Establishment and follow-up of bifidobacterial species in the gut of healthy bottle-fed infants of 1-4 months age.

Twenty-one healthy bottle-fed infants were screened monthly (1-4 months) for bifidobacteria in their stools. Bifidobacteria were detected by culture and isolates specified by PCR. Alternatively, direct PCR in undiluted fecal suspensions was carried out for detection of bifidobacteria under the cultural detection level. All infants harbored cultivable bifidobacteria throughout the study period. Beerens medium was shown to permit a better recovery of bifidobacteria than MRS and horse blood Columbia agar. Direct PCR detection proved valuable in detecting species for which no cultural isolate could be recovered since the species were under the cultural detection level. B. bifidum, B. longum-infantis and B. breve were confirmed as dominant and stable species in infant stools while B. adolescentis and B. catenulatum group exhibited unstable colonization profiles. A trend towards B. breve decrease began at month 3 while carriage of the B. catenulatum group and B. adolescentis was rising. This observation warrants further analysis to assess a possible switch occurring at month 3 in bottle-fed infants, between so-called infant and adult bifidobacterial species.

[Spondylodiscitis due to Salmonella enteritica serotype Typhi]. / Spondylodiscite à Salmonella enteritica sérotype Typhi.

We reported a case of lombar spondylodiscitis caused by Salmonella enteritica serotype Typhi in an immunocompetent patient. Salmonella is a rare causative agent of spondylodiscitis. Early bacteriological diagnosis is essential to avoid longterm sequelae.

Pseudomonas brenneri sp. nov., a new species isolated from natural mineral waters.

The vernacular name 'fluorescent Pseudomonas group 97-391' was coined for a group of 11 strains isolated from two French natural mineral waters. All these strains were Gram-negative, rod-shaped and motile by means of a single polar flagellum. They produced fluorescent pigment (pyoverdin) on King B medium, catalase and cytochrome oxidase. They were not able to accumulate poly-beta-hydroxybutyrate. They were capable of respiratory but not fermentative metabolism. DNA-DNA hybridization results and DNA base composition analysis revealed that strains of the 'fluorescent Pseudomonas group 97-391' were members of a new species, for which the name Pseudomonas brenneri sp. nov. (type strain CIP 106646T) is proposed. The levels of DNA-DNA relatedness within this group ranged from 70 to 100% with DeltaTm below 1 degree C. The G+C content of the DNA of the type strain was 58 mol%. DNA relatedness with 72 strains representing well-known or partially characterized species of the genus Pseudomonas (sensu stricto) was below 48%. The complete 16S rRNA sequence of the type strain CIP 106646T was determined and compared with those of the type strains of Pseudomonas species. Finally, a phylogenetic tree was inferred from sequence analysis and demonstrated that the new species fell into the 'Pseudomonas fluorescens intrageneric cluster'. The clinical significance of P. brenneri is unknown.

Taxonomic study of bacteria isolated from Lebanese spring waters: proposal for Pseudomonas cedrella sp. nov. and P. orientalis sp. nov.

Deoxyribonucleic acid relatedness studies (S1 nuclease method) have shown that 15 strains isolated from three Lebanese spring waters, belonging to the genus Pseudomonas, formed two homogeneous DNA groups, with a within-group DNA relatedness ranging from 70 to 100%. These groups are referred to as Pseudomonas cedrella sp. nov. and Pseudomonas orientalis sp.nov. These strains were previously grouped on the basis of a numerical analysis in phenons Ve, Vd, Vg, and VI. DNA relatedness with 65 strains representing 24 species of the genus Pseudomonas sensu stricto was below 50%. The highest DNA binding value (50%) was found with P. marginalis species. A comparison of the complete 16S rRNA gene sequences of the strains representing the two new deoxyribonucleic acid hybridization groups, i.e., strains CFML 96-198T and CFML 96-170T, and the sequence of other strains of the genus Pseudomonas revealed that these strains (CFML 96-198T and CFML 96-170T) fell within the 'Pseudomonas fluorescens intrageneric cluster'. The G+C contents of the DNA of P. cedrella CIP 105541T and P. orientalis CIP 105540T were 59 and 60 mol%, respectively. The two species can be differentiated from each other by the fact that P. cedrella strains hydrolyze erythritol and D-lyxose. P. cedrella grouped together a total of nine strains from phenotypic groups Ve, Vg, and VI. P. orientalis grouped together six strains from both phenotypic groups Vd and Ve.

The medical and economic consequences of automation in bacteriology: a case study in a French university hospital.

In a bacteriology laboratory where automated and non-automated procedures co-existed during the study period (1 year), patients were randomly assigned to each type of procedure and we observed the physicians behaviour as well as patients well-being in a surgical service using the results from the laboratory. Contrary to our expectations, the reduction in the time delay necessary to obtain information did not alter either the prescribing behaviour of physicians nor the welfare of patients. Besides, the gain in time was significantly lower than expected. We also discuss in detail the meaning and relevance of the results.

Taxonomic study of bacteria isolated from natural mineral waters: proposal of Pseudomonas jessenii sp. nov. and Pseudomonas mandelii sp. nov.

The taxonomic position of 23 strains isolated from mineral waters and previously grouped in the authentic pseudomonads on the basis of a phenotypic analysis (cluster IX, subclusters XIIIa and XIIIc of VERHILLE, S., ELOMARI, M., COROLER, L., IZARD, D., LECLERC, H. (Syst. Appl. Microbiol, 20, 137-149, 1997) has been genotypically further studied in the present work. On the basis of hybridization results, these strains were gathered into two new genomic groups for which we propose the names of Pseudomonas jessenii sp. nov. (Type strain CIP 105274) and Pseudomonas mandelii sp. nov. (Type strain CIP 105273). Deoxyribonucleic acid relatedness levels showed homologies ranging from 78 to 100% for Pseudomonas jessenii and from 77 to 100% for Pseudomonas mandelii. Furthermore, hybrization rates with 66 representative well characterized species or only partially characterized species of the genus Pseudomonas were below 53%, with delta Tm values of 7 degrees C and more. The mol% G + C content ranged from 57 to 58. The two new species presented basic morphological characteristics common to all pseudomonads. Various phenotypic features, such as denitrification, growth at 4 degrees C or 41 degrees C, trigonelline assimilation, alpha-L-glutamyl-L-histidine arylarmidase activity, growth on benzoate and meso-tartrate were found to differentiate Pseudomonas jessenii from Pseudomonas mandelii and from other Pseudomonas species. Pseudomonas jessenii encompassed a total of 9 strains from both phenotypic groups IX and XIIIa. Pseudomonas mandelii clustered a total of 13 strains from both phenotypic groups IX and XIIIc. Their clinical significance is unknown. The 16S rDNA of each type strain was sequenced and compared with the known sequences of the representative strains of the genus Pseudomonas. A phylogenetic tree was constructed to determine the intrageneric relationships within the genus Pseudomonas.

Rahnella aquatilis, a potential contaminant in lager beer breweries.

The species Rahnella aquatilis has been isolated mostly from water, soil, and, in a few cases, from human clinical specimens; little is known about its ecological role. The application of polyacrylamide gel electrophoresis of soluble proteins, DNA-DNA hybridizations and API 20 E systems has shown that Rahnella aquatilis might also be encountered as a contaminant in lager beer breweries.

Antimicrobial activity of cefotiam combinations against Enterobacteriaceae moderately susceptible or resistant to this new cephalosporin.

The interaction of cefotiam with each of the four aminoglycosides gentamicin, tobramycin, netilmicin and amikacin were studied by the broth microdilution method ("checkerboard" technique) against 36 strains of Enterobacteriaceae chosen for their moderate susceptibility (MIC: 4-32 mg/l) or resistance (MIC: 64-512 mg/l) to cefotiam. A high rate of synergistic combinations was found with all the aminoglycosides: 81% with gentamicin, 76% with amikacin, 67% with tobramycin and netilmicin. The therapeutic value of these interactions appeared excellent.

In vitro activity of pefloxacin.

A broth dilution method was used to measure the minimal inhibitory concentrations of pefloxacin, a new quinolone derivative. Because this new agent could be used in systemic infections, mezlocillin, cefotaxime, ceftazidime and amikacin were used as comparative agents. Pefloxacin was found to be more active than mezlocillin and as active as or more active than cefotaxime, ceftazidime and amikacin. With the exception of Serratia marcescens, only 7% of Enterobacteriaceae strains were resistant to pefloxacin with a MIC greater than 8 micrograms/ml. Pefloxacin was also found to be the best agent against the non-fermenters Staphylococcus aureus and St. epidermidis. Resistance to ampicillin, carbenicillin and gentamicin did not affect the effectiveness of pefloxacin. However, pefloxacin appeared more susceptible to nalidixic acid resistance, with MICs four to sixfold higher.

[Rapid, automatic methods for identifying enterobacteria]. / Méthodes rapides et automatiques d'identification des entérobactéries.

The biochemical testing of Enterobacteriaceae can be rapidly and automatically performed with commercial identification systems. There are three groups of identification systems: the kits, the microtiter plates and laboratory apparatus. The ratios of agreement between these new systems and the conventional tests are equivalent. The two first categories of systems are inexpensive unlike apparatus which must be utilized only in important laboratories.

[Rapid diagnosis of purulent meningitis]. / Diagnostic rapide des méningites purulentes.

The cerebrospinal fluid of 589 subjects, 78 of whom were suffering from a purulent meningitis were examined. Comparatively by classical bacteriological techniques (direct examination and culture) and by electro-immunodiffusion, latex agglutination, and Limulus endotoxin assay. Soluble bacterial Haemophilus influenzae type B, Neisseria meningitidis group A, C, and Streptococcus pneumoniae antigens, were tested by electro-immunodiffusion and latex agglutination, and soluble bacterial N. meningitidis group B, Listeria monocytogenes and Streptococcus agalactiae antigens by electro-immunodiffusion. Specific antigens and endotoxin were found in 75.8 per cent of the specimens with a rapid answer (120 min). The three tests revealed also only the diagnosis in 29.1 per cent of cases of pneumococcal meningitis, in 33.3 per cent of meningococcal meningitis and in 47 per cent of Gram-negative bacteria meningitis. Only five cerebrospinal fluid from the 589 specimens tested were given a non-specific reaction. These two advantages--sensitivity and specificity--of these three tests render them techniques of the future in the diagnosis of purulent meningitis.

[Epidemiology of infections associated to "Burkholderia cepacia complex" in the course of cystic fibrosis]. / Epidémiologie des infections provoquées par les bactéries du "complexe Burkholderia cepacia" au course de la mucoviscidose.

Within a few years bacteriological knowledge on Burkholderia cepacia species has progressed considerably. Within bacterial classification (taxonomy), B. cepacia gathers eight species and one species on standby of nomenclature (genomovar VI); the whole of these species constitutes the "B. cepacia complex" or B. cepacia "sensu lato" and the denomination B. cepacia "sensu stricto" is attributed to the genomovar I. These new data call into question the knowledge on the clinic and the epidemiology of B. cepacia "sensu lato" infection in the course of cystic fibrosis. Among these newly described species, B. cenocepacia (formerly genomovar III) and B. multivorans (formerly genomovar II) are the most frequent species and together they represent more than 90% of infections associated to "B. cepacia complex" in the course of cystic fibrosis. B. cenocepacia is often associated to the "cepacia syndrome" which is characterized as a fatal necrotizing pneumonia with bacteremia. The progress of molecular epidemiology allowed the description of bacterial clones of which some are highly transmissible from person-to-person. Their distribution varies according to the species and the geography. The identification of these new species appears particularly difficult and, by the fact, the data on taxonomy and molecular epidemiology can be provided only by highly specialized reference centers.

[Pharmacokinetics of cefotaxime in cirrhotic patients with or without ascites]. / Pharmacocinétique du céfotaxime chez les malades cirrhotiques avec ou sans ascite.

Pharmacokinetic values of cefotaxime were measured in 12 cirrhotic male patients (6 without ascites and 6 with ascites) after intravenous injection of a single 2 gram dose of the antibiotic. In patients without ascites elimination of the drug was about the same as in normal subjects or control patients, although clearance was increased. In patients with ascites, the drug elimination half-life was significantly more prolonged (7.5 +/- 3.9 h versus 1.3 +/- 0.4 h, P less than 0.01) and the drug clearance was significantly lower (193.6 +/- 92.4 ml/min versus 475.8 +/- 152.2 ml/min, P less than 0.01) than in the other group. The accumulation of cefotaxime in these patients produced concentration in the ascites fluid that were above the critical therapeutic values for about 20 hours.

[Clinical value of rapid bacteriological results in nosocomial infections. Comparison with traditional methods]. / Intérêt clinique des résultats rapides de bactériologie au cours de l'infection nosocomiale. Comparaison avec les méthodes traditionnelles.

With automated analysers, bacterial identification and susceptibility testing can be performed in 4-5 hours instead of 12-18 hours with conventional methods. A controlled trial was carried out in the surgical ward of a university hospital to evaluate the clinical repercussions of these rapid methods. The automated analyser reduced delays in the laboratory by about 25%, and optimizing information transfer from laboratory to ward brought the reduction up to 50%. It was found that earlier results of susceptibility testing modified prescriptions and this may be expected to result in a more rational use of antibiotics.

[A method of bacterial count by epifluorescence with acridine orange. Application to skin biopsies performed in burnt patients]. / Etude d'une méthode de dénombrement des bactéries par épifluorescence à l'acridine orange. Application aux biopsies de peau effectuées chez les brûlés.

Acridine orange was used for staining and counting micro-organisms obtained from 136 skin biopsies performed in burned patients. The number of organisms per gram of tissue was compared to the number of colony-forming-units (CFU) calculated from cultures of the same biopsies. The staining method was positive in 97 per cent of septic samples, and in 25 per cent of these it proved more sensitive than bacterial cultures, with a 100 to 1000-fold greater number of pathogens detected. Acridine orange also demonstrated bacteria in 69 biopsies which remained sterile after culture. In some cases, the same bacterial species was found in other samples taken a few days later.

[A 4-year study of Pseudomonas aeruginosa susceptibility to antibiotics (1998-2001) in northern Lebanon]. / Sensibilité de Pseudomonas aeruginosa aux antibiotiques: étude sur quatre ans (1998-2001) au nord du Liban.

Four hundred and sixty-four Pseudomonas aeruginosa strains were isolated in northern Lebanon at the Islami Hospital Microbiology department, in Tripoli. The purpose of this study was to evaluate the susceptibility of these strains to antibiotics, to compare this susceptibility according to the nature of the sample and the year of sampling. The results show that urinary samples were the most frequent (39.3%), followed by wound samples (21.2%), and ear samples (16.5%). The average rate of susceptible strains was 39.8% to ticarcillin, 56.9% to piperacillin, 58.2% to piperacillin + tazobactam, 74.1% to imipenem, 63.3% to ceftazidime, 60.4% to cefepime, 62.1% to aztreonam, 60.3% to netilmicin, 57.5% to gentamicin, 62.2% to tobramycin, 69% to amikacin, 100% to colistin, 45.4% to pefloxacin and ofloxacin, 57.7% to ciprofloxacin and 1.3% to rifampicin. The study showed that the strains isolated from pulmonary secretions were the most resistant to antibiotics.

[Sensitivity of Pseudomonas aeruginosa to amikacin and to isepamicin in surgery and in intensive care]. / Sensibilité de Pseudomonas aeruginosa à l'amikacine et à l'isépamicine en chirurgie et en réanimation.

To evaluate the respective interest of amikacin and isepamicin in P. aeruginosa infection, the resistance level was ascertained using the disk method. Susceptibility was also tested for gentamicin, tobramycin and netilmicin. Isolates came from three surgical units and from two intensive care units. Serotyping was proceeded, and isolates coming from the same patient, with the same susceptibility pattern and the same serotype was included once only: 197 strains were thus obtained. Resistance level was 22.3% for amikacin, and 28.4% for isepamicin. Discrepancies were found in 14.7% of cases (major: 8.1%; minor: 6.6%). Discordant strains were more susceptible to amikacin than to isepamicin in 30/37 cases, and more susceptible to isepamicin in 7/37 cases. This difference was highly significant (paired Chi-2 test: p < 10(-4). The best susceptibility to amikacin was found in all serotypes and units.

DNA relatedness between Enterobacter sakazakii and other members of the genus Enterobacter.

A DNA-DNA hybridization study (nitrocellulose filter method) was carried out with 13 strains of Enterobacter sakazakii and 38 strains belonging to other Enterobacter species (E. cloacae, E. amnigenus, E. intermedium and E. gergoviae). E. sakazakii strains were highly related (mean relative binding ratio +/- standard deviation: 89% +/- 10) to the strain R 16-76 (labelled DNA). No close relationship was found with the other Enterobacter species. The low relative binding ratios were not due to a difference in genome size (as shown by reciprocal binding experiments).