Evaluation of the MALDI-TOF mass spectrometry technique for the identification of dermatophytes: Use of an extended database.

BACKGROUND: Identification of dermatophytes is usually performed through morphological analyses. However, it may be hindered due to the discovery of new species and complexes and, with some isolates, by the absence of fructification. Matrix-assisted laser desorption/ionisation-time-of-flight mass spectrometry (MALDI-TOF MS) seems to be an option for improving identification. AIMS: To develop a database (DB) for the identification of dermatophytes with MALDI-TOF MS, including 32 isolates from the Red de Micología de la Ciudad Autónoma de Buenos Aires [Mycology Network of the Autonomous City of Buenos Aires] (RMCABA) and one reference isolate (RMCABA DB), and evaluate its performance when added to the DB from the supplier, Bruker (Bruker DB). METHODS: All the isolates in the RMCABA DB were identified based on morphology and sequencing. To evaluate the performance of the extended DB (Bruker DB plus RMCABA DB), 136 clinical isolates were included. RESULTS: The percentages of identification at the species level increased from 45% to 88%, but the identification at the genus level decreased from 23% to 7%. CONCLUSIONS: MALDI-TOF MS yielded better performance in the identification of dermatophytes after including the RMCABA DB, which encompassed local isolates.

Evaluation of the MALDI-TOF mass spectrometry technique for the identification of dermatophytes: Use of an extended database / Evaluación de la técnica de espectrometría de masas MALDI-TOF para la identificación de dermatofitos: utilización de una base de datos ampliada

Background: Identification of dermatophytes is usually performed through morphological analyses. However, it may be hindered due to the discovery of new species and complexes and, with some isolates, by the absence of fructification. Matrix-assisted laser desorption/ionisation-time-of-flight mass spectrometry (MALDI-TOF MS) seems to be an option for improving identification. Aims: To develop a database (DB) for the identification of dermatophytes with MALDI-TOF MS, including 32 isolates from the Red de Micología de la Ciudad Autónoma de Buenos Aires [Mycology Network of the Autonomous City of Buenos Aires] (RMCABA) and one reference isolate (RMCABA DB), and evaluate its performance when added to the DB from the supplier, Bruker (Bruker DB). Methods: All the isolates in the RMCABA DB were identified based on morphology and sequencing. To evaluate the performance of the extended DB (Bruker DB plus RMCABA DB), 136 clinical isolates were included. Results: The percentages of identification at the species level increased from 45% to 88%, but the identification at the genus level decreased from 23% to 7%.Conclusions: MALDI-TOF MS yielded better performance in the identification of dermatophytes after including the RMCABA DB, which encompassed local isolates.(AU)

Antecedentes: La identificación de dermatofitos se realiza, usualmente, a través del análisis morfológico, pero puede verse dificultada por la aparición de nuevas especies y complejos de especies y, en algunos aislamientos, por la falta de elementos de fructificación. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) aparece como una alternativa para mejorar la identificación. Objetivos: Construir una base de datos (BaDa) para la identificación de dermatofitos mediante MALDI-TOF MS, con 32 aislamientos de la Red de Micología de la Ciudad Autónoma de Buenos Aires (RMCABA) y una cepa de referencia (BaDa RMCABA), y evaluar su desempeño al incorporarla a la BaDa del proveedor Bruker (BaDa Bruker). Métodos: Todos los aislamientos incorporados a la BaDa RMCABA se identificaron por morfología y secuenciación. Para la evaluación de la BaDa ampliada (BaDa Bruker más BaDa RMCABA) se incluyeron 136 aislamientos clínicos. Resultados: El porcentaje de identificación a nivel de especie se incrementó del 45 al 88%, mientras que la identificación a nivel de género disminuyó del 23 al 7%. Conclusiones: Con la incorporación de la BaDa RMCABA, que contó con aislamientos regionales, se observó un mejor desempeño en la identificación de dermatofitos por MALDI-TOF MS.(AU)

Determinación de la concentración mínima inhibitoria de lotes de OLEOZON® tópico frente a Trichophyton rubrum, Trichophyton mentagrophytes y Epidermophyton floccosum

Introducción: En la actualidad las infecciones por hongos afectan entre el 20 y el 25 % de la población. Objetivo: Determinar la concentración mínima inhibitoria de dos lotes de OLEOZON® almacenados a temperaturas de 5 y 30°C en envases de vidrio y polietileno de alta densidad, durante 24 meses como parte del estudio de estabilidad del OLEOZON® tópico. Métodos: Mediante el método dilución en agar fueron evaluadas cinco concentraciones del producto frente a los dermatofitos trichophyton rubrum, trichophyton mentagrophytes y epidermophyton floccosum. Resultados: Se obtuvo que el 8,9 mg/mL fue el valor de la CMI para las cepas evaluadas en el estudio "vida de estante"; se observó en el estudio acelerado el mismo valor frente a las cepas de trichophyton mientras que para epidermophyton floccosum fue de 17,8 mg/mL a excepción del envase frasco de vidrio del lote 803295 donde se obtuvo 8,9 mg/mL. El análisis estadístico, tanto del estudio acelerado como en vida de estante, mostró que existe diferencia estadísticamente significativa entre el primer y el último mes de ensayo, estas son las diferencias más apreciables en los lotes almacenados en frasco de vidrio. El OLEOZON® tópico almacenado en frasco de polietileno de alta densidad presentó mejores valores de actividad frente a los dermatofitos. Conclusiones: Todos los valores de la concentración mínima inhibitoria encontrados, independiente del tipo de envase, el tiempo o la temperatura de almacenamiento, muestran que el producto mantiene su actividad antifúngica. Se evidenció una similitud entre las cepas del género trichophyton en comparación con el género epidermophyton.

Introduction: Currently fungal infections affect between 20 and 25% of the population. Objective: To determine the minimum inhibitory concentration of two batches of OLEOZON® stored at temperatures of 5 and 30 0C in glass and high-density polyethylene containers, for 24 months as part of the stability study of topical OLEOZON®. Methods: Using the agar dilution method, five concentrations of the product were evaluated against the dermatophytes Trichophyton rubrum, Trichophyton mentagrophytes and Epidermophyton floccosum. Results: It was found that 8.9 mg/mL was the MIC value for the strains evaluated in this shelf life study; and the same value was observed in the accelerated study against the trichophyton strains, while for epidermophyton floccosum it was 17.8 mg/mL with the exception of the glass bottle container of batch 803295 where 8.9 mg/mL was obtained. The statistical analysis, both in the accelerated study and in shelf life, showed that there is a statistically significant difference between the first and the last month of the trial, these are the most appreciable differences in the batches stored in glass jars. Topical OLEOZON® stored in a high-density polyethylene bottle presented better activity values against dermatophytes. Conclusions: All the values of the minimum inhibitory concentration found, regardless of the type of container, the time or the storage temperature, show that the product maintains its antifungal activity during the months of study. A similarity was apparent between the strains of the genus trichophyton compared to the genus epidermophyton.

Trichophyton benhamiae, un dermatofito zoofílico emergente en Argentina con reservorio en cobayos: descripción de 7 casos en un hospital de la Ciudad Autónoma de Buenos Aires / Trichophyton benhamiae, an emergent zoonotic pathogen in Argentina associated with Guinea pigs: Description of 7 cases in Buenos Aires

Resumen Trichophyton benhamiae es un dermatofito zoofílico. Puede causar tinea corporis, tinea faciei y tinea capitis. Se caracteriza por producir lesiones inflamatorias, sobre todo en niños. El objetivo de esta publicación es describir 7 casos clínicos de pacientes pediátricos atendidos entre julio del 2019 y enero del 2020 en nuestra institución. A los pacientes se les solicitó estudio micológico convencional, con posterior confirmación con MALDI-TOF MS y secuencia-ción del ADN ribosomal. Se aisló e identificó T. benhamiae como agente etiológico; el nexo epidemiológico fue el contacto con cobayos. Estas son las primeras descripciones de infecciones causadas por T. benhamiae en Argentina. Al realizar estudios micológicos convencionales, este agente puede confundirse con otros dermatofitos, por lo tanto, se requieren herramientas como MALDI-TOF MS o la secuenciación para llegar a un diagnóstico definitivo. Es importante contar con datos epidemiológicos, como el contacto con mascotas no tradicionales, para una presunción diagnóstica adecuada.

Trichophyton benhamiae is a zoonotic dermatophyte that can cause tinea corporis, tinea faciei and tinea capitis, producing inflammatory lesions, especially in children. In this publication, we describe 7clinical cases of pediatric patients that occurred in our institution between July 2019 and January 2020. All patients underwent a conventional mycological study. The identification of fungi isolates was confirmed by MALDI-TOF MS and sequencing of the ribosomal DNA. T. benhamiae was identified as the etiological agent, whose epidemiological link in all cases was the contact with Guinea pigs. This is the first description of infections caused by T. benhamiae in Argentina. This dermatophyte can be misidentified as other more frequent dermatophytes when performing conventional studies. Molecular technology should be used to reach a definitive diagnosis. It is important to have epidemiological data from patients such as contact with non-traditional pets, especially Guinea pigs, for an adequate presumptive diagnosis of this dermatophytosis.

Tiña capitis por Trichophyton tonsurans en un paciente pediátrico / Tinea capitis by Trichophyton tonsurans in a pediatric patient

La tiña capitis (TC) es una micosis superficial del cuero cabelludo, considerada una de las infecciones más frecuentes por dermatofitos en niños. Hasta la actualidad, las especies descritas con mayor frecuencia en nuestro medio son el Microsporum y, en segundo término, Trichophyton, los cuales se contraen principalmente por contacto directo con animales o seres humanos infectados, respectivamente. Se presenta el caso de una paciente de 8 años con alopecia y lesiones inflamatorias de 2 años de evolución, en la que finalmente se llegó al diagnóstico de TC inflamatoria causada por T. tonsurans, un hongo antropofílico considerado poco frecuente en Argentina y emergente en la provincia de Buenos Aires. Este patógeno presenta alta transmisibilidad; son varios los países que han reportado brotes escolares y comunitarios. Se debe resaltar la importancia de su sospecha clínica temprana para un tratamiento adecuado.

Tinea capitis (TC) is a superficial mycosis of the scalp, considered one of the most common dermatophyte infections in children. Until now, the species mainly described in our environment are Microsporum and secondly Trichophyton, which are contracted mainly by direct contact with infected animals or humans, respectively. We present the case of an 8-year-old patient with alopecia and inflammatory lesions of 2 years of evolution, finally reaching the diagnosis of inflammatory tinea capitis caused by T. tonsurans, an anthropophilic fungus considered rare in Argentina and emerging in the province of Buenos Aires. This pathogen has high transmissibility; several countries have reported school and community outbreaks. The importance of its early clinical suspicion for adequate treatment should be emphasized.

Chemical prospecting and antioxidant, anticholinesterase and antifungal activity of ethanol extracts from Senna Mill. species. (Fabaceae) / Prospecção química, atividade antioxidante, anticolinesterásica e antifúngica de extratos etanólicos de espécies de Senna Mill. (Fabaceae)

The objective of this work was to evaluate the chemical profile, total phenol content and the antioxidant, antiacetylcholinesterase and antifungal activities of ethanol extracts from leaves (EEF) and stem (EEC) of five species of Senna, as well as to evaluate the correlation between the content of total phenols with the DPPH free radical scavenging activity and with the inhibition of the acetylcholinesterase AChEI enzyme. The phytochemical test showed the presence of secondary metabolites in all extracts. The EEF and EEC of Senna trachypus, stood out for presenting the highest values of total phenols. S. trachypus EEF and EEC showed the best action on the acetylcholinesterase enzyme. All species showed antioxidant activity, especially the EEF from S. trachypus. A strong correlation was observed for the variables total phenols and antioxidant activity (r = -0.71), very strong for total phenols and anti-acetylcholinesterase (r = -0.84) and moderate activity for antioxidant and anti-acetylcholinesterase activity (r = 0.43). Only S. trachypus extracts showed activity against all dermatophytes, when compared to the other species. The species S. trachypus was considered the most promising plant for further studies, in order to find compounds with pharmacological actions.

O objetivo deste trabalho foi avaliar o perfil químico, teor de fenóis totais e as atividades antioxidante, antiacetilcolinesterásica e antifúngica dos extratos etanólicos das folhas (EEF) e do caule (EEC) de cinco espécies de Senna, bem como avaliar a correlação entre o conteúdo de fenóis totais com a atividade sequestradora do radical livre DPPH e com a inibição da enzima acetilcolinesterase AChEI. No teste fitoquímico, evidenciou-se a presença de metabólitos secundários em todos os extratos. Os EEF e EEC de Senna trachypus, destacaram-se por apresentar os maiores valores de fenóis totais. Os EEF e EEC de S. trachypus mostraram a melhor ação sobre a enzima acetilcolinesterase. Todas as espécies apresentaram atividade antioxidante, em especial o EEF de S. trachypus. Foi observado correlação forte para as variáveis fenóis totais e atividade antioxidante (r = -0,71), muito forte para fenóis totais e antiacetilcolinesterase (r = -0,84) e atividade moderada para atividade antioxidante e antiacetilcolinesterase (0,43). Apenas os extratos de S. trachypus apresentaram atividade contra todos os dermatófitos, quando comparado às demais espécies. A espécie S. trachypus foi considerada a planta mais promissora para estudos subsequentes, a fim de encontrar compostos com ações farmacológicas.

[Trichophyton benhamiae, an emergent zoonotic pathogen in Argentina associated with Guinea pigs: Description of 7cases in Buenos Aires]. / Trichophyton benhamiae, un dermatofito zoofílico emergente en Argentina con reservorio en cobayos: descripción de 7 casos en un hospital de la Ciudad Autónoma de Buenos Aires.

Trichophyton benhamiae is a zoonotic dermatophyte that can cause tinea corporis, tinea faciei and tinea capitis, producing inflammatory lesions, especially in children. In this publication, we describe 7clinical cases of pediatric patients that occurred in our institution between July 2019 and January 2020. All patients underwent a conventional mycological study. The identification of fungi isolates was confirmed by MALDI-TOF MS and sequencing of the ribosomal DNA. T. benhamiae was identified as the etiological agent, whose epidemiological link in all cases was the contact with Guinea pigs. This is the first description of infections caused by T. benhamiae in Argentina. This dermatophyte can be misidentified as other more frequent dermatophytes when performing conventional studies. Molecular technology should be used to reach a definitive diagnosis. It is important to have epidemiological data from patients such as contact with non-traditional pets, especially Guinea pigs, for an adequate presumptive diagnosis of this dermatophytosis.

Anti-dermatophyte activity of the aqueous extracts of Thai medicinal plants

Abstract Medicinal plants have long been prescribed in Thailand for centuries. Different constituents of extracts have been used for treating of various infectious diseases. However, there is even less information available regarding the use in fungal skin infection. In order to assess traditional Thai claims about the therapeutic potential, this study is focused on exploring the anti-dermatophyte property of the plants that are currently used as traditional medicines. The potential of four different plant species were selected for investigate in vitro anti-dermatophyte activity. Ethanolic extracts of Chromolaena odorata (L.), Ageratina adenophora (Spreng.), Eclipta prostrate (Linn.), and Acorus calamus (L.). were analysed for their total phenolic content as well as total flavonoid content and were then subjected to test of their anti-dermatophyte properties using agar well diffusion method. Qualitative flavonoids and phenolics analysis of the extracts showed their biologically active constituents. Among the species examined, the result indicated that most of the extracts demonstrated anti-dermatophyte activity. In particular, A. calamus showed the highest efficacy against test organisms. The experiment confirmed the chemical constituents and efficacy of some selected plants and provides a scientific confirmation of the use of Thai plants in traditional medicine for fungal skin infections.

Resumo As plantas medicinais são prescritas há muito tempo na Tailândia, há séculos. Diferentes constituintes de extratos têm sido usados para o tratamento de várias doenças infecciosas. No entanto, existem ainda menos informações disponíveis sobre o uso em infecções fúngicas da pele. A fim de avaliar as alegações tradicionais tailandesas sobre o potencial terapêutico, este estudo está focado em explorar a propriedade antidermatófita das plantas que são usadas atualmente como medicamentos tradicionais. O potencial de quatro espécies de plantas diferentes foi selecionado para investigar a atividade antidermatófita in vitro. Extratos etanólicos de Chromolaena odorata (L.), Ageratina adenophora (Spreng.), Eclipta prostrate (Linn.) e Acorus calamus (L.) foram analisados quanto ao seu conteúdo fenólico total, bem como ao conteúdo de flavonoides totais. E então submetidos ao teste de suas propriedades antidermatófitas usando o método de difusão em ágar bem. A análise qualitativa de flavonoides e fenólicos dos extratos mostrou seus constituintes biologicamente ativos. Entre as espécies examinadas, o resultado indicou que a maioria dos extratos demonstrou atividade antidermatófita. Em particular, A. calamus mostrou a maior eficácia contra organismos de teste. O experimento confirmou os constituintes químicos e a eficácia de algumas plantas selecionadas e fornece uma confirmação científica do uso de plantas tailandesas na medicina tradicional para infecções fúngicas da pele.

Anti-dermatophyte activity of the aqueous extracts of Thai medicinal plants / Atividade antidermatófita dos extratos aquosos de plantas medicinais tailandesas

Medicinal plants have long been prescribed in Thailand for centuries. Different constituents of extracts have been used for treating of various infectious diseases. However, there is even less information available regarding the use in fungal skin infection. In order to assess traditional Thai claims about the therapeutic potential, this study is focused on exploring the anti-dermatophyte property of the plants that are currently used as traditional medicines. The potential of four different plant species were selected for investigate in vitro anti-dermatophyte activity. Ethanolic extracts of Chromolaena odorata (L.), Ageratina adenophora (Spreng.), Eclipta prostrate (Linn.), and Acorus calamus (L.). were analysed for their total phenolic content as well as total flavonoid content and were then subjected to test of their anti-dermatophyte properties using agar well diffusion method. Qualitative flavonoids and phenolics analysis of the extracts showed their biologically active constituents. Among the species examined, the result indicated that most of the extracts demonstrated anti-dermatophyte activity. In particular, A. calamus showed the highest efficacy against test organisms. The experiment confirmed the chemical constituents and efficacy of some selected plants and provides a scientific confirmation of the use of Thai plants in traditional medicine for fungal skin infections.

As plantas medicinais são prescritas há muito tempo na Tailândia, há séculos. Diferentes constituintes de extratos têm sido usados para o tratamento de várias doenças infecciosas. No entanto, existem ainda menos informações disponíveis sobre o uso em infecções fúngicas da pele. A fim de avaliar as alegações tradicionais tailandesas sobre o potencial terapêutico, este estudo está focado em explorar a propriedade antidermatófita das plantas que são usadas atualmente como medicamentos tradicionais. O potencial de quatro espécies de plantas diferentes foi selecionado para investigar a atividade antidermatófita in vitro. Extratos etanólicos de Chromolaena odorata (L.), Ageratina adenophora (Spreng.), Eclipta prostrate (Linn.) e Acorus calamus (L.) foram analisados quanto ao seu conteúdo fenólico total, bem como ao conteúdo de flavonoides totais. E então submetidos ao teste de suas propriedades antidermatófitas usando o método de difusão em ágar bem. A análise qualitativa de flavonoides e fenólicos dos extratos mostrou seus constituintes biologicamente ativos. Entre as espécies examinadas, o resultado indicou que a maioria dos extratos demonstrou atividade antidermatófita. Em particular, A. calamus mostrou a maior eficácia contra organismos de teste. O experimento confirmou os constituintes químicos e a eficácia de algumas plantas selecionadas e fornece uma confirmação científica do uso de plantas tailandesas na medicina tradicional para infecções fúngicas da pele.

Commercial real time PCR implementation for rapid diagnosis of onychomycosis: A new workflow in a clinical laboratory.

INTRODUCTION: Onychomycosis is a frequent and underdiagnosed condition. Approximately 90% of toenail onychomycosis infections are caused by dermatophytes, but classical diagnosis based on culture and microscopy observation is slow and has low sensitivity. Both limitations can be solved incorporating molecular techniques to routine diagnosis of onychomycosis. OBJECTIVE: Prospective evaluation of the utility of incorporating in the clinical laboratory workflow a commercial real time PCR (qPCR) for dermatophytes detection in nails after potassium hydroxide direct observation screening. MATERIALS AND METHODS: 152 nail samples were included (34 KOH negative and 118 KOH positive) and processed by culture and qPCR. RESULTS: In the negative KOH group, only one dermatophyte grew in culture and three were detected by qPCR. In the group of positive KOH, 57 dermatophytes grew in culture and 81 were detected by qPCR. In this group, 25% of diagnosed dermatophytes were detected only by qPCR. The sensitivity of qPCR compared to culture is 92.8% and time of response decreases from days to hours. CONCLUSION: Based in our results, we propose a workflow algorithm for a clinical laboratory that eliminates culture for qPCR positive samples.

PCR a tiempo real comercial para diagnóstico rápido de las onicomicosis: un nuevo algoritmo de trabajo en el laboratorio clínico / Commercial real time PCR implementation for rapid diagnosis of onychomycosis: A new workflow in a clinical laboratory

Introduction: Onychomycosis is a frequent and underdiagnosed condition. Approximately 90% of toenail onychomycosis infections are caused by dermatophytes, but classical diagnosis based on culture and microscopy observation is slow and has low sensitivity. Both limitations can be solved incorporating molecular techniques to routine diagnosis of onychomycosis. Objective: Prospective evaluation of the utility of incorporating in the clinical laboratory workflow a commercial real time PCR (qPCR) for dermatophytes detection in nails after potassium hydroxide direct observation screening. Materials and methods: 152 nail samples were included (34 KOH negative and 118 KOH positive) and processed by culture and qPCR. Results: In the negative KOH group, only one dermatophyte grew in culture and three were detected by qPCR. In the group of positive KOH, 57 dermatophytes grew in culture and 81 were detected by qPCR. In this group, 25% of diagnosed dermatophytes were detected only by qPCR. The sensitivity of qPCR compared to culture is 92.8% and time of response decreases from days to hours. Conclusion: Based in our results, we propose a workflow algorithm for a clinical laboratory that eliminates culture for qPCR positive samples.(AU)

Introducción: La onicomicosis es una patología frecuentemente infradiagnosticada. Aproximadamente el 90% de las infecciones en las uñas del pie están causadas por dermatofitos, pero el diagnóstico microbiológico clásico basado en cultivo y microscopia es lento y tiene una baja sensibilidad. Ambas limitaciones pueden resolverse incorporando técnicas moleculares al diagnóstico de la onicomicosis. Objetivo: Evaluación prospectiva de la utilidad de incorporar en un laboratorio clínico una PCR a tiempo real (qPCR) comercial para detección de dermatofitos en uñas tras cribado por examen directo con hidróxido de potasio (KOH). Materiales y métodos: Se incluyeron 152 muestras de uñas (34 KOH negativas y 118 KOH positivas) y se procesaron mediante cultivo y qPCR. Resultados: En el grupo KOH negativo, solo un dermatofito creció en cultivo y 3 se detectaron mediante qPCR. En el grupo KOH positivo, 57 dermatofitos crecieron en cultivo y 81 se detectaron por qPCR. En este grupo, el 25% de los dermatofitos diagnosticados se detectaron únicamente mediante qPCR. La sensibilidad de la qPCR comparada con el cultivo es del 92,8% y el tiempo de respuesta disminuye de días a horas. Conclusión: En base a nuestros resultados, proponemos un algoritmo de flujo de trabajo para los laboratorios de microbiología clínica, que elimina el cultivo para aquellas muestras con qPCR positiva.(AU)

Dermatophytoses in patients attended from a private clinical analysis laboratory in João Pessoa-PB, between 2015 to 2019 / Dermatofitoses em pacientes atendidos em um laboratório privado de João Pessoa-PB, entre 2015 a 2019

Introduction: dermatophytoses or "tineas" are characterized by being mycoses caused by fungi of the genera Epidermophyton, Trichophyton and Microsporum. These mycotic infections can present themselves as a form of lesions that affect the skin, hair and nails of individuals of both genders and all ages. Objective: to elucidate the epidemiological profile of dermatophytoses in patients examined by a private clinical analysis laboratory in João Pessoa-PB, between 2015 and 2019. Methodology: this is an epidemiological, analytical, retrospective and documentary study, in which data collection took place at the Clinical Pathology Laboratory ­ "HEMATO", located in João Pessoa ­ PB. Results: the profile of those affected was predominantly female (58.5%), 18 to 59 years old (38.4%), white (53.6%) and with lesions, mainly in skin glabrous (38.5%), feet (33.3%) and nails (12.8%). When relating the age group to the injury site, it was noticed that injuries on glabrous skin, feet and nails, were more frequent in individuals aged 18 to 59 years, while injuries to the scalp were mostly found in individuals younger than 18 years old. The most prevalent species were M. canis (31.9%) and T.rubrum (31.9%). When correlating the fungal species with the lesion site, it was noted that M. canis was the main agent responsible for lesions in glabrous skin, scalp and hands, while T. rubrum was predominantly observed in nails and T. mentagrophytes in feet. Conclusion: it is concluded that the data present in this research can promote the development of indicators and public policies for the population most susceptible to dermatophytosis.

Introdução: dermatofitoses ou tineas se caracterizam por serem micoses causadas por fungos dos gêneros Epidermophyton, Trichophyton e Microsporum. Essas infecções micóticas podem se apresentar na forma de lesões que acometem pele, pelo e unhas de indivíduos de ambos os gêneros e todas as idades. Objetivo: elucidar o perfil epidemiológico de dermatofitoses de pacientes atendidos por um laboratório privado de análises clínicas em João Pessoa-PB, entre 2015 a 2019. Metodologia: trata-se de um estudo epidemiológico, analítico, retrospectivo e documental, em que a coleta de dados ocorreu no Laboratório de Patologia Clínica ­ HEMATO, localizado em João Pessoa ­ PB. Resultados: o perfil de acometidos foi predominantemente de indivíduos do sexo feminino (58,5%), com 18 a 59 anos de idade (38,4%), brancos (53,6%) e com lesões, principalmente, em pele glabra (38,5%), pés (33,3%) e unhas (12,8%). Ao relacionar a faixa etária com o local da lesão, percebeu-se que lesões em pele glabra, pés e unhas, foram mais frequentes em indivíduos de 18 a 59 anos, enquanto que lesões no couro cabeludo foram majoritariamente encontradas em indivíduos menos de 18 anos. As espécies mais prevalentes foram M. canis (31,9%) e T. rubrum (31,9%). Ao correlacionar a espécie fúngica com o local da lesão, notou-se que M. canis foi o principal agente responsável por lesões em pele glabra, couro cabeludo e mãos, enquanto T. rubrum foi predominantemente observado em unhas e T. mentagrophytes em pés. Conclusão: os dados obtidos nesta pesquisa podem fomentar o desenvolvimento de indicadores e políticas públicas para a população mais susceptível às dermatofitoses.

Onicomicosis en pediatría: actualización y tratamiento / Pediatric onychomycosis: update and management

Resumen: La onicomicosis (OM) es una infección fúngica de las uñas, cuyo principal agente causal es el Tricophytum rubrum. Si bien es una patología infrecuente en niños, se ha observado un aumento en la prevalencia en el último tiempo. Hasta la fecha, existen diversos estudios y guías clínicas de OM en adultos. Sin embargo, la literatura en edad pediátrica es escasa, lo que dificulta el tratamiento en pediatría. En el presente articulo se revisa la literatura actual, los métodos diagnosticos de OM, datos epidemiológicos locales y globales, y se presentan las opciones de tratamiento disponibles conside rando su eficacia y perfil de seguridad en población pediátrica.

Abstract: Onychomycosis (OM) is a fungal infection of the nails, whose main etiologic agent is Trichophytum rubrum. Although, it is an unusual pathology in children, in the last years an increase in its preva lence has been observed. To date, there are several studies and clinical guidelines for OM in adults. However, literature in children is scarce, which makes pediatric treatment difficult. The objective of this publication was to review the current literature in order to establish diagnostic methods for OM, national and international epidemiological data, and to provide treatment options taking into account their efficiency and safety profile in the pediatric population.

Significativo aumento de onicomicosis por hongos filamentosos no dermatofitos en red de salud privada en Santiago de Chile entre los períodos 2008-2009 y 2016-2017 / Significant increase in onychomycosis due to Filamentous Fungi Non Dermatophytes in Network of Private Health in Santiago de Chile among the periods 2008-2009 and 2016-2017

INTRODUCCION: La onicomicosis corresponde a una patología prevalente causada por hongos dermatofitos, levaduras y en menor proporción, hongos filamentosos no dermatofitos (HFND). Se ha reportado un aumento a nivel mundial en la incidencia de onicomicosis por HFND1 , sin embargo, en Chile no hay registros de ello. OBJETIVO: Comparar la epidemiología local de onicomicosis por HFND en una red privada de Santiago de Chile entre dos períodos de tiempo. Material y métodos: Se realizó un estudio transversal de análisis de los registros de laboratorio de la Red de Salud UC-Christus entre los períodos 2008- 2009 y 2016-2017. RESULTADOS: Se obtuvieron un total de 9.579 muestras en ambos períodos, 4.985 entre 2008-2009 y 4.594 entre 2016-2017, con 3.442 (36%) cultivos positivos para hongos, 1.831 (36,7%) en el primer período y 1.611 (35%) en el segundo. Del total 40 muestras (1,2%) fueron de HFND, destacando que la prevalencia aumentó significativamente entre ambos períodos: 8 (0,4%) y 32 (2,0%) cultivos, respectivamente (p<0,0001). Del total, las especies de HFND más prevalentes fueron: 23 Fusarium sp (57,5%), 8 Scopulariopsis sp (20%) y 7 Acremonium sp (17,5%). En pacientes con cultivo positivo para HFND, se buscaron asociaciones probables en los antecedentes a través de la ficha clínica, sin encontrar resultados significativos. CONCLUCION: Los HFND son la tercera causa de onicomicosis y su prevalencia fue 1,2%. Las especies más frecuentes fueron Fusarium sp, Scopulariopsis sp, Acremonium sp. La prevalencia de HFND aumentó significativamente en un período de 6 años.

INTRODUCTION: Onychomycosis is a frequent nail disease caused mainly by dermatophytes, in less proportion yeast and last non-dermatophyte molds (NDM). NDM onychomycosis is an increasing problem worldwide, though in Chile there is no epidemiological registry about it. OBJETIVE: The aim of the study was to determine the local epidemiology of NDM onychomycosis. MATERIAL Y METHODS: We did a transversal cohort study in the Red de Salud UC-Christus between 2008-2009 and 2016-2017. RESULTS: Of the 9,579 clinically suspected cases of onychomycosis, 4,985 in 2008-2009 and 4,594 in 2016-2017, 3,448 (36%) cultures were positive in total, 1,831 (36.7%) in the first period and 1,611 (35%) in the second. Only 40 cultures (1.2%) were NDM in total, but the prevalence significantly increased between both periods with 8 (0.45%) and 32 (2.0%) cultures, respectively (p y (p<0.0001). Among the total of NDM, the most prevalent species were: 23 Fusarium sp (57.5%), 8 Scopulariopsis sp (20%) and 7 Acremonium sp (17.5%). We searched for possible associations of patients with NDM onychomycosis and morbid history, but found no significant result. COMCLUSION: NDM are the third cause of onychomycosis and its prevalence of 1.2% was predominantly from toe nails. The most frequent species were Fusarium sp, Scopulariopsis sp. and Acremonium sp. The prevalence of NDM increased significantly in a period of 6 years.

Dermatofitos y hongos levaduriformes causantes de micosis superficiales de piel lampiña en un centro dermatológico, San Lorenzo-Paraguay / Dermatophytes and levaduriform fungi causing superficial mycoses of skin in a dermatological center, San Lorenzo-Paraguay

Introducción: las micosis superficiales o dermatomicosis se encuentran entre las enfermedades infecciosas de piel más comunes. Se localizan en las capas superficiales de la piel y sus anexos (pelo y uñas), por esta razón son motivo de consulta dermatológica. De estas micosis, las dermatofitosis, las malasseziosis y las candidiasis cutáneas son las más frecuentes. Objetivo: describir las características epidemiológicas de las micosis superficiales de piel lampiña. Metodología: estudio observacional, retrospectivo y de corte transverso, donde se analizaron fichas epidemiológicas de pacientes que acudieron al Centro de Especialidades Dermatológicas en el periodo comprendido entre julio del 2017 hasta agosto del 2019, con diagnóstico presuntivo de micosis superficiales de piel lampiña. Resultados: se observaron 341 casos de micosis superficiales de piel lampiña. Entre los agentes causales, los dermatofitos fueron los responsables del 56 % (190/341) de los casos; Malassezia spp. 42 % (142/341) y Candida spp. 2 % 9/341. Se observaron 3 casos de micosis mixtas, donde Candida spp. y dermatofitos fueron los agentes causales. Conclusión: entre las micosis superficiales que afectan piel lampiña, se encuentran en primer lugar los dermatofitos seguidos por las malasseziosis. Este reporte constituye un aporte importante para el conocimiento de la epidemiología de las micosis superficiales en nuestro país.

Introduction: superficial mycoses or dermatomycosis are among the most common skin infectious diseases. These are located in the superficial layers of the skin and its annexes (hair and nails), for this reason it´s a frecuent cause of medical consult. Of these mycoses, dermatophytosis, malasseziosis and cutaneous candidiasis are the most frequent. Objectives: describe the epidemiological characteristics of superficial mycoses of skin. Methodology: observational, retrospective and cross-sectional study, where epidemiological records of patients who were attended at the Dermatological Specialties Center were analyzed in the period from July 2017 to August 2019, with presumptive diagnosis of superficial mycoses of skin. Results: 341 cases of superficial mycosis of skin were observed. Among the causative agents, dermatophytes were responsible for 56 % (190/341) of the cases; Malassezia sp. 42 % (142/341) and Candida sp. 2 % (9/341). Three cases of mixed mycosis were observed; Candida spp. and Dermatophytes were the causative agents. Conclusion: among the superficial mycoses of skin, dermatophytes followed by malaseziosis are in the first places. This report constitutes an important contribution to the knowledge of the epidemiology of superficial mycoses in our country.

Tiña de la cabeza: características epidemiológicas de 132 casos / Tinea capitis: epidemiological characteristics of 132 cases

Introducción: la tiña de la cabeza es una micosis superficial producida por dermatofitos, más común en niños y excepcional en los adultos. Objetivo: conocer las características epidemiológicas y micológicas de la tiña de la cabeza en pacientes que acudieron al Centro de Especialidades Dermatológicas en la ciudad de San Lorenzo. Metodología: estudio observacional retrospectivo de corte transverso, donde se analizaron fichas y registros de todos los pacientes con sospecha clínica de tiña de la cabeza remitidos al laboratorio del Centro de Especialidades Dermatológicas en el periodo julio 2017 - agosto 2019. Los pacientes fueron sometidos a estudios micológicos directos y cultivos para la confirmación del diagnóstico e identificación del agente etiológico. Se consideró el sexo, edad, residencia, posesión de mascotas domésticas y agente etiológico aislado. Resultados: se estudiaron 132 pacientes con diagnóstico de tiña de la cabeza, 69 (52.3 %) varones y 63 (47.7 %) mujeres. La edad de presentación fue desde 1 año hasta 14 años, con una media de 5 años. El principal agente etiológico aislado fue Microsporum canis 99/132 (75 %) seguido de Trichophyton tonsurans 29/132 (22 %). Conclusiones: los hallazgos epidemiológicos obtenidos coinciden con otros trabajos científicos, destacándose M. canis y T. tonsurans como los principales agentes etiológicos.

Introduction: tinea capitis is a superficial mycosis produced by dermatophytes, more common in children and exceptional in adults. Objectives: to know the epidemiological and mycological characteristics of tinea capitis in patients who were attended in the Dermatological Specialties Center of San Lorenzo. Methodology: retrospective, observational and cross-sectional study, in which the files and records of all patients with clinical suspicion of tinea capitis, referred to the laboratory of the Dermatological Specialties Center in the period july 2017 - august 2019 were analyzed. The patients underwent direct mycological studies and cultures for confirmation of the diagnosis and identification of the etiologic agent. The sex, age, residence, possession of domestic pets and isolated etiological agent were considered. Results: 132 patients with a diagnosis of tinea capitis, 69 (52.3 %) men and 63 (47.7 %) women were studied. The age of presentation was from 1 year to 14 years, with an average of 5 years. The main etiological agent isolated was Microsporum canis 99/132 (75 %) followed by Trichophyton tonsurans 29/132 (22 %). Conclusions: the epidemiological findings obtained are agree with other scientific studies, highlighting M. canis and T. tonsurans as the main etiological agents.

Prevalência de infecções fúngicas em um laboratório de análises clínicas da cidade de Veranópolis, Rio Grande do Sul / Prevalence of the fungal infecctions in a clinical analysis laboratory in the city of Veranópolis, Rio Grande do Sul

Objetivo: As infecções fúngicas, atualmente, constituem um problema de saúde pública, devido à elevada prevalência e frequência com que acometem a população. Fatores predisponentes como condições climáticas, região geográfica e características culturais e socioeconômicas favorecem a ocorrência destas infecções. O objetivo do presente estudo foi determinar a prevalência das infecções fúngicas em pacientes atendidos em um laboratório de análises clínicas na cidade de Veranópolis, Rio Grande do Sul (RS). Métodos: Foi realizado um estudo retrospectivo, por meio da consulta ao banco de dados, de 403 laudos de exames micológicos, direto e cultural, realizados no laboratório durante o período de abril de 2014 a abril de 2017. Resultados: Dentre os laudos analisados no estudo, houve positividade em 251 (62,28%) exames micológicos diretos (EMD) e em 226 (56,08%) exames culturais. As micoses superficiais e cutâneas constituíram as principais infecções diagnosticadas, sendo os fungos dermatófitos os agentes etiológicos mais isolados (69,47%), representados majoritariamente pela espécie Trichophyton mentagrophytes, seguidos por leveduras (20,35%), especialmente as pertencentes ao gênero Candida. Houve predomínio de infecções em pacientes do sexo feminino (64,54% EMD e 61,5% cultura) e adultos de 31 a 60 anos foram os mais acometidos pelas micoses diagnosticadas (58,96% EMD e 60,62% cultura). Conclusão: O conhecimento do perfil epidemiológico pode servir como guia na correta identificação do patógeno no diagnóstico laboratorial, influenciando diretamente no prognóstico e na escolha da conduta terapêutica adequada.

Objective: The fungal infections are a public health issue, because ofthe high prevalence and frequency that they victimize the population. Predisposal factors as bioclimatic conditions, geographic region and cultural and socioeconomic characteristics favor the occurrence of this infections, generating, this way, variation in the prevalence of species in each region. The goal of the present study was to determinate the prevalence of fungal infections in patients in a clinical analysis laboratory in the city of Veranópolis, Rio Grande do Sul (RS). Methods: A retrospective study was made, using the consult of data basis, of 403 direct and culture mycological examination reports made in the laboratory during the period of April 2014 to April 2017. Results: In the analyzed examination reports in the study, there was positivity in 251 (62,28%) mycological direct exams (EMD) and, in 226 (56,08%) cultural exams. The superficial and cutaneous mycosis constituted the mainly diagnosed infections, being the dermatophytic fungi the most isolated etiological factors (69,47%), represented mainly, by the Trichophyton mentagrophytes species, followed by yeasts (20,35%), especially those who belong to the Candida genre. There was prevalence of infections in women patients (64,54% EMD and 61,5% culture). Adults among 31 and 60 years old were the most victimized by the diagnosed mycosis (58,96% EMD e 60,62% cultural). Conclusion: Knowledge of the epidemiological profile can serve as guide in the correct identification of the pathogen in the laboratory diagnosis, influencing directly in the prognosis and in the choice of the right therapeutic conduct

Padronização de uma PCR para diagnóstico molecular de Microsporum canis em amostras de pelos e crostas de cães e gatos / Standardization of a pcr for molecular diagnosis of microsporum canis in samples of fur and crusts of dogs and cats

Objetivou-se neste estudo padronizar um protocolo de reação em cadeia da polimerase (PCR) para detecção de Microsporum canis em amostras de pelos e/ou crostas de cães e gatos. Foram selecionadas 48 amostras previamente identificadas por meio de cultura. Destas, 23 foram positivas para dermatófitos no cultivo. Padronizou-se a PCR a partir de primers desenhados para o alvo M. canis. Sessenta e um por cento (14/23) das amostras positivas para dermatófitos foram identificadas como M. canis em cultura. Desse total, 71,4% (10/14) apresentaram um fragmento de 218pb compatível com o esperado para a espécie fúngica alvo dessa reação. Observou-se uma sensibilidade de 71,4% e especificidade de 100% na PCR, além de uma boa concordância entre essas técnicas de diagnóstico (Kappa: 0,78; P<0,0001). O protocolo utilizado neste estudo apresentou alta especificidade na detecção de M. canis diretamente de amostras de pelos e/ou crostas de cães e gatos, viabilizando um diagnóstico mais rápido e específico, podendo esse protocolo ser empregado como um método confirmatório para agilizar a detecção de M. canis.(AU)

The aim of this study was to standardize a Polymerase Chain Reaction protocol (PCR) for the detection of Microsporum canis in fur and/or crusts of dogs and cats. 48 samples previously identified by culture were selected. Of these, 23 were positive for dermatophytes in culture. PCR was standardized from drawn primers whose target is M. canis. A total of 61% (14/23) of the dermatophyte positive samples were identified as M. canis in culture. Of this total, 71.4% (10/14) presented a fragment of 218bp compatible with that expected for the fungal species target of the reaction. A sensitivity of 71.4% and specificity of 100% in the PCR were observed, in addition to a good agreement between the techniques (Kappa: 0.78; P<0.0001). The protocol used in this study showed high specificity in the detection of M. canis directly from fur and/or crusts of dogs and cats, making possible a faster and more specific diagnosis. This protocol could be used as a confirmatory method, speeding the detection of M. canis.(AU)

Padronização de multiplex PCR para detecção de dermatófitos em pelos e crostas de cães e gatos / Standardization of a PCR multiplex for the detection of dermatophytes in dogs and cats fur and crusts

Objetivou-se padronizar uma reação do tipo multiplex PCR (mPCR) para detectar Microsporum canis, Microsporum gypseum e o complexo Trichophyton mentagrophytes em amostras de pelos e/ou crostas de cães e gatos. 250 amostras de pelos e/ou crostas de cães e gatos foram analisadas por meio de exame direto e cultura, o DNA das mesmas foi extraído para mPCR. Primers foram desenhados e como controle positivo da reação utilizou-se o DNA extraído de colônias de M. canis (URM 6273), M. gypseum (URM 6921) e T. mentagrophytes (URM 6211), provenientes da Coleção de Culturas (Micoteca URM), Departamento de Micologia, Centro de Ciências Biológicas da Universidade Federal de Pernambuco (CCB/UFPE). Como controles negativos de reação, utilizou-se água destilada esterilizada e DNA extraído de Alternaria sp. para verificar a especificidade dos primers. Do total de amostras analisadas, 15 (6%) foram identificadas, em cultura, como dermatófitos, e destas, 10 foram M. canis, três M. gypseum e dois T. mentagrophytes (complexo). Destas 15 amostras positivas, 11 (73,3%) foram detectadas por meio da mPCR. Além destas, seis outras, negativas em cultura, foram identificadas como M. gypseum. Verificou-se uma boa concordância entre os resultados da cultura e mPCR (Kappa: 0,66). O protocolo padronizado neste estudo pode ser utilizado como um método de triagem, por apresentar uma sensibilidade maior que a da cultura, usado paralelamente aos exames de rotina, permitindo um diagnóstico em menor tempo.(AU)

The aim of this study was to standardize a multiplex PCR (mPCR) reaction to detect Microsporum canis, Microsporum gypseum and the Trichophyton mentagrophytes complex in dog and cat fur and/or crusts. 250 fur and/or crusts samples from dogs and cats were analyzed by direct examination and culture, DNA from them was extracted for mPCR. Primers were designed and the DNA extracted from colonies of M. canis (URM 6273), M. gypseum (URM 6921) and T. mentagrophytes (URM 6211) from the Collection of Cultures - URM Micoteca - Department of Mycology, Biological Sciences Center of the Federal University of Pernambuco (CCB / UFPE). As negative controls, sterile distilled water and DNA extracted from Alternaria sp., were used to verify the specificity of the primers. Of the total samples analyzed, 15 (6%) were identified in culture as dermatophytes, and of these, 10 were M. canis, three M. gypseum and two T. mentagrophytes (complex). Of these 15 positive samples, 11 (73.3%) were detected by mPCR. Besides these, six others, negative in culture, were identified as M. gypseum. There was good agreement between culture results and mPCR (Kappa: 0.66). The protocol standardized in this study can be used as a screening method, because it has a sensitivity greater than that of the culture, used in parallel to the routine exams, allowing a diagnosis in a shorter time.(AU)

Frequency of mixed onychomycosis with total nail dystrophy in patients attended in a Guatemalan Dermatology Center / Frecuencia de onicomicosis mixta con distrofia ungueal total en pacientes atendidos en un Centro de Dermatología de Guatemala

Introduction: Onychomycosis are fungal nail infections that can be caused by dermatophytes, non-dermatophytic molds and yeasts, which are capable of breaking down keratin. Mixed onychomycosis are a controversial subject and they are the outcome of the combination of two dermatophytes, dermatophytes/nondermatophytic molds or dermatophytes/yeast. Objetives: To determine the frequency of total dystrophic onychomycosis caused by more than one etiological agent (mixed onychomycosis) in outpatients from a Dermatologic Center in Guatemala and to establish the characteristics associated with this fungal infection. Methods: Prospective observational study from August to December of 2012. Nail samples were obtained from patients with total dystrophic onychomycosis to identify the causal agents by culture in Sabouraud dextrose and Mycosel® agar. Results: 32 of 130 patients had mixed onychomycosis. 68.5% were associated to tinea pedis. The most common association was between T. rubrum + Candida, T. rubrum + M. canis and T. rubrum + opportunist fungi. Conclusions: Mixed onychomycosis represent 25% of the total dystrophic onychomycosis in Guatemala. We observed an important relationship between diabetes and the main association was T. rubrum with Candida spp.

Introducción: Las onicomicosis son infecciones fúngicas de las uñas que pueden ser causadas por dermatofitos, mohos no dermatofitos y levaduras, que son capaces de degradar la queratina. Las onicomicosis mixtas son un tema polémico y es el resultado de la combinación de dos dermatofitos, dermatofitos / mohos no dermatofitos o dermatofitos / levadura. Objetivos: Determinar la frecuencia de la onicomicosis distrófica total causada por más de un agente etiológico (onicomicosis mixta) en pacientes ambulatorios de un Centro Dermatológico en Guatemala y establecer las características asociadas a esta infección fúngica. Métodos: Estudio observacional prospectivo de agosto a diciembre de 2012. Se obtuvieron muestras de uñas de pacientes con onicomicosis distrófica total para identificar los agentes causales en cultivo de agar dextrosa Sabouraud y Mycosel®. Resultados: 32 de 130 pacientes tenían onicomicosis mixta. 68.5% se asociaron a tinea pedis. La asociación más común fue entre T. rubrum + Candida, T. rubrum + M. canis y T. rubrum + hongos oportunistas. Conclusiones: La onicomicosis mixta representa el 25% de la onicomicosis distrófica total en Guatemala. Observamos una relación importante entre la diabetes y la asociación principal fue T. rubrum con Candida spp.