aerobic and anaerobic bacteria to ofloxacin (MIC90s, 2.0 and 8 micrograms/ml, respectively). Occasional strains of aerobic streptococci were resistant to oxytetracycline, but all anaerobes were susceptible. Tinidazole was active against all anaerobes (MIC90, 2.0 micrograms/ml). beta-Lactamase was produced only by the B. fragilis isolate.

Atrial natriuretic peptide and its mRNA in heart and brain of vasopressin-deficient Brattleboro rats.

To understand the secretion and synthesis of atrial natriuretic peptide we measured immunoreactive atrial natriuretic peptide from plasma, heart tissues and brain areas, and ANP mRNA was determined from heart auricles and ventricles of vasopressin-deficient Brattleboro rats (DI) and from desmopressin treated Brattleboro rats (DI+DDAVP). Long-Evans rats (LE) served as controls. DI+DDAVP rats were given for 3 days sc. injections of 0.5 micrograms 1-desamino-8-D-arginine vasopressin in 1 mL saline twice a day. The rats were housed in single metabolic cages and urinary output and water intake were measured daily. All the body and organ weight parameters were similar in the three groups when the rats were killed. No change was seen in the plasma ANP level between the groups. The right ventricle of DI+DDAVP rats had significantly (P < 0.05) higher concentration of ANP than LE rats (15.8 +/- 4.4 vs. 3.4 +/- 0.6 ng mg-1 tissue). The left ventricle of DI and DI+DDAVP had significantly (P < 0.05) lower amounts of ANP mRNA than LE rats (0.5 +/- 0.2 vs. 1.3 +/- 0.2 and 0.5 +/- 0.1 vs. 1.3 +/- 0.2 arbitrary units). In the hypothalamus, the ANP concentration was significantly (P < 0.05) lower both in DI and in DI+DDAVP rats than in LE rats (9.3 +/- 1.3 vs. 14.5 +/- 1.6 and 6.1 +/- 0.6 vs. 14.5 +/- 1.6 pg mg-1 tissue). To conclude, although the water intake and urinary output of DI rats were changed towards normal with desmopressin treatment, the heart ventricular and hypothalamic ANP did not parallel the change.(ABSTRACT TRUNCATED AT 250 WORDS)

Phylogenetic analysis of members of the genus Porphyromonas and description of Porphyromonas cangingivalis sp. nov. and Porphyromonas cansulci sp. nov.

The partial 16S rRNA gene sequences of representative strains of two groups of anaerobic, gram-negative, pigmented, asaccharolytic, rod-shaped bacteria isolated from subgingival plaque of dogs with naturally occurring periodontal disease were determined. A comparative analysis of the rRNA sequence data revealed that the two groups of organisms represent previously unknown lines of descent within the genus Porphyromonas. On the basis of our phylogenetic findings and the phenotypic distinctiveness of the organisms, two new species, Porphyromonas cangingivalis and Porphyromonas cansulci, are proposed.

Porphyromonas canoris sp. nov., an asaccharolytic, black-pigmented species from the gingival sulcus of dogs.

A new species, Porphyromonas canoris, is proposed for black-pigmented asaccharolytic strains isolated from subgingival plaque samples from dogs with naturally occurring periodontal disease. This bacterium is an obligately anaerobic, nonmotile, non-spore-forming, gram-negative, rod-shaped organism. On laked rabbit blood or sheep blood agar plates, colonies are light brown to greenish brown after 2 to 4 days of incubation and dark brown after 14 days of incubation. Colonies on egg yolk agar and on nonhemolyzed sheep blood agar are orange. The cells do not grow in the presence of 20% bile and have a guanine-plus-cytosine content of 49 to 51 mol%. The type strain is VPB 4878 (= NCTC 12835). The average levels of DNA-DNA hybridization between P. canoris strains and other members of the genus Porphyromonas are as follows: Porphyromonas gingivalis ATCC 33277T (T = type strain), 6.5%; Porphyromonas gingivalis cat strain VPB 3492, 5%; Porphyromonas endodontalis ATCC 35406T, 1%; Porphyromonas salivosa NCTC 11362T, 5%; and Porphyromonas circumdentaria NCTC 12469T, 6%. The level of hybridization between P. canoris NCTC 12835T DNA and Porphyromonas asaccharolytica ATCC 25260T DNA is 3%. P. canoris cells produce major amounts of acetic, propionic, isovaleric, and succinic acids and minor amounts of isobutyric and butyric acids as end products of metabolism in cooked meat medium. The major cellular fatty acid is 13-methyltetradecanoic acid (iso-C15:0). Glutamate and malate dehydrogenases are present, as are glucose-6-phosphate dehydrogenase activity (65.7 nmol mg of protein-1 min-1) and 6-phosphogluconate dehydrogenase activity (63.0 nmol mg of protein-1 min-1).(ABSTRACT TRUNCATED AT 250 WORDS)

The efficacy of tinidazole in naturally occurring periodontitis in dogs: bacteriological and clinical results.

The efficacy of tinidazole in addition to a single course of scaling was studied in 14 dogs with periodontitis. Three test teeth, two with periodontitis and one with healthy periodontium, were selected per dog. Subgingival bacterial samples were taken, and clinical examination was carried out at each of four visits (0, 14, 90 and 180 days). The bacterial samples were cultured anaerobically using selective and nonselective media. All teeth in the dentition, except one diseased test tooth with periodontitis were scaled at the first visit and tinidazole or placebo administered twice a day for 7 days. The mean probing depth of the diseased test pockets was significantly more reduced in tinidazole+scaling (T1S1) than scaling alone (T0S1) group at all visits compared to baseline values. The most significant decreases of bacterial counts in T1 or T0 groups were found in agar corroding Gram-negative rods in both diseased and healthy T1S1 pockets and in slimy Gram-negative rods in diseased T1S0 pockets between visits 1 and 2. The intergroup comparisons showed that Gram-positive cocci decreased significantly more in S1 pockets as well as sporeforming Gram-positive rods in diseased S0 pockets of T1 than T0 group. The highest number of isolates was found in the group of pigmented Gram-negative anaerobe rods, mainly Porphyromonas spp. The percentage of Porphyromonas gingivalis-like isolates decreased to zero and Porphyromonas endodontalis-like isolates increased in all test teeth of T1 group at 14 days. It is concluded that tinidazole has good efficacy against P. gingivalis-like bacteria which seem to be periodontal pathogens in dogs.

Porphyromonas-like gram-negative rods in naturally occurring periodontitis in dogs.

A total of 259 Gram-negative Porphyromonas-like rods isolated from subgingival plaque samples of 16 family-owned dogs with naturally occurring periodontitis were characterized phenotypically by biochemical reactions, metabolic end products and enzymatic activities (API-ZYMTM, RoscoTM). Four distinct groups were found. Group A isolates (63) were asaccharolytic, lipase negative, trypsin positive and produced phenylacetic acid (PAA) from peptone-yeast extract glucose broth. Unlike P. gingivalis strains they were catalase positive. Group B isolates (42) differed from those of group A by a positive lipase reaction and from those of group D by failing to ferment sugars. Group C isolates (88) were asaccharolytic and did not produce PAA. They were alpha-fucosidase, N-acetyl-beta-glucosaminidase (beta-NAG) and trypsin negative, resembling P. endodontalis, but unlike human isolates, they were catalase positive. Subgroup C.1 isolates (6) differed from those of parent group C by producing minor amounts of PAA, and subgroup C.2 isolates (12) were beta-NAG positive. Group D isolates (46) were weakly fermentative, lipase, catalase and trypsin positive, and produced PAA. They resembled the B (P.) salivosus type strain which, in our hands, fermented weakly glucose, lactose and mannose. Two isolates could not be assigned to any of the previous groups.

Plasma ANP during hypertonic NaCl infusion in man.

To determine the relationship between hyperosmolality and immunoreactive atrial natriuretic peptide of heart atrial plasma six healthy men were given 0.06 ml kg-1 min-1 855 mmol l-1 NaCl, i.v., for 2 h. The right atrial pressure and atrial plasma atrial natriuretic peptide were measured. During the infusion, right atrial pressure was kept constant by lowering the legs of the subject in a supine position downwards if any increase in the pressure was seen. There was a significant and linear increase in atrial serum osmolality, from 288 +/- 3.3 to 307 +/- 3.2 mOsm kg-1 (P less than 0.001). No statistically significant changes in right atrial pressure were seen. Regression analysis revealed that there was a statistically significant correlation between serum osmolality and plasma ANP in three subjects (responders) (r2: 0.5241, 0.8965, 0.6695). In three other subjects (nonresponders), there was no correlation between osmolality and ANP. The mean basal osmolality of responders was 280 mOsm kg-1 and the mean basal osmolality of nonresponders was 295 mOsm kg-1. In contrast, all subjects responded with an increase in plasma ANP (P less than 0.05) after RAP had been increased by tilting the legs of the subject upwards for 30 min. We conclude that the right atrial pressure regulates the release of atrial natriuretic peptide. Serum hyperosmolality may also contribute to the regulation of atrial natriuretic peptide independently of the right atrial pressure in man.

Are certain oral pathogens part of normal oral flora in denture-wearing edentulous subjects?

A complete loss of teeth may eliminate some pathogenic bacteria for lack of a suitable habitat for colonization. The occurrence of Actinobacillus actinomycetemcomitans, black-pigmented Bacteroides, mutans streptococci, lactobacilli and yeasts was investigated in 51 edentulous subjects with complete denture(s); mean age 74 years (range 44-92 years). None of the subjects had received antibiotics during the preceding 6 months. Four swab samples per subject were taken from the fitting surface of the upper denture, from the palate, buccal mucosae and dorsum of the tongue. At least 0.5 ml of stimulated saliva was collected into a sterile tube. The samples were cultivated aerobically and anaerobically using nonselective and selective media. A. actinomycetemcomitans was not found in any of the samples. Black-pigmented Bacteroides were found in 49 (96%) and yeasts in 25 (49%) of the subjects. Of the 49 saliva samples, 41 (84%) yielded mutans streptococci and 45 (92%) lactobacilli. In contrast to black-pigmented Bacteroides, mutans streptococci and lactobacilli, A. actinomycetemcomitans may not belong to the normal oral flora of edentulous subjects wearing dentures.

Daily patterns of plasma atrial natriuretic peptide, serum osmolality and haematocrit in the rat.

The present study documents daily rhythms of plasma atrial natriuretic peptide, serum osmolality and haematocrit in the rat. One-hundred and twenty-five Sprague-Dawley rats were used. They were bred under a cycle of 12 h light/12 h dark starting at 07.00 h. Fifty-three rats were decapitated between 09.00 and 16.00 h (study I) and 72 rats in groups of six were decapitated at 2-h intervals for a period of 24 h (study II). In study I, plasma atrial natriuretic peptide was 156 +/- 11 pg mg-1 (mean +/- SEM). In study II, atrial natriuretic peptide was at a control level from 08.00 to 18.00 h and then began to increase. At 22.00 h, atrial natriuretic peptide was 420 +/- 105 pg ml-1, which was significantly higher than at 08.00 h (P less than 0.05). The serum osmolality was over 300 mosmol kg-1 during the day. The highest mean osmolalities (315, 317, 312 mosmol kg-1) were found from 18.00 to 22.00 h. These were significantly different (P less than 0.05) from other groups during the day. The haematocrit was highest at 14.00 h (49.5 +/- 0.7%) and lowest at 24.00 h (43.6 +/- 0.8%) (P less than 0.05). In conclusion, we have shown that there are significant daily rhythms of plasma atrial natriuretic peptide, serum osmolality and haematocrit during a 24-h period and 12 h light/12 h dark cycle in the rat.

The long-term efficacy of systemic doxycycline medication in the treatment of localized juvenile periodontitis.

The presence of Actinobacillus actinomycetemcomitans in the deep periodontal pockets of patients with localized juvenile periodontitis has been causally associated with active periodontal destruction. Thus, eradication of this microorganism has become the goal of treatment. It has been postulated that such eradication cannot be achieved without systemic antimicrobial treatment. The efficacy of a semisynthetic tetracycline (doxycycline) in a double-blind follow-up study of 14 patients with localized juvenile periodontitis was evaluated. For assessment of the periodontal status, probing depth and bleeding after probing at 4 sites of all teeth were recorded. The treatment consisted of instruction in oral hygiene, scaling and root planing, periodontal surgery and systemic medication for 2 weeks with either doxycycline (Doximycin) or placebo. A. actinomycetemcomitans was cultivated from subgingival samples taken from 4 sites. The periodontal condition and the prevalence of A. actinomycetemcomitans were monitored at the baseline and at 2, 8 and 20 months. The periodontal condition improved in both groups; the only significant difference was a greater reduction in the prevalence of A. actinomycetemcomitans 8 months after treatment with doxycycline as compared with the placebo.

The immediate efficacy of adjunctive doxycycline in treatment of localized juvenile periodontitis.

A randomized, double-blind, placebo-controlled study on the immediate clinical and microbiological efficacy of doxycycline (100 mg for 14 days) was carried out to determine the benefit of adjunctive medication in 16 patients with localized juvenile periodontitis. Measurements of gingival fluid flow, probing depths, bleeding on probing and suppuration were determined at 2 periodontal sites with and 2 without radiographic attachment loss, at weeks 0, 1, 3 and 8. Subgingival bacterial samples were taken with curettes from the same sites. Spirochaetes were searched for by dark-field microscopy. Actinobacillus actinomycetemcomitans, pigmented and non-pigmented Bacteroides spp., Capnocytophaga, Fusobacterium and Actinomyces spp. were cultured on various selective and non-selective media. Bacterial species found at least in 50% of the patients and comprising on average 5% or more of the cultivable flora were included in the analysis. Neither short-term clinical nor microbiological efficacy beyond that of a course of mechanical debridement alone was found by using systemic medication with doxycycline in patients with localized juvenile periodontitis.

Subgingival microflora, dental and periodontal conditions in patients with hereditary fructose intolerance.

Hereditary fructose intolerance (HFI) is a rare autosomally recessive disease which leads to severe hypoglycemia. The fructose-free diet of these patients apparently influences dental health. Half of the patients are free of caries, but there are no reports on their periodontal condition nor on the composition of their subgingival microflora. Therefore 18 patient with HFI were examined for the following parameters: radiographic bone loss, caries rate, gingival bleeding, occurrence of calculus, pocket depths, salivary flow rate, buffer capacity and pH from paraffin wax-stimulated saliva. Salivary S. mutans and lactobacilli were also enumerated. Specific antibody titers from whole saliva and serum to Actinobacillus actinomycetemcomitans gamma 4, Bacteroides gingivalis, and Capnocytophaga ochracea were determined with ELISA. Finally the subgingival plaque was analyzed by culture. Altogether 196 anaerobic or microaerophilic isolates representing 29 different species were obtained from the HFI patients and 164 isolates of 26 species from the controls. The frequency of Actinomyces odontolyticus, Veillonella parvula, and Wolinella recta in the HFI patients was significantly high The DMF-induces were lower in the study group than in the controls and so was the occurrence of S. mutans and lactobacilli in the saliva. Of the periodontal parameters examined, the only difference found was an increased incidence of gingival bleeding and calculus formation in the HFI patients. The results suggest that in the HFI patients the common gingival bleeding is associated with the more frequent occurrence of Actinomyces odontolyticus, Veillonella parvula, and Wolinella recta.

Certain bacterial species and morphotypes in localized juvenile periodontitis and in matched controls.

Subgingival microflora in localized juvenile periodontitis (LJP) was examined by culture of certain bacterial species and by direct dark-field microscopy in affected and unaffected periodontal pockets of 11 patients and in their age- and sex-matched controls. Only Actinobacillus actinomycetemcomitans (A.a.) correlated to periodontal destruction and occurred most frequently at the affected LJP sites. A.a., Bacteroides intermedius and Fusobacterium nucleatum were isolated in proportions greater than 5% of the flora at the affected LJP sites, but various Actinomyces spp. at the unaffected and control sites. The proportions of straight rods, fusiforms and small spirochetes correlated to gingival bleeding tendency and were higher at the affected LJP sites than at the control sites. Certain nonpigmented Bacteroides spp. and Eikenella corrodens were never isolated when only cocci were seen at dark-field microscopy. The more complex the flora, the more frequently B. intermedius occurred. The results support a significant role for A.a. in LJP and indicate that certain bacterial species appear to prefer occurring in the complex composition of subgingival microflora.

Actinobacillus actinomycetemcomitans and clinical periodontal status in Finnish juvenile periodontitis patients.

The relationship between the clinical periodontal status and the occurrence of Actinobacillus actinomycetemcomitans (A.a.) in 19 Finnish patients with localized juvenile periodontitis (LJP) was studied. Clinical examination included the Plaque Index, Gingival Index, suppuration, probing depth and bleeding on probing. The subgingival bacterial samples were taken from two diseases periodontal pockets with radiographic bone loss and two periodontal pockets exhibiting no radiographic alveolar bone loss. The results indicate that A.a. was isolated in 17 (89%) patients, in 68% of the diseased and in 32% of the control periodontal sites. Supragingival plaque, marginal gingival inflammation, gingival bleeding on probing, and suppuration were found as frequently in A.a.-positive as in A.a.-negative diseased LJP pockets. It was concluded that A.a. was frequently, but not always, detected in diseased LJP lesions. No association was found between the clinical status and the occurrence of A.a.