THE PREVALENCE OF ONYCHOMYCOSIS IN THE ZAGREB AREA - THE ROLE OF SABOURAUD AGAR WITHOUT CYCLOHEXIMIDE IN DIAGNOSIS AND THERAPY.

- Mycological analysis is considered the gold standard in the diagnosis of very common fungal diseases of nails. The purpose of the article is to present the prevalence of onychomycosis obtained by using direct microscopic examination (DME) and cultivation on Sabouraud media without cycloheximide. The quantitative retrospective research was conducted on 2706 patients of both sexes (mostly middle-aged to elderly) with various toenail lesions, who had undergone mycological analyses in the Mycological Laboratory of our Department of Dermatovenereology in 2013 and 2014. Analysis of the results obtained revealed that there were 35.4% of positive DME results; of these, there were 39.2% of dermatophytes, 59.1% of yeasts and 1.7% of molds. The percentage of dermatophytes (Trichophyton spp.) was twice as high in men as in women. A considerable percentage (12%) of mixed isolates was also observed. Particularly worrisome was the high percentage (one-third) of suppressed and insufficiently developed dermatophytes (a possible indicator of inappropriate preparation of patients for mycological analysis). Cultivation on culture media without cycloheximide requires frequent inspections because of the fast-growing molds while not allowing sporulation for a great number of dermatophytes. However, it has certain advantages, i.e. it offers an insight into the wide range of the present fungi. As part of the biofilm, they can be potentially pathogenic in dystrophic nails under certain circumstances because, according to our results, the percentage of dermatophytes does not change substantially with aging; however, what does change is the percentage of molds, yeasts and mixed isolates.

Diagnosis of Dermatophytosis Using Molecular Biology.

Identification of fungi in dermatological samples using PCR is reliable and provides significantly improved results in comparison with cultures. It is possible to identify the infectious agent when negative results are obtained from cultures. In addition, identification of the infectious agent can be obtained in 1 day. Conventional and real-time PCR methods used for direct fungus identification in collected samples vary by DNA extraction methods, targeted DNA and primers, and the way of analysing the PCR products. The choice of a unique method in a laboratory is complicated because the results expected from skin and hair sample analysis are different from those expected in cases of onychomycosis. In skin and hair samples, one dermatophyte among about a dozen possible species has to be identified. In onychomycosis, the infectious agents are mainly Trichophyton rubrum and, to a lesser extent, Trichophyton interdigitale, but also moulds insensitive to oral treatments used for dermatophytes, which renders fungal identification mandatory. The benefits obtained with the use of PCR methods for routine analysis of dermatological samples have to be put in balance with the relative importance of getting a result in a short time, the price of molecular biology reagents and equipment, and especially the time spent conducting laboratory manipulations.

Micosis superficiales: casuística del Departamento de Micología del Instituto Nacional de Higiene “Rafael Rangel”, Caracas,Venezuela (2001-2014) / Superficial mycoses: casuistry of the Mycology Department of the Instituto Nacional de Higiene “Rafael Rangel”, Caracas, Venezuela (2001-2014)

Las micosis superficiales son muy comunes y por ello son motivo de consulta médica frecuente. El objetivo de este trabajo fue conocer la frecuencia de diagnóstico de las micosis superficiales en el Departamento de Micología del Instituto Nacional de Higiene “Rafael Rangel” en Caracas, Venezuela, durante 14 años (2001-2014). Se realizó un estudio transversal y retrospectivo de revisión de historias micológicas de pacientes con diagnóstico presuntivo de micosis superficial. Las muestras procesadas fueron uñas, pelos y escamas epidérmicas. La identificación de los hongos se realizó mediante observación macro y microscópica de las colonias y pruebas de identificación bioquímicas y fisiológicas, según requerimiento del agente aislado. Para la investigación de Malassezia spp. solo se realizó examen directo. De las 3228 muestras procesadas, 1098 (34%) resultaron positivas y su distribución según el agente etiológico fue: 79,5% dermatofitos; 10,9% levaduras; 5,1% hongos no dermatofitos y 4,5% Malassezia spp. El dermatofito más aislado fue el Complejo Trichophyton rubrum (70,1%), seguido del Complejo T. mentagrophytes (15,1%), Microsporum canis (9,4%) y Epidermophyton floccosum (4%). Las tiñas más frecuentes fueron: Tinea unguium (66,8%), seguida de Tinea pedis (16,4%) y Tinea capitis (8,1%). En el grupo de levaduras el Complejo Candida parapsilosis (37,5%) fue el más aislado y entre los hongos no dermatofitos el más frecuente fue Fusarium spp. (53,6%), seguido de Aspergillus spp. (19,6%) y Acremonium spp. (10,7%). La identificación del agente etiológico es fundamental para orientar un tratamiento adecuado. Esta casuística constituye un aporte importante para el conocimiento de la epidemiología de las micosis superficiales en nuestro país.

The superficial mycoses are very common infectious diseases and therefore are a frequent reason for medical consultation. The aim of this study was to determine the diagnostic frequency of superficial mycoses in the Mycology Department of the Instituto Nacional de Higiene “Rafael Rangel” during 14 years (2001-2014). A retrospective cross-sectional study was performed to review the mycological records of patients with presumptive diagnosis of superficial mycosis. Nails, hairs and epidermal scales were the processed samples. The identification of fungi was performed by macro and microscopic observation of colonies and biochemical and physiological tests, as required of the isolated agent. For the investigation of Malassezia spp. only direct examination was performed. Of the 3 228 samples processed, 1 098 (34%) were positive and their distribution according to the etiological agent was: dermatophytes 79.5%; 10.9% yeasts; non-dermatophytes fungi 5.1% and 4.5% Malassezia spp. The most frequently isolated dermatophyte was Trichophyton rubrum Complex (70.1%), followed by T. mentagrophytes complex (15.1%), Microsporum canis (9.4%) and Epidermophyton floccosum (4%). The most frequent ringworms were: Tinea unguium (66.8%), followed by Tinea pedis (16.4%) and Tinea capitis (8.1%). Candida parapsilosis complex (37.5%) was the most frequently isolated yeast and Fusarium spp. (53.6%) was the most isolated among non-dermatophyte fungi, followed by Aspergillus spp. (19.6%) and Acremonium spp. (10.7%). The identification of the etiological agent is essential to guide appropriate treatment. This study constitutes an important contribution to the knowledge of the epidemiology of superficial mycoses in our country.

Advances in Microbiology, Infectious Diseases and Public Health: Fungal Occurrence in the Hair and Skin of Symptomatic Pets in Turin, Italy.

Companion animals, often asymptomatic reservoir of fungi, can be important sources of infection in humans, due to the close contact with their owners. The present study was aimed to assess the occurrence of dermatophytes and other fungi isolated from pet dermatological lesions in Turin, Italy. Dermatological specimens were examined for fungal elements by direct microscopy and cultured to detect dermatophytes, other filamentous fungi and yeasts: 247 pets (118 cats, 111 dogs and 18 dwarf rabbits) were positive for fungal detection in culture. Microsporum canis was the most frequent dermatophyte in cats and dogs, whereas Trichophyton mentagrophytes was the most common in rabbits. Among the other fungi, for all examined pets, dematiaceous fungi were the most isolated, followed by Mucorales, penicilli, yeasts and yeast-like fungi, and aspergilli. No gender predisposition was detected for dermatophyte growth; on the contrary, for the other fungi male cats were more susceptible than female. The highest fungal occurrence was recorded in <1-year-old cats for dermatophytes, and in <5-year-old cats and dogs for the other fungi. Autumn was the period associated with a relevant incidence of fungal infection. Finally, fungi were more frequent in non pure-breed cats and in pure-breed dogs. These data underline the importance to timely inform pet owners about the potential health risk of infection caused not only by dermatophytes but also by non-dermatophyte fungi, routinely considered to be contaminants or harmless colonizers, since their role as source of zoonotic infections is not to be excluded.

Primer caso de onicomicosis causada por Cylindrocarpon lichenicola, en un paciente inmunosuprimido en Costa Rica / First case of onychomycosis caused by Cylindrocarpon lichenicola in an immunosuppressed patient un Costa Rica

Antecedentes: la onicomicosis es una infección fúngica de las uñas, que puede ser causada por dermatofitos, levaduras y hongos filamentosos no dermatofitos. El objetivo del presente trabajo fue identificar-morfológica y molecularmente- un aislamiento realizado a partir de la uña del dedo medio de la mano derecha de un paciente inmunosuprimido con trasplante renal. Métodos: a la muestra se le realizó un examen microscópico directo en KOH 40 por ciento, y las características de la colonia fueron evaluadas en medio de cultivo Agar Sabouraud Glucosado y Agar Mycosel. El hongo aislado se identificó molecularmente mediante la amplificación, por reacción en cadena de la polimerasa (PCR) y secuenciación de la región espaciadora transcrita (ITS) de los genes ribosomales y de una porción del gen del factor de elongación 1 alfa (EF-1). La secuencia de nucleótidos de cada región fue comparada con las secuencias publicadas en la base de datos del Anco de Genes del Centro Nacional para la Información Biotecnológica (NCBI). Resultados: las características macro y microscópicas de la colonia concordaron con las directrices para Cylindrocarpon lichenicola (sinónimo Fusarium lichenicola). La secuencia de nucleótidos, tanto de la región ITS (544 pb) como de la porción del gen EF-1alfa (710 pb), presentó un 100 por ciento de similitud con secuencias de F. lichenicola. Conclusión: se identificó por primera vez en Costa Rica C. lichenicola asociado con onicomicosis en un paciente inmunosuprimidos. Este tipo de pacientes deben ser instruidos sobre las posibles fuentes de agentes infecciosos, particularmente los riesgos que pueden generar la jardinería y la agricultura...