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Biomedical subjects

J Van Eldere

Publications and source records attributed to J Van Eldere.

At least 19 recordsLinked to original sources

Macrolide resistance in Belgian Streptococcus pneumoniae.

The genetic basis of macrolide resistance was characterized in 59 Streptococcus pneumoniae isolates. All isolates were collected in 1995 and 1997 and were from invasive infections. The majority of the isolates (54 of 59 isolates) were erythromycin and clindamycin resistant (MLS(B)-phenotype) and carried the ermAM gene. Five isolates were erythromycin resistant but clindamycin susceptible (M-phenotype). Using PCR the mefE gene was detected in these five isolates. Contrary to the situation found in Canada and the USA, mefE-mediated erythromycin resistance in S. pneumoniae is uncommon in Belgium.

Anti-Bacterial Agents

Increase in penicillin resistance rates in Belgium due to clonal spread of a penicillin-resistant 23F Streptococcus pneumoniae strain.

In 1994 a sudden increase in penicillin resistance was observed in Belgium among invasive pneumococci. To determine whether this increase was due to clonal spread of a resistant strain or to de novo acquisition of penicillin resistance, pneumococci of capsular types 23F, 19, 14, 9, and 6 isolated in 1993 and 1994, were analyzed by capsular serotyping and DNA macrorestriction analysis, resolved by pulsed-field gel electrophoresis. Furthermore, pneumococcal isolates from northern France, a region with a high prevalence of penicillin resistance, and from southern Belgium, a region with a low but increasing prevalence of penicillin resistance, were analyzed. The rate of resistance of invasive pneumococci to penicillin increased from 2.3% in 1993 to 7.6% in 1994. Pneumococcal serotype 23F represented 26.7% of the penicillin-resistant isolates in 1993 and 40.4% in 1994, while the prevalence of serotype 23F decreased from 10.9% in 1993 to 8.8% in 1994. In 1994 up to 35.8% of serotype 23F isolates were penicillin resistant. The Belgian penicillin-resistant 23F isolates from 1994 were genetically closely related to the French 23F penicillin-resistant isolates and, as clones were clearly distinct from the other serotypes as well as from the penicillin-susceptible 23F isolates. These data demonstrate the important contribution of the clonal spread of a penicillin-resistant pneumococcal strain in the overall increase of penicillin resistance in our country.

Belgium

Staphylococcus aureus adherence to nasal epithelial cells in a physiological in vitro model.

Nasal carriage of Staphylococcus aureus represents a risk factor for subsequent invasive infections and interpatient transmission of strains. No physiological in vitro model of nasal epithelial cells is available to study both patient- and bacteria-related characteristics and their interaction, leading to adherence and colonization. Starting with tissues from human nasal polyps, a confluent, squamous, nonkeratinized epithelium in collagen-coated 96-well microtiter plates was obtained after 14 d. This in vitro cell-layer was characterized histologically, ultrastructurally, and immunohistochemically and showed features that were indistinguishable from those observed in the squamous nonkeratinized epithelium found in the posterior part of the vestibulum nasi. Adherence experiments were performed with four different 3H-thymidine-labeled Staphylococcus aureus strains. The effect of bacterial inoculum size, temperature of incubation, and incubation medium were studied. The adherence results were found to be reproducible, reliable and sensitive, allowing detection of small quantitative differences in adherence between the Staphylococcus aureus strains. There was no significant difference in adherence at 23 degrees C and 37 degrees C, nor between the incubation medium M199 and phosphate-buffered saline. Plastic adherence could be reduced and standardized with use of siliconized tips and a constant bacterial inoculum volume of 100 microl/well. This physiological and reliable in vitro cell-culture model offers a unique opportunity to study Staphylococcus aureus adherence to squamous, nonkeratinized nasal epithelial cells and both patient and bacterial characteristics involved in this interaction.

Bacterial Adhesion

Foreign body infection: a new rat model for prophylaxis and treatment.

A subcutaneous catheter model in the rat was developed that allowed the study of prevention and treatment strategies for foreign body infection. In contrast to earlier models, the foreign body was inoculated with a low inoculum of Staphylococcus epidermidis just before implantation, thus mimicking intraoperative contamination with skin flora. Reproducible infection of all catheters followed if no prophylaxis was given. However, foreign body infection could be prevented or treated with antibiotics such as teicoplanin, which was marginally effective, and rifampicin, which proved very effective.

Animals

Formation of hyodeoxycholic acid from muricholic acid and hyocholic acid by an unidentified gram-positive rod termed HDCA-1 isolated from rat intestinal microflora.

From the rat intestinal microflora we isolated a gram-positive rod, termed HDCA-1, that is a member of a not previously described genomic species and that is able to transform the 3alpha,6beta, 7beta-trihydroxy bile acid beta-muricholic acid into hyodeoxycholic acid (3alpha,6alpha-dihydroxy acid) by dehydroxylation of the 7beta-hydroxy group and epimerization of the 6beta-hydroxy group into a 6alpha-hydroxy group. Other bile acids that were also transformed into hyodeoxycholic acid were hyocholic acid (3alpha, 6alpha,7alpha-trihydroxy acid), alpha-muricholic acid (3alpha,6beta, 7alpha-trihydroxy acid), and omega-muricholic acid (3alpha,6alpha, 7beta-trihydroxy acid). The strain HDCA-1 could not be grown unless a nonconjugated 7-hydroxylated bile acid and an unidentified growth factor produced by a Ruminococcus productus strain that was also isolated from the intestinal microflora were added to the culture medium. Germfree rats selectively associated with the strain HDCA-1 plus a bile acid-deconjugating strain and the growth factor-producing R. productus strain converted beta-muricholic acid almost completely into hyodeoxycholic acid.

Animals

Autopsy-controlled prospective evaluation of serial screening for circulating galactomannan by a sandwich enzyme-linked immunosorbent assay for hematological patients at risk for invasive Aspergillosis.

Efforts to improve the diagnosis of invasive aspergillosis (IA) have been directed towards the detection of fungal antigens, including galactomannan (GM). However, previous evaluations of GM detection have been hampered by a lack of proven cases of IA and by a nonserial study design. This prospective study assessed the diagnostic value of serial screening for circulating GM by using a recently developed sandwich enzyme-linked immunosorbent assay (ELISA) for prolonged-neutropenic and/or steroid-treated patients with hematological disorders. Serum GM levels were monitored twice weekly for 186 consecutive patients at increased risk for IA. The patients were stratified according to the likelihood of IA (proven, probable, possible, and no evidence of IA) by using stringent criteria. Proven IA was defined by characteristic histopathological findings together with a positive culture for Aspergillus species. Autopsy and culture from autopsy specimens was used to verify both positive and negative test results. A total of 2,172 serum samples were tested from 243 episodes (mean, 9 samples/episode). Based on the analysis of 71 patients with confirmed disease status (culture and histology), the sensitivity and specificity of serial GM monitoring were 92.6 and 95.4%, respectively. The positive predictive value was almost 93%, the negative predictive value was 95%, and the efficacy was 94%. False-positive reactions occurred at a rate of nearly 8%, although this figure might have been overestimated. Less than 1% of all tested sera were considered inconclusive. In more than half of the cases, antigenemia was detected before clinical suspicion of IA (median, 6 days before). Serial determination of serum GM by the sandwich ELISA technique is a sensitive tool for the diagnosis of IA in hematological patients at risk. This approach may substantially influence clinical management with regard to preemptive and empirical antifungal therapy.

Adolescent

Comparison of fluorescence microscopy and culture assays to quantitate adhesion of Porphyromonas gingivalis to mono- and multi-layered pocket epithelium cultures.

BACKGROUND: The present study compared 2 different methods (direct versus indirect evaluation) for the quantification of the adhesion of Porphyromonas gingivalis strains to in vitro cultured mono-layers of pocket epithelium. METHODS: The indirect culture viability assay (calculation of colony forming units) was compared to a direct microscopic evaluation using a novel fluorescent stain. The fluorescent kit was found to stain both bacteria and epithelial cells and enabled a differentiation between dead and living cells. RESULTS: Comparing the visual to the culture data, a high and significant correlation was found (Pearson's correlation = 0.75; P <0.001). The adhesion capacity was in general higher for dead epithelial cells than for living cells (P <0.01). Although comparable numbers of bacteria of 2 P. gingivalis strains (Pg 4 and Pg 5) were applied, Pg 4 showed a significantly lower adhesion capacity. This intra-strain variability was observed by the culture assay (2.3 x 10(6) versus 7.8 x 10(6)+/-2.7 x 10(6); P <0.01) and by the direct microscopy (P <0.01) for both live and dead epithelial cells. A second goal was to see whether there was a difference in the amount of bacterial adherence to mono- and multi-layers of in vitro cultured epithelium. No significant differences were found for the 5 examined P. gingivalis strains. However, interstrain differences in adhesion capacity were evident for both tissues. CONCLUSIONS: This study highlights the reproducibility of a direct microscopic evaluation of bacterial adhesion to in vitro cultured epithelial cells, and suggests both intrastrain (P. gingivalis) and inter-cell (live versus dead) variation in adhesion capacity. Studies are needed to determine the extent to which P. gingivalis strain variation is reflected in variation of other strains in humans.

Bacterial Adhesion

One stage full- versus partial-mouth disinfection in the treatment of chronic adult or generalized early-onset periodontitis. II. Long-term impact on microbial load.

BACKGROUND: Recent studies showed the clinical benefits of a one stage full-mouth disinfection, when compared to the worldwide standard treatment strategy of consecutive root planings per quadrant without proper disinfection of the remaining intraoral niches. The purpose of this study was to investigate the microbiological benefits of such a one stage full-mouth disinfection with special attention to all intraoral niches for periodontopathogens and to evaluate the perception by the patients of the new treatment strategy. METHODS: Sixteen patients with early-onset periodontitis and 24 patients with severe adult periodontitis were randomly assigned to test and control groups. The control group was scaled and root planed, per quadrant, at 2-week intervals and given oral hygiene instructions. The test group received the one stage full-mouth disinfection treatment. At baseline and after 1, 2, 4, and 8 months, microbiological samples were taken from all niches (tongue, mucosa, saliva, and pooled samples from single- and multi-rooted teeth). The samples were cultured on selective and non-selective media. Patient perception of the treatment was evaluated using a questionnaire. RESULTS: In comparison to the standard therapy, the one stage full-mouth disinfection resulted in significant additional microbial improvements. The test group showed larger reductions in the proportions of spirochetes and motile organisms in the subgingival flora, and more significant reductions in the density of key pathogens, with even the eradication of P. gingivalis. The beneficial effects in the other niches were primarily restricted to the number of colony-forming units/ml of black-pigmented bacteria, especially on the mucosa and in the saliva and to a lesser extent on the tongue. Both treatments were well tolerated by the patients and the overall severity rating for both therapies was comparable, although 4 quadrants were treated within 24 hours in the test group versus only 1 in the control group. The full-mouth disinfection approach resulted more frequently in a slight increase of body temperature, especially after the second day. CONCLUSIONS: These findings support the benefit of a one stage full-mouth disinfection in the treatment of patients with either chronic adult or early-onset periodontitis.

Adult

In vitro volatile sulfur compound production of oral bacteria in different culture media.

OBJECTIVE: The purpose of this study was to detect the relative contribution of Porphyromonas gingivalis, Fusobacterium nucleatum, and Prevotella intermedia in the production of oral malodor. METHOD AND MATERIALS: The volatile sulfur compounds produced by these bacteria in vitro were measured semiquantitatively by a portable sulfide monitor. RESULTS: Samples from the tongue, tonsils, and pharynx showed a significantly higher production (550 ppb) of volatile sulfur compounds during the first 6 hours after anaerobic incubation in broths (brain-heart infusion, Columbia, and Trypticase Soy) than after incubation in agar media (300 ppb) (P < 0.001). After 24 hours, values in broths and agars leveled off at 350 ppb (P = 0.3) and remained constant during the next 6 days. Measurement of separate pure cultures showed that maximal volatile sulfur compound production was reached 6 hours after incubation (450 ppb for the 3 bacteria). Higher volatile sulfur compound values were measured in brain-heart infusion. When measurements of mixed cultures of the 3 pathogens were performed every 15 minutes, the maximal value was reached after only 30 minutes of incubation (nearly 500 ppb). CONCLUSION: The in vitro volatile sulfur compound production of oral samples is preferably measured in broths. Maximal sulfur production from mixed cultures is reached after 30 minutes of incubation. Samples should always be inoculated at the same dilution.

Culture Media

Toxic shock syndrome without rash in a young child: link with syndrome of hemorrhagic shock and encephalopathy?

A diffuse macular erythroderma and subsequent desquamation after 1 to 2 weeks are two of the five major diagnostic criteria of toxic shock syndrome (TSS). We present the case of a 15-month-old girl with TSS, but without erythroderma or desquamation. She was admitted with high fever, shock, and multiorgan involvement. Minimal or no cutaneous signs were present. Initially the diagnosis of the syndrome of hemorrhagic shock and encephalopathy was made. After 7 days, a TSS toxin 1-producing strain of Staphylococcus aureus was cultured from an inguinal lymph node, where inflammation had already been noticed on admission. Moreover, the girl had no antibodies against this toxin. The serum cytokine profile during the acute phase of her illness showed high levels of tumor necrosis factor-alpha, interleukin-6 and interferon-gamma, as is seen during activation of the immune system by TSS toxin 1. Other possible causes for the patient's illness were excluded. We conclude that the patient had TSS without rash. Without the evidence implicating a TSS toxin 1-producing strain of S. aureus as the cause of her disease, a diagnosis of syndrome of hemorrhagic shock and encephalopathy would have been made. It is possible that some cases of syndrome of hemorrhagic shock and encephalopathy represent a variant of TSS in small children.

Bacterial Toxins

Origin and transmission of methicillin-resistant Staphylococcus aureus in an endemic situation: differences between geriatric and intensive-care patients.

Imported vs. hospital-acquisition of MRSA was assessed in > 6000 patients at a large tertiary care teaching hospital. About five percent (5.1%) of patients carried MRSA on admission, mostly without clinical symptoms; the highest percentage (11.6%) being in geriatric patients. Hospital-acquisition of MRSA occurred in 1.7% of patients and was particularly high in intensive-care units (5.2%). Phenotype and genotype analysis of 158 MRSA strains isolated from 61 patients revealed a cluster of closely related strains in the hospital-acquired MRSA infections and the close relationship of this cluster to the regional epidemic MRSA strain. The MRSA strains imported by geriatric patients were genetically different, did not spread between geriatric patients and were only a minor source of nosocomial infection.

Age Distribution

Polyclonal staphylococcus endocarditis.

Coagulase-negative staphylococcus (CNS) is the most frequent cause of nosocomial bacteremia and prosthetic valve endocarditis. CNS bacteremia can be polyclonal. No data exist on the clonality of CNS causing endocarditis. We present a case of CNS aortic homograft endocarditis in which at least five different genotypes of CNS were identified in initial blood-culture isolates by genomic macrorestriction enzyme analysis and pulsed field gel electrophoresis. Since the polyclonality was accompanied by differences in antibiotic susceptibility, this observation may have important consequences for the treatment of CNS endocarditis. Because of the parallels in the pathogenesis of CNS prosthetic valve endocarditis and CNS infections of a variety of other prosthetic devices, it might also have consequences for CNS prosthetic device infections in general. We suggest that antibiotic susceptibility testing of just one blood-culture isolate may be insufficient.

Coagulase

The possibilities and limitations of nucleic acid amplification technology in diagnostic microbiology.

Nucleic acid amplification technology is examined from the critical viewpoint of a clinical microbiologist working in a routine diagnostic bacteriology laboratory. Widely recognised limitations of amplification technology include those of false-positive and false-negative results, the difficulty of obtaining quantitative results, the problem of using this technology for susceptibility testing, and the difficulty of detecting routinely the wide range of possible pathogens contained in a clinical sample. On the positive side, amplification technology brings welcome new possibilities for rapid detection of specific pathogens in a sample, including viruses, slowly growing bacteria, fastidious or uncultivable bacteria, fungi and protozoa. Other possible applications include screening normally sterile clinical samples for non-specific bacterial contamination and the use of amplification-based DNA fingerprinting methods for identification and typing of microorganisms. Nevertheless, it is predicted that-in contrast to research and reference facilities-routine bacteriology laboratories will continue to rely on culture as the preferred 'amplification method' for most diagnostic applications.

Bacteria

Clonal analysis and identification of epidemic strains of methicillin-resistant Staphylococcus aureus by antibiotyping and determination of protein A gene and coagulase gene polymorphisms.

Forty-three methicillin-resistant Staphylococcus aureus (MRSA) isolates with known genetic and epidemiological relatedness and different degrees of transmission were analyzed by antibiotyping, protein A gene polymorphism analysis, and coagulase gene polymorphism analysis. The three typing systems were evaluated for their performance and convenience to define clones and to discriminate between epidemic MRSA (EMRSA) and sporadic MRSA (SMRSA). Antibiotyping and AluI restriction fragment length polymorphism analysis of the coagulase gene were able to define clones in the same way as DNA macrorestriction analysis (SmaI). However, both techniques presented disadvantages, making neither of them useful as a single typing method. Protein A gene polymorphism analysis appeared to be of no value for clonal analysis. None of the three typing methods was able to differentiate between EMRSA and SMRSA.

Bacterial Typing Techniques

The influence of abutment surface roughness on plaque accumulation and peri-implant mucositis.

Bacterial adhesion to intra-oral, hard surfaces is firmly influenced by the surface roughness to these structures. Previous studies showed a remarkable higher subgingival bacterial load on rough surfaces when compared to smooth sites. More recently, the additional effect of a further smoothening of intra-oral hard surfaces on clinical and microbiological parameters was examined in a short-term experiment. The results indicated that a reduction in surface roughness below R(a) = 0.2 microns, the so-called "thresholds R(a)", had no further effect on the quantitative/qualitative microbiological adhesion or colonisation, neither supra- nor subgingivally. This study aims to examine the long-term effects of smoothening intra-oral hard transgingival surfaces. In 6 patients expecting an overdenture in the lower jaw, supported by endosseus titanium implants, 2 different abutments (transmucosal part of the implant): a standard machined titanium (R(a) = 0.2 microns) and one highly polished and made of a ceramic material (R(a) = 0.06 microns) were randomly installed. After 3 months of intra-oral exposure, supra- and subgingival plaque samples from both abutments were compared with each other by means of differential phase-contrast microscopy (DPCM). Clinical periodontal parameters (probing depth, gingival recession, bleeding upon probing and Periotest-value) were recorded around each abutment. After 12 months, the supra- and subgingival samples were additionally cultured in aerobic, CO2-enriched and anaerobic conditions. The same clinical parameters as at the 3-month interval were recorded after 12 months. At 3 months, spirochetes and motile organisms were only detected subgingivally around the titanium abutments. After 12 months, however, both abutment-types harboured equal proportions of spirochetes and motile organisms, both supra- and subgingivally. The microbial culturing (month 12) failed to detect large inter-abutment differences. The differences in number of colony- forming units (aerobic and anaerobic) were within one division of a logarithmic scale. The aerobic culture data showed a higher proportion of Gram-negative organisms in the subgingival flora of the rougher abutments. From the group of potentially "pathogenic" bacteria, only Prevotella intermedia and Fusobacterium nucleatum were detected for anaerobic culturing and again the inter-abutment differences were negligible. Clinically, the smoothest abutment showed a slightly higher increase in probing depth between months 3 and 12, and more bleeding on probing. The present results confirm the findings of our previous short-term study, indicating that a further reduction of the surface roughness, below a certain "threshold R(a)" (0.2 microns), has no major impact on the supra- and subgingival microbial composition.

Adult

Full- versus partial-mouth disinfection in the treatment of periodontal infections. A pilot study: long-term microbiological observations.

A standard periodontal treatment consists of 4 to 6 scalings and rootplanings at a 1- to 2-week interval, which allows reinfection of a previously disinfected area before completion of the treatment. The present pilot study aims to examine the microbiological long-term effects of a full-mouth disinfection. 10 patients with advanced chronic periodontitis were randomly allocated to a test and control group. The patients from the control group received scaling and rootplaning and oral hygiene instructions at a 2-week interval. The full-mouth disinfection (test group) consisted of a full-mouth scaling and rootplaning in 2 visits within 24 h in combination with: tongue brushing with 1% chlorhexidine gel for 1 min, mouth rinsing with 0.2% chlorhexidine solution for 2 min and subgingival irrigation of all pockets (3x in 10 min) with 1% chlorhexidine gel. The patients of the test group were instructed to rinse 2x daily with 0.2% chlorhexidine. Plaque samples were taken at baseline and after 1, 2, 4 and 8 months. Differential phase-contrast microscopy showed a significantly larger reduction of spirochetes and motile organisms in the test group up to month 2 for the single-rooted and up to month 8 for the multi-rooted teeth. Furthermore, the culture data supported the effectiveness of the new treatment strategy. In both groups, the number of anaerobic CFU decreased 1 log around single- and 0.5 log around multi-rooted teeth. The number of anaerobic CFU remained low in the test group, in contrast to the control group. At 1 month, the test group harboured a significantly (p<0.01) lower proportion of pathogenic organisms, but this difference disappeared with time. Moreover, the test sites showed a significantly higher (p<0.02) increase in the proportion of beneficial micro-organisms up to 4 months. These findings suggest that a full-mouth disinfection leads to a significant microbiological improvement up to 2 months, which could be consolidated, although not significant, for the next 6 months.

Adult

Tauroconjugation of cholic acid stimulates 7 alpha-dehydroxylation by fecal bacteria.

We examined the effect of the type of cholic acid conjugation (taurine-conjugated, glycine-conjugated, or unconjugated cholic acid) on cholic acid 7 alpha-dehydroxylation by intestinal flora. Cholic acid 7 alpha-dehydroxylation in fecal cultures, in cultures of a defined limited flora consisting of a mixture of seven bacterial species isolated from the intestinal tract, and in a binary culture of a 7 alpha-dehydroxylating Clostridium species plus a cholic acid-deconjugating Bacteroides species was studied. We found that tauroconjugation of cholic acid significantly (P < 0.05) increased bacterial 7 alpha-dehydroxylation of cholic acid into deoxycholic acid from 34 to 55% in fecal cultures, from 45 to 60% in defined limited fecal cultures, and from 75 to 100% in binary cultures. Equimolar concentrations of free taurine did not stimulate 7 alpha-dehydroxylation in fecal cultures or in the defined limited flora, but free taurine did stimulate 7 alpha-dehydroxylation in the binary culture. In the binary culture of Clostridium species strain 9/1 plus Bacteroides species strain R1, the minimal flora capable of increased 7 alpha-dehydroxylation of taurocholic acid, strain R1 deconjugated taurine and rapidly reduced it to H2S. Bacteroides species strain R1 did not grow unless taurine or another appropriate reducible sulfur source was present. Clostridium species strain 9/1 did not grow or 7 alpha-dehydroxylate unless H2S or another source of reduced sulfur was present. We conclude that the increased 7 alpha-dehydroxylation of tauroconjugated cholic acid depends on the reduction of taurine to H2S, which is a necessary growth factor for the 7 alpha-dehydroxylating bacteria.

Bacteria

Prospective clinical evaluation of Amplicor Mycobacterium tuberculosis PCR test as a screening method in a low-prevalence population.

Of 656 respiratory samples analyzed for Mycobacterium tuberculosis by microscopy, culture, and the Amplicor PCR method, 25 were positive by culture, 12 were positive by microscopy, and 17 were positive by the Amplicor PCR method; 16 samples were Amplicor PCR positive and culture negative. No patient except one with culture-negative, Amplicor PCR-positive samples had clinical indications of tuberculosis. The sensitivity and specificity of the Amplicor PCR compared with those of culture were 68 and 97.4%, respectively. For culture-positive, smear-negative samples, the sensitivity of the Amplicor PCR was 46%.

Adult