Bacteriologic findings associated with chronic bacterial maxillary sinusitis in adults.

An open-label, multicenter study was performed to assess bacteriologic findings associated with chronic bacterial maxillary sinusitis in adults. Seventy aerobic (52.2%) and 64 anaerobic (47.8%) pathogens were recovered from clinically evaluable patients at baseline (before therapy). The most commonly isolated anaerobes were Prevotella species (31.1%), anaerobic streptococci (21.9%), and Fusobacterium species (15.6%). The aerobes most frequently recovered included Streptococcus species (21.4%), Haemophilus influenzae (15.7%), Pseudomonas aeruginosa (15.7%), and Staphylococcus aureus and Moraxella catarrhalis (10.0% each). Recurrences of signs or symptoms of bacterial maxillary sinusitis associated with anaerobes were twice as frequent as were those associated with aerobes when counts of anaerobes were > or =10(3) cfu/mL. A pathogenic role for Granulicatella species in cases of chronic sinusitis was documented for the first time.

Hypochlorhydria induced by a proton pump inhibitor leads to intragastric microbial production of acetaldehyde from ethanol.

BACKGROUND: Acetaldehyde, produced locally in the digestive tract, has recently been shown to be carcinogenic in humans. AIM: To examine the effect of iatrogenic hypochlorhydria on intragastric acetaldehyde production from ethanol after a moderate dose of alcohol, and to relate the findings to the changes in gastric flora. METHODS: Eight male volunteers ingested ethanol 0.6 g/kg b.w. The pH, acetaldehyde level and microbial counts of the gastric juice were then determined. The experiment was repeated after 7 days of lansoprazole 30 mg b.d. RESULTS: The mean (+/- S.E.M.) pH of the gastric juice was 1.3 +/- 0.06 and 6.1 +/- 0.5 (P < 0.001) before and after lansoprazole, respectively. This was associated with a marked overgrowth of gastric aerobic and anaerobic bacteria (P < 0. 001), by a 2.5-fold (P=0.003) increase in gastric juice acetaldehyde level after ethanol ingestion, and with a positive correlation (r=0. 90, P < 0.001) between gastric juice acetaldehyde concentration and the count of aerobic bacteria. CONCLUSIONS: Treatment with proton pump inhibitors leads to hypochlorhydria, which associates with intragastric overgrowth of aerobic bacteria and microbially-mediated acetaldehyde production from ethanol. Since acetaldehyde is a local carcinogen in the concentrations found in this study, long-term use of gastric acid secretory inhibitors is a potential risk-factor for gastric and cardiac cancers.

Bacteriology of histopathologically defined appendicitis in children.

BACKGROUND: Acute appendicitis is the most common surgical emergency in childhood. However, the pathogenesis and detailed microbiology are obscure. OBJECTIVE: To determine in detail the bacterial etiology of appendicitis in children in relation to the histologic tissue pathology. STUDY DESIGN: Tissue samples obtained at surgery from 41 children with suspected acute appendicitis were examined histologically and by culture for aerobic and anaerobic bacteria. The patients were analyzed according to histopathologic and clinical findings. RESULTS: Aerobic and anaerobic species were isolated from 40 of 41 (98%) samples; on average, 14.1 isolates per specimen (10.4 anaerobes and 3.7 aerobes). Specimens from patients with gangrenous appendices yielded significantly higher numbers of anaerobic isolates per specimen than did specimens from patients with healthy appendices (11.7 vs. 7.7; P < 0.01). Bacteria belonging to the Bacteroides fragilis group were the most frequently isolated anaerobic microorganisms (95%). Other organisms frequently isolated in all histology groups were Peptostreptococcus micros (66%), Bilophila wadsworthia (63%), Fusobacterium nucleatum (44%), Eggerthella lenta (44%) and a hitherto undescribed bile-resistant, pigment-producing Gram-negative rod (41%). Of the aerobes Escherichia coli (88%) and Streptococcus anginosus group (former Streptococcus "milleri" group) organisms (61%) were the most frequent findings. CONCLUSIONS: The shift from histologically normal toward gangrenous appendices was clearly associated with markedly elevated anaerobic bacterial counts in terms of species. The unusually high frequencies of B. wadsworthia (75%) and the hitherto undescribed bile-resistant, pigment-producing Gram-negative rod (56%) in gangrenous appendices represent unique and different findings from those reported in adults.

Current perspectives on anaerobic infections: diagnostic approaches.

This article discusses when to look for anaerobes, anaerobic infections as clues to other problems in patients, and underlying clinical conditions as clues to the nature of anaerobic infections. Diagnostic approaches, identification methods, and susceptibility testing are reviewed.

Age-related frequency of penicillin resistance of oral Veillonella.

Veillonella spp. are early colonizing inhabitants in the mouth. As part of studies on penicillin resistance among oral indigenous anaerobic microbiota in childhood, the aim of the present longitudinal study was to examine the emergence of resistant strains in Veillonella populations. Altogether 305 Veillonella isolates from saliva of 49 healthy infants followed from 2 to 24 months of age were examined for their in vitro susceptibility to penicillin G and, further, 20 penicillin-resistant isolates representing 5 MIC categories to ampicillin, amoxicillin, amoxicillin/clavulanate, cefoxitin, and beta-lactamase production. In infants positive for oral Veillonella, the recovery rate of penicillin-resistant (MIC >/=2 microg/ml) strains increased with age up to 68%, however, most infants simultaneously harbored penicillin-susceptible strains. During the follow-up, the MIC(50) increased from 0.5 microg/ml to 2 microg/ml. In addition to penicillin G, 8/20 strains also showed reduced susceptibility to ampicillin and/or amoxicillin but none produced beta-lactamase. Our study suggests other mechanisms than enzymatic degradation of beta-lactam ring for resistance of oral Veillonella to penicillin.

Establishment of streptococci in the upper respiratory tract: longitudinal changes in the mouth and nasopharynx up to 2 years of age.

As part of a series of longitudinal studies on the development of the indigenous microflora of the upper respiratory tract, the establishment of streptococci in the oral cavity and nasopharynx and IgA1 protease production by the early streptococcal flora was examined in 50 healthy Caucasian infants at the ages of 2, 6, 12, 18 and 24 months. In the oral cavity, streptococci were found in all infants on every sampling occasion, Streptococcus mitis biovar 1 being the main finding in each age group. S. salivarius and S. mitis biovar 2 reached their highest prevalence during the first year of life, whereas the prevalence of S. oralis and S. sanguis showed no significant increase before 12 months of age. Salivary streptococci mainly consisted of the above-mentioned species during the follow-up period. In contrast to the oral cavity, no stable colonisation pattern was observed for viridans streptococci in the nasopharynx. S. mitis biovar 1 and S. pneumoniae, a traditional respiratory pathogen, were the principal streptococcal species among nasopharyngeal isolates. IgA1 protease production by early streptococci was common in infancy. Among the oral streptococcal microflora, S. mitis biovar 1 (especially during the first year of life) and S. oralis and S. sanguis constituted the main species responsible for this enzyme activity. In the nasopharynx, IgA1 protease was produced by S. mitis biovar 1, S. oralis and S. pneumoniae. In conclusion, streptococcal colonisation differs in these two close habitats in the upper respiratory tract.

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.

beta-Lactamase production and in vitro antimicrobial susceptibility of Porphyromonas gingivalis.

beta-Lactamase production by 98 Porphyromonas strains was investigated by the nitrocefin (chromogenic cephalosporin) test. Human isolates of P. gingivalis (91), P. endodontalis (2), and P. asaccharolytica (1) were tested, with four closely related Porphyromonas spp. of animal origin and four reference strains. The in vitro susceptibility of 64 P. gingivalis strains was investigated on Brucella blood agar by the E test. None of the human Porphyromonas isolates tested produced beta-lactamase, but one Porphyromonas strain of animal origin, most closely resembling P. endodontalis, produced beta-lactamase. P. gingivalis was susceptible to almost all of the drugs tested: benzylpenicillin, ampicillin, cefaclor, cefuroxime, erythromycin, clindamycin, tetracycline, doxycycline, metronidazole and ciprofloxacin; all strains were inhibited at 0.016 microgram/ml, 0.023 microgram/ml, 0.315 microgram/ml, 0.064 microgram/ml, 0.19 microgram/ml, 0.016 microgram/ml, 0.094 microgram/ml, 0.047 microgram/ml, 0.023 microgram/ml, and 0.75 microgram/ml of these drugs, respectively. Cotrimoxazole exhibited variable efficacy against P. gingivalis; the range of MICs was 0.1095-32.0 micrograms/ml. The results indicate that beta-lactamase production is currently not a problem amongst clinical isolates of P. gingivalis and strains are susceptible to most antimicrobial agents.

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.

Intra- and inter-individual comparison of Porphyromonas gingivalis genotypes.

Genetic analysis of 31 clinical strains of Porphyromonas gingivalis isolated from nine subjects, 2-6 strains per subject, was performed by Southern hybridization. Chromosomal DNA was extracted by the method of Moncla et al. [1] and digested to completion with restriction endonucleases PstI, ClaI and BglI. The DNA fragments were separated electrophoretically on agarose gels, transferred to nylon membranes and hybridized to the non-radioactively labelled plasmid pKK 3535 which contains the rmB ribosomal RNA operon of the Escherichia coli chromosome. Of the three enzymes, BglI was the most suitable for the genetic analysis of P. gingivalis. With this enzyme, the intra-individual strains were shown to be identical in eight of the nine subjects, whereas inter-individual strains were different.

Altered antigenicity is seen in the lipopolysaccharide profile of non-serotypeable Actinobacillus actinomycetemcomitans strains.

Non-serotypeable Actinobacillus actinomycetemcomitans strains may be derived from the serotypeable ones. In the present study, we compared the outer membrane proteins (OMPs) and lipopolysaccharides (LPSs) of serotypeable and non-serotypeable A. actinomycetemcomitans strains (n=24) of the same genotype in the same subject (n=6) to find out if alterations on the cell-surface contribute to the non-serotypeability. Serotypeable and non-serotypeable A. actinomycetemcomitans strains showed great similarity in the OMP patterns both within and between subjects. Using serotype-specific antisera, clear immunoblotting LPS profiles in the O-antigenic region were seen in serotype b and c strains but not in non-serotypeable strains from the same subjects. The results suggest that changes in LPS lead to the altered antigenicity of non-serotypeable A. actinomycetemcomitans strains.

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.

Salivary levels of suspected periodontal pathogens in relation to periodontal status and treatment.

The primary ecological niche for suspected periodontal pathogens seems to be the subgingival area, even though periodontal pathogens are also frequently recovered from saliva. The interrelationship of different periodontal conditions and the salivary levels of suspected periodontal pathogens is not known. In the present study, salivary levels of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Campylobacter rectus, and Peptostreptococcus micros were determined by bacterial culture and related to clinical periodontal status in 40 subjects with either advanced, moderate, or initial/no periodontitis. Culture-positive subjects harbored the 5 bacterial species in mean numbers ranging from 2 x 10(5) to 6 x 10(7) colony-forming units (CFU)/mL saliva. A. actinomycetemcomitans was found in none and P. gingivalis in one of the subjects with initial periodontitis, whereas both species were found in 33% and 44%, respectively, of the subjects with moderate periodontitis and in 60% and 40%, respectively, of the subjects with advanced periodontitis. The mean numbers of CFU/mL of P. intermedia, C. rectus and P. micros were significantly higher in subjects with advanced periodontitis than in subjects with initial/no periodontitis. Ten patients with advanced periodontitis were treated mechanically and with adjunctive systemic metronidazole, and were re-examined 1 and 6 months after treatment. Periodontal treatment eradicated or significantly reduced the levels of salivary periodontal pathogens for half a year, whereas in untreated subjects, the levels and the detection frequencies generally remained fairly stable. In conclusion, the results showed that the salivary levels of periodontal pathogens reflect the periodontal status of the patient.

Bacteremia following surgical dental extraction with an emphasis on anaerobic strains.

Our aim was to investigate bacteremia caused by surgical extraction of partly erupted mandibular third molars. From 16 young adults, bacterial samples were taken from the third-molar pericoronal pocket and post-operatively from the extraction socket, and blood samples were drawn from the ante-cubital vein up to 30 min after surgery. Of the subjects, 88% had detectable bacteremia-50% 1 min after the incision, 44% immediately after extraction. The respective percentages at 10, 15, and 30 min were 44%, 25%, and 13%. Blood cultures contained 31 species (74% anaerobes), with 3.9 +/- 2.6 species isolated per subject. Most prevalent were the anaerobes Prevotella, Eubacterium, and Peptostreptococcus sp. and the aerobes viridans-group streptococci and Streptococcus milleri group. Any species found in the blood was also isolated from the mouth, from 93% of the pericoronal pockets and from 43% of the extraction sockets. Surgical dental extraction clearly causes bacteremia of a high frequency and lasting longer than thus far assumed.

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.

Age, dental infections, and coronary heart disease.

Epidemiological and intervention studies have suggested that infections are risk factors for coronary heart disease (CHD). Dental infections have appeared as cardiovascular risk factors in cross-sectional and in follow-up studies, and the association has been independent of the "classic" coronary risk factors. This case-control study aimed at detailed assessment of the dental pathology found in various CHD categories (including elderly patients). Altogether, 85 patients with proven coronary heart disease and 53 random controls, matched for sex, age, geographic area, and socio-economic status, were compared with regard to dental status, assessed blindly with four separate scores, and to the "classic" coronary risk factors (seven of the controls had CHD, and they were not included in the analyses). The dental indices were higher among CHD patients than in the controls, but, contrary to previous studies, the differences were not significant (between the CHD patients and their matched controls or among the different CHD categories). This result could not be explained by potential confounding factors. The participants in the present study were older and had more often undergone recent dental treatment in comparison with subjects in our earlier studies. Age correlated with the severity of dental infections only in the random controls but not in the coronary patients who, although young, already had high dental scores. We believe that the higher age of the participants in the present study is the most likely reason for the results. Other possible explanations include an age-related selection bias among older CHD patients, and the fact that those participating in studies like this may have better general health and thus also less severe dental infections. Thus, the role of dental infections as a coronary risk factor varies according to the characteristics of the population studied.

Matrilysin (matrix metalloproteinase-7) expression in human junctional epithelium.

Matrilysin is a matrix metalloproteinase expressed in exocrine and mucosal epithelium in many human tissues. Immunohistochemical staining showed that matrilysin is expressed in suprabasal cells of junctional epithelium facing the teeth and in epithelial cell rests of Malassez. No matrilysin expression was seen in the periodontal pocket tissue. In a tissue culture model mimicking junctional epithelium, matrilysin expression was also observed in suprabasal epithelial cells. Of 13 anaerobic oral bacterial species tested, F. nucleatum, F. necrophorum, P. endodontalis, and P. denticola stimulated matrilysin expression in porcine periodontal ligament epithelial cells from 2.5- to 5.7-fold, compared with untreated cells. The enzyme was localized in intracytoplasmic vesicles that also reacted with antibodies against lysosomal membrane protein h-lamp-1. The results indicate that matrilysin may play an important role in the normal physiology of junctional epithelium.

Inhibition of bacteriocolonic pathway for ethanol oxidation by ciprofloxacin in rats.

Many colonic bacteria possess marked alcohol dehydrogenase (ADH) activity and are capable of oxidizing ethanol to acetaldehyde both in vitro and in vivo. We have recently shown that part of ingested ethanol is metabolized to acetaldehyde in the colon during normal alcohol metabolism. To assess the contribution of this bacteriocolonic pathway for ethanol oxidation to total ethanol metabolism, the elimination rate of ethanol, faecal aerobic flora and faecal ADH activity were determined in rats before and after the treatment with ciprofloxacin (200 mg/kg/day) for four days. Ciprofloxacin treatment decreased ethanol elimination rate from 310+/-9 to 282+/-13 mg/kg/h (mean+/-SE; p<0.02), markedly reduced faecal aerobic flora, and also lowered faecal ADH activity from 63+/-17 to 17+/-7 nmol/min/mg faeces (p<0.05). Neither hepatic ADH nor microsomal ethanol oxidizing system activities were affected by the ciprofloxacin treatment. On the contrary, an acute intraperitoneal dose of ciprofloxacin had no effect on the rate of ethanol elimination. These results support the significant role of the bacteriocolonic pathway in total ethanol elimination, and open a new, microbiological, perspective for studies on ethanol metabolism and pathogenesis of alcohol related organ damages.

Penicillin G and penicillin G-tinidazole treatment of experimentally induced summer mastitis--effect on elimination rates of bacteria and outcome of the disease.

Two udder hind quarters of ten pregnant heifers were inoculated experimentally with a combination of Actinomyces pyogenes, Peptostreptococcus indolicus and Fusobacterium necrophorum at a dose of 9 x 10(8) colony forming units (CFU) of each species of bacteria. The development of aerobic-anaerobic mastitis was followed-up clinically and through analysis of udder secretion samples. All heifers developed acute clinical mastitis. Thick, purulent secretions with foul odour were already present 24 h after infusion. At 32 h post inoculation, 19 of 20 quarters harboured all infused bacteria, the viable counts of bacteria varying from 10(4) to 10(5) CFU/ml. No bacteria were isolated from blood samples taken simultaneously. NAGase values of udder secretions, indicating tissue damage, were high. The heifers were treated systemically with penicillin G alone, or in combination with tinidazole. Treatment started 32 h after inoculation and was continued for three days. The anaerobic bacteria (F. necrophorum and P. indolicus) were usually eliminated within four days, while A. pyogenes infections often persisted in the quarters. Penicillin G with tinidazole was not more effective than penicillin G alone: of the four heifers which recovered from infection, two were treated with penicillin G alone and two with the drug combination. The most important factor affecting the recovery seemed to be the host response of individual heifers. The six heifers which did not recover from the infection were continuously infected after calving, harbouring A. pyogenes in ten quarters, and P. indolicus and F. necrophorum each in three quarters.

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.