Candida nivariensis as a New Emergent Agent of Vulvovaginal Candidiasis: Description of Cases and Review of Published Studies.

Candida nivariensis is a new emergent agent related to human infections in the vaginal tract and other localizations, but the phenotypic characteristics are very similar to Candida glabrata and can be misidentified and underdiagnosed. We described four cases of vulvovaginitis identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and confirmed the results with PCR amplification and sequencing of the entire ITS genomic region (ITS1, ITS2 and 5.8 rRNA). We reinforce the need for new diagnostic tools for the correct identification of yeast infections.

[Infections due to Mycobacterium simiae]. / Infecciones por Mycobacterium simiae.

Mycobacterium simiae is a slow-growing photochromogenic environmental mycobacterium, first described in 1965. Rarely associated with human infections, possibly due to its limited pathogenicity, it mainly produces lung infection in immunocompetent elderly patients with underlying lung disease, and in disseminated infections in immunosuppressed young patients with AIDS. A microbiological culture is needed to confirm the clinical suspicion, and genetic sequencing techniques are essential to correctly identify the species. Treating M. simiae infections is complicated, owing to the multiple resistance to tuberculous drugs and the lack of correlation between in vitro susceptibility data and in vivo response. Proper treatment is yet to be defined, but must include clarithromycin combined with other antimicrobials such as moxifloxacin and cotrimoxazole. It is possible that M. simiae infections are undiagnosed.

[Evaluation of mass spectrometry for the identification of clinically interesting yeasts]. / Evaluación de la espectrometría de masas en la identificación de levaduras de interés clínico.

INTRODUCTION: Identification of yeasts is based on morphological, biochemical and nutritional characteristics, and using molecular methods. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, a new method for the identification of microorganisms, has demonstrated to be very useful. The aim of this study is to evaluate this new method in the identification of yeasts. METHODS: A total of 600 strains of yeasts isolated from clinical specimens belonging to 9 genera and 43 species were tested. Identification was made by sequencing of the ITS regions of ribosomal DNA, assimilation of carbon compounds (ID 32C), and mass spectrometry on a Microflex spectrometer (Bruker Daltonics GmbH, Germany). RESULTS: A total of 569 strains (94.8%) were identified to species level by ID 32C, and 580 (96.7%) by MALDI-TOF. Concordance between both methods was observed for 553 strains (92.2%), with 100% in clinically relevant species: C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, and almost 100% in C. krusei. MALDI-TOF identified species requiring molecular methods: Candida dubliniensis, C. nivariensis, C. metapsilosis and C. orthopsilosis. Some irregularities were observed in the identification of arthroconidia yeast and basidiomycetes. CONCLUSION: MALDI-TOF is a rapid, effective and economic method, which enables the identification of most clinically important yeasts and the differentiation of closely related species. It would be desirable to include more species in its database to expand its performance.

Candida galli as a cause of tinea unguium-molecular characterization of a rare clinical fungal entity.

We present the first clinical report of an infection caused by Candida galli, an anamorphic yeast species in the Yarrowia clade. C. galli has been described in the literature only four times, but never before it has been isolated from clinical samples. The colony morphology on Sabouraud medium and morphotype on CHROMagar Candida medium were similar to C. lipolytica as well as the carbon assimilation profile. The phenotypic differences with C. lipolytica were the non-assimilation of N-acetyl glucosamine, the absence of urease activity, growth in 10 % NaCl with 5 % glucose and in vitamin-free medium. MALDI-TOF MS could not generate reliable identification of the strain. Molecular analysis based on amplification of the ITS1-5.8S-ITS2 rDNA regions confirmed the identity as C. galli. Antifungal susceptibility test clearly demonstrated high MICs to 5-fluorocytosine, amphotericin B and fluconazole, as in the species belonging to the Yarrowia clade.

[Infections due to rapidly growing mycobacteria]. / Infecciones por micobacterias de crecimiento rápido.

Rapidly growing mycobacteria (RGM) are ubiquitous in nature and widely distributed in water, soil and animals. During the past three decades we have observed a notable increment of infections caused by RGM, both localized and disseminated, as well as nosocomial outbreaks of contaminated medical equipment. The microbiological diagnosis of RGM infections includes direct microscopic observation and culture. The taxonomic identification is performed by phenotypic, biochemical, chromatographic and molecular biology techniques. The treatment differs from that of other mycobacteriosis like tuberculosis, owing to the variable in vitro susceptibility of the species of this group. The RGM are resistant to conventional antituberculous drugs, but can be susceptible to broad spectrum antimicrobial agents. In this study we comment on the significant aspects of human infections by RGM, including their biology, epidemiology, pathology, microbiological diagnosis, taxonomic identification, antimicrobial susceptibility and treatment.

Urethritis due to Corynebacterium glucuronolyticum.

We describe a case of urethritis in a young man caused by Corynebacterium glucuronolyticum. This bacterium is a synonym of Corynebacterium seminale, a known agent of non-gonococcal urethritis, that cannot be regarded as commensal flora in the urogenital region when is isolated in a symptomatic clinical context. Accuracy in diagnosis and correct treatment is important for avoiding probable complications, for example prostatitis. Bacterial isolation is convenient, and an antimicrobial susceptibility test should be conducted to discover antimicrobial resistance. In our case the patient was successfully treated with fluoroquinolones and was symptom-free in 2 weeks.

Otomycosis due to filamentous fungi.

Otomycosis is common throughout the world but barely studied in Spain. Our objective was to determine the microbiological and epidemiological characteristics of this pathology in Cadiz (Spain) between 2005 and 2010. Samples from patients with suspicion of otomycosis underwent a direct microscopic examination and culture on different media for fungi and bacteria. Mycological cultures were incubated at 30°C for at least seven days. Identification of fungi was based on colonial morphology and microscopic examination of fungal structure. From a total of 2,633 samples, microbial growth was present in 1,375 (52.2%) and fungal isolation in 390 (28.4%). We identified 228 yeasts and 184 filamentous fungi (13.4% of positive cultures and 47.2% of otomycosis), associated with yeasts in 22 cases (5.6%). The most frequent species were Aspergillus flavus (42.4%), A. niger (35.9%), A. fumigatus (12.5%), A. candidus (7.1%), A. terreus (1.6%), and Paecilomyces variotii (0.5%). Infection was predominant in men (54.9%) and patients beyond 55 years old (46.8%). The most common clinical symptoms were itching (98.9%), otalgia (59.3%), and hypoacusis (56.0%). Fall season reported the lowest number of cases (20.1%). Incidence of otomycosis and fungi producing otomycosis vary within the distinct geographical areas. In Cadiz, this infection is endemic due to warm temperatures, high humidity, sea bathing, and wind, which contributes to disseminate the conidia. Despite Aspergillus niger has been reported as the main causative agent, A. flavus is predominant in Cadiz. Although infection is usually detected in warm months, we observed a homogeneous occurrence of otomycosis in almost all the seasons.

Evaluation of INNO-LiPA mycobacteria v2 assay for identification of rapidly growing mycobacteria.

A total of 54 rapidly growing mycobacteria (RGM) isolated from patients attended in the two hospitals of Cádiz Bay (Spain) were selected during a seven-year-period (2000-2006) in order to evaluate the INNO-LiPA Mycobacteria v2 assay for mycobacterial identification, based on the reverse hybridization principle. The strains were cultured in Löwenstein-Jensen and Middlebrook 7H9 media and identified to the species level by sequencing of the 16S rRNA, PCR-restriction enzyme analysis of the hsp65 gene, conventional tests and INNO-LiPA Mycobacteria v2 assay. By the molecular methods we identified a total of 12 different species: 23 Mycobacterium fortuitum, 11 M. chelonae, 10 M. abscessus, 2 M. senegalense, 1 M. alvei, 1 M. brumae, 1 M. mageritense, 1 M. mucogenicum, 1 M. neoaurum, 1 M. peregrinum, 1 M. septicum and 1 M. smegmatis. Fifty two strains (96.3%) were correctly identified by conventional techniques and 47 strains (87.0%) by INNO-LiPA Mycobacteria v2 assay. We find INNO-LiPA Mycobacteria v2 assay simple to perform but it provides few advantages in comparison with conventional methods and sometimes needs complementary tests to identify Mycobacterium fortuitum complex, M. chelonae complex and specific species due to the great heterogeneity in the RGM group.

[Fungemia caused by Candida lipolytica: Speaking of two cases]. / Fungemia por Candida lipolytica: a propósito de 2 casos.

Candida lipolytica has rarely been reported as a human pathogen. We observed two cases of fungemia caused by C. lipolytica, one of them in a 12-year-old child with cystic pancreatic fibrosis in advanced phase and another in a 86-year-old woman who presented vesical neoformation with peritoneal fibrosis, bilateral hydronephrosis and recurrent urinary tract infections. After antifungal treatment and catheter removal, the fungemia appeared to be finished and blood cultures were negative.

[Kodamaea ohmeri fungemia associated with surgery]. / Fungemia por Kodamaea (Pichia) ohmeri asociada a cirugía.

We report a case of fungemia caused by Kodamaea (Pichia) ohmeri associated with surgery in a patient with a history of diabetes and chronic renal failure. Kodamaea ohmeri is a yeast rarely involved in human infections. Since the first report of a case of fungemia in 1998, only thirteen cases of K. ohmeri infection have been so far described in the medical literature.

[Human infections by black yeasts of genus Exophiala]. / Infecciones humanas por levaduras negras del género Exophiala.

Clinical and laboratory aspects of the infections caused by Exophiala species are reviewed with regard to its epidemiology, diagnosis and treatment. Exophiala is a genus of dematiaceous hyphomycetes whose taxonomy and nomenclature undergo constant revision. Exophiala species are widely distributed in nature, and they are uncommon human pathogens. In recent years it appears to have increased its frequency as a cause of human infections, mainly in immunocompromised patients. They have been associated with phaeohyphomycosis, chromoblastomycosis, eumycotic mycetoma and disseminated infection. The procedures recommended for diagnosis consist of detection of fungal elements in tissue and growth of the organism in culture. Identification is mostly based upon microscopic observation of morphological characteristics and conidiogenesis, combined with the evaluation of physiological tests and nitrate and carbohydrate assimilations. Antifungal agents such as amphotericin B, itraconazole and voriconazole showed in vitro activity to most of the Exophiala species of clinical interest. The therapeutic recommendations are mainly deduced from the observation of single cases.

[Dermatophytoses due to Microsporum gypseum: report of eight cases and literature review]. / Dermatofitosis por Microsporum gypseum: descripción de ocho casos y revisión de la literatura.

Microsporum gypseum is a geophilic fungus infrequent agent of human dermatophytoses and world-wide in distribution. In Cadiz, Spain, between 1997 and 2003, a study of 133 positive cases showed that the fifth more isolated dermatophyte was M. gypseum (6.0%), followed by Trichophyton mentagrophytes (24,8%), Microsporun canis (24,6%), Trichophyton rubrum (21,8%) y Trichophyton violaceum (6,8%). During 2003 the infection due to this fungus has been repeatedly observed in our area (17.5%). We report herein eight new cases of infection by M. gypseum. Our epidemiological data were compared with those obtained by other authors in other regions of Spain and in those reported in other countries.

[Candida ciferrii in an immunocompromised patient]. / Aislamiento de Candida ciferrii en un paciente inmunodeficiente.

A case of possible infection due to Candida ciferrii in an immunocompromised patient is presented. This fungal species has been rarely reported as cause of human infection. The isolate showed in vitro resistance to fluconazole.

[Carbohydrate assimilation by clinical and environmental Rhodotorula glutinis strains]. / Asimilación de carbohidratos por cepas de Rhodotorula glutinis de origen clínico y ambiental.

This study was carried to determine the carbohydrate assimilation patterns of Rhodotorula strains isolated from clinical and environmental specimens. We have tested the commercial system ID 32C (bioMerieux, France) on 80 different strains of Rhodotorula glutinis: 47 strains from clinical samples and 33 strains from environmental samples. The assimilation percentages obtained in our study for galactose, cellobiose, gluconate and sorbose were lower than those showed in the identification table of the method. However, the assimilation percentages for mannitol and esculin were higher. According to our results, we conclude that the numerical profiles and the identification software of the commercial system present limitations for the characterization of some R. glutinis strains.

[Susceptibility to antifungal agents of Cryptococcus species of clinical interest]. / Sensibilidad a antifúngicos de especies de Cryptococcus de interés clínico.

BACKGROUND: Studies on the susceptability of Cryptococcus species to antifungal agents are scarce. METHOD: We investigate the susceptibility of 54 clinical and environmental strains from six different species using the Sensititre and the NCCLS methods. RESULTS: Environmental strains were less susceptible than clinical strains. We found MICs >= 64 mg/l for fluconazole, MICs >= 1 mg/l for itraconazole and MICs >= 32 mg/l for 5-fluorocytosine in C. albidus, C. neoformans and C. uniguttulatus. CONCLUSIONS: These findings point at the importance of determining the antifungal susceptibility of Cryptococcus species.

[Susceptibility to fluconazole of clinical interest yeasts: new breakpoints]. / Sensibilidad a fluconazol de levaduras de interés clínico: nuevos puntos de corte.

INTRODUCTION: Recently, Pfaller et al (Drug Resist Update 2010; 13:180-95), have proposed new breakpoints for determining the in vitro susceptibility to fluconazole of Candida albicans, C. parapsilosis and C. tropicalis. The aim of this study was to establish the variations in sensitivity of these species applying these breakpoints, in relation to those of the Clinical and Laboratory Standards Institute (CLSI). METHODS: We analyzed 112 strains of Candida: 49 C. albicans, 40 C. parapsilosis and 23 C. tropicalis. Susceptibility to fluconazole was performed by the method Sensititre YeastOne. The breakpoints used to determine the minimum inhibitory concentration (MIC) were identified by CLSI and the ones proposed by Pfaller et al. RESULTS: According to the CLSI criteria, all isolates were susceptible to fluconazole. MIC50 and MIC90 were 0.5 mg/L and 2 mg/L for C. albicans and C. parapsilosis, 0.5 mg/L and 1 mg/L for C. tropicalis. With the new criteria, 109 (97%) strains were susceptible. Variations were seen in C. albicans, with 3 strains (6%) susceptible dose-dependent. CONCLUSIONS: When applying the breakpoints recommended by Pfaller et al, and EUCAST, the number of fluconazole-susceptible strains decreased according to the CLSI criteria, especially C. albicans.