In vitro susceptibility testing of Candida and Aspergillus spp. to voriconazole and other antifungal agents using Etest: results of a French multicentre study.

Minimum inhibitory concentrations (MICs) of the antifungal agent voriconazole were determined using the Etest and compared with those of amphotericin B, itraconazole and fluconazole using 1986 clinical isolates of Candida spp. Voriconazole MICs were also compared with those of amphotericin B and itraconazole using 391 clinical isolates of Aspergillus spp. Voriconazole was found to have more potent activity and lower MIC values than amphotericin B, itraconazole and fluconazole against C. albicans, C. tropicalis, C. parapsilosis and C. kefyr. Against C. glabrata and C. krusei, voriconazole was more active than either of the other two azole antifungals but had similar activity to amphotericin B. For species of Aspergillus, MIC values of voriconazole were lower than those of amphotericin B and itraconazole against A. fumigatus and A. flavus, and were similar to those of amphotericin B against A. niger. Against A. terreus, MIC values for voriconazole and itraconazole were similar. A. terreus is known to be resistant to amphotericin B, and this was reflected in higher MIC values compared with those of voriconazole and itraconazole. Voriconazole therefore compares very favourably with other antifungal agents against a large number of clinical isolates of Candida and Aspergillus spp.

Case report of three Candida albicans infections detected at delivery.

We report three similar cases of Candida albicans infections in neonates, at delivery. A retrospective study of the isolates was conducted to define the diversity of infective strains and their susceptibility to amphotericin B and fluconazole. Three neonates with fever, 'not doing well' at delivery had positive cultures for C. albicans. Samples were then taken from the mothers who did not exhibit any clinical symptoms of infection. Candida albicans strains isolated from both neonates and mothers were cultured, six colonies of each were typed by multilocus enzyme electrophoresis. The E-test method was used to determine the susceptibility of each colony to the two antifungals commonly used in this unit: amphotericin B and fluconazole. The initial isolates were composed of different types of strains. In the three cases, one of the mother types was found in the neonate isolates, leading us to suggest a vertical transmission of strains. All of the other types were distinct. All of the types were susceptible to amphotericin B, although three of them, one type isolated from a neonate and two types isolated from the mother, were resistant to fluconazole. The diversity of infective strains remains alarming and encourages the consideration of several colonies per isolate or several isolates, when it is possible, per infection case. This study also points out the need to survey the susceptibility of infective strains, since some of them appear soon to be resistant to fluconazole.

[Bactericidal power's assessment of eight antiseptic products intended to surgeon's hand-washing (author's transl)]. / Evaluation du pouvoir bactéricide de huit antiseptiques destinés au lavage de type chirurgical des mains.

A standard hand-washing technique was used in order to test the relative effectiveness of eight both detergent and alcoholic preparations intended to surgeon's hands disinfection. A single four or seven minutes washing with alcoholic solutions was shown to eliminate a much larger proportion of the skin flora than could be removed by a single four or seven minutes hand-washing with detergent antiseptic preparations. Authors have also determined effect of wearing surgical rubber gloves after skin disinfection and compared viable bacterial counts in hand washings immediately after the antiseptic treatment and when gloves had been worn for one hour.

[Synergic effect of 5-fluorocytosine and imidazole derivatives against yeast strains resistant to 5-fluorocytosine]. / Effet synergique entre la 5-flurocytosine et les dérivés imidazolés vis-à-vis de souches de levures résistantes à la 5-fluorocytosine.

Antifungal agents associations are widely used in therapy of deep mycotic diseases, particularly amphotericin B-5-fluorocytosine association. Synergistic effect has also been described between 5-fluorocytosine and imidazole derivates. The authors have tested here eventual synergy between 5-fluorocytosine and imidazole derivatives (miconazole, ketoconazole, fluconazole, itraconazole) against 57 years isolates resistant to 5-fluorocytosine by a semi-automated methods in liquid medium (Yeast Nitrogen Base and Brain Heart Infusion). The synergistic effect between 5-fluorocytosine and antifungal imidazoles varies widely with the drug tested. It's more frequent with ketoconazole. Itraconazole and fluconazole present very little synergistic effects in vitro.

[Optimization of sensitivity tests to ketoconazole. Value of experience planning in the choice of a medium]. / Optimisation des tests de sensibilité au kétoconazole. Intérêt des plans d'expériences dans le choix d'un milieu.

Using factorial design, the authors evaluate the factors which influence in vitro, the anti-fungal activity of ketoconazole against a strain of Candida albicans (CBS 562). With the four mediums most frequently used, (Sabouraud, Casitone, MEM, YNB) they precise the results concerning inoculum, pH, incubation temperature, growth time, serum concentration, and shaking. This method allows the evaluation of each factor in optimal conditions of test. In most cases, the best results are obtained with a 10(4) UFC/ml inoculum, a pH = 7, in absence of serum and without shaking. The influence of temperature is very small. In these conditions, the Casitone medium seems the most satisfactory.

[Fusarium pleural effusion after a ventricular assist device]. / Pleurésie à Fusarium compliquant une implantation de ventricule artificiel.

We report the case of a 36-year-old man with a pleural effusion that complicates the postoperative period after the implantation of a ventricular assist device (VAD). The epidemiological, etiologic and therapeutic features of Fusarium infections were reviewed. Complete recovery of the infection was obtained after a treatment by liposomal amphotericine B (AmBisome) and 5 fluorocytosine.

Carbon assimilation tests: substrates assimilation profiles.

Liquid medium assays for yeasts carbon assimilation tests are the more precise but longer methods. For rapid and automated yeasts identification purposes we analysed the assimilation of 34 carbon compounds by 149 reference strains. Assays were carried in liquid shaken medium (Autobac system) and readings were nephelemetric. Valuable results are obtained in 72 hours and their analysis allowed us to classify substrates for their ability to minimize the number of doubtful results.

Multicentric genetic study of Candida albicans isolates from non-neutropenic patients using MLEE typing: population structure and mode of reproduction.

A mycological survey was conducted on non-neutropenic patients in three distinct intensive care units in two hospitals in Marseille (France) from November 1993 to November 1995. Candida albicans positive cultures from 62 patients were included in this study. Every first isolate of each patient was typed by multilocus enzyme electrophoresis (MLEE). The enzyme profiles obtained from 15 polymorphic loci were then compared. This analysis demonstrated a strong population differentiation of C. albicans infective strains within and between the different care units and confirmed the probable preponderant clonal mode of reproduction of this yeast.

[Laboratory's controls of antifungal treatment by the fluconazole of the candidiasis in immunocompromised patients]. / Apport du laboratoire dans la surveillance du traitement antifongique par le fluconazole des candidoses chez les immunodéprimés.

The use of antifungal drugs in immunocompromised patients impose to the laboratory the control of the efficacy of these therapy. With fluconazole, one of the most recent antifungal agents these control use a special method (Central Research Pfizer) different of those they are used with others antifungals. We have comparatively tested, using MIC technic four broth mediums (High Resolution medium (Oxoid) YMB (Difco) YNB dextrose (Autobac) and Casitone) and three agar mediums (HR, YMA, Casitone) incubated at 28 degrees C for 24 and 48 h. The strains of yeasts are isolated from oro-pharyngeal prelevements on HIV antibody positive patients observed during six to twelve months and eventually treated by fluconazole. Sixty patients are controlled, 33 give one or more positive cultures with 74 strains of C. albicans and four other yeasts. By determination of the MICs with seven different methods we find any resistant strains with the MICs range from 3.12 to 12.5 or 25 micrograms/ml.

[In vitro evaluation of the sensitivity to fluconazole of different species of yeasts isolated in pathology]. / Evaluation de la sensibilité in vitro au fluconazole de différentes espèces de levures rencontrées en pathologie.

In a previous study, the authors developed a technique for evaluating the in vitro susceptibility (or resistance) of Candida albicans to fluconazole, using casitone broth and agar. Photometric readings of growth in liquid media proved more accurate for evaluating antifungal activity and consistently agreed with clinical findings in all studied cases. This method was consequently extended from C. albicans to other yeasts recovered from high-risk patients (C. glabrata, C. famata, C. tropicalis, C. krusei, C. pseudotropicalis, C. parapsilosis, etc...). High resolution antifungal assay medium (broth and agar) and casitone medium (broth and agar) with or without agitation (Autobac System) were used to study the activity of fluconazole against approximately one hundred yeast strains. MICs above 3.12 micrograms/ml were found for several strains, particularly belonging to the C. glabrata and C. krusei species. These values are equal to or greater than serum levels achieved during treatment with fluconazole, a fact which raises practical questions concerning fluconazole therapy and may explain the failure of fluconazole to eradicate yeasts in some patients.

Multilocus enzyme electrophoresis analysis of Candida albicans isolates from three intensive care units. An epidemiological study.

To evaluate the mechanism and risk factor associated with the nosocomial acquisition of Candida albicans, a 3-month prospective study was conducted on non-neutropenic patients in three distinct intensive care units in distinct hospitals. A total of 43 samples from 19 patients has been typed by multilocus enzyme electrophoresis (MEE). Samples (24) from the deep pharynx of hospital staff members were also cultured and typed. Thirteen of the 19 enzyme loci studied were polymorphic. The 52 electrophoretic types were assigned to 67 isolates, each type was represented by one to five isolates; this proved the great diversity of the isolates. It appeared that most patients were colonized or infected with different C. albicans strains. This study pointed out a possible cross-infection between patients and hospital staff and between healthy members of the hospital staff, and also showed the successive stages of infection by different electrophoretic types. Intubation and respiratory equipment as surgery intervention were possible sources of observed infections.

[Amphotericin B into parenteral lipid emulsions. Galenic stability of mixtures. Preliminary results]. / L'amphotéricine B dans des émulsions lipidiques parentérales. Stabilité galénique des mélanges. Résultats préliminaires.

The amphotericin B in glucose solution is the reference's treatment of the major systemic mycoses. Because of its bad tolerance, certain authors put forward others vehicules such as fat emulsions. The aim of our work has been to study the physico-chemical stability of two regeneration's methods; P1: direct regeneration. P2: regeneration of 50mg of Fungizone in 5 ml of glucose serum and laced with 45 ml of Ivelip 20% versus PR: 50 mg of Fungizone in 50 ml of glucose serum. Our results seem to show that the P1 is totally incompatible with a parenteral administration, because 23% of the granulometric population has a diameter superior to 5 microns. The solution P2 seems better even if the pH is different from PR and a light depot comes after reconstitution.

[Mycological monitoring of Candida albicans infections in various hospital care units. Molecular typing of isolated strains and epidemiological survey]. / Suivi mycologique d'infections á Candida albicans dans divers services hospitaliers. Typage moléculaire des souches isolées et enquête épidémiologique.

To evaluate risk factors associated with the nosocomial infection of Candida albicans, a prospective study is conducted twice for three months in three intensive care units. Samples from patients HIV negatives, non neutropenic and non immunodepressive are collected as they came in the unit, on several anatomic sites. Every C. albicans carriers are included in a mycological monitoring. Samples from environmental surfaces, hands and deep pharynx from hospital personnel were also cultured. Strains genetic profile are defined by isoenzyme electrophoresis technique. Thirteen polymorphic loci allowed samples classement into 52 electrophoretic types (ET). If only one crossed contamination is described, strains regroupment into some ET incites us to extend this study. C. albicans strains from patients closed environment have never been isolated.

Evaluation of internal transcribed spacer region of ribosomal DNA sequence analysis for molecular characterization of Candida albicans and Candida dubliniensis isolates from HIV-infected patients.

Molecular typing systems have been needed to study Candida colonization in HIV-infected patients, particularly for investigating virulence and fluconazole resistance. Three methods--electrophoretic karyotyping (EK), detection of restriction fragment length polymorphisms (RFLP) and randomly amplified polymorphic DNA analysis (RAPD)--have been most frequently used. In this study, comparative sequence analysis of the internal transcribed spacer (ITS) region of rDNA was evaluated for delineation of Candida isolates from 14 HIV-infected patients. EK, ITS sequence analysis, RFLP and RAPD resulted in 11, 10, 9 and 8 DNA genotypes, respectively, from 39 Candida albicans isolates. The 10 genotypes observed using ITS sequence analysis were defined by six variation sites in the sequence. Molecular typing of sequential oral isolates showed the persistence of the same genotype of C. albicans in nine patients, and genotype variation in one patient. EK and RAPD showed that another patient was co-infected by two distinct genotypes and ITS analysis identified one of the two genotypes as Candida dubliniensis. Comparative ITS sequence analysis is a quick and reproducible method that provides clear and objective results, and it also identifies C. dubliniensis. The discriminatory power of this new typing approach could be improved by concomitant analysis of other DNA polymorphic sequences.

[Results of a year's mycologic and immunologic surveillance of Candida in an intensive care ward]. / Bilan d'une année de surveillance mycologique et immunologique des Candida dans un service de réanimation.

Patients hospitalized in an intensive care unit must be considered to be exposed to a "high risk" in terms of candida infection. They often combine several of the factors favouring the development of "candida disease": generally deficient state, multiple antibiotics, iatrogenic or pathological immune depression. While certain patients who are admitted are already suffering from "candida infestation" (endogenous localization), the possibilities of exagenous infection are numerous. This study involved 63 patients spending at least one week in the department, from September 1977 to September 1978. Mycological (pharyngeal swab, urine culture, blood culture) and immunological (hemagglutination, immunoelectrophoresis, immunofluorescence) studies were carried out routinely on admission and then every week. Amongst the 63 patients studied, 12 were admitted with negative serology, which became positive during the first 10 days. 5 already had positive serology at the time of admission. From a mycological standpoint, the pharyngeal swab was frequently positive on admission (17 times out of 63). In 63 patients, the infection was acquired during the course of the hospital stay and, in this case, positive urine cultures were more regularly associated with a positive pharyngeal culture.

Specific antibody detection in human aspergillosis: a GEMO* multicentre evaluation of a rapid immunoelectrophoresis method (Paragon). Group d'Etude des Mycoses Opportunistes.

A new immunoelectrophoresis system, the Paragon system, was evaluated in three different hospital centres with the aim of improving standardization of the serodiagnosis of human aspergillosis. To select the most efficient antigen, various commercial and home-made antigens were first tested on 19 sera from 19 patients with highly probable aspergillosis. The value measured using the Paragon anti-Aspergillus antibody detection system was then compared with the results obtained by conventional serological diagnostic methods (conventional immunoelectrophoresis, enzyme linked-immunosorbent assay, indirect immunofluorescence): this step was performed using the first 19 sera as well as 16 other sera from 13 patients with suspected aspergillosis. Concordant results were obtained in 28 cases. The discrepancies observed with seven sera were probably related to differences in the nature of the antigens. Paragon immunoelectrophoresis proved to be a practicable technique requiring only a small amount of serum and giving results within a shorter time than competitive methods (24-48 h). Its major drawbacks compared with conventional immunoelectrophoresis are some difficulties in reading, fewer precipitin lines and the relatively high cost of routine analysis.

Report of a working group for the European Pharmacopoeia. Collaborative assay on human albumin of different origin.

A collaborative assay was conducted by 9 laboratories on 31 samples of human albumin which were in clinical use. It was the object of the study to establish test systems which would differentiate between albumins of venous or placental origin. The properties examined for this purpose were: appearance, total protein, haem, polymers, alkaline phosphatase and blood group substances. Additional tests such as for beta-thromboglobulin and citrate were included; pyrogenicity, however, was excluded because this was under study for all plasma proteins at that time. Results obtained were in satisfactory agreement both between laboratories and between samples. They, therefore, enabled the verification of a number of correlations in the test systems. The evaluation did not allow, however, the differentiation of the samples in relation to their origin. The results were, therefore, regarded as a tool to define the upper limits of acceptance for human albumins corresponding to the quality prescribed by the European Pharmacopoeia.