Biomedical subjects
Publications and source records attributed to C Branger.
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To differentiate imported and acquired strains of methicillin-resistant Staphylococcus aureus (MRSA), a 48-hour delay from hospital admission to the first MRSA-positive culture is usually considered. To assess if taking into account this delay without any other consideration is an accurate method, we defined 3 situations for whom we considered the MRSA acquisition status as questionable. The other situations were defined as either acquired MRSA or imported MRSA. We determined the acquisition status of MRSA (acquired, imported, or questionable) isolated during a 20-month period by considering or not considering screening samples performed on admission. The ratio "imported MRSA/acquired MRSA" (I/A) was calculated according to (1) the consideration of MRSA with questionable status as imported or acquired, and (2) the consideration of screening samples or not in the calculation of the ratio. The acquisition status in our hospital was questionable in 3.6% of patients when all samples were considered and in 12,0% when only clinical samples were taken into account (p = 0,01). The ratio I/A was 4-fold higher by considering both clinical and screening cultures and questionable status as imported than by considering only clinical samples and questionable status as acquired. Using a 48-hour delay without any other consideration is probably an accurate method to differentiate acquired and imported MRSA when a selective screening programme at admission in operational. Conversely, this definition seems to be more hazardous in the absence of screening.
Identification of the meticillin-resistant Staphylococcus aureus (MRSA) reservoir by active screening followed by the implementation of contact precautions is one of the major components of MRSA control programmes. The objective of this study was to evaluate the results of a programme of selective screening in an emergency department (ED) and the appropriateness of the contact precautions implemented. This was estimated by distinguishing necessary and unnecessary days of contact precautions. This estimation was performed for all days of contact precautions and, more specifically, for days of preventive contact precautions implemented before the availability of screening results. During a three-year period, screening of MRSA carriers was performed on 0.95% (N=605) of patients visiting the emergency ward. Among the 193 (31.9%) MRSA carriers identified, 159 were hospitalized in the short-length-hospitalization area (SLHA) of the ED and/or in other wards. Among the 140 patients admitted to the SLHA, 44 were hospitalized for at least 48 h, with a mean length of hospitalization of 5.9 days. The cumulative duration of hospitalization of carriers identified by screening was 1897 days. In total, 2370 days of contact precautions (including 924 days of preventive precautions) were implemented for patients screened in the ED. Considering the whole hospital, the appropriateness of this entire programme of contact precautions for patients screened in the ED was 80.0% (52.1% for the SLHA), whereas the specific appropriateness of preventive isolation days was 48.6% (43.6% for the SLHA). This study underscores the risk of MRSA cross-transmission in the SLHA, and the usefulness of implementing a control programme of screening carriers in the ED.
INTRODUCTION: Simplification of combined antiretroviral therapy in HIV-infected patients is possible, but virological success can be compromised by the development or emergence of resistant viruses. CASE: Worsening renal functioning in a patient under successful combination antiretroviral therapy resulted led to the replacement of indinavir by abacavir. Eight weeks later, his viral load rose and he developed a mutant virus resistant to all the nucleoside analogs. DISCUSSION: Our case report illustrates the danger of streamlining combined antiretroviral therapy composed only of nucleoside analogs in patients already successfully treated with nucleoside analogs, by exposing them to the risk of the emergence of a mutant virus.
The use of DNA constructs encoding leptospiral proteins is a promising new approach for vaccination against leptospirosis. In previous work we determined that immunization with hemolysis-associated protein 1 (Hap1) (LipL32) expressed by adenovirus induced significant protection against a virulent Leptospira challenge in gerbils. To avoid the use of the adenovirus vector, we checked for clinical protection against lethal challenge by DNA vaccination. A DNA vaccine expressing Hap1 was designed to enhance the direct gene transfer of this protein into gerbils. A challenge was performed 3 weeks after the last immunization with a virulent strain of serovar canicola. Our results show that the cross-protective effect with pathogenic strains of Leptospira, shared by Hap1, could be mediated by the DNA plasmid vector. This finding should facilitate the design and development of a new generation of vaccines against bacteria, particularly Leptospira interrogans sensu lato.
Nocardia cyriacigeorgica is a recently characterized species within the genus of Nocardia. We report a brain abscess, following a primary pulmonary colonization, due to this species in a human immunodeficiency virus-infected patient. This case confirms that isolation of Nocardia in sputum is associated with a high risk of disseminated infection in immunocompromised patients.
A fluorimetric procedure for the determination of aluminium with matrix removal in drinking water is proposed. The system is based both on the solid phase extraction of aluminium on a new chelating resin (XAD-4 modified by grafting salicylic acid) and the fluorimetric detection of a complex formed between 8-hydroxyquinoline-5-sulfonic acid (HQS) and Al(III), after elution of the resin by hydrochloric acid. The sorption and elution of aluminium were studied in both competitive and noncompetitive conditions, varying pH, flow-rates, volume and concentration of reagents, as well as time contact. The optimised procedure allows determination of Al3+ at the sub-ppb level (LOD: 0.2 microg L(-1) for 1 ml of sample) within a working range of 0.2-500 microg L(-1). The analytical procedure was successfully employed for the determination of aluminium in drinking water during and after flocculation/coagulation treatment processes.
Twenty-four specific pathogen-free beagles were randomly allocated into four groups (three vaccinated groups and one control group) and inoculated at nine and 12 weeks of age with one of three commercial inactivated Leptospira vaccines: A (Vanguard 7; Pfizer Santé Animale), B (Dohyvac 7L; Fort Dodge), and C (Nobivac DHPPi + Lepto; Intervet International); the control group received Nobivac DHPPi (Intervet International). Seven weeks after the second vaccination all the dogs were challenged with Leptospira interrogans serogroup canicola. All the vaccinated dogs developed a mild serological response (microscopic agglutination titres) after the booster vaccination. A significant serological response after the challenge was observed, particularly in the controls. The challenge induced fever and clinical disorders in the control group, whereas in the vaccinated groups the clinical signs were mild. Blood cultures became positive in all control dogs, and in one of six dogs vaccinated with vaccine A and two of four dogs vaccinated with vaccine B; none of the six dogs vaccinated with vaccine C was leptospiraemic at any stage of the experiment. Urine cultures were positive in all the control dogs two weeks after the challenge. One of six dogs vaccinated with vaccine A and two of four dogs vaccinated with vaccine B shed bacteria in their urine after the challenge, but none of the dogs vaccinated with vaccine C shed bacteria in their urine at any time during the experiment.
In order to compare the microbiological characteristics of nosocomial and community-acquired episodes of bacterial peritonitis, 95 consecutive, spontaneous episodes were reviewed. Seventy of these episodes were bacteriologically documented. Fifty-three (55.8%) episodes were nosocomial and 42 (44.2%) were community acquired. A total of 78 pathogens were isolated, including 40 gram-positive cocci (34 streptococci, 6 Staphylococcus aureus), 35 gram-negative bacilli (including 23 Escherichia coli), 2 gram-positive bacilli and 1 yeast. Streptococci were found more frequently in community-acquired episodes (53.8%) than in nosocomial episodes (33.3%). Gram-negative bacilli were significantly more frequent in nosocomial episodes than in community-acquired episodes (56.4% vs. 33.3%, P<0.05). Nosocomial isolates were significantly more resistant to amoxicillin-clavulanic acid (48.7% vs. 18.4%, P<0.01) and cefotaxime (33.3% vs. 13.2%, P<0.05) than community-acquired isolates, but no difference was detected regarding resistance to ciprofloxacin. The results indicate that the empirical treatment of spontaneous bacterial peritonitis should differ for nosocomial and community-acquired cases.
New vaccine strategies are needed for the prevention of leptospirosis, a widespread human and animal disease caused by pathogenic leptospires. Our previous work determined that a protein leptospiral extract conferred cross-protection in a gerbil model of leptospirosis. The 31- to 34-kDa protein fraction of Leptospira interrogans serovar autumnalis was shown sufficient for this purpose. In the present study, N-terminal sequencing of a 32-kDa fraction and Southern blotting of genomic DNA with corresponding degenerated oligonucleotide probes identified two of its constituents: hemolysis-associated protein 1 (Hap1) and the outer membrane Leptospira protein 1 (OmpL1). Adenovirus-mediated Hap1 vaccination induces significant protection against a virulent heterologous Leptospira challenge in gerbils, whereas a similar OmpL1 construct failed to protect the animals. These data indicate that Hap1 could be a good candidate for developing a new generation of vaccines able to induce broad protection against leptospirosis disease.
Killed whole-cell preparations were used as bacterins against leptospirosis. As this type of protection is considered to be serogroup-specific, several serogroups were added to the usual vaccines, and the most pathogenic serovar was chosen for each group. Different leptospire extracts were evaluated for their protective capacity against acute lethal leptospirosis in gerbils (Meriones unguiculatus). Total extracts induced complete protection against homologous challenges and partial protection against heterologous challenges. LPS fractions protected against homologous but not heterologous challenges, whereas protein extract induced significant protection against both types of challenge. Thus, cross-protection within L. interrogans was related to the protein extract.
OBJECTIVE: To investigate interstrain relatedness of TEM-24-producing Enterobacter aerogenes clinical strains isolated between 1993 and 1998 in 10 French hospitals from nine areas by pulsed-field gel electrophoresis (PFGE) and plasmid patterns. METHODS: Fifteen TEM-24-producing strains and a set of 16 control strains having various other antibiotic resistance phenotypes were genotyped by PFGE. Plasmid DNA from TEM-24-producing strains and transconjugants was analyzed. RESULTS: Analysis of XbaI macrorestriction patterns revealed only minor variations, and showed that all 15 TEM-24-producing strains were closely related. Some isolates originating from distant areas had indistinguishable patterns. According to their clustering correlation coefficients, they were also genomically distant from the control strains. Two plasmid patterns were observed in TEM-24-producing strains, one of them in 13 of the strains. Large plasmids of 85 kb encoding TEM-24 beta-lactamase were present in all isolates and, in all except one strain, could be transferred with high frequency by conjugation. CONCLUSIONS: These results confirm that the spread of the TEM-24 extended-spectrum beta-lactamase in France was essentially due to the dissemination of a single clone.
An 80-year-old debilitated patient developed purulent pleurisy caused by a Campylobacter lari isolate. The patient underwent surgical drainage and received antibiotic therapy with amoxicillin/clavulanic acid and ofloxacin. Antibiotic susceptibility data showed that the isolate was fully sensitive to clarithromycin, tetracycline, aminoglycosides. and ciprofloxacin. Imipenem and amoxicillin plus clavulanic acid were the most active beta-lactam agents.
Seventy isolates of Klebsiella pneumoniae with extended-spectrum beta-lactamases (ESBLs) were compared. These were isolated from 51 patients on 10 separate wards in one hospital over an 18-month period between 1992 and 1994. Antibiograms were determined and the isolates were typed by pulsed-field gel electrophoresis of their DNA digestion with XbaI. The isolates were compared to three genotypically different epidemic strains responsible for previous outbreaks at the hospital between 1988 and 1991. Isolates from 84% of the present patients had closely related XbaI patterns, and most (74%) produced an ESBL with an iso-electric point (pI) of 7.0. A similar pattern was found for one of the previous epidemic strains, but it produced an ESBL with a pI of 7.8; isolates with this latter enzyme variant were found only in six of the present patients. The two other previous epidemic strains had ESBLs with a pI of 6.3 and organisms related to them were found in one and two of the present patients, respectively.
The genetic diversity among 54 human isolates and 33 animal isolates belonging to the species Streptococcus dysgalactiae (20 alpha-haemolytic Streptococcus dysgalactiae, 23 Streptococcus equisimilis, 43 group G streptococci and one group L streptococcus) was evaluated by macrorestriction analysis of chromosomal DNA with SmaI and resolution by pulsed-field gel electrophoresis. This technique revealed a high degree of intraspecies polymorphism, leading to the differentiation of 80 distinct banding patterns, and identified the presence of two major clusters, one containing isolates of human origin and the other isolates of animal origin. These results suggest than human and animal isolates of S.dysgalactiae are genetically distinct, and support the recent proposal of the subspecies S. dysgalactiae subsp. equisimilis for human isolates. The heterogeneity revealed within isolates from the same host type indicates that pulsed-field gel electrophoresis is a powerful epidemiological tool for studying S. dysgalactiae infections.
The SmaI restriction endonuclease digestion patterns of chromosomal DNAs from 99 pyogenic streptococci belonging to Lancefield group A (41 Streptococcus pyogenes), group C (seven S.dysgalactiae, 11 "S. equisimilis", three S. equi, eight S. zooepidemicus) and group G (25 human group G Streptococcus, four S. canis) were analyzed by pulsed-field gel electrophoresis (PFGE), and the results were compared with those previously obtained by multilocus enzyme electrophoresis (MLEE) and random amplified polymorphic DNA analysis (RAPD). PFGE revealed 93 distinct types among the 99 strains, and no patterns were common to strains of different species. The discriminatory power of PFGE was greater than that of MLEE and RAPD for groups A and G streptococci. The polymorphism among group C streptococci was similar with the three techniques. PFGE is, therefore, the most efficacious method for epidemiological typing of pyogenic streptococci.
Thirty-seven isolates of extended-spectrum beta-lactamase-producing (ESBL) Klebsiella pneumoniae implicated in five nosocomial outbreaks (I-V) on three distinct wards of our hospital were compared using capsular typing, biotyping, antibiotyping, enzyme electrophoresis typing and DNA macrorestriction analysis with Xba I resolved by pulsed-field gel electrophoresis. The isolates from each outbreak had common phenotypic and genotypic characteristics indicating that they were related epidemiologically. Isolates from outbreaks I (four patients) and V (13 patients), although they occurred in two different wards (neurology and surgery) and three years apart, produced the same ESBL with a pI of 7.8 (SHV-4) and were of serotype K25. The Xba I patterns were closely related. The isolates of outbreaks II (seven patients), III (four patients) and IV (seven patients), which occurred in a single surgical intensive care unit, produced an ESBL with a pI of 6.3 (TEM-3). Isolates from outbreaks III and IV, which occurred six months apart, were of serotype K68 and had similar Xba I patterns suggesting that the two outbreaks were due to a single strain which persisted endemically in the ward. The isolates from outbreak II were of serotype K62, and had distinct characteristics from the two later outbreaks. The Xba I patterns of the isolates from outbreaks "I and V', II and "III and IV' had Dice similarity coefficients under 40% showing that the three groups were genetically distant. DNA macrorestriction analysis was a useful complement to phenotypic methods for identifying K. pneumoniae strains responsible for outbreaks harbouring a common ESBL.
Eight out of 20 (40%) patients with haematological malignancies hospitalized in the same unit of our hospital from 24 January to 24 April 1995, suffered from diarrhoea due to Clostridium difficile. The C. difficile isolates were characterized by serotyping and by arbitrary primed polymerase chain reaction (AP-PCR) using three different 10-mer oligonucleotides. It was found by serotyping that five patients had non-typeable isolates and three had serogroup H isolates. The AP-PCR typed all the isolates and yielded various patterns suggesting that there had been no cross-transmission between the patients. Control faecal sample cultures showed that two patients were still carrying the same isolates after specific treatment with vancomycin or metronidazole, and that one patient had acquired an isolate with a new AP-PCR type. AP-PCR was found to be a rapid, effective discriminative method for the immediate epidemiological tracking of hospital-acquired infections due to C difficile.