Search

Aims: The effect of different concentrations of 2-hydroxyethyl methacrylate (HEMA) was evaluated on biofilm formation and preformed biofilm of Streptococcus mitis, Streptococcus mutans and Streptococcus oralis, alone or combined to each other. Methods and Results: Twofold serial dilution of HEMA ranged from 12 to 0·75 mmol l−1 was added to Streptococcal broth cultures and mature biofilms in 96-well-microtitre plates to evaluate bacterial biomass and cell viability. HEMA affected the Streptococcal population in a strain-specific way producing few significant effects. A reduction on biofilm formation and a detachment of preformed biofilm was recorded in Strep. mitis ATCC 6249, whereas in mixed cultures, the monomer expressed a general aggregative effect on mature biofilms. A reduction in cell viability was also recorded in an HEMA-concentration-dependent way in each experimental condition studied. Conclusions: These results suggest that the HEMA prevalent effects are both the reduction of bacterial adhesion to a polystyrene surface and the increase in dead cells also characterized by an aggregative status. Significance and Impact of the Study: Understanding the potential effect of HEMA, released from resin-based materials, on oral bacteria may furnish information for surveillance of the risk reduction in secondary caries via hindering biofilm generation.

Stenotrophomonas maltophilia is a pathogen that causes infections mainly in immunocompromised patients. Despite increased S. maltophilia isolation from respiratory specimens of patients with cystic fibrosis (CF), the real contribution of the microorganism to CF pathogenesis still needs to be clarified. The aim of the present study was to evaluate the pathogenic role of S. maltophilia in CF patients by using a model of acute respiratory infection in DBA/2 mice following a single exposure to aerosolized bacteria. The pulmonary bacterial load was stable until day 3 and then decreased significantly from day 3 through day 14, when the bacterial load became undetectable in all infected mice. Infection disseminated in most mice, although at a very low level. Severe effects (swollen lungs, large atelectasis, pleural adhesion, and hemorrhages) of lung pathology were observed on days 3, 7, and 14. The clearance of S. maltophilia observed in DBA/2 mouse lungs was clearly associated with an early and intense bronchial and alveolar inflammatory response, which is mediated primarily by neutrophils. Significantly higher levels of interleukin-1β (IL-1β), IL-6, IL-12, gamma interferon (IFN-γ), tumor necrosis factor alpha (TNF-α), GROα/KC, MCP-1/JE, MCP-5, macrophage inflammatory protein 1α (MIP-1α), MIP-2, and TARC were observed in infected mice on day 1 with respect to controls. Excessive pulmonary infection and inflammation caused systemic effects, manifested by weight loss, and finally caused a high mortality rate. Taken together, our results show that S. maltophilia is not just a bystander in CF patients but has the potential to contribute to the inflammatory process that compromises respiratory function.

Background: The use of locally delivered antibacterials containing chlorhexidine (CHX) was proposed to improve the effectiveness of non-surgical periodontal treatment. The present multicenter randomized study investigated the effects of a xanthan-based chlorhexidine (Xan-CHX) gel used as an adjunct to scaling and root planing (SRP) in the treatment of chronic periodontitis. Methods: Ninety-eight systemically healthy subjects with moderate to advanced periodontitis were recruited in four centers (59 females and 39 males; aged 24 to 58 years). For each subject, two experimental sites located in two symmetric quadrants were chosen with probing depths (PD) ‡5 mm and positive for bleeding on probing (BOP). These two sites were randomized at the split-mouth level with one receiving a single SRP treatment and the other receiving a single SRP + Xan-CHX gel treatment. Supragingival plaque, modified gingival index, PD, clinical attachment level (CAL), and BOP were evaluated at baseline (prior to any treatment) and after 3 and 6 months. At the same times, subgingival microbiologic samples and gingival crevicular fluid (GCF) were collected for the analysis of total bacterial counts (TBCs), including the identification of eight putative periodontopathogens, and alkaline phosphatase (ALP) activity, respectively. Results: The Xan-CHX treatment group showed greater improvements compared to the SRP group for PD and CAL at 3a nd 6m onths (P

We tested 40 clinical Stenotrophomonas maltophilia strains to investigate the possible correlation between adherence to and formation of biofilm on polystyrene, and cell surface properties such as hydrophobicity and motility. Most of the strains were able to adhere and form biofilm, although striking differences were observed. Eleven (27.5%) of the strains were hydrophobic, with hydrophobicity greatly increasing as S. maltophilia attached to the substratum. A positive correlation was observed between hydrophobicity and levels of both adhesion and biofilm formation. Most of the isolates showed swimming and twitching motility. A highly significant negative correlation was observed between swimming motility and level of hydrophobicity. Hydrophobicity is thus a significant determinant of adhesion and biofilm formation on polystyrene surfaces in S. maltophilia.

Aims: To assess the ability of Listeria monocytogenes to form biofilm on different food-contact surfaces with regard to different temperatures, cellular hydrophobicity and motility. Methods and Results: Forty-four L. monocytogenes strains from food and food environment were tested for biofilm formation by crystal violet staining. Biofilm levels were significantly higher on glass at 4, 12 and 22°C, as compared with polystyrene and stainless steel. At 37°C, L. monocytogenes produced biofilm at significantly higher levels on glass and stainless steel, as compared with polystyrene. Hydrophobicity was significantly (P

Background: The main therapeutic approach for periodontal diseases is mechanical treatment of root surfaces via scaling and root planing (SRP). Multicenter clinical trials have demonstrated that the adjunctive use of a chlorhexidine (CHX) chip is effective in improving clinical results compared to SRP alone. However, some recent studies failed to confirm these clinical results, and conflicting results were reported regarding the effects of the CHX chip on subgingival microflora. The aim of this study was to provide further data on the clinical and microbiologic effects of CHX chips when used as an adjunct to SRP. Methods: A total of 116 systemically healthy individuals with moderate to advanced periodontitis, aged 33 to 65 years, were recruited from the Departments of Periodontology of four Italian universities. For each subject, two experimental sites were chosen that had probing depths (PD) >= 5 mm and bleeding on probing (BOP) and were located in two symmetric quadrants. These two sites were randomized at the split-mouth level, with one receiving SRP treatment alone and the other receiving treatment with SRP plus one CHX chip (SRP + CHX). PD, relative attachment level (RAL), and BOP were evaluated at baseline, prior to any treatment, and after 3 and 6 months. Supragingival plaque and the modified gingival index were evaluated at baseline and after 15 days and 1, 3, and 6 months. Subgingival microbiologic samples were harvested at baseline and after 15 days and 1, 3, and 6 months, cultured for total bacterial counts (TBCs), and investigated by polymerase chain reaction analysis for the identification of eight putative periodontopathogens. Results: When all of the pockets were considered, the PD and RAL were significantly less at 3 and 6 months compared to the baseline scores (P= 7 mm were considered. The percentage of sites positive for BOP was similar between the treatments at each time point. At 15 days and I month, the TBC for the SRP + CHX treatment group was significantly lower than for the SRP treatment group (P

Trichosporon asahii is the most common cause of fatal disseminated trichosporonosis, frequently associated with indwelling medical devices. Despite the use of antifungal drugs to treat trichosporonosis, infection is often persistent and is associated with high mortality. This drove our interest in evaluating the capability of T. asahii to form a biofilm on biomaterial-representative polystyrene surfaces through the development and optimization of a reproducible T. asahii -associated biofilm model. Time course analyses of viable counts and a formazan salt reduction assay, as well as microscopy studies, revealed that biofilm formation by T. asahii occurred in an organized fashion through four distinct developmental phases: initial adherence of yeast cells (0 to 2 h), germination and microcolony formation (2 to 4 h), filamentation (4 to 6 h), and proliferation and maturation (24 to 72 h). Scanning electron microscopy and confocal scanning laser microscopy revealed that mature T. asahii biofilms (72-h) displayed a complex, heterogeneous three-dimensional structure, consisting of a dense network of metabolically active yeast cells and hyphal elements completely embedded within exopolymeric material. Antifungal susceptibility testing demonstrated a remarkable rise in the MICs of sessile T. asahii cells against clinically used amphotericin B, caspofungin, voriconazole, and fluconazole compared to their planktonic counterparts. In particular, T. asahii biofilms were up to 16,000 times more resistant to voriconazole, the most active agent against planktonic cells (MIC, 0.06 μg/ml). Our results suggest that the ability of T. asahii to form a biofilm may be a major factor in determining persistence of the infection in spite of in vitro susceptibility of clinical isolates.

Time-kill and postantifungal effect (PAFE) of amphotericin B, caspofungin, fluconazole, and voriconazole were determined against clinical isolates of Candida guilliermondii , Candida kefyr , and Candida lusitaniae . Azoles displayed fungistatic activity and no measurable PAFE, regardless of the concentration tested. Amphotericin B and caspofungin demonstrated concentration-dependent fungicidal activity, although amphotericin B only produced a significant dose-dependent PAFE against all isolates tested.

Objectives: The present controlled, single-blind study was performed to assess and compare the clinical healing and the microbiological findings following repeated intrasulcular applications of 1% metronidazole or 1% chlorhexidine gels in persistent periodontal pockets previously treated by scaling and root planing (SRP). Material and Methods: Sixty-three systemically healthy subjects, 25 males and 38 females (mean age 48.4±7.2 years), diagnosed for chronic periodontitis were enrolled in this study. They underwent SRP and received oral hygiene instructions (OHI). Three months later, at baseline, a single persistent pocket with a probing depth (PD) of 5–9 mm was chosen as the experimental site in each patient; the subjects were stratified into three matched experimental groups on the basis of the treatment to be performed, which consisted of the subgingival administration of 1% metronidazole gel (MG, n=19), 1% chlorhexidine gel (CG, n=20) or placebo gel (PG, n=24). The treatments consisted of four repeated administrations of subgingival gels, each separated by 7 days, starting at the baseline. Clinical assessment was performed at the baseline and at the 180-day follow-up, after the end of treatment. For microbiological evaluations, subgingival plaque was sampled from the experimental sites at baseline, prior to the first subgingival gel administration, and at 7, 15, 30 and 90 days after the end of the treatment (days 28, 36, 51 and 111 from baseline). Results: Plaque accumulation did not change significantly in all three groups. Bleeding on probing and clinical attachment levels reduced in the MGs and CGs only. PD was significantly reduced by the same amount in all experimental groups. In the MGs and CGs a remarkable reduction in the frequencies of detection of several periodontopathic micoorganisms was recorded after the treatment. The same was not seen for the PGs. Conclusions: Subgingival administration of MG or CG, both at 1%, may have a role in the management of persistent pockets during chronic periodontitis.

Objectives: The relationship between subgingival dental restorations and periodontal health has been thoroughly investigated for many years. However, longitudinal data on the subgingival microflora features after the placement of well-finished subgingival restorations are still lacking. Therefore, this study compares the short-term clinical and microbiological features occurring in the gingiva after the completion of different subgingival restorations. Material and Methods: Sixteen systemically healthy subjects, 10 males and six females (ages: 31.7-45.8 years; mean age 39.3 ′ 5.1 years), who were non-smokers and were positive for the presence of three cervical abrasion/erosion defects to be restored in three different adjacent teeth were enrolled in this study. The cervical abrasion/erosion defects were each restored by using one of three different materials: amalgam, glass ionomer cement, or composite resin. Immediately before class V cavity preparations and restorations (baseline), clinical monitoring and subgingival plaque sampling were performed in the mid-buccal aspect of each experimental restored tooth and in one adjacent sound, non-treated, control tooth. These procedures were repeated every 4 months over the following 1 year. Results: Throughout the study, the clinical parameters recorded did not change significantly in any of the experimental groups, and no differences were detected among them at each clinical session. Over this time, no significant changes in the composition of the subgingival microflora were observed in amalgam, glass ionomer cement, and control groups. Conversely, in the composite resin group, there was a significant increase in the total bacterial counts, and a significant (p

Summary Background : The resistance of Helicobacter pylori to antibiotics has been advocated as a major cause of treatment failure, and antimicrobial sensitivity testing has been proposed to improve efficacy; however, its role before first-line therapy has not been investigated in detail. Aim : To assess whether antimicrobial sensitivity testing improves the eradication rate of first-line anti-Helicobacter treatments and to compare the effectiveness of ranitidine bismuth citrate and omeprazole in the presence of H. pylori resistance to antibiotics. Methods : Two hundred and forty-two patients were assigned to either empirical or antimicrobial sensitivity testing-based treatment; within each group, subjects were further randomized to receive ranitidine bismuth citrate, 400 mg b.d., tinidazole, 500 mg b.d., and clarithromycin, 500 mg b.d., or omeprazole, 20 mg b.d., clarithromycin, 500 mg b.d., and amoxicillin, 1 g b.d., for 1 week, with substitution of the resistant antibiotic in the antimicrobial sensitivity testing-based treatment group. Results : Eradication rates were 67% [confidence interval (CI), 55–79%] in the empirical treatment group and 76% (CI, 65–87%) in the antimicrobial sensitivity testing-based group (P = N.S.). The overall success rate was 60% (CI, 51–69%) with omeprazole and 82% (CI, 73–91%) with ranitidine bismuth citrate (P

The susceptibility to levofloxacin of 194 consecutive staphylococcal (45 Staphylococcus aureus and 149 coagulase-negative staphylococci) isolates from neutropenic patients was determined by Etest and the results compared with those obtained using NCCLS-methods (broth microdilution, agar dilution and disk diffusion). Overall agreement at +/- 1log(2) dilution for Etest compared with broth microdilution and agar dilution was 99.0 and 83.5%, respectively. The Etest category agreement with broth microdilution and disk diffusion was 95.9 and 89.7%, respectively. Comparison of categories with Etest and agar dilution method gave only 67.0% absolute categorical agreement, with 29.9% minor errors and 10.7% major errors. No very major errors occurred by the four methods tested. Our results show that Etest is a valid alternative to the reference NCCLS-methods for monitoring the clinical usefulness of levofloxacin against staphylococci isolates from neutropenic patients.

Background: Novel fluoroquinolones have been recently introduced in the management of neutropenic patients because of their increased activity against gram-positive and gram-negative micro-organisms. Methods: The activities of levofloxacin and ciprofloxacin were determined by the E test against 223 bacterial isolates from patients with haematological malignancies. Results: In general, the activity of levofloxacin was comparable to that of ciprofloxacin. Levofloxacin was somewhat more active against methicillin-resistant Staphylococcus aureus isolates. All methicillin-susceptible S. aureus isolates were inhibited by ciprofloxacin and levofloxacin at a concentration of ≤0.5 and ≤0.25 µg/ml, respectively. Among gram-negative isolates tested, levofloxacin was significantly (p < 0.001) more active than ciprofloxacin against Stenotrophomonas maltophilia, inhibiting 68 and 53% of these isolates, respectively. Conclusions: Levofloxacin was not superior to ciprofloxacin in its overall antibacterial activity, although small differences between these agents were seen depending on the species tested. In particular, our data suggested that levofloxacin may potentially be used for the management of S. maltophilia infections in neutropenic patients.

It has been recently observed that in implants with screw-retained abutments, in in vitro as well as in vivo conditions, bacteria can penetrate inside the internal cavity of the implant as a consequence of leakage at the implant-abutment interface. An alternative to screw-retained abutments is represented by implants that can receive cemented abutments. In this case, the abutment goes through a transmucosal friction implant extension (collar) and is cemented inside the internal hexagonal portion of the implant. The aim of the present research was to compare fluids and bacterial penetration in 2 different implant systems, one with cement-retained abutments (CRA) and the other with screw-retained abutments (SRA).Twelve CRA dental implants and 12 SRA implants were used in this study. The research was done in 3 steps: scanning electron microscopic (SEM) analysis, fluid penetration analysis, and bacterial penetration analysis.1) Under SEM it was possible to observe in the SRA implants a mean 2 to 7 micron gap between implant and abutment, while in the CRA implants, the gap was 7 micron. In the latter group, however, the gap was always completely filled by the fixation cement. All the spaces between abutment and implant were filled by the cement. 2) With SRA implants, it was possible to observe the presence of toluidine blue at the level of the fixture-abutment interface and the internal threads; the absorbent paper was stained in all cases. With CRA implants, the absorbent paper inside the hollow portion of the implants was never stained by toluidine blue. No penetration of toluidine blue was observed at the implant-abutment interface and inside the hollow portion of the implants. 3) In all the SRA implant assemblies, bacterial penetration was observed at the implant-abutment interface. No bacteria were detected in the hollow portion of the CRA implants.On the basis of the results obtained in the present study using 2 different implant systems, we conclude that CRA implants offer better results relating to fluid and bacterial permeability compared to SRA implants.

Background: The aim of this study was to report on the prevalence of Actinobacillus actinomycetemcomitans (Aa) and the periodontal clinical conditions in children and adolescents from a rural area of central Italy compared with the ones from an urban area of the same region. Method: The study population consisted of 780 systemically healthy children, aged 6–14 years inhabiting the county of Chieti. 505 children attended 3 primary and 2 secondary schools from a rural area whereas 275 individuals attended 1 primary and 1 secondary school from the city of Chieti. The 2 provincial areas present a great difference in socioeconomic level and cultural background. Clinical examination consisted of recording the % of gingival sites positive for the presence of plaque (Pl+), bleeding on probing (BOP+), mean probing depth (PD) from each primary or permanent tooth fully erupted in the oral cavity. Loss of periodontal attachment (AL+) was evaluated only in interproximal sites. AL+ subjects were distinguished in juvenile periodontitis (JP) prepubertal periodontis and early periodontitis (EP) patients. 8 gingival sites were microbiologically sampled in each subject and cultured, after pooling, for the presence of Aa. Results: 30.3% of rural subjects, were positive for the presence of Aa, the difference from urban children (16%) being statistically significant (p=0.01) irrespective of gender and age. Aa showed a significantly (p=0.006) higher mean proportion in subgingival plaque samples from rural children (0.13% versus 0.02%). Loss of periodontal attachment in at least one site was found in 18 rural children (3.56%) (3 JP; 15 EP) and 2 urban girls (0.72%) (1 JP; 1 EP). No significant differences for AL were observed within the rural group according to the gender and age differentiation. In the urban group, both AL+ subjects were Aa+, while among children from rural areas all 3 JP and 13 EP subjects were Aa+. Rural subjects evidenced significantly worse clinical parameters with respect to urban children (% Pl+ sites: p=0.000; % BOP+ sites: p=0.010; mean PD: p=0.000.) The relative risk for AL+ sites was significantly greater (2.42) in rural subjects harboring Aa in subgingival plaque. Similarly, the presence of Aa in subgingival plaque was related to a greater risk of more than 50% of BOP+ gingival sites in both rural and urban subjects (1.45 and 8.40, respectively). Conclusions: Results of this study suggest that Aa colonization in children and adolescents from central Italy is affected by socioeconomic and cultural factors; these factors also affect the periodontal condition of the subjects.

A high prevalence of Actinobacillus actinomycetemcomitans (Aa) in subgingival plaque in patients for orthodontia already has been observed. The present study had the following aims: 1) to ascertain a direct relationship between the orthodontic appliance placement and the subgingival colonization by Aa, and 2) to determine whether the Aa growth specifically occurred on teeth with braces attached or whether the presence of orthodontic appliances could also cause the isolation of Aa in teeth free from therapeutic appliances. Twenty-four young systemically and periodontally healthy subjects with malaligned and crowded teeth in the anterior sextants of both dental arches participated in this study. After 1 session of ultrasonic scaling with oral hygiene instructions during the first experimental session, the mesiobuccal sites of the first molars and the distobuccal sites of the lateral incisors in both dental arches in each participant were subjected to clinical and microbiologic examination for the recovery of Aa. Clinical examination consisted of recording the presence of plaque and the examination of gingival bleeding on probing and probing depth. Microbiologic sampling was obtained with the insertion of 3 sterile paper points at the deepest part of each gingival sulcus. Altogether, 192 periodontal sites were examined. After the examinations, the patients received fixed orthodontic appliances in only 1 dental arch (test sites) and the other one was left free from appliances (control sites). Clinical examination and microbiologic sampling were repeated in the same experimental test and control sites after 4, 8, and 12 weeks. At the 12-week session, the orthodontic appliance was removed from the test arch, and, 4 weeks later, a further clinical and microbiologic examination was performed. The results showed that, during the period with orthodontic appliances, the presence of plaque scores and the gingival bleeding on probing scores were increased significantly and that Aa, initially absent from all but 1 subject, was isolated in 19 and 20 subjects after 4 and 8 weeks, respectively. Furthermore, no gingival sites from the control teeth (free from Aa colonization at baseline) showed positive results for the sought-after bacterium throughout the entire length of the study. It was concluded that the placement of orthodontic appliances promotes the subgingival growth of Aa; this specific microbial change is specifically restricted to subgingival plaque from orthodontic appliance-bearing teeth. The presence of orthodontic bands and brackets therefore cannot affect the microbiologic condition of the whole mouth.

The aim of the present study was: (1) to assess longitudinally the occurrence of Actinobacillus actinomycetemcomitans (Aa) in young subjects wearing fixed orthodontic appliances compared to matched appliance-free controls; (2) to determine whether the presence of the micro-organism at baseline could influence the periodontal status assessed 3 years later. 70 subjects, 27 male and 43 female, aged between 12 and 20 years participated in the study: 35 subjects under orthodontic treatment with fixed appliances for at least 6 months, and 35 appliance-free individuals matched for age and gender. All subjects were free of clinically demonstrable loss of attachment. They all received oral hygiene instructions 2x during the 2 months preceding the first clinical and microbiological examination. No subgingival instrumentation was performed between baseline and the 3-year examination. Clinical parameters included gingival bleeding index (GBI), pocket probing depth (PPD) and measurements of attachment level (AL). Statistically significant differences were reported regarding frequency of detection of Aa between both groups at each examination. The %s of orthodontic subjects infected with Aa at the baseline and at the 3-year examination were 86% and 80%, respectively, while the corresponding figures for control subjects were 16.6% and 26.6%. The frequency distribution of %s of Aa in the total anaerobic subgingival flora among control subjects remained fairly stable, whereas the proportion of orthodontic subjects yielding Aa at a concentration > or = 1.0% dropped significantly from 32% at baseline to 19% at the 3-year visit. Calculations of the relative risk for increasing GBI and PPD in both groups when Aa was present at baseline, revealed that the orthodontic subjects positive for Aa had a negligible relative risk of experiencing worse periodontal conditions compared to orthodontic patients where Aa was not detected at baseline. In contrast, control subjects initially infected with Aa presented with a risk for increased GBI 6.6x higher than that for subjects without Aa. In conclusion, the present study confirmed previous cross-sectional findings reporting that young individual with an integer periodontium wearing fixed orthodontic appliances harbor Aa with a statistically significant greater frequency than appliance-free matched controls. However, although orthodontic patients exhibited more inflammation, their deteriorated clinical conditions could not be accounted for by the sole presence of Aa in their sulci. In contrast, appliance-free young subjects initially infected with Aa had a higher risk of experiencing more gingival inflammation than subjects without the bacterium during a 3-year observation period.

The aim of the present study was to assess: (1) the occurrence of Actinobacillus actinomycetemcomitans (Aa) in subgingival plaque from young patients undergoing orthodontic treatment with fixed appliances; (2) a possible relationship between the presence of Aa and the clinical conditions; (3) a relation between the duration of orthodontic treatment and the microbiological and clinical parameters; (4) whether differences exist when taking into consideration the different type of appliances, i.e., bands or brackets. 34 subjects aged between 12 and 20 years participated in the study. Of these, 20 subjects had worn orthodontic appliances (test group), while the remaining 14 subjects served as matched control (control group). 4 to 8 sites in each patient were available for clinical and microbiological examination. Clinical parameters consisted of presence/absence of plaque and gingival bleeding index (GBT), Microbiological sampling was performed in the same sites as in the clinical examination. A statistically significant difference was present when comparing %s of GB1 positive scores between teeth from the test group (57.5%) and teeth from the control group (25%). Plaque was present in 53% of test sites and 37% of control sites, but this difference was not statistically significant. Aa was detected, from at least one site in 85% of test subjects and in 15% of the control subjects (p

In order to expand the present knowledge of the pathogenic potential of Blastoschizomyces capitatus in central venous catheter (CVC)-related bloodstream infections, six strains of the organism recovered from three leukemic patients with CVC-related fungemia in different years were investigated. Isolates and control strains were tested for their genetic relatedness and for their ability to produce slime in glucose-containing solutions. DNA restriction enzyme analysis revealed that all strains of B. capitatus were identical, whereas slime production assays and examination of ex vivo material showed that they were able to produce large amounts of slime. Slime production may therefore play a relevant pathogenic role in cases of CVC-related fungemia caused by B. capitatus.

The efficacy of oral prophylaxis with ciprofloxacin, ofloxacin or pefloxacin was assessed in preventing bacterial infection in neutropenic patients with treatment being allocated randomly before beginning chemotherapy. Bacteraemia developed in six of 78 episodes (8%) treated with ciprofloxacin, in eight of 80 (10%) allocated to ofloxacin and in 12 of 77 (16%) when pefloxacin was given. However, there were no episodes involving Gram-negative bacilli among those given ciprofloxacin whereas three and seven episodes occurred in patients given ofloxacin or pefloxacin respectively (P = 0.013). With the exception of Pseudomonas aeruginosa, all potential pathogens isolated were resistant to all three fluoroquinolones. Faecal anaerobes were not affected by treatment with pefloxacin whereas their total numbers were reduced in 12 cases who had received ofloxacin and in nine cases who had been given ciprofloxacin (P = 0.002). Fourteen patients (18%) were colonized with pefloxacin resistant P. aeruginosa at the end of treatment with this agent compared with only two and five of those given ciprofloxacin or ofloxacin respectively. A similar trend was seen with other resistant Gram-negative bacilli colonizing 14%, 20% and 23% of patients for ciprofloxacin, ofloxacin and pefloxacin, respectively. Ciprofloxacin was therefore superior to the other two fluoroquinolones in preventing infections due to Gram-negative bacteria in this population of neutropenic patients.

The complete amino acid sequence of glutathione transferase from Proteus mirabilis was determined. The sequence was reconstructed by analysis of peptides obtides obtained after cleavabge by trypsin, Glu-C and Asp-N endoproteinases. The enzyme subunit is composed of 203 amino acid residues corresponding to a molecular mass of 22856 Da. Comparison of this sequence with other known primary structures of the corresponding enzyme from different sources shows a low level of identity (17–26%) with only seven conserved residues in all the sequences considered. This novel glutathione transferase could represent the prototype of a new class, possibly including other bacterial enzymes.

Stenotrophomonas maltophilia is an emerging nosocomial bacterial pathogen that is currently isolated with increasing frequency from the airways of cystic fibrosis (CF) patients. In this study the effect of subinhibitory concentrations (subMICs) of moxifloxacin on adhesion, biofilm formation and cell-surface hydrophobicity of two strains of S. maltophilia isolated from CF patients were evaluated. Adhesion and biofilm formation assays were carried out on polystyrene and quantified by colony counts. Cell-surface hydrophobicity was determined by a test for adhesion to n-hexadecane. Moxifloxacin at 0.03× and 0.06× MIC caused a significant decrease in adhesion and biofilm formation by both strains tested. A significant reduction in cell-surface hydrophobicity following exposure to subMICs of moxifloxacin was observed for one strain only. The results of the present study provide an additional rationale for the use of moxifloxacin in CF patients and more generally in biofilm-related infections involving S. maltophilia.

An affinity binding protein from the cytosolic fraction of Bacteroides fragilis was purified by using epoxy activated-Sepharose 6B resin immonobilized with GSH or with hexyl-GSH. This protein showed a subunit molecular mass (22 kDa) similar to that of glutathione transferase purified from Proteus mirabilis (22.5 kDa). However, the affinity binding protein of Bacteroides fragilis , unlike the GSH-affinity binding protein of Proteus mirabilis , was devoid of the capacity to conjugate GSH to the most commonly used glutathione transferase substrates. The GSH-affinity binding protein of Bacteroides fragilis was also antigenically different from the GSH-affinity bound protein of Proteus mirabilis . It was concluded that the anaerobic microorganism is not able to express glutathione transferase even though it contains a GSH-affinity binding protein with a structural characteristic reminiscent of aerobic glutathione transferase.

Five immunocompromised patients, four with AIDS and one who had undergone bone marrow transplantation, showing ocular signs of cytomegalovirus retinitis, were treated with 9-(2-hydroxy-1-(hydroxymethyl)ethoxymethyl) guanine (Ganciclovir), given intravenously at the dose of 5 mg/kg twice daily for a period ranging from 10 to 20 days. At the end of the treatment, in 4 of 5 patients, the ophthalmoscopic picture had improved, with reduced exudation and an arrest in the progression of retinal necrosis, the pattern clearly indicating a trend towards organization and scarring. Complete resolution of the retinitis without subsequent relapse was observed only in the bone marrow transplant patient, who recovered immunologically, whereas improvement of the eye involvement was only transient in the three AIDS patients.

This case report is a case history of a femoral prosthesis infection caused by Rhodotorula mucilaginosa in a human immunodeficiency virus patient. Though the pathogenicity of this organism for bone tissue has been previously reported, this is the first reported case of an orthopedic prosthesis infection by this species of the genus Rhodotorula .

Microbial penetration inside an implant's internal cavity results in a bacterial reservoir that has been associated with an area of inflamed connective tissue facing the fixture-abutment junction. The aim of the present clinical trial was to evaluate the effectiveness of a 1% chlorhexidine gel on the internal bacterial contamination of implants with screw-retained abutments.Thirty subjects (age range: 27.3 to 54.2 years) underwent single implant restoration. Three months after prosthodontic restoration, the modified sulcus bleeding index, modified plaque index, full-mouth plaque score, and full-mouth bleeding score were recorded. Microbiologic samples were also collected from the internal part of each fixture. Subjects were then divided into two equal groups: control and test groups (CG and TG, respectively). The CG had the abutment screwed and the crown cemented without any further intervention. Conversely, the TG had the internal part of the fixture filled with a 1% chlorhexidine gel before the abutment placement and screw tightening. Six months later, microbiologic and clinical procedures were repeated in both groups. Total bacterial count and multiplex polymerase chain analysis were performed to detect specific pathogens.Clinical parameters remained stable throughout the study. From baseline to the 6-month examination, the total bacterial counts underwent a significant reduction in the TG (P0.05). Detection of the single pathogen species did not show any significant differences. However, periopathogens were detected more frequently in the CG.The application of a 1% chlorhexidine gel seemed to be an effective method to reduce bacterial colonization of the implant cavity over a 6-month period.

Most of the current knowledge of the complex microbiology of oral biofilms, which initiates and maintains periodontal lesions, has been facilitated by the introduction of molecular techniques. Several studies exalt the high sensitivity and specificity of molecular tests in the detection and quantification of periodontal pathogens. Although they have large a diffusion, the old method of bacterial culture remains nowadays the gold standard when determining the utility of a new microbial test. Moreover, cultures have the important advantage of allowing an antibiotic sensitivity test and this is much more important during the treatment of patients with aggressive periodontitis.

Invasive aspergillosis (IA) is the leading direct or contributory cause of death in patients with haematological malignancies. Early diagnosis remains difficult and often elusive due the heterogeneity of clinical presentations and the low sensitivity of both histological examination and cultures of specimens obtained from patients at risk. We report two cases of IA, both of which lacked both histological and cultural evidence of IA from pulmonary specimens. In both patients, detection of galactomannan (GM) by enzyme immunoassay (EIA) on pulmonary tissue homogenates led to the diagnosis of IA, which was confirmed by Aspergillus DNA (real time PCR). In conclusion, we provide preliminary evidence that lung homogenates may be prepared for GM EIA assays, which may contribute to quick diagnosis of IA on otherwise negative samples. We feel that our results open up the opportunity of a prospective and comparative evaluation of this diagnostic technique.

Type-specific persistent infection with Human Papillomavirus (HPV) is a significant risk factor for the development of cervical diseases. Persistent infection could be further refined by a sequencing approach to detect early cervical lesions that are at high risk of developing an invasive squamous cervical cancer. The aim of the present study is to investigate the clinical utility of detecting mRNA transcripts of HPV oncogenes E6/E7 by using a Real-time NASBA technology (mRNA test) and to identify women with low-grade cytological disease but with an increased risk of developing high-grade cervical abnormalities or invasive squamous cervical cancer. Our preliminary results show that E6/E7 is detected in only a subset of HR-HPV-positive cases. Since viral persistence is considered to be the true precursor of neoplastic progression, only the detection of E6/E7 mRNA can identify the infection which is more likely to persist and induce neoplasia in future. For these reasons we believe that this test would be useful for the characterization of women with HR-HPV DNA positivity who should be effectively treated because at high-risk of developing a high grade cervical lesion or an invasive squamous cervical cancer.

The influence of environmental factors (temperature, aerobiosis-anaerobiosis, static-dynamic conditions, pH) was determined on biofilm formation by 51 S. maltophilia clinical isolates. The strains produced more biofilm at 32 degrees C than at 37 or 18 degrees C. Aerobic and 6% CO2 atmosphere yielded comparable biofilm amounts, higher than under anaerobic conditions. Biofilm production was not affected by static vs. agitated culture conditions. Biofilm production at pH 7.5 and 8.5 was comparable but significantly higher than at pH 5.5. The capacity of individual strains to form biofilm and thus contribute to the severity of some diseases is influenced by host traits and environmental conditions at the site of infection, and play an important role in the pathogenesis of biomaterial-related disease caused by S. maltophilia.

Patients with deep periodontal pockets were treated with either Vector System (TG) or manual instruments (CG). Clinical assessments by supragingival plaque (PL+), gingival index (GI), bleeding on probing (BOP), probing depth (PD), clinical attachment level (CAL) and subgingival plaque collection for microbiological analysis were made prior to and after treatment. Multiplex Polymerase Chain Reaction was used to determine the presence of Actinobacillus actinomycetemcomitans, Campylobacter rectus, Eikenella corrodens, Fusobacterium nucleatum, Prevotella intermedia, Porphyromonas gingivalis, Tannerella forsythensis and Treponema denticola. GI, PD, CAL and the number of BOP+ sites underwent a significant reduction over time in both groups. When compared to baseline, the pair-wise analyses showed significantly lower PD and CAL at 6 months in the CG and significant reductions in the GI, PD, CAL and a number of BOP+ sites at 3 and 6 months in the TG. For microbiological results, significant reductions were seen for C. rectus and R. gingivalis in the CG and for T. forsythensis, E. corrodens and T. denticola in the TG. The total bacterial count underwent a reduction in both groups. Both ultrasonic and manual debridement are equally effective in non-surgical periodontal therapy of severe periodontitis in terms of clinical and microbiological effects.

The aims of the present study were: (i) to assess whether H. pylori could be succesfully detected by PCR from the same biopsy sample used for CPtest; and ii) to evaluate CPtest comparatively to both PCR and histology for detection of H. pylori infection in dyspeptic patients. Three antral gastric biopsies were collected from each of 80 consecutive dyspeptic patients undergoing oesophagogastroduodenoscopy. Two biopsies were for histology (gold standard), one for CPtest, scored at 20min, 1h and 24h for the presence of urease activity. Gastric biopsy was then removed from CPtest and used for ureC-targeted PCR. Fifty-five (68.7%) patients were positive for H. pylori infection by histology. CPtest yielded an overall diagnostic accuracy of 93.8% (95% CI: 91–96.4%), regardless of observation period. No erroneous categorization of H. pylori status occurred using PCR, yielding sensitivity, specificity, positive and negative predictive values, and overall diagnostic accuracy of 100%. Our results suggest that H. pylori can be detected by PCR in gastric biopsies previously taken for CPtest, so reducing the workload of the endoscopist by saving additional biopsies for culture analysis and susceptibility tests.

We investigated the in vitro effects of seven fluoroquinolones (ciprofloxacin, grepafloxacin, levofloxacin, moxifloxacin, norfloxacin, ofloxacin, and rufloxacin), compared to those of trimethoprim-sulfamethoxazole (SXT) and ceftazidime on total biomass and cell viability of Stenotrophomonas maltophilia biofilm. S. maltophilia attached rapidly to polystyrene, within 2 h of incubation, and then biofilm formation increased over time, reaching maximum growth at 24 h. In the presence of fluoroquinolones at one-half and one-fourth the MIC, biofilm biomass was significantly ( P < 0.01) reduced to 55 to 70% and 66 to 76% of original mass, respectively. Ceftazidime and SXT did not exert any activity. Biofilm bacterial viability was significantly reduced by all antibiotics tested at one-half the MIC. At one-fourth the MIC all antibiotics, except levofloxacin, significantly reduced viability. Treatment of preformed biofilms with bactericidal concentrations (500, 100, and 50 μg/ml) of all fluoroquinolones caused, except for norfloxacin, significant reduction of biofilm biomass to 29.5 to 78.8, 64.1 to 83.6, and 70.5 to 82.8% of original mass, respectively. SXT exerted significant activity at 500 μg/ml only. Ceftazidime was completely inactive. Rufloxacin exhibited the highest activity on preformed biofilm viability, significantly decreasing viable counts by 0.6, 5.4, and 17.1% at 500, 100, and 50 μg/ml, respectively. Our results show that (i) subinhibitory (one-half and one-fourth the MIC) concentrations of fluoroquinolones inhibit adherence of S. maltophilia to polystyrene and (ii) clinically achievable concentrations (50 and 100 μg/ml) of rufloxacin are able to eradicate preformed S. maltophilia biofilm.

A prospective, randomized, double-blind trial was conducted on 124 febrile patients with hematological malignancies to compare teicoplanin with vancomycin as an addition to the initial empiric amikacin-ceftazidime regimen after documented bacteremia due to gram-positive cocci. At enrolment, patients in both groups were comparable with respect to age, sex, underlying hematologic disorders and duration of neutropenia. Rates of therapeutic success were 55/63 (87.3%) in the teicoplanin group and 56/61 (91.8%) in the vancomycin group (p = 0.560). The mean duration of treatment was similar, being 12.2 and 11.4 days, respectively (p = 0.216). Patients treated with teicoplanin remained febrile for slightly longer than those treated with vancomycin (4.9 vs. 4.0 days) (p = 0.013). Thirteen patients experienced an adverse drug reaction, but without any significant difference in the two arms. Isolated staphylococci showed a progressive and significant decrease in susceptibility to both glycopeptides during the 8 study years. The economic analysis performed showed that the addition of vancomycin is cost-saving.

The aim of this study was to evaluate the efficacy of the Pumped Diodium-Nd:YAG laser in sterilizing contaminated root canals. After hand instrumentation, 30 teeth were inoculated with Actinomyces naeslundii CH-12 and 30 teeth with Pseudomonas aeruginosa ATCC 27853 and incubated for 24 h. The teeth were divided into three subgroups: subgroup A received no treatment; subgroup B was irradiated with laser (5 Hz for 15 s or 10 Hz for 15 s); and subgroup C was irrigated with 5.25% NaOCl. The number of viable bacteria in each group was evaluated by using the surface-spread plate technique. The results indicated an average of 34.0% decrease in colony-forming units for A. naeslundii CH-12 and 15.7% for P. aeruginosa ATCC 27853 with the 5 Hz/15 s laser treatment, and for the 10 Hz laser frequency, a decrease of the 77.4% for A. naeslundii CH-12 and 85.8% for P. aeruginosa ATCC 27853. No bacteria were detected in the canals treated with 5.25% NaOCl. The results show an antibacterial effect of the Pumped Diodium TM -Nd:YAG laser, depending on the radiation frequency. However, 5.25% NaOCl was more effective than either laser application.

Coagulase-negative staphylococci are frequently isolated from clinical specimens and they represent the most common cause of bacteraemia in hospitalised patients. Particularly, venous catheter-related bloodstream infections are often due to non- aureus staphylococci. These are opportunistic pathogens in immunocompromised hosts and may behave as reservoirs of antibiotic resistance determinants.1 A 75-year-old woman was admitted to hospital because of diffuse bone pain. A relapse of myeloid acute leukaemia was diagnosed (the first diagnosis had been made 5 months earlier), and she was admitted to the haematology department. Nine days later she developed fever (to 39°C) and chills. A Gram-negative infection was suspected, and blood samples (the set included two BacT/Alert aerobe/yeast bottles (bioMerieux, Marcy l’Etoile, France) plus one anaerobe bottle) were taken for culture. A second set (including two BacT/Alert aerobe/yeast bottles, without the anaerobe bottle) was taken after 30 min, and meropenem treatment was started (1 g every 8 h, intravenous). Due to persistence of fever, meropenem was replaced with piperacillin–tazobactam (4.5 g every 8 h, intravenous) 2 days later. After 48 h incubation, all of the samples were detected as positive by the BacT/Alert; Gram staining showed Gram-positive cocci, and cultures yielded Staphylococcus pasteuri as a single organism; typical yellow colonies were observed after 24 h incubation under aerobic and anaerobic conditions. The species was initially identified as Staphylococcus …

The aim of this study was to verify the in vitro influence of various dilutions of a viscoelastic substance containing 2% hydroxy-propyl-methylcellulose (HPMC) on the viability of some microbial strains representative of the normal flora of the external eye. Pure reference strain cultures of Candida albicans, Pseudomonas aeruginosa, Propionibacterium acnes, Staphylococcus aureus, Staphylococcus epidermidis and a fresh clinical isolate of Proteus mirabilis were selected for this study. Serial twofold dilutions of 2% HPMC, prepared so as to obtain a final concentration ranging from 50 to 0.78% of the product in sterile saline solution (0.85% NaCl), were taken out with a pipette that delivered 1.0 ml per tube. One hundred microliters of the abovementioned microbial inocula, used for the evaluation of the positive control of the test organism, were dispensed into each tube. After 24 h of incubation, 100 μl of samples were taken from each tube and plated into the specific medium for the growth of the test organism. After 24–48 h of incubation, these agar plates were examined and the colony-forming-unit count of each test organism was compared to the corresponding total colony count, acting as a positive control, in order to determine the quantitative variation of the test organism grown in the presence of the viscoelastic compounds. C. albicans and P. aeruginosa showed a statistically significant increase in growth with HPMC dilutions varying from 1:2 to 1:16. P. acnes and P. mirabilis growth was significantly reduced by all dilutions except for the 1:128 one. S. epidermidis growth was also significantly reduced in the presence of HPMC dilutions varying from 1:2 to 1:64. S. aureus growth was not significantly influenced. The viability of P. aeruginosa in HPMC dilutions needs to be carefully considered because of the ability of this organism to induce endophthalmitis, and the possibility that during cataract surgery, a small amount of HPMC may be left in the eye, trapped behind the intraocular lens optic.

The capacity of clinical isolates and type strains of Actinobacillus actinomycetemcomitans to survive in a new transport medium (AaTM), phosphate-buffered saline (PBS) and Ringer's solution (RS) was evaluated. The effects of exposure to air, transportation time and temperature on viability were also studied. In addition, the culture of A. actinomycetemcomitans from subgingival plaque of patients with different forms of periodontitis was quantified. The results following storage in AaTM, PBS and RS showed that A. actinomycetemcomitans survived better in AaTM than in PBS or RS when transportation times exceeded 20-22 h, and that survival was enhanced by storage at below 12 degrees C. Serotype b strains of A. actinomycetemcomitans were able to survive better than either serotype a or c. In the clinical study the optimal transportation conditions for subgingival plaque containing A. actinomycetemcomitans were AaTM at a temperature of 8 degrees C for 24 h under anaerobic conditions. These conditions resulted in a high survival and isolation rate for A. actinomycetemcomitans without inhibition of the other periodontopathic bacteria isolated from deep periodontal pockets. These findings have practical implications for future multicentre clinical trials in which the transportation of oral specimens over relatively long distances and at different ambient temperatures during various periods of the year are required.

The Leifson staining method was used to diagnose Helicobacter pylori infection and was compared to histology, culture, and the rapid urease test (RUT). Histology gave the best sensitivity (98%), compared to Leifson staining (97%), culture (92%), and RUT (85%) ( P < 0.005). Leifson staining is a sensitive, rapid, economical method for diagnosis of H. pylori infection in dyspeptic patients.

The E test was evaluated in comparison with reference agar methods (National Committee for Clinical Laboratory Standards) for the susceptibility testing of 248 Pseudomonas aeruginosa isolates from bladder-catheterized patients against nine antibiotics. The E-test MICs correlated well with those determined by the agar dilution and disk diffusion reference methods (88 and 92.5% within 1 log 2 dilution step, respectively), confirming that the E test is a reliable method for the determination of MICs of antibiotics for catheterization-associated P. aeruginosa isolates.

The accuracy and efficiency of Staf-Sistem 18-R (Liofilchem s.r.l., Roseto degli Abruzzi, Teramo, Italy) were compared with those of conventional biochemical methods to identify 523 strains belonging to 16 different human Staphylococcus species. Overall, 491 strains (93.9%) were correctly identified (percentage of identification, > or = 90.0), with 28 (5.4%) requiring supplementary tests for complete identification. For 14 isolates (2.8%), the strains did not correspond to any key in the codebook and could not be identified by the manufacturer's computer service. Only 18 isolates (3.4%) were misidentified. The system is simple to use, is easy to handle, gives highly reproducible results, and is inexpensive. With the inclusion of more discriminating tests and adjustment in supplementary code numbers for some species, such as Staphylococcus lugdunensis and Staphylococcus schleiferi, Staf-Sistem 18-R is a suitable alternative for identification of human coagulase-positive and coagulase-negative Staphylococcus species in microbiological laboratories.