Nosocomial Candiduria in the Elderly: Microbiological Diagnosis.

Candiduria is associated with high morbidity, mortality, and long hospitalization, involving high costs for the healthcare system. The use of increasingly aggressive treatments has prolonged the lives of patients susceptible to candiduria, namely the immunosuppressed, the premature, and the elderly. Our objective was to evaluate the incidence of nosocomial candiduria and the implicated species in hospitalized patients aged over 80 years old from three Spanish centers during 2012 and 2013. Urine samples received from these patients were cultured and analyzed by flow cytometry in search of leukocyturia, hematuria, proteinuria, and microbial nitrate reductase activity. The isolated yeast species were identified microscopically, by germ tube formation in serum, colony morphology after subculture onto CHROMagar Candida (Becton-Dickinson, UK), assimilation of carbon compounds ID32C (bioMérieux, France), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDITOF) (Bruker Daltonics, Germany) and, in case of inconsistency, by sequencing of the ITS regions of ribosomal DNA (ITS1-5, 8S-ITS2). Susceptibility tests were also performed. The incidence of candiduria in the elderly population was 10.3%. A total of 155 strains of yeasts were isolated. The predominant species was Candida albicans, followed by Candida glabrata and then Candida tropicalis. Several infrequent species were found; among them, the first isolate of candiduria-producing Candida pulcherrima described in the literature. Our finding should raise concerns about the elderly population, which is probably the most important risk group for candiduria in the present moment, and the emergence of unusual yeast species producing candiduria, which are resistant against the commonly used antifungal agents.

[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.

[Maternal chorioamnionitis and neonatal conjunctive infection due to an infrequent pathogen]. / Corioamnionitis materna e infección conjuntival neonatal por un patógeno infrecuente.

Chorioamnionitis generates significant neonatal mortality and morbidity. Its incidence in premature birth can reach 30% and up to 30-40% of cases of premature rupture of membranes is due to this entity. We describe a case of chorioamnionitis by a commensal of the oral flora (Eikenella corrodens) in a pregnant woman with premature rupture of membranes and preterm delivery, which caused conjunctivitis in the newborn. On occasion of this case, we review the issue, delving into the diagnosis and clinical significance of this pathogen.

[Diagnosis of vaginitis-vaginosis by hibridization with DNA strands]. / Diagnóstico de vaginitis-vaginosis mediante hibridación con sondas de ADN.

BACKGROUND: Vaginal infections lie among the most common causes women ask for medical advice. In order of frequency bacterial vaginosis, vulvovaginal candidiasis, and trichomoniasis are responsible for 90% of vaginitis/vaginosis. OBJECTIVE: To evaluate a DNA hybridization test for simultaneous molecular detection of Gardnerella vaginalis, Candida species and Trichomonas vaginalis, as an alternative to conventional microbiological methods. MATERIAL AND METHODS: Cohort, cross-sectional, and comparative study of 1,003 vaginal samples from symptomatic women from our health-care area. Two swabs were obtained from each woman, one for routine microbiological diagnosis of vaginal infection (wet mount, Gram stain, and mycological culture) and the other for the DNA hybridization test (Affirm VPIII, Becton Dickinson). This method detects clinically significant levels of G. vaginalis (2 x 10(5) CFU/ml), Candida spp. (1 x 10(4) cells) and T. vaginalis (5x103 trichomonads). RESULTS: Out of the 1,003 women studied, 30.6% tested positive for bacterial vaginosis, 23.3% for vulvovaginal candidiasis, and 0.5% for trichomoniasis. The Affirm VPIII method turned out positive in 27.5%, 27.4% and 0.5% of cases, respectively. No statistically significant differences were found between the molecular technique and conventional methods for microbiological diagnosis of vaginitis/ vaginosis (p < 0.05). CONCLUSION: The Affirm VPIII test correlated well with wet mount, Gram stain and mycological culture. Although its cost is relatively high, it is fast, reproducible, easy, and can be done in either clinical laboratories or Gynecology offices, which permits prescribing a specific early treatment.

[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.

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.

[Tinea capitis by Microsporum gypseum, an infrequent species]. / Tiña capitis por Microsporum gypseum, una especie infrecuente.

Tinea capitis is considered the most frequent dermatophyte infection in children. The etiological agents vary from time to time and by geographical area, although they normally are zoophilic dermatophytes and in the last years also anthropophilic species. We report a clinical case of inflammatory tinea capitis in a 6-year-old child caused by Microsporum gypseum, a geophilic fungus pathogenic to humans and animals. The sources of human infection are soil, cats, dogs and small mammals. This species is less frequent as a cause of dermatophytosis in humans, described mainly in tinea corporis and rarely in tinea capitis. In the diagnosis of tinea capitis identifying the causative species is a determinant of the treatment.

La tiña capitis es considerada la infección por dermatofitos más frecuente en los niños. Los agentes etiológicos varían con el tiempo y según la zona geográfica, aunque, normalmente, son dermatofitos de origen zoofílico y, en los últimos años, también dermatofitos antropofílicos. Se presenta un caso de tiña capitis inflamatoria en un niño de 6 años de edad causada por Microsporum gypseum, un hongo geofílico patógeno para humanos y animales. Las fuentes de infección humana son el suelo, los gatos, los perros y pequeños mamíferos. Esta especie es poco frecuente como causa de dermatofitosis en el hombre, descrita, sobre todo, en tiña corporis y, raramente, en tiña capitis. En el diagnóstico de tiña capitis, identificar la especie causal es un factor determinante para el tratamiento.

[Fungemia due to Trichosporon asahii in a patient with hematological malignancy]. / Fungemia por Trichosporon asahii en un paciente con neoplasia hematológica.

BACKGROUND: Trichosporonosis is an opportunistic infection caused by the genus Trichosporon. The majority of cases of invasive trichosporonosis occurs in immunocompromised individuals. CASE REPORT: We describe a case of disseminated infection by Trichosporon asahii in a hematology patient. A 52-year-old man diagnosed with acute lymphoblastic leukemia developed a febrile episode during the third cycle of the induction chemotherapy. The blood cultures were positive after 24h incubation, showing elongated structures compatible with fungal elements in the Gram stain. The identification of the fungus as Trichosporon asahii was carried out by the assimilation of compounds of carbon and the amplification and sequencing of the D1/D2 domain and the internal transcribed spacer of the ribosomal DNA. The fungus was also isolated from the pustular lesions that the patient had in the chest. After treatment with amphotericin B, the patient progressed satisfactorily. CONCLUSIONS: Trichosporon asahii is an emergent pathogen in immunosupressed patients and its presence should not be considered as colonization, as there is risk of invasive infection.

[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.

Tiña capitis por Microsporum gypseum, una especie infrecuente / Tinea capitis by Microsporum gypseum, an infrequent species

La tiña capitis es considerada la infección por dermatofitos más frecuente en los niños. Los agentes etiológicos varían con el tiempo y según la zona geográfica, aunque, normalmente, son dermatofitos de origen zoofílico y, en los últimos años, también dermatofitos antropofílicos. Se presenta un caso de tiña capitis inflamatoria en un niño de 6 años de edad causada por Microsporum gypseum, un hongo geofílico patógeno para humanos y animales. Las fuentes de infección humana son el suelo, los gatos, los perros y pequeños mamíferos. Esta especie es poco frecuente como causa de dermatofitosis en el hombre, descrita, sobre todo, en tiña corporis y, raramente, en tiña capitis. En el diagnóstico de tiña capitis, identificar la especie causal es un factor determinante para el tratamiento.

Tinea capitis is considered the most frequent dermatophyte infection in children. The etiological agents vary from time to time and by geographical area, although they normally are zoophilic dermatophytes and in the last years also anthropophilic species. We report a clinical case of inflammatory tinea capitis in a 6-year-old child caused by Microsporum gypseum, a geophilic fungus pathogenic to humans and animals. The sources of human infection are soil, cats, dogs and small mammals. This species is less frequent as a cause of dermatophytosis in humans, described mainly in tinea corporis and rarely in tinea capitis. In the diagnosis of tinea capitis identifying the causative species is a determinant of the treatment.