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Clinical microbiologists and microbiology laboratories are experiencing changes due to evolving views on‘healthcare delivery’ as an economic activity, due to changes in the medical environment and the demographics of the workforce, and technical evolution. Cost-effectiveness of laboratory procedures has been achieved through consolidation and integration of laboratories. Consolidation offers economy of scale and reduction in numbers of on-site staff, but also leads to separation of microbiologists from their clinical colleagues. Integration puts different laboratory disciplines under a single management, and leads to reorganisation of laboratories along common work-lines. Cost-savings combined with on-site availability of laboratories are achieved at the expense of a reduction in the influence of microbiologists in the daily running of the laboratory. Medically, there is growing emphasis on evidence-based diagnostics. Because of time-delays inherent in culturing, microbiology has a limited impact on patient outcomes. Increased clinical relevance of microbiological testing through rapid testing is mandatory. There is an increasing shortage in Europe and the USA of trained microbiology laboratory technicians and microbiologists. This reinforces the trend towards more automation and integration. Technological advances, particularly in molecular diagnostics, offer the possibility of rapid reporting and improvement of the impact of clinical microbiology on patient management. Molecular tests, however, fit perfectly the concept of an integrated laboratory and may further loosen the link between microbiologist and microbiology tests. The challenge for clinical microbiology will be to use new techniques to improve its cost-effectiveness and impact on infectious disease management. The future organisation of microbiology laboratories must support this but is itself of secondary importance. The training of future microbiologists must prepare them for this changing environment. [ABSTRACT FROM AUTHOR]

We report a case of cryptococcal meningitis in an immunocompetent female patient with exposure to a pet magpie (Pica pica). Genetically indistinguishable isolates were cultured from the cerebrospinal fluid of the patient and excreta of the bird. Our data strongly suggest zoonotic transmission of Cryptococcus neoformans var. grubii from a magpie to this patient. [ABSTRACT FROM AUTHOR]

Lagrou K, Van Eldere J, Keuleers S, Hagen F, Merckx R, Verhaegen J, Peetermans WE, Boekhout T (University Hospital Leuven, Leuven, Belgium; and Centraalbureau voor Schimmelcultures, CT Utrecht, the Netherlands) Zoonotic transmission ofCryptococcus neoformansfrom a magpie to an immunocompetent patient (Case report).J Intern Med2005;257:385–388.We report a case of cryptococcal meningitis in an immunocompetent female patient with exposure to a pet magpie (Pica pica). Genetically indistinguishable isolates were cultured from the cerebrospinal fluid of the patient and excreta of the bird. Our data strongly suggest zoonotic transmission ofCryptococcus neoformansvar.grubiifrom a magpie to this patient. [ABSTRACT FROM AUTHOR]

Evaluates the need for changes in the approaches in clinical microbiology in Belgium. Impact of clinical microbiology on the management of infectious diseases; Models for enhancement in diagnostic microbiology; Emphasis on point-of-care testing.

Describes the presence of multiple coagulase-negative staphylococci clones in a patient with prosthetic heart valves, which was probably caused by genetic variability in the infecting strain. Genome macrorestriction and amplified fragment-length polymorphism analyses of Staphylococcus epidermidis strains; Changes in antibiotic susceptibility during an infection; Distinction between resistant and susceptible clones.

Of 233 erythromycin-resistant pneumococcal isolates collected in Belgium in 1999–2000, 89.7% carried theerm(B) gene, 6% themef(A) gene, and 3.5%erm(B) plusmef(A). Two isolates contained neithererm(B) normef(A); one contained anerm(A) subclasserm(TR) gene, while the other contained an A2058G mutation in domain V of the 23S rRNA gene. Of 209erm(B)-positive isolates, 191 had clindamycin MICs> 16 mg/L and 18 had MICs≤ 16 mg/L.Mef(A)-positive isolates all displayed the M resistance phenotype. Telithromycin remained active against erythromycin-resistant isolates, with the highest telithromycin MIC50 being found inmef(A)-positive isolates. No difference in the prevalence of different resistance mechanisms was observed compared to isolates collected in 1995–1997. [ABSTRACT FROM AUTHOR]

Presents an introduction of the article "Progress Towards Meeting the Challenges in Clinical Microbiology and Infectious Diseases," published in the April 2, 2005 issue of "Clinical Microbiology & Infection."

Highlights the multidisciplinary workshop of the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) that aims to meet the challenges in clinical microbiology and infectious disease in Europe. Appraisal of the significant progress towards improving the preparedness of Europe to respond to the threats of emerging infections, antibiotic resistance and bioterrorism; Importance of urgent studies to measure the health and economic burden of nosocomial infections and antibiotic resistance; Role of ESCMID in achieving better-coordinated surveillance and prevention of infection.

Abstract: We describe the development and application of a rapid and universal molecular technique for direct identification of multiple bacteria in clinical samples. Amplification of the 16S–23S rRNA spacer-region using universal primers led to fragment patterns distinct for different bacterial species and that were analyzed with fluorescent amplicon length analysis (FALA). 136 pure cultures of clinical isolates and 20 culture collection strains belonging to 22 different medically important species were used to create a primary database of fragments with sizes between 100 and 1000 bp. Subsequently, 127 respiratory samples were analyzed with culture-based techniques and via FALA of the 16S–23S rRNA spacer-region. Two DNA extraction methods were evaluated: Instagene® (FALA-I) and Fastprep™ (FALA-P). Of the 127 samples, 26 culture-negative samples were also negative with FALA-P. Of 18 samples with growth of commensal oral flora, 10 gave a mixed oral flora pattern with FALA-P and 8 gave a negative result. For 54 samples with growth of a single bacterial species, FALA-P gave an identical result for 46. For 29 samples with growth of more than one bacterial species, identical results were obtained in 19 samples. False-negative results with FALA-P were mostly due to paucity (less than 103 CFU/ml) of bacteria (12 out of 18 false-negatives) or difficulties with homogenization of viscous samples (6 out of 18 false-negatives).With regard to identification of all significant pathogens of clinical samples tested, the sensitivity of FALA-P was 77% and its specificity was 100%. With FALA-I, the number of false-negative results was higher than with FALA-P due to less efficient extraction of DNA, particularly with Staphylococcal species. FALA-P allows rapid and direct identification of multiple species directly from clinical samples; pauci-cellular samples may give false-negative results. [Copyright &y& Elsevier]

Clinical microbiology should have a physical presence, but not necessarily on-site diagnostic laboratory facilities, in each hospital to ensure a quality laboratory-based infection service and strong professional interaction with clinicians. The adoption of industrial practices and the introduction of new costly molecular techniques raise the possibility that non-microbiological functions of laboratory management could be left to management professionals. This remains highly controversial; the advantages must be contrasted with the potential to disrupt the traditional managerial responsibility of the microbiologist and the links between the laboratory and clinical staff. Managers and healthcare professionals must resolve this issue, perhaps with the support of the ESCMID. Views varied, according to current professional arrangements and size of the laboratory and population served, on whether there should be a common laboratory for microbiology and other pathology disciplines with joint access to new high-technology techniques, or whether microbiology must continue as a separate facility. Clinical microbiology and infection control were viewed as core services that must be present even in smaller hospitals. Larger community hospitals and teaching centres require a full complement of expertise in laboratory and clinical practice. Integration of these disciplines within a department of infection is an emerging concept. A concern was the shortfall in trained expertise because of the ageing nature of current specialists. The importance of recruiting talented new graduates was emphasised. The importance of this topic led to a recommendation that an ESCMID working party be established to investigate the current arrangements of infection services in Europe and to make recommendations for the future organisation. [ABSTRACT FROM AUTHOR]

Quinolones are one of the largest classes of antimicrobial agents used worldwide. This review considers the quinolones that are available currently and used widely in Europe (norfoxacin, ciprofloxacin, ofloxacin, levofloxacin and moxifloxacin) within their historical perspective, while trying to position them in the context of recent and possible future advances based on an understanding of: (1) their chemical structures and how these impact on activity and toxicity; (2) resistance mechanisms (mutations in target genes, efflux pumps); (3) their pharmacodynamic properties (AUC/MIC andCmax/MIC ratios; mutant prevention concentration and mutant selection window); and (4) epidemiological considerations (risk of emergence of resistance, clonal spread). Their main indications are examined in relation to their advantages and drawbacks. Overall, it is concluded that these important agents should be used in an educated fashion, based on a careful balance between their ease of use and efficacy vs. the risk of emerging resistance and toxicity. However, there is now substantial evidence to support use of the most potent drug at the appropriate dose whenever this is required. [ABSTRACT FROM AUTHOR]

Abstract Objectives: Porphyromonas gingivalis , one of the key pathogens in the development of periodontitis, produces a number of virulence factors that might explain its pathogenicity. One of them is the ability to adhere and invade pocket epithelium. The aim of this study was to follow, over time, the association of P. gingivalis and consequent morphological changes of the pocket epithelium cells. Material and Methods: The association capacity of four P. gingivalis serotypes [K1, K2, K4, K- (nonencapsulated)] with in vitro cultured mono-layers from periodontal pocket epithelial cells of patients with periodontitis, was followed by fluorescence microscopy and bacterial culture. The contact time between bacteria and epithelium cells ranged from 45 min to 8 h. The microscopic evaluation allowed differentiation between dead and living cells (bacteria as well as epithelium) and description of the morphological changes after association. Results: A highly significant difference in the number of associating bacteria was found between dead and living epithelium cells, and between non-capsulated and capsulated strains. A significant increase in the proportion of dead pocket epithelium cells was found with prolonged association time. The morphological changes (rounding of the epithelial cell, detachment from the glass cover-slip and loss of intercellular contact) occurred faster for mono-layers inoculated with the non-encapsulated P. gingivalis strain. Conclusions: This study indicates that dead pocket epithelium cells harbor more P. gingivalis cells, and that a positive correlation exists between contact time and cell death. For the P. ginigvalis species, non-encapsulated strains associate in higher number. As a result, the damage they cause to the host cell seems to occur faster than occurs in encapsulated strains. As such, cell death can be seen as the end-result of bacterial association. [ABSTRACT FROM AUTHOR]

A challenge panel of bacterial strains useful for clinical laboratories to validate their European Committee on Antimicrobial Susceptibility Testing (EUCAST) antimicrobial susceptibility test (AST) system was established. A total of 117 strains, obtained from Belgian Reference Centres ( n = 57) and from routine clinical samples ( n = 60) was selected based on resistance pattern. These strains were analysed in seven different laboratories by three different automated AST systems (Vitek ( n = 2), Phoenix ( n = 2) and Microscan ( n = 2)) and by disc diffusion from five different manufacturers (Rosco ( n = 2), Becton-Dickinson ( n = 2), Biomérieux ( n = 1), Bio-rad ( n = 1) and i2a ( n = 1)). To select the challenge panel, selection criteria were set for categorical agreement between the different systems and the number of very major errors, major errors and minor errors. Very major and major errors for at least two antibiotics were observed in 43% of all strains, leading to the exclusion of these strains from the selected panel. In only 10% of all tested strains was there 100% categorical agreement for all antibiotics. Finally, 28 strains (14 Gram-positive and 14 Gram-negative) covering a wide spectrum of resistance mechanisms were selected. Pilot-testing of this challenge panel in 20 laboratories mainly confirmed the results of the validation study. Only six strains withheld for the pilot study could not be used as challenge strain due to an overall (very) major error rate of >5% for a particular antibiotic ( n = 5) or for two antibiotics ( n = 1). To conclude, this challenge panel should facilitate the implementation and use of EUCAST breakpoints in laboratories. [ABSTRACT FROM AUTHOR]

Biofilm-associated bacteria display a decreased susceptibility towards antibiotics. Routine assessment of antibiotic susceptibility of planktonic bacteria therefore offers an insufficient prediction of the biofilm response. In this study, in vitro biofilms of eight clinical Staphylococcus epidermidis strains were subjected to treatment with vancomycin, teicoplanin, oxacillin, rifampicin and gentamicin. In addition, the biofilms were subjected to combinations of an antibiotic with rifampicin. The effects on the biofilms were assessed by crystal violet staining to determine the total biofilm biomass, staining with XTT to determine bacterial cell viability, and microscopy. Combining these methods showed that treatment of S. epidermidis biofilms with glycopeptides increased the total biofilm biomass and that these antibiotics were not effective in killing bacteria embedded in biofilms. The decreased killing efficacy was more pronounced in biofilms produced by strains that were classified as ‘strong’ biofilm producers. Rifampicin, oxacillin and gentamicin effectively killed biofilm-associated bacteria of all tested strains. Combining antibiotics with rifampicin increased the killing efficacy without influencing the total biofilm biomass. When vancomycin or teicoplanin were combined with rifampicin, the increase in biofilm biomass was neutralised and also the killing efficacy was influenced in a positive way. We conclude that the combined methodology used in this study showed that glycopeptides were not effective in eradicating S. epidermidis biofilms but that combination with rifampicin improved the killing efficacy in vitro. [ABSTRACT FROM AUTHOR]

The purpose of this investigation was to determine the proportion of influenza-like illness (ILI) attributable to specific viruses during the influenza A(H1N1)2009 pandemic and to describe the demographic and clinical characteristics of ILI due to respiratory viruses in Belgium. Nasopharyngeal swabs were collected from ILI patients by general practitioners (GPs) and paediatricians (PediSurv) and analysed for viruses. Of 139 samples collected from children <5 years of age by PediSurv, 86 were positive, including 28 influenza (20%), 27 respiratory syncytial virus (RSV) (19%), 21 rhinovirus (17%), 12 human metapneumovirus (hMPV) (9%) and ten parainfluenza virus (PIV) (7%). Of 810 samples received from GPs, 426 were influenza (53%). Of 312 influenza-negative samples, 41 were rhinovirus (13%), 13 RSV (4%), 11 PIV (4%) and three hMPV (1%). Influenza mostly affected the 6-15 years old age group. Other respiratory viruses were commonly detected in the youngest patients. Similar clinical symptoms were associated with different respiratory viruses. Influenza A(H1N1)2009 was the most detected virus in ILI patients during the 2009-2010 winter, suggesting a good correlation between ILI case definition and influenza diagnosis. However, in children under 5 years of age, other respiratory viruses such as RSV were frequently diagnosed. Furthermore, our findings do not suggest that the early occurrence of the influenza A(H1N1)2009 epidemic impacted the RSV epidemic in Belgium. [ABSTRACT FROM AUTHOR]

Background: Low serum 25-hydroxyvitamin D (25-[OH]D) levels have been associated with lower FEV(1), impaired immunologic control, and increased airway inflammation. Because many patients with chronic obstructive pulmonary disease (COPD) have vitamin D deficiency, effects of vitamin D supplementation may extend beyond preventing osteoporosis.Objective: To explore whether supplementation with high doses of vitamin D could reduce the incidence of COPD exacerbations.Design: Randomized, single-center, double-blind, placebo-controlled trial. (ClinicalTrials.gov registration number: NCT00666367)Setting: University Hospitals Leuven, Leuven, Belgium.Patients: 182 patients with moderate to very severe COPD and a history of recent exacerbations.Intervention: 100,000 IU of vitamin D supplementation or placebo every 4 weeks for 1 year.Measurements: The primary outcome was time to first exacerbation. Secondary outcomes were exacerbation rate, time to first hospitalization, time to second exacerbation, FEV(1), quality of life, and death.Results: Mean serum 25-(OH)D levels increased significantly in the vitamin D group compared with the placebo group (mean between-group difference, 30 ng/mL [95% CI, 27 to 33 ng/mL]; P < 0.001). The median time to first exacerbation did not significantly differ between the groups (hazard ratio, 1.1 [CI, 0.82 to 1.56]; P = 0.41), nor did exacerbation rates, FEV(1), hospitalization, quality of life, and death. However, a post hoc analysis in 30 participants with severe vitamin D deficiency (serum 25-[OH]D levels <10 ng/mL) at baseline showed a significant reduction in exacerbations in the vitamin D group (rate ratio, 0.57 [CI, 0.33 to 0.98]; P = 0.042).Limitation: This was a single-center study with a small sample size.Conclusion: High-dose vitamin D supplementation in a sample of patients with COPD did not reduce the incidence of exacerbations. In participants with severe vitamin D deficiency at baseline, supplementation may reduce exacerbations.Primary Funding Source: Applied Biomedical Research Program, Agency for Innovation by Science and Technology (IWT-TBM). [ABSTRACT FROM AUTHOR]

Recently, the predation of Bdellovibrio bacteriovorus on a periodontal pathogen has been described. The current study explores the potential antimicrobial activity of a range of predatory bacteria against key periodontal pathogens. A number of representatives from the Bdellovibrio, Bacteriovorax and Peredibacter lineages (called 'BALOs') were tested for their activity towards a group of key periodontal pathogens and an optimal multiplicity of infection was established. As the oral cavity contains a wide variety of bacteria that are not preyed upon, it was investigated if they can have an effect on the predation efficiency of BALOs. It was concluded that a number of important variables involved in bacterial predation are found to be compatible with the composition of the oral microbiota. This finding makes the case for continued study of the potential for BALOs to combat periodontal pathogens. [ABSTRACT FROM AUTHOR]

OBJECTIVES: To investigate the presence of extended-spectrum beta-lactamases (ESBLs) among Pseudomonas aeruginosa clinical isolates referred to two Belgian reference laboratories. METHODS: Antibiograms were analysed for P. aeruginosa isolates referred between 2004 and 2008. Isolates resistant to ceftazidime (MIC > 8 mg/L) and with a positive double-disc synergy test between ceftazidime and clavulanate were serotyped and screened for the presence of ESBL-encoding genes. Genes encoding metallo-beta-lactamases (bla(MBL)) were sought by PCR in ESBL-producing isolates with positive imipenem/EDTA synergy tests. PFGE of SpeI-digested genomic DNA was used to compare isolates and selected strains were characterized by multilocus sequence typing. RESULTS: Forty-eight (2.2%) of 2150 P. aeruginosa isolates were confirmed as class A ESBL-producing isolates by molecular testing. bla(BEL) and bla(PER) alleles were detected, respectively, in 39 and 10 P. aeruginosa isolates originating from 16 hospitals (two isolates were simultaneously positive for BEL and PER). Fifteen of the isolates were found to co-produce ESBLs and VIM carbapenemases. These strains were pan-resistant and remained susceptible only to colistin (MICs

Objectives: To investigate the presence of extended-spectrum β-lactamases (ESBLs) among Pseudomonas aeruginosa clinical isolates referred to two Belgian reference laboratories. Methods: Antibiograms were analysed for P. aeruginosa isolates referred between 2004 and 2008. Isolates resistant to ceftazidime (MIC.8 mg/L) and with a positive double-disc synergy test between ceftazidime and clavulanate were serotyped and screened for the presence of ESBL-encoding genes. Genes encoding metallo-b-lactamases (blaMBL) were sought by PCR in ESBL-producing isolates with positive imipenem/EDTA synergy tests. PFGE of SpeI-digested genomic DNA was used to compare isolates and selected strains were characterized by multilocus sequence typing. Results: Forty-eight (2.2%) of 2150 P. aeruginosa isolates were confirmed as class A ESBL-producing isolates by molecular testing. blaBEL and blaPER alleles were detected, respectively, in 39 and 10 P. aeruginosa isolates originating from 16 hospitals (two isolates were simultaneously positive for BEL and PER). Fifteen of the isolates were found to co-produce ESBLs and VIM carbapenemases. These strains were pan-resistant and remained susceptible only to colistin (MICs≤2 mg/L). The majority of the ESBL-producing isolates belonged to the same PFGE clone and were identified as ST235; serotype O11. Conclusions: BEL enzymes were produced by 80% of P. aeruginosa isolates with phenotypic evidence of ESBL production. BEL or PER ESBLs co-existed with VIM carbapenemases in 15 isolates and caused outbreaks in four hospitals. Our data further highlight the epidemic potential of the international clone ST235, which may have acquired blaBEL-1 gene cassettes from a yet unidentified local gene reservoir. [ABSTRACT FROM AUTHOR]

Abstract: Objectives: The aim of the study was to evaluate the antibiotic resistance in noninvasive clinical isolates of Streptococcus pneumoniae collected in Belgium during winter 2008–2007. Method: Four hundred and forty eight unduplicated isolates collected by 15 laboratories were tested by microdilution following CLSI. Results: Insusceptibility rates (I+R) were as follows: penicillin G (PEN) 11.6% (4.0% R), ampicillin 11.4% (4.0% R), amoxicillin+/–clavulanic acid 0, cefaclor 10.3% (9.6% R), cefuroxime 9.2% (8.7% R), cefuroxime-axetil 8.7% (7.8% R), cefotaxime, ceftazidime and cefepime 2.0% (0% R), imipenem 2.5% (0% R), ciprofloxacin and ofloxacin 5.1% (0.4% R), levofloxacin 0.7% (0.4% R), moxifloxacin 0.4% (0.2% R), erythromycin (ERY) 29.7% (29.2% R), azithromycin 29.7% (28.8% R), telithromycin 0%, clindamycin 26.3% (25.4% R) and tetracycline (TET) 21.9% (16.5% R). From 2001 to 2008, a significant decrease in penicillin-insusceptibility (21.0% to 11.6%), penicillin-resistance (9.7% to 4.0%) and ciprofloxacin-insusceptibility (11.2% to 5.1%) was found. Cross-resistance between penicillin and other betalactams in penicillin-insusceptible isolates was incomplete: all these isolates remained fully susceptible to amoxicillin. Erythromycin-insusceptibility was significantly higher in children than in adults (43.9%/27.4%), while penicillin-insusceptibility significantly higher in Brussels than in the Flanders (22.9%/8.1%). The commonest resistance phenotype was ERY-TET (12.7%) followed by ERY (7.4%) and PEN-ERY-TET (5.8%). Capsular types 19 (25%), 14 (19.3%), 23 (15.4%) and 15 (13.5%) were the most important in penicillin-insusceptible. Conclusion: We noted a decrease in resistance to the majority of the compounds. Insusceptibility rates were higher in children than in adults and the difference between the north and the south of Belgium became less marked. [Copyright &y& Elsevier]

Introduction: Microbial interactions are considered to be important for bacterial colonization. Interactions that inhibit colonization of pathogens could possibly be used as a new treatment approach for periodontitis. The aim of this study was to test this hypothesis on soft surfaces in vitro, taking into account the hydrodynamic forces continuously present in vivo. Methods: Cultured epithelial cells were precolonized with Streptococcus sanguinis KTH-4, Streptococcus cristatus CC5A, Streptococcus salivarius TOVE and Streptococcus mitis BMS before Aggregatibacter actinomycetemcomitans colonization. Experiments were performed in a modified Robbins-device-type flow cell. Bacterial colonization and the number of epithelial cells were evaluated by microbial culturing and quantitative polymerase chain reaction. Results: The streptococci were able to inhibit A. actinomycetemcomitans colonization on soft tissue surfaces under flow conditions. Statistically significant differences were found between streptococcal pretreatments and the controls, with the most pronounced effect caused by S. sanguinis. Conclusion: These data confirm the possibility of applying beneficial bacteria in periodontal treatment. [ABSTRACT FROM AUTHOR]

Quantification of Bdellovibrio-and-like organisms (BALOs) by microbial culturing has a number of substantial drawbacks. Therefore a quantitative PCR (qPCR) assay was designed for the culture-independent enumeration of the Bdellovibrionaceae. After optimization, the dynamic range of the qPCR assay was assessed, the specificity was evaluated and a comparison with quantitative microbial culturing was made. To evaluate the suitability of the qPCR assay for analysing environmental samples, fresh water samples were investigated by microbial culturing and by the qPCR assay. The results revealed a substantial difference between the two techniques and indicate that most Bdellovibrionaceae cells are left undetected in environmental samples when only current microbial culturing techniques are used. The application of this new technique is therefore likely to confirm the hitherto underestimated sizes and roles of predatory bacterial populations in nature. [ABSTRACT FROM AUTHOR]

Introduction: Oral bacteria must attach to hard and soft tissues to colonize the oral cavity in the presence of a variety of forces caused by shear and flow. In vitro models mimicking this dynamic process are indispensable to study factors that might interfere with the first step towards infection. For extrapolation purposes the comparability between the dynamics of colonization on hard vs. soft surfaces needs to be evaluated. Methods: The colonization of glass and epithelial cell surfaces by the periodontal pathogen Aggregatibacter actinomycetemcomitans was followed in time with two flow cell models: a modified Robbins device (MRD) and an in situ image analysis system. Results: The number of A. actinomycetemcomitans recovered from the soft surfaces in the MRD experiments was higher than on glass. The amount of bacteria on the hard surfaces kept increasing with time, while on soft surfaces saturation was reached. The microscope-mounted flow cell allowed real-time in situ monitoring of the colonization process of both surfaces. Conclusion: These experimental models may have a great contribution to make in the development of new treatment approaches for periodontal diseases. Colonization by A. actinomycetemcomitans could be studied under flow conditions and its dynamics showed important surface-dependent characteristics. [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa is a major cause of nosocomial infections. This organism shows a remarkable capacity to resist antibiotics, either intrinsically (because of constitutive expression of β-lactamases and efflux pumps, combined with low permeability of the outer-membrane) or following acquisition of resistance genes (e.g., genes for β-lactamases, or enzymes inactivating aminoglycosides or modifying their target), over-expression of efflux pumps, decreased expression of porins, or mutations in quinolone targets. Worryingly, these mechanisms are often present simultaneously, thereby conferring multiresistant phenotypes. Susceptibility testing is therefore crucial in clinical practice. Empirical treatment usually involves combination therapy, selected on the basis of known local epidemiology (usually a β-lactam plus an aminoglycoside or a fluoroquinolone). However, therapy should be simplified as soon as possible, based on susceptibility data and the patient's clinical evolution. Alternative drugs (e.g., colistin) have proven useful against multiresistant strains, but innovative therapeutic options for the future remain scarce, while attempts to develop vaccines have been unsuccessful to date. Among broad-spectrum antibiotics in development, ceftobiprole, sitafloxacin and doripenem show interesting in-vitro activity, although the first two molecules have been evaluated in clinics only against Gram-positive organisms. Doripenem has received a fast track designation from the US Food and Drug Administration for the treatment of nosocomial pneumonia. Pump inhibitors are undergoing phase I trials in cystic fibrosis patients. Therefore, selecting appropriate antibiotics and optimising their use on the basis of pharmacodynamic concepts currently remains the best way of coping with pseudomonal infections. [ABSTRACT FROM AUTHOR]

Objectives The aim of this study was to examine the accuracy of the presence of high-risk human papillomavirus (HR-HPV) DNA (HR-HPV DNA test) postconisation as prediction of recurrent or residual cervical intraepithelial neoplasia (CIN) after treatment of high-grade cervical intraepithelial lesions (CIN2+) in a prospective study and to compare this with follow-up cytology and the marginal status of the excised tissue. Design Prospective follow-up study. Setting Unselected women presenting at colposcopy clinic of University Hospital Gasthuisberg, Leuven. Population Seventy-two women treated with conisation for CIN2 or CIN3. Methods Women were followed by HR-HPV DNA test (Hybrid Capture II test of Digene®) every 3 to 6 months. The same vial was used for cytology and the HR-HPV DNA test (SurePath™). All women were further followed by colposcopy and cytology for 24 months at 6-month intervals. The outcome of the study was presence of >CIN2, proven with colposcopy-directed biopsy occurring within 24 months after treatment. HR-HPV status was correlated with recurrent or residual CIN2+. Main outcome measures Sensitivity, specificity, predictive values and diagnostic odds ratios to predict treatment failure or cure were computed for HR-HPV testing, marginal status and follow-up cytology. HR-HPV status was also correlated with section margins postconisation and with the first cervical smear. Results In 6 of the 72 treated women (8%), residual or recurrent CIN occurred. Women with recurrence were significantly older than women without a recurrence (51.5 ± 9.6 versus 39.8 ± 12.2 years, P= 0.007). All six women with recurrence were HR-HPV positive, four had a positive follow-up smear (≥atypical squamous cells of uncertain significance = ASCUS+) and only two had involved section margins. Among the 66 cured women, 15 were HR-HPV positive, 6 had an abnormal smear and 12 had positive section margins. Sensitivity of cytology, positive section margins and HR-HPV DNA positivity was 66.7, 33.3 and 100% to predict treatment failure. Specificity of the three tests was, respectively, 90.9, 81.8 and 77.3%. Women with HR-HPV DNA at 3 to 6 months showed recurrent or residual CIN in 15% (2/13) if they had normal follow-up Pap smears and in 50% (4/8) if they had abnormal Pap smears. Margin status was not statistically significantly associated with human papillomavirus status. Conclusion Persistence or clearance of HR-HPV DNA is an early valid prognostic marker of failure or cure after treatment for CIN2+ and is more accurate than cytology or section margin status at the time of conisation. The absence of HR-HPV DNA has a 100% negative predictive value. Higher age at conisation may be a previously unrecognised risk factor for recurrence. [ABSTRACT FROM AUTHOR]

During a 4-month period we prospectively investigated the frequency of polyclonal catheter infections with Staphylococcus epidermidis. Of each catheter with pure growth of S. epidermidis, six colonies were genotypically analyzed with pulsed-field gel electrophoresis. Two out of 12 patients with catheter infection had a polyclonal infection. Both clones of each catheter had a clearly different antibiotic susceptibility. This study shows that polyclonal catheter infections are not exceptional. Further studies are needed to define the clinical consequences of polyclonal catheter infection. [ABSTRACT FROM AUTHOR]

Abstract Background The aim of this project was to study the influence of pneumococci on nasopharyngeal epithelial integrity as a function of time and pneumococcal density. Materials and methods Cell layers of an in vitro model of human nasopharyngeal epithelium were inoculated with different pneumococcal strains. The transepithelial electrical resistance (TEER), a measure of the integrity of the cell layers, and the pneumococcal concentration in the apical fluid on the epithelial cells were measured at different times after inoculation. Results Pneumococci caused a decrease in the TEER when a density of 1 × 107 CFU mL -1 was reached. The growth rate of pneumococci in our in vitro model differed between the strains tested and, for the same strain, between in vitro culture on the epithelial cells and broth culture. Differences in timing of the onset of decrease in the TEER between strains were the result of differences in growth rate on the epithelial cells. Antibiotic-induced lysis of pneumococci caused an immediate decrease in the TEER of the cell layers. Conclusion Pneumococci cause a decrease in the TEER at a density of 1 × 107 CFU mL -1 . Our hypothesis is that this decrease in the TEER is the result of quorum-induced lysis of the pneumococci. [ABSTRACT FROM AUTHOR]

Objectives: To evaluate the usefulness of detecting two genes involved in biofilm formation (icaA and aap ) and one gene involved in initial adhesion (atlE ) for discrimination between contaminant, colonizing and invasive Staphylococcus epidermidis isolates involved in catheter-related infections. Patients: The first group contained 29 isolates that were isolated from the skin of healthy volunteers (contaminant isolates). The second group contained 16 isolates recovered from catheters (>1000 CFUs on quantitative catheter culture) from asymptomatic patients without bacteremia. These isolates were considered to be colonizing isolates. The third group contained 34 isolates grown in ≥2 different blood cultures from patients with a systemic inflammatory response. These isolates were considered to be invasive isolates. Results: The prevalence of atlE did not differ between the three groups. The icaA and aap genes were significantly more prevalent in colonizing isolates (88% aap ; 88% icaA ) than in invasive isolates (68% aap , P = 0.179; 59% icaA , P = 0.055) and than in skin isolates (52% aap , P = 0.02; 38% icaA , P = 0.002). Conclusions: The high prevalence of aap and icaA in skin isolates and their higher prevalence in colonizing than in invasive isolates led to a low specificity when these genes were used to differentiate between contamination, colonization and invasive infection. We conclude that, although the prevalence of these genes differs in the three groups, their presence cannot be used for clinical decision-making. [ABSTRACT FROM AUTHOR]

An internal RNA standard proved less suitable in bacterial gene expression experiments. We therefore developed a method for simultaneous RNA and gDNA (genomic DNA) isolation from in vitro and in vivo samples containing staphylococci and combined it with quantitative PCR. The reliability of gDNA for bacterial quantification and for standardisation in gene expression experiments was evaluated. Quantitative PCR proves equivalent to quantitative culture for in vitro samples, and superior for in vivo samples. In gene expression experiments, gDNA permits a good standardisation for the initial amount of bacteria. The average interassay variability of the in vitro expression is 20.1%. The in vivo intersample variability was 73.3%. This higher variability can be attributed to the biological variation of gene expression in vivo. This method permits exact quantification of the number of bacteria and the expression of genes in staphylococci in vivo (e.g., in biofilms, evolution in time) and in vitro. [Copyright &y& Elsevier]

Objective To follow the evolution of capsular types and resistance of Streptococcus pneumoniae, isolated from deep sites. Methods More than 100 Belgian laboratories permanently collect S. pneumoniae strains isolated from puncture specimens (blood, cerebrospinal fluid, middle ear fluid, etc.) and forward them to the reference center in Leuven, in order to determine the capsular serogroups and types (SGTs) and their resistance. Results From 1994 to 1998, the 5486 S. pneumoniae strains examined belonged to 39 of the 46 currently identified SGTs. The 10 most frequent SGTs accounted for 78.9% of the isolates, and 97% of all isolates belonged to SGTs included in the 23-valent vaccine. Overall mortality of patients with pneumococcal bacteremia or meningitis was 9.7%, and 23.8% in patients over 80 years. From 1994 to 1998, resistance to penicillin (P) increased from 7.6% to 14.2%, to tetracycline (T) from 14.9% to 28.0%, and to erythromycin (E) from 22.9% to 31%. Triple resistance (PTE) increased from 0.9% in 1994 to 6.6% in 1998. Five SGTs (6, 9, 14, 19 and 23) accounted for 50% of the isolates, but for > 90% of the penicillin-resistant or erythromycin-resistant isolates. Conclusions Resistance of S. pneumoniae to penicillin, erythromycin and tetracycline is steadily increasing and is concentrated in five serotypes included in the 23-valent pneumococcal vaccine. Increasing resistance and high mortality of invasive infections are an incentive to vaccinate vulnerable groups. [ABSTRACT FROM AUTHOR]

Introduction/Background Documentation of the International Classification of Functioning, Disability and Health (ICF) components in physiotherapeutic electronic health records may improve interdisciplinary productivity and efficiency We examine the effect of targeted teaching and personalized feedback on physiotherapists’ completion of physiotherapeutic patient records and description of these components. Material and method A randomized controlled trial in a tertiary care hospital in Belgium Ten physiotherapists were included in the intervention group and five physiotherapists were in the control group The intervention (2015–2016) included targeted teaching and weekly personalized feedback for a period of four weeks regarding reporting of ICF components in electronic patient records At baseline, after targeted training, after each round of personalized feedback, and at long-term follow-up, electronic patient records for 10 random patients per physiotherapist in the intervention group were reviewed for completion of a physiotherapeutic patient record and reporting of ICF components Data for the control group were collected at baseline, after the third round of feedback, and at long term follow-up 670 and 140 patient records from 729 unique patients were reviewed for the intervention and control group, respectively. Results In the intervention group, all outcomes improved (Table 1 and Fig. 1) Although reporting declined for several components between personalized feedback and at long-term follow-up, at both occasions patient record completion and reporting of ICF components of activity, participation, and personal factors were significantly higher compared to the control group For environmental factors, the significant effect after personalized feedback disappeared at long-term follow-up Reporting of body functions and structures improved similarly across groups. Conclusion Targeted teaching and personalized feedback improved completion of physiotherapeutic patient records and reporting of ICF components Intervention strategies should factor in regular reminders to ensure continued reporting of ICF components in the long term. [ABSTRACT FROM AUTHOR]

A response by J. Maertens, K, Lagrou and J. Van Eldere to the letter to the editor about their article on preemptive antifungal therapy among neutropenic patients is presented.