Posaconazole MIC Distributions for Aspergillus fumigatus Species Complex by Four Methods: Impact of cyp51A Mutations on Estimation of Epidemiological Cutoff Values.

Estimating epidemiological cutoff endpoints (ECVs/ECOFFS) may be hindered by the overlap of MICs for mutant and nonmutant strains (strains harboring or not harboring mutations, respectively). Posaconazole MIC distributions for the Aspergillus fumigatus species complex were collected from 26 laboratories (in Australia, Canada, Europe, India, South and North America, and Taiwan) and published studies. Distributions that fulfilled CLSI criteria were pooled and ECVs were estimated. The sensitivity of three ECV analytical techniques (the ECOFFinder, normalized resistance interpretation [NRI], derivatization methods) to the inclusion of MICs for mutants was examined for three susceptibility testing methods (the CLSI, EUCAST, and Etest methods). The totals of posaconazole MICs for nonmutant isolates (isolates with no known cyp51A mutations) and mutant A. fumigatus isolates were as follows: by the CLSI method, 2,223 and 274, respectively; by the EUCAST method, 556 and 52, respectively; and by Etest, 1,365 and 29, respectively. MICs for 381 isolates with unknown mutational status were also evaluated with the Sensititre YeastOne system (SYO). We observed an overlap in posaconazole MICs among nonmutants and cyp51A mutants. At the commonly chosen percentage of the modeled wild-type population (97.5%), almost all ECVs remained the same when the MICs for nonmutant and mutant distributions were merged: ECOFFinder ECVs, 0.5 µg/ml for the CLSI method and 0.25 µg/ml for the EUCAST method and Etest; NRI ECVs, 0.5 µg/ml for all three methods. However, the ECOFFinder ECV for 95% of the nonmutant population by the CLSI method was 0.25 µg/ml. The tentative ECOFFinder ECV with SYO was 0.06 µg/ml (data from 3/8 laboratories). Derivatization ECVs with or without mutant inclusion were either 0.25 µg/ml (CLSI, EUCAST, Etest) or 0.06 µg/ml (SYO). It appears that ECV analytical techniques may not be vulnerable to overlap between presumptive wild-type isolates and cyp51A mutants when up to 11.6% of the estimated wild-type population includes mutants.

Multicenter Study of Method-Dependent Epidemiological Cutoff Values for Detection of Resistance in Candida spp. and Aspergillus spp. to Amphotericin B and Echinocandins for the Etest Agar Diffusion Method.

Method-dependent Etest epidemiological cutoff values (ECVs) are not available for susceptibility testing of either Candida or Aspergillus species with amphotericin B or echinocandins. In addition, reference caspofungin MICs for Candida spp. are unreliable. Candida and Aspergillus species wild-type (WT) Etest MIC distributions (microorganisms in a species-drug combination with no detectable phenotypic resistance) were established for 4,341 Candida albicans, 113 C. dubliniensis, 1,683 C. glabrata species complex (SC), 709 C. krusei, 767 C. parapsilosis SC, 796 C. tropicalis, 1,637 Aspergillus fumigatus SC, 238 A. flavus SC, 321 A. niger SC, and 247 A. terreus SC isolates. Etest MICs from 15 laboratories (in Argentina, Europe, Mexico, South Africa, and the United States) were pooled to establish Etest ECVs. Anidulafungin, caspofungin, micafungin, and amphotericin B ECVs (in micrograms per milliliter) encompassing ≥97.5% of the statistically modeled population were 0.016, 0.5, 0.03, and 1 for C. albicans; 0.03, 1, 0.03, and 2 for C. glabrata SC; 0.06, 1, 0.25, and 4 for C. krusei; 8, 4, 2, and 2 for C. parapsilosis SC; and 0.03, 1, 0.12, and 2 for C. tropicalis The amphotericin B ECV was 0.25 µg/ml for C. dubliniensis and 2, 8, 2, and 16 µg/ml for the complexes of A. fumigatus, A. flavus, A. niger, and A. terreus, respectively. While anidulafungin Etest ECVs classified 92% of the Candida fks mutants evaluated as non-WT, the performance was lower for caspofungin (75%) and micafungin (84%) cutoffs. Finally, although anidulafungin (as an echinocandin surrogate susceptibility marker) and amphotericin B ECVs should identify Candida and Aspergillus isolates with reduced susceptibility to these agents using the Etest, these ECVs will not categorize a fungal isolate as susceptible or resistant, as breakpoints do.

Multicenter study of anidulafungin and micafungin MIC distributions and epidemiological cutoff values for eight Candida species and the CLSI M27-A3 broth microdilution method.

Since epidemiological cutoff values (ECVs) using CLSI MICs from multiple laboratories are not available for Candida spp. and the echinocandins, we established ECVs for anidulafungin and micafungin on the basis of wild-type (WT) MIC distributions (for organisms in a species-drug combination with no detectable acquired resistance mechanisms) for 8,210 Candida albicans, 3,102 C. glabrata, 3,976 C. parapsilosis, 2,042 C. tropicalis, 617 C. krusei, 258 C. lusitaniae, 234 C. guilliermondii, and 131 C. dubliniensis isolates. CLSI broth microdilution MIC data gathered from 15 different laboratories in Canada, Europe, Mexico, Peru, and the United States were aggregated to statistically define ECVs. ECVs encompassing 97.5% of the statistically modeled population for anidulafungin and micafungin were, respectively, 0.12 and 0.03 µg/ml for C. albicans, 0.12 and 0.03 µg/ml for C. glabrata, 8 and 4 µg/ml for C. parapsilosis, 0.12 and 0.06 µg/ml for C. tropicalis, 0.25 and 0.25 µg/ml for C. krusei, 1 and 0.5 µg/ml for C. lusitaniae, 8 and 2 µg/ml for C. guilliermondii, and 0.12 and 0.12 µg/ml for C. dubliniensis. Previously reported single and multicenter ECVs defined in the present study were quite similar or within 1 2-fold dilution of each other. For a collection of 230 WT isolates (no fks mutations) and 51 isolates with fks mutations, the species-specific ECVs for anidulafungin and micafungin correctly classified 47 (92.2%) and 51 (100%) of the fks mutants, respectively, as non-WT strains. These ECVs may aid in detecting non-WT isolates with reduced susceptibility to anidulafungin and micafungin due to fks mutations.

Multilaboratory study of epidemiological cutoff values for detection of resistance in eight Candida species to fluconazole, posaconazole, and voriconazole.

Although epidemiological cutoff values (ECVs) have been established for Candida spp. and the triazoles, they are based on MIC data from a single laboratory. We have established ECVs for eight Candida species and fluconazole, posaconazole, and voriconazole based on wild-type (WT) MIC distributions for isolates of C. albicans (n=11,241 isolates), C. glabrata (7,538), C. parapsilosis (6,023), C. tropicalis (3,748), C. krusei (1,073), C. lusitaniae (574), C. guilliermondii (373), and C. dubliniensis (162). The 24-h CLSI broth microdilution MICs were collated from multiple laboratories (in Canada, Brazil, Europe, Mexico, Peru, and the United States). The ECVs for distributions originating from ≥6 laboratories, which included ≥95% of the modeled WT population, for fluconazole, posaconazole, and voriconazole were, respectively, 0.5, 0.06 and 0.03 µg/ml for C. albicans, 0.5, 0.25, and 0.03 µg/ml for C. dubliniensis, 8, 1, and 0.25 µg/ml for C. glabrata, 8, 0.5, and 0.12 µg/ml for C. guilliermondii, 32, 0.5, and 0.25 µg/ml for C. krusei, 1, 0.06, and 0.06 µg/ml for C. lusitaniae, 1, 0.25, and 0.03 µg/ml for C. parapsilosis, and 1, 0.12, and 0.06 µg/ml for C. tropicalis. The low number of MICs (<100) for other less prevalent species (C. famata, C. kefyr, C. orthopsilosis, C. rugosa) precluded ECV definition, but their MIC distributions are documented. Evaluation of our ECVs for some species/agent combinations using published individual MICs for 136 isolates (harboring mutations in or upregulation of ERG11, MDR1, CDR1, or CDR2) and 64 WT isolates indicated that our ECVs may be useful in distinguishing WT from non-WT isolates.

Multicenter study of isavuconazole MIC distributions and epidemiological cutoff values for Aspergillus spp. for the CLSI M38-A2 broth microdilution method.

Epidemiological cutoff values (ECVs) were established for the new triazole isavuconazole and Aspergillus species wild-type (WT) MIC distributions (organisms in a species-drug combination with no detectable acquired resistance mechanisms) that were defined with 855 Aspergillus fumigatus, 444 A. flavus, 106 A. nidulans, 207 A. niger, 384 A. terreus, and 75 A. versicolor species complex isolates; 22 Aspergillus section Usti isolates were also included. CLSI broth microdilution MIC data gathered in Europe, India, Mexico, and the United States were aggregated to statistically define ECVs. ECVs were 1 µg/ml for the A. fumigatus species complex, 1 µg/ml for the A. flavus species complex, 0.25 µg/ml for the A. nidulans species complex, 4 µg/ml for the A. niger species complex, 1 µg/ml for the A. terreus species complex, and 1 µg/ml for the A. versicolor species complex; due to the small number of isolates, an ECV was not proposed for Aspergillus section Usti. These ECVs may aid in detecting non-WT isolates with reduced susceptibility to isavuconazole due to cyp51A (an A. fumigatus species complex resistance mechanism among the triazoles) or other mutations.

Wild-type MIC distributions and epidemiological cutoff values for amphotericin B, flucytosine, and itraconazole and Candida spp. as determined by CLSI broth microdilution.

Clinical breakpoints (CBPs) and epidemiological cutoff values (ECVs) have been established for several Candida spp. and the newer triazoles and echinocandins but are not yet available for older antifungal agents, such as amphotericin B, flucytosine, or itraconazole. We determined species-specific ECVs for amphotericin B (AMB), flucytosine (FC) and itraconazole (ITR) for eight Candida spp. (30,221 strains) using isolates from 16 different laboratories in Brazil, Canada, Europe, and the United States, all tested by the CLSI reference microdilution method. The calculated 24- and 48-h ECVs expressed in µg/ml (and the percentages of isolates that had MICs less than or equal to the ECV) for AMB, FC, and ITR, respectively, were 2 (99.8)/2 (99.2), 0.5 (94.2)/1 (91.4), and 0.12 (95.0)/0.12 (92.9) for C. albicans; 2 (99.6)/2 (98.7), 0.5 (98.0)/0.5 (97.5), and 2 (95.2)/4 (93.5) for C. glabrata; 2 (99.7)/2 (97.3), 0.5 (98.7)/0.5 (97.8), and 05. (99.7)/0.5 (98.5) for C. parapsilosis; 2 (99.8)/2 (99.2), 0.5 (93.0)/1 (90.5), and 0.5 (97.8)/0.5 (93.9) for C. tropicalis; 2 (99.3)/4 (100.0), 32 (99.4)/32 (99.3), and 1 (99.0)/2 (100.0) for C. krusei; 2 (100.0)/4 (100.0), 0.5 (95.3)/1 (92.9), and 0.5 (95.8)/0.5 (98.1) for C. lusitaniae; -/2 (100.0), 0.5 (98.8)/0.5 (97.7), and 0.25 (97.6)/0.25 (96.9) for C. dubliniensis; and 2 (100.0)/2 (100.0), 1 (92.7)/-, and 1 (100.0)/2 (100.0) for C. guilliermondii. In the absence of species-specific CBP values, these wild-type (WT) MIC distributions and ECVs will be useful for monitoring the emergence of reduced susceptibility to these well-established antifungal agents.

Evaluation of the Etest method for susceptibility testing of Aspergillus spp. and Fusarium spp. to three echinocandins.

We performed Etest and broth microdilution (BMD) susceptibility testing of caspofungin, micafungin and anidulafungin against 67 clinical isolates of Aspergillus spp. and 10 Fusarium spp. Minimal effective concentrations (MECs) by BMD were read after 24 h of incubation at 35 degrees C and Etest MICs were read at 24 and 48 h. MECs < or =0.25 mg/l were obtained with caspofungin for all Aspergillus spp. tested but Etest MICs were < or =1 mg/l at 24 h. The agreement between caspofungin Etest and broth microdilution was good for all Aspergillus species tested (range 82.4-100%) except for A. niger and A. glaucus at 24 h of incubation. Micafungin and anidulafungin MEC and MIC results were lower than those of caspofungin (< or =0.015 mg/l) at 24 and 48 h for all Aspergillus tested. The agreement between the methods was excellent (100%) for micafungin and anidulafungin for all Aspergillus species tested. The three echinocandins were inactive against all isolates of Fusarium spp. showing MECs and MICs >8 mg/l. The Etest method could be a suitable procedure to test the susceptibility of most Aspergillus species to caspofungin, micafungin and anidulafungin; the best agreement was at 24 h.

Activity of BAL 4815 against filamentous fungi.

OBJECTIVES: BAL 4815 is a new antifungal drug and it is the active component of the antifungal triazole BAL 8557 (the water-soluble precursor). We studied the in vitro fungistatic and fungicidal activities of BAL 4815 against 103 clinical isolates of filamentous fungi, including 51 isolates of Aspergillus spp. and 52 isolates of non-Aspergillus filamentous fungi. METHODS: We evaluated the in vitro activity of BAL 4815 against 51 isolates of Aspergillus spp., 20 isolates of dematiaceous fungi, 18 isolates of hyaline Hyphomycetes and 14 isolates of Zygomycetes. MICs were determined following the CLSI M38-A broth microdilution method, using RPMI 1640 medium buffered to pH 7.0 with MOPS. Microdilution plates were incubated at 35 degrees C and read at 24 and 48 h (Mucorales were read at 24 h). Minimal fungicidal concentrations were also determined. RESULTS: For all isolates, geometric mean MICs, MIC(50)s, MIC(90)s and MIC ranges (mg/L) were: Aspergillus spp., 1.67, 2, 4 and 0.5-4; dematiaceous fungi, 1.62, 1, >8 and 0.03 to >8; hyaline Hyphomycetes, 2.41, 2, >8 and 0.03 to >8; and Zygomycetes, 6.81, 8, >8 and 0.03 to >8. Differences in susceptibility between genera were noted. Scedosporium prolificans, Fusarium spp., Mucor spp. and Rhizopus spp. (MIC(90) > 8 mg/L) were less susceptible than Aspergillus spp. (MIC(90) = 4 mg/L). CONCLUSIONS: BAL 4815 has excellent in vitro activity against Aspergillus spp. and variable activity against other filamentous fungi.

Comparison of the Sensititre YeastOne colorimetric antifungal panel with the modified Clinical and Laboratory Standards Institute broth microdilution (M38-A) method for antifungal susceptibility testing of dermatophytes.

BACKGROUND: To compare the susceptibility of different dermatophyte species to itraconazole (I), fluconazole (F) and voriconazole (V) by the modified reference (microdilution, CLSI M38-A) and the colorimetric method Sensititre YeastOne. The microdilution method is not very practical for use in routine susceptibility testing in the clinical laboratory, thus necessitating the use of other methods. METHODS: We studied a total of 46 dermatophyte strains isolated from clinical specimens. The microdilution reference susceptibility testing was performed following the CLSI M38-A document, using I, F and V drugs. The MIC were defined as the lowest drug concentration that produced 100% (I and V) and 50% inhibition (F) after 72 h incubation at 35 degrees C. The Sensititre MIC were detected by a change in color from pink to blue or purple. RESULTS: Agreement levels between the 2 methods (+/-2 dilutions) for I, F and V were 30, 53.3 and 83.3%, 0, 12.5 and 66.6% and 37.5, 44.4 and 75% for Trichophyton mentagrophytes, Trichophyton rubrum and Microsporumgypseum, respectively. The MIC(50/90) (mg/l) of I, F and V for T. mentagrophytes were 0.25/0.5, 16/64 and 0.12/0.25 by the microdilution method and 0.016/0.06, 8/16 and 0.03/0.06 by the Sensititre method. The MIC for I, F and V for T. rubrum were 0.25/1, 8/64 and 0.25/0.5 by the microdilution and 0.008/0.03, 2/8, 0.016/0.03 by the Sensititre method. For M. gypseum, MIC were 0.5/1 (I), /256 (F) and 0.25/1 (V) as well as 0.016/0.25 (I), 16/256 (F) and 0.06/0.25 (V) by the microdilution and Sensititre methods, respectively. CONCLUSIONS: The MICs obtained were lower by the Sensititre than the microdilution method. The best correlation between both methods was obtained for V in T. mentagrophytes (>80%), but was low for T. rubrum. Although the Sensititre method is easy to use in a clinical laboratory, it shows poor agreement with the reference method for dermatophytes.

Antifungal susceptibility of Cryptococcus neoformans isolates in HIV-infected patients to fluconazole, itraconazole and voriconazole in Spain: 1994-1996 and 1997-2005.

The antifungal drug susceptibility of 70 Cryptococcus neoformans isolates obtained in Spain from 1994 to 1996 (23 isolates) and from 1997 to 2005 (47 isolates) was investigated. The MICs of fluconazole, itraconazole and voriconazole were determined by the modified Clinical and Laboratory Standards Institute (CLSI; formerly National Committee for Clinical Laboratory Standards) broth microdilution method. The MIC(50) and MIC(90) for itraconazole and voriconazole did not change significantly from 1994 to 2005. The MIC(50) of fluconazole remained stable and the MIC(90) decreased by 2 log(2) dilution in the isolates collected from 1997 to 2005. We conclude that the in vitro resistance to fluconazole decreased over an 11-year period. In addition, a tendency for the development of possible cross-resistance between the three triazoles was observed.

Performance of the NG OligoGen kit for the diagnosis of Neisseria gonorrhoeae: comparison with cobas 4800 assay.

PCR assays are nowadays between the most sensitive and reliable methods for screening and diagnosing sexually transmitted infections (STIs). The aim of this study was to analyze the reliability, accuracy, and usefulness of the new NG OligoGen kit in comparison with the cobas 4800 assay for the detection of Neisseria gonorrhoeae in clinical samples. A prospective study was designed for detection of N. gonorrhoeae including urine samples (n=152), rectal (n=80), endocervical (n=67), pharyngeal (n=41), and urethral swabs (n=5) that were sent from a regional STI clinic in Seville, Spain. Samples were collected from 255 (73.9%) men and 90 women. Sensitivity, specificity, positive and negative predicative values, and kappa value for N. gonorrhoeae detection using the NG OligoGen kit were 99.6%, 100%, 100%, 99.1%, and 0.99, respectively. Statistical data obtained in this study confirm the usefulness and reliable results of this new assay.

Evaluation of the new API Candida system for identification of the most clinically important yeast species.

The API Candida system (bioMérieux) a new yeast identification system, was evaluated for its reliability in identifying 198 clinical yeast isolates in comparison with the API 20C system (bioMérieux) that was used as reference standard. The API Candida system correctly identified 91.4% and 71.7% of the isolates, with and without additional tests, respectively. The API Candida system identified 96.3% of common isolates studied, and 66.6% of uncommon isolates. We think that API Candida system is an easy and good yeast identification system and it could be used in a routine clinical microbiology laboratory.

Evaluation of CHROMagar Candida medium for the isolation and presumptive identification of species of Candida of clinical importance.

CHROMagar candida is a new differential culture medium that allows the isolation and presumptive identification of species of yeast of clinical importance. We tested 618 strains of yeast, including 128 direct isolates from clinical specimens. After two days of incubation at 37 degrees C, 339 of 341 Candida albicans, 98 of 99 Candida glabrata, all the Candida tropicalis, and all the Candida krusei were identified correctly. The sensitivity and specificity in these cases were both superior to 99%. Easy to prepare, with low cost, CHROMagar Candida proves to be a useful medium for the identification of species of yeast that are isolated with greater frequency in clinical material and for the identification of mixed cultures.

[Evaluation of a new chromogenic medium (Candida ID) for the isolation and presumptive identification of Candida albicans and other medically important yeasts]. / Evaluación micológica de un nuevo medio de cultivo cromógeno (Candida ID) para el aislamiento e identificación presuntiva de Candida albicans y otras levaduras de interés médico.

Candidiasis is a frequent human infection caused mainly by Candida albicans. However, other species are emerging as important pathogens, as Candida glabrata, Candida parapsilosis, Candida tropicalis, Candida krusei or Candida guilliermondii. Rapid identification of clinical isolates could facilitate diagnosis and treatment. Candida ID (bioMerieux, Spain) is a new medium for the isolation and presumptive identification of yeasts: C. albicans grows as blue colonies, and C. tropicalis, C. guilliermondii, Candida kefyr and Candida lusitaniae as pink ones. The utility of Candida ID was evaluated with more than 700 clinical isolates and type culture collection strains from different genera including Candida, Cryptococcus, Saccharomyces, and Rhodotorula. Presumptive identification was confirmed by germ tube test, microscopic morphology and chlamydoconidia production on corn meal agar and carbohydrate assimilation on API-ATB ID 32C or Vitek (bioMerieux). Growth on Candida ID was rapid (18-24 h) for most of the yeast strains tested. Sensitivity and specificity of identification of C. albicans was significantly high (>98%), since a very low number of isolates were found to be false negative or false positive. A better result was obtained for species growing as pink colonies (>99.5%). Detection of different species of medical important yeasts was easy with Candida ID, as perfectly distinct colors and textures of colonies were observed on this medium. Candida ID allowed the discrimination between C. glabrata (creamy and smooth) and C. krusei (rough and white) colonies. Other species showed different colony textures and colours, white being the predominant colour. Candida ID was very useful for the presumptive identification C. albicans isolates.

[Epidemiological study of cryptococcosis in Spain: first results]. / Estudio epidemiologico de la criptococosis en España: primeros resultados.

The study constitutes an approach to the knowledge of the epidemiology of cryptococosis in Spain. For detection of cases 167 Spanish hospitals were contacted. All cases included were accompanied by the correspondent isolate of Cryptococcus neoformans, together with clinical, demographic and mycological data. Results obtained from January 1998 to end of December 1999 are analysed and presented here. Fifty-six Spanish hospitals reported 58 cases of cryptococcosis; only 43 of them were adequately documented and accompanied by the clinical isolate. The results showed a higher incidence in males (88.4%) than in females (11.6%); being most frequently affected those between 30 and 40 years old (48.8%). The 84.6% (33) corresponded to new cases and 15.4% (6) to relapses of the disease. The HIV infection was the most frequent risk factor reported (86%) and, for 29.7% (11) of them, cryptococcosis was the AIDS defining disease. For the diagnosis, CSF analysis showed the best results (India ink; culture and antigen detection). All strains collected (100%) corresponded to C. neoformans variety neoformans. Serotypes distribution was 45.5% for serotype A and 22.7% for each of serotypes D and AD.

[In vitro activity of a liposomal nystatin formulation (Nyotran) against Cryptococcus neoformans]. / Actividad in vitro de una formulación liposómica de nistatina (Nyotran) frente a Cryptococcus neoformans.

The in vitro antifungal activity of a new liposomal nystatin formulation (NISTL, Nyotran, Aronex Ltd., EE.UU.) was evaluated by a microdilution method with RPMI based on the M27A document of the National Committee for Clinical Laboratory Standards (NCCLS) against 22 isolates of Cryptococcus neoformans. This antifungal activity was compared with those of other seven antifungal agents, such as nystatin (NIST), amphotericin B deoxycholate, liposomal amphotericin B, amphotericin B lipid complex, amphotericin B colloidal dispersion, fluconazole, and itraconazole. NISTL was more active in vitrothan NIST, showing MIC values 2-3 fold smaller in 90% of the isolates. The results obtained suggest that this new formulation would be very helpful for the treatment of cryptococcosis.

Multicenter evaluation of Neo-Sensitabs, a standardized diffusion method for yeast susceptibility testing.

The agar diffusion method Neo-Sensitabs for sensitivity testing, was evaluated with 33 reference strains by fourteen laboratories. Tablets with 5-fluorocytosine, amphotericin B, nystatin, fluconazole, itraconazole, ketoconazole and tioconazole were used on Shadomy modified medium. These tests classify each strain as susceptible, intermediate or resistant to all tested antifungals by measuring the inhibition zone diameters. Intra and interlaboratory reproducibility was studied. Neo-Sensitabs sensitivity for fungi was easy to perform and reliable method with a reproducibility of 97.1% and superior to other commercialized methods, being specially interesting for antifungal susceptibility in vitro testing of triazole derivatives fluconazole and itraconazole.

Multicenter survey of in vitro antifungal resistance in yeasts of medical importance isolated from Spanish patients.

Twelve Spanish laboratories collected 325 yeast clinical isolates during a 30 day's period, among them 224 Candida albicans, 30 Candida glabrata, and 27 Candida parapsilosis. In vitro antifungal susceptibility to amphotericin B, ketoconazole, fluconazole and itraconazole was determined by an agar diffusion test (Neo-Sensitabs, Rosco, Denmark). All the isolates tested were susceptible in vitroto amphotericin B and nearly all (97.2%) to itraconazole. In vitrosusceptibility to fluconazole and ketoconazole was high (90.2% and 91.4% of isolates, respectively) but showed variations depending on the species tested. Resistance to fluconazole and ketoconazole was low in C. albicans (4% and 3%, respectively), but 30% of Candida guilliermondii and 36% of C. glabrata isolates were resistant to fluconazole. Ketoconazole resistance was observed in 40% of C. glabrata, and 17% of Candida tropicalis. Resistance to antifungal drugs is very low in Spain and it is related to non-C. albicans isolates.

[Activity of trovafloxacin and ten other antimicrobials against gram negative anaerobic bacilli]. / Actividad de trovafloxacino y otros diez antimicrobianos frente a bacilos gramnegativos anaerobios.

As an increase in anaerobes resistant to a great variety of antimicrobials have been seen in the last few years, a search for new agents with activity against these microorganisms is needed. One of these agents is trovafloxacin. In our study, all strains were susceptible to imipenem, metronidazole, chloramphenicol and piperacillin/tazobactam. A total of 97% of the strains were inhibited with 2 mg/ml of trovafloxacin. The microorganisms Bacteroides fragilis and B. uniformis showed the least susceptibility against the antimicrobials studied, with a MIC90 for trovafloxacin of 1 and 2 mg/ml, respectively. Fusobacterium spp. were the most susceptible, with an MIC90 for trovafloxacin of 0.5 mg/ml. Trovafloxacin showed good activity against Gram-negative anaerobe rods, and could therefore be considered as an alternative in the treatment of the infections produced by these microorganisms.

[Seroepidemiologic study of human herpesvirus-6 in intravenous drug addicts, with and without infection caused by human immunodeficiency virus type 1]. / Estudio seroepidemiológico del virus herpes humano 6 en adictos a drogas por vía intravenosa, con y sin infección por el virus de la inmunodeficiencia humana tipo 1.

BACKGROUND: The human herpesvirus-6 (HHV-6) may be a cofactor of infection by the human immunodeficiency virus type 1 (HIV-1). However, there are discrepancies with respect to the possible epidemiological relation between both viruses. The aim of the present study was to study the prevalence of infection by the HHV-6 in intravenous drug addicts (IVDA) with and without HIV-1 infection. METHODS: IgG antibodies vs HHV-6 (anti-HHV-6-IgG) were determined by indirect immunofluorescence in 100 IVDA (29 seronegative and 71 seropositive for HIV-1 of which 45 were in stage II and 26 in IV-C1 of CDC) as well as in 100 healthy subjects of a similar age (control group). RESULTS: The prevalence of anti-HHV-6-IgG was much higher in the whole group of IVDA than in the control group and was equal in the IVDA with HIV-1 infection and in those patients without infection. There was no significant difference between the latter and the control group with the same being seen between the IVDA in different stages of HIV-1 infection. CONCLUSIONS: The results of this study suggest the existence of an epidemiological relation between human herpes virus-6 (HHV-6) infection and human immunodeficiency virus -1 (HIV-1). However, infection by the HHV-6 has no relation with the evolutive degree of the HIV-1 infection nor with intravenous drug addiction.