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Biomedical subjects

J Fleurette

Publications and source records attributed to J Fleurette.

At least 19 recordsLinked to original sources

Cadmium-resistance plasmid in Staphylococcus lugdunensis.

Plasmids that confer resistance to cadmium (MIC > or = 125 microM), were found in 18 out of 30 independent Staphylococcus lugdunensis strains from clinical specimens. Variants that were cured of their plasmid were cadmium sensitive. Restriction endonuclease sites of a 3.2-kb cadmium-resistance plasmid of S. lugdunensis, designated pLUG10, were similar to those of pOX6, a S. aureus cadmium-resistance plasmid containing the cadB gene. Southern-blot hybridisation was performed with a probe intragenic to cadB. Hybridisation was found with the cadB probe in the cadmium-resistant S. lugdunensis isolates to the 2.9-, 3.2- and 3.7-kb plasmids. These findings suggest that cadmium-resistance in some S. lugdunensis strains is due to a gene sharing homology with the cadB gene of S. aureus.

Cadmium

Fosfomycin enhances the expression of penicillin-binding protein 2 in methicillin-sensitive and methicillin-resistant Staphylococcus aureus strains.

The effects of fosfomycin on penicillin-binding proteins (PBPs) were studied on the methicillin-resistant Staphylococcus aureus strain CIP (Collection de l'Institut Pasteur, Paris, France) 65-25 and on a methicillin-susceptible S. aureus strain CIP 65-6. The combinations of fosfomycin and oxacillin were synergistic, additive or antagonistic, depending on antibiotic concentrations. Fosfomycin induced modifications of the PBP profile of the two strains studied. In particular, it increased the expression of PBP2. This suggested that this protein is inducible; the only PBP not affected by fosfomycin was PBP3.

Bacterial Proteins

Comparison of coagulase-negative staphylococci by pulsed-field electrophoresis.

Five pathogenic strains each of Staphylococcus epidermidis, S. haemolyticus, S. lugdunensis and S. schleiferi were analysed by conventional electrophoresis and field inversion gel electrophoresis. For these coagulase-negative staphylococci, the restriction endonuclease SmaI emerged as the most suitable enzyme for pulsed-field electrophoresis by providing an adequate number of clearly separated DNA fragments. Field inversion gel electrophoresis confirmed the differences among strains already discriminated by conventional electrophoresis, and furthermore, differentiated strains which had previously appeared identical. Among the species that were studied, S. epidermidis showed great genomic diversity with a few common bands. On the contrary, S. haemolyticus, S. lugdunensis and S. schleiferi showed less diversity. Although these minor variations may be epidemiologically significant, this question has to be investigated on a larger number of strains.

Coagulase

[Acute pharyngitis: value of the bacteriological diagnosis].

The aetiological diagnosis of acute pharyngitis relies upon laboratory examinations, but these are difficult to obtain in routine general practice. While taking throat swabs is mandatory in some clinical situations (membranous or ulcerous pharyngitis, special investigations in some patients) where it provides essential diagnostic data, it is in fact seldom performed in patients with common acute pharyngitis. Penicillin is systematically administered for fear of the severe sequelae of group A streptococci pharyngitis, but this type of infection occurs in only 10 to 20% of the cases so that on statistical grounds routine antibiotic therapy is not fully justified. For this reason, rapid diagnostic tests without cultures and feasible at the patient's bedside have been devised. These tests have excellent specificity, but their sensitivity is not so good. In addition, their use by general practitioners raises a problem of medical responsibility, and they are not reimbursed by the french Sécurité sociale. The future of these fast methods will depend on the solution of these two problems.

Acute Disease

Identification of coagulase negative staphylococci isolated from lambs as Staphylococcus caseolyticus.

A group of 17 strains of coagulase negative staphylococci isolated from slaughtered lambs, and which could not be identified with the conventional methods, exhibited high levels of DNA homology (92%) with the S. caseolyticus reference strain. The isolates described in this study provide a more extensive comprehension of S. caseolyticus. The original description of this species was based on only two strains isolated from milk. To our knowledge, S. caseolyticus had never been previously associated with animal microflora.

Abscess

An analysis of the microbial flora of premature neonates.

An analysis of the microbial flora of 10 premature neonates hospitalized in a neonatal intensive care unit (NICU) was made. The babies had received neither antibiotics nor antiseptics and nine out of 10 were born by caesarean section. Samples were collected on the fourth or fifth day of life from 18 skin or mucosal sites. Detailed bacterial counts were obtained by plating out suitable dilutions of the samples on to selective media. Representative samples of each colony type were then subcultured and identified, using standard laboratory methods. Two hundred and fifty-six isolates of staphylococci were obtained and their susceptibility to 23 antibiotics tested. Only 11% of the samples were sterile. Coagulase-negative staphylococci (CNS) were the commonest species isolated and were predominant in every site studied. They were found in 79% of the samples and represented almost 81% of the neonates' flora. Eight species and biotypes of CNS were identified. In decreasing order of frequency, they comprised S. epidermidis (biotypes 1 and 2), S. hominis (biotype 1), S. warneri, S. haemolyticus, S. capitis, S. cohnii and S. hominis (biotype 2). CNS distribution appeared to be highly heterogeneous with no significant specificity of any species for a particular body site. The main quantitative and qualitative variations seemed to relate to the method of delivery, and the intensity and nature of exposure of the neonate to its local environment. A high level of antibiotic resistance was found among the CNS isolates (especially S. epidermidis and S. haemolyticus): penicillin G (96%), oxacillin (31%), erythromycin (52%) and gentamicin (28%). Moreover, multiresistant strains were numerous, supporting the nosocomial origin of CNS.

Bacterial Typing Techniques

A collaborative study of the in-vitro sensitivity to RP 59500 of bacteria isolated in seven hospitals in France.

The in-vitro activity of RP 59500 was determined against 1051 recent clinical bacterial isolates. The susceptibility to RP 59500 was determined with an agar dilution technique for all the isolates, while MICs and MBCs were determined for 82 selected strains in broth. Isolates of both Staphylococcus aureus and coagulase-negative staphylococci appeared to be potentially susceptible to RP 59500, independent of susceptibility to methicillin or MLS resistance. (S. aureus: methicillin-sensitive, MIC90, 1.0 mg/L; methicillin-resistant, MIC90 1.0 mg/L; coagulase-negative staphylococci: methicillin-sensitive, MIC90 0.5 mg/L). Lancefield group A, B, C and G streptococci (MIC50 0.5 and MIC90 1.0 mg/L) and Streptococcus pneumoniae (MIC50 0.5 and MIC90 1.0 mg/L) appeared to be susceptible to RP 59500. Some Streptococcus spp. and enterococci as well as Listeria monocytogenes were inhibited by a higher concentration of RP 59500 (enterococci: MIC90 4 mg/L, range 0.125-16 mg/L). Comparatively low MICs were seen when Legionella spp., Neisseria gonorrhoeae and Gardnerella vaginalis were tested. Broth dilution MIC/MBC determinations showed no evidence of tolerance, as MIC values were within two dilutions of MBC values. RP 59500 might be a useful compound in the treatment of infections caused by a range of Gram-negative and Gram-positive bacteria, including those resistant to methicillin and/or macrolides.

Coagulase

Synthetic DNA probes for detection of genes for enterotoxins A, B, C, D, E and for TSST-1 in staphylococcal strains.

A dot blot hybridization technique with oligonucleotide probes was developed for the specific detection of the TSST-1 gene and the staphylococcal enterotoxin (SE) genes A, B, C, D and E. For each toxin gene a probe sequence was chosen from the previously determined sequence. A total of 145 staphylococcal strains (133 Staphylococcus aureus and 12 coagulase-negative staphylococci (CNS) were studied by this genotypic method and by two phenotypic assays (gel immunodiffusion and ELISA). An excellent correlation (96%) was observed between the genotypic and phenotypic assays. DNA from two CNS strains hybridized with a probe without detection of the corresponding toxin (SEB for one strain and SEC for the other strain). One Staph. aureus strain was shown to be an SEC producer, but was not detected by the corresponding probe. Gene probe and immunological assays seem to be complementary methods for studies of staphylococcal strains producing (or potentially producing) TSST-1 or enterotoxins.

Bacterial Toxins

Description and evaluation of the semiautomated 4-hour rapid ID 32 Strep method for identification of streptococci and members of related genera.

The rapid ID 32 Strep system (bioMérieux, La Balme les Grottes, France) is a new system which allows the identification in 4 h of most streptococci and members of related genera encountered in medical and veterinary bacteriology. Four hundred thirty-three isolates first identified by conventional methods and belonging to the genera Streptococcus, Lactococcus, Enterococcus, Aerococcus, Gemella, Leuconostoc, Erysipelothrix, Gardnerella, and Listeria were tested. Overall, rapid ID 32 Strep correctly identified 413 (95.3%) of the strains, with 109 (25.1%) requiring extra tests for complete identification. Sixteen strains (3.7%) were not identified, and 4 (1.0%) were misidentified. The rapid ID 32 Strep system is a suitable alternative for rapid identification of members of the genus Streptococcus and of related genera.

Bacterial Typing Techniques

Detection of Legionella spp. in bronchoalveolar lavage fluids by DNA amplification.

By using Taq polymerase, DNA amplification of a specific fragment of the macrophage infectivity potentiator (mip) gene from Legionella pneumophila was used to detect Legionella spp. in bronchoalveolar lavage (BAL) fluid specimens. We were able to detect DNAs from all 30 L. pneumophila strains tested (serogroups 1 to 14), L. micdadei, and L. bozemanii serogroup 1. DNA from bacteria of other species tested and DNA from human leukocytes were not amplified by this procedure. After optimization of the conditions for DNA extraction from BAL fluid, a 2-ml sample of BAL fluid seeded with 25 CFU/ml tested positive after DNA amplification. A total of 68 frozen BAL fluid specimens sent to the laboratory because of suspected legionellosis were tested in a retrospective study. The eight culture-positive samples were all positive after specific DNA amplification. Among 60 culture-negative samples, 7 were positive after amplification. Of these seven samples, four were from patients who had presented a typical clinical history of legionellosis; the samples had antibody titer increases of 2 dilutions. For the three remaining samples, serological diagnosis of legionellosis in the patients from whom the samples were obtained could not be documented, and although the causative agent of these pulmonary infections was not determined, the clinical features of the patients were in accordance with legionellosis.

Bacteriological Techniques

Study of cross-reaction between Coxiella burnetii and Legionella pneumophila using indirect immunofluorescence assay and immunoblotting.

Patients with Q fever and legionellosis may present identical clinical symptom. Differentiation of these diseases is made by serology, mainly the indirect immunofluorescence assay (IFA). Using IFA the authors tested 154 Q fever positive sera from 55 patients with acute Q fever and 28 patients with chronic Q fever for Legionella pneumophila antibodies and 57 sera from 57 patients with legionellosis for Coxiella burnetii antibodies. Of the 211 sera tested, four sera from different patients had antibodies to both C. burnetii and L. pneumophila. Using cross-adsorption studies and protein immunoblotting, no cross-reaction between C. burnetii and L. pneumophila antibodies could be identified. The moderate antibody titers against L. pneumophila in two Q fever patients and vice versa for one legionellosis patient are consistent with the incidence of seroprevalence in healthy blood donors and were not due to cross-reactivity. One patient was identified with concurrent Q fever and legionellosis.

Antibodies, Bacterial

Streptococcal bacteremia in neutropenic adult patients.

In order to define the circumstances of occurrence, clinical presentation and prognostic factors of streptococcal bacteremia in neutropenic patients, we retrospectively reviewed 60 cases of streptococcal bacteremia following intensive chemotherapy for acute myeloblastic leukemia (AML), acute lymphoblastic leukemia (ALL) or allogeneic bone marrow transplantation. Causative streptococcal species included streptococcus viridans (42 cases), streptococcus faecalis (12), streptococcus pneumoniae (4) and other streptococci (2). All patients were febrile and 32% presented diffuse pneumopathy with frequent isolation of streptococci from bronchoalveolar washing fluids. There were no statistical differences in presentation between the bacteremia caused by different streptococcal species. Death occurred in 22% of the patients and factors favoring poor prognosis included pneumopathy (p less than 0.001), more than 2 positive blood cultures (p less than 0.01) and initial chemotherapy for AML (p less than 0.01). Pneumopathy occurred more frequently after chemotherapy for AML. It is concluded that streptococcal bacteremia is to be prevented in patients undergoing intensive chemotherapy, particularly if cytarabine is used.

Adolescent

[Iodized polyvidone (betadine) and prevention of postoperative infection. A multicenter study].

A multicentre study of a new preparation of povidone iodine (5% betadine) was carried out on 150 patients undergoing cataract surgery. Pre-operatively the eye was prepared by daubing the peri-ocular skin and a two-minute instillation into the conjunctival fornices. No other antiseptic was applied. Tolerance to the product was evaluated in all the patients: clinical evaluation postoperatively and during a 60-day follow-up period confirmed that the preparation was harmless to the eye. Microbiological investigations (comprising conjunctival swabbing, bacterial culture and count of the bacterial species isolated, before and after application of the product) performed on 60 patients showed a significant fall in the number of colonies after treatment.

Administration, Topical

Evaluation of a blood culture medium BHI-S-Lysis for BCB Release System Roche.

The new blood culture medium BHI-S Lysis for the BCB Release System (F. Hoffmann-La Roche AG., Basel, Switzerland), which lyses blood cells, was compared with the Hémoline performance Duo (BioMérieux, Marcy l'Etoile, France) including a diphasic medium and a modified Wilkins Chalgren broth. A total of 1998 sets were each inoculated with 7 ml of blood. In the laboratory, agar-coated paddles were attached to the BHI-S-Lysis. Overall, there was a statistically significant difference between the BHI-S-Lysis and the diphasic medium, or Wilkins Chalgren broth, in the recovery of clinically important microorganisms. The yield of pathogens with the BHI-S-Lysis, diphasic medium and Wilkins Chalgren broth was 115, 68 and 58, respectively. Staphylococci were detected by the BHI-S-Lysis significantly more frequently. Out of 50 bacteraemias, 12 were diagnosed with the BHI-S-Lysis only, three with the diphasic medium only, and three with the Wilkins Chalgren broth only. Contaminant isolates occurred with significantly greater frequency in the BHI-S-Lysis than in the two BioMérieux systems, in particular with coagulase-negative staphylococci. No speed advantage was present for all groups of micro-organisms and blood culture media. This good performance of the BHI-S-Lysis medium in detecting a larger number of pathogens should be confirmed by further evaluations.

Bacteremia

Characterization of staphylococcal plasmids hybridizing with the fosfomycin resistance gene fosB.

The distribution of the fosB gene, coding for fosfomycin resistance, in 105 fosfomycin-resistant isolates of Staphylococcus from various geographical areas, was studied by Southern blot hybridization. Nucleotide sequences related to fosB were detected in 36 strains belonging to five species. Plasmids bearing fosB were often of a size similar to that of pIP1842 (2.54 kb) in S. epidermidis, most often small (2.4 to 4.1 kb) in other species including S. aureus where a 2.7-kb plasmid was found in 16 out of the 18 strains studied. The fosB gene was geographically dispersed since it was present in six different locations in France and also in Japan. The weak hybridization observed with plasmid DNA of certain strains of S. aureus, S. epidermidis, S. haemolyticus, S. saprophyticus, and S. warneri may indicate gene heterogeneity for fosfomycin resistance in Staphylococcus spp.

Blotting, Southern

Pefloxacin and vancomycin vs. gentamicin, colistin sulphate and vancomycin for prevention of infections in granulocytopenic patients: a randomised double-blind study.

To test the value of pefloxacin for the prevention of infections in patients with chemotherapy-induced neutropenia, oral pefloxacin plus vancomycin (PV) (n = 76) or gentamicin, colistin sulphate and vancomycin (GCV) (n = 74) were administered in a randomised double-blind study. Infections were significantly less severe in the PV than in the GCV group. Patients receiving PV had significantly fewer episodes of bacteraemia and central venous line infections than patients treated with GCV. Gram-positive and gram-negative infections were significantly less frequent in patients receiving PV, because of fewer infections with Staphylococcus species and enterobacteriaceae. Stool culture detected significantly more gram-positive organisms in the PV group and more gram-negative organisms in the GCV group. Thus, PV was more efficacious than GCV for the prevention of gram-positive and gram-negative infections in the neutropenic patients, despite lower efficacy in eradicating gram-positive organisms from the lower intestinal tract.

Adult

Delta-like haemolysin produced by Staphylococcus lugdunensis.

Thirty independent clinical isolates of Staphylococcus lugdunensis were tested for expression of haemolysin characteristics of Staphylococcus aureus and for the presence of DNA sequences with homology to S. aureus toxin genes. Twenty-six strains produced a thermostable haemolysin with greatest activity against rabbit erythrocytes. DNA-DNA hybridisation was performed with S. lugdunesis DNA and probed with the alpha, beta, delta and gamma toxin gene probes of S. aureus as well as that for coagulase. Hybridisation was found with the delta probe in 28 strains including 6 which did not express haemolysin. Two strains did not hybridise with any of the probes. These findings suggest that S. lugdunensis strains produce a delta-like toxin which shares homology with the delta toxin of S. aureus.

Bacterial Proteins

Difficulties in identifying Klebsiella strains of clinical origin.

Two hundred and four strains of Gram-negative bacteria of clinical origin, initially identified as Klebsiella using the API 20 E system, and 10 reference strains were further analysed with the API 20 EC test system and the API 50 CH, API 50 AO, API 50 AA assimilation systems. Four clusters corresponding to the species Klebsiella pneumoniae subsp. pneumoniae, K. oxytoca, K. planticola, and K. terrigena were formed after numerical analysis of 155 selected tests and the 26 most discriminating tests were determined. A comparison was made between conventional identification using the API 20 E system and the results of the numerical analysis. The conventional method resulted in incorrect identification of 13% of the strains tested, especially for the new species: K. planticola and K. terrigena. After numerical analysis, 17 out of 204 strains (8.3%) of clinical origin were identified as K. planticola. Only 1 strain of clinical origin was identified as K. terrigena, and 1 strain as K. ornithinolytica.

Humans