The activity of propolis against pathogenic fungi isolated from human infections

Abstract Propolis is a resinous hive product collected by bees from the buds or other parts of plants. It is known for having various biological properties, including antifungal activity. Among the substances present in propolis, flavonoids and phenolic acids and their esters are responsible for its antifungal properties. This means that propolis is ideal for use as an antifungal agent in alternative medicine to treat a number of both topical and systemic infections caused by Candida species and other yeast-like fungi, dermatophyte and nondermatophyte moulds, without the serious side effects typical of synthetic treatment. It is also active against strains of fungi that are resistant to polyenes and azoles, the classes of drugs most commonly used to treat fungal infections. In this article, we review current knowledge about the activity of propolis from different parts of the world and its components in vitro and in vivo against pathogenic fungi isolated from human infections. The article also indicates the possible mechanism of antifungal activity of propolis and its components.

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.

Dermatofitosis por Microsporum gypseum en un niño con inmunodeficiencia primaria / Microsporum gypseum dermatophytosis in a child with primary immunodeficiency

El Microsporum gypseum es un hongo geofílico que puede producir lesiones cutáneas inflamatorias en personas sanas. Se han descripto lesiones más extensas en pacientes inmunocomprometidos. Se presenta el caso de un paciente con dermatofitosis, con exámenes micológicos positivos para Candida sp, Epidermophytom floccosum y Trichophyton tonsurans, al que, ante la mala respuesta al tratamiento con griseofulvina e itraconazol a dosis habituales, se le realizó biopsia cutánea para cultivo que evidenció la presencia de M. gypseum. Debido a la extensión y a la mala respuesta al tratamiento, se realizó evaluación inmunológica y se diagnosticó un defecto en STAT1 con ganancia de función (STAT1-GOF). Los pacientes que tienen esta inmunodeficiencia primaria son susceptibles a las infecciones micóticas, especialmente por Candida, pero también, aunque en menor medida, a virus y bacterias. El paciente aquí presentado recibió tratamiento prolongado con antimicóticos imidazólicos sistémicos, con resolución de las lesiones.

Microsporum gypseum is a geophilic fungus that can cause inflammatory skin lesions in heathy people. More extensive lesions have been described in immunocompromised patients. We present a patient with extensive dermatophytosis, which mycological examination led the identification of Candida sp, Epidermophyton Floccosum and Trichophyton tonsurans and showed poor response to treatment with griseofulvina and itraconazol at usual doses. When skin biopsy was performed, it had positive culture for M. gypseum. Due to the extension and poor response to treatment, immunological assessment was performed and it showed a defect of STAT1 with gain of function (STAT 1-GOF). Patients with primary immunodeficiency are susceptible to fungal infections, especially Candida but also virus and bacteria, although to a lesser extent. The patient received long-term treatment with systemic imidazole antifungal recovering for the lesions.

Phytochemical characterization and biological activities of Plectranthus barbatus Andrews / Caracterização fitoquímica e atividades biológicas de Plectranthus barbatus Andrews

Plectranthus barbatus Andrews (Lamiaceae) is widely distributed in the world and has a range of popular therapeutic indications. This work aimed to evaluate the phytochemical characterization of two leaf extracts of P. barbatus, and their antimicrobial, antineoplastic and immunomodulatory potential. After collection, herborization and obtainment of the P. barbatus aqueous extract (PBA) and acetone:water 7:3 P. barbatus organic extract (PBO), the phytochemical characterization was performed by high-performance liquid chromatography (HPLC). The antimicrobial activity was performed to determine the minimum inhibitory concentration (MIC) against eight bacterial strains using the microdilution test and the fungus Trichophyton rubrum by disc diffusion assay and microdilution test. Cytotoxicity was assessed by MTT and trypan blue methods in normal peripheral blood mononuclear cells (PBMCs) at concentrations ranged between 0.1 to 100 µg.mL-1 and in neoplastic cell lines Toledo, K562, DU-145 and PANC-1 at 1, 10 and 100 µg.mL-1 . Immunomodulatory activity, was evaluated by sandwich ELISA of proinflammatory cytokines at BALB/c mice splenocytes cultures supernatant. Both extracts presented flavonoids, cinnamic derivatives, steroids and ellagic acid. PBO showed bacteriostatic activity against Acinetobacter baumannii (MIC = 250 µg.mL-1) clinical isolate and PBA fungistatic activity against Trichophyton rubrum (MIC = 800 µg.mL-1). The extracts did not exhibit toxicity to PBMCs and neoplastic cells (IC50 > 100 µg.mL-1). Additionally, PBO at 100 µg.mL-1 significantly inhibited IFN-γ and IL-17A cytokines (p = 0.03). Plectranthus barbatus is a potential candidate for therapeutic use due to its low toxicity in healthy human cells and exhibits biological activities of medical interest as bacteriostatic, fungistatic and immunomodulatory.

Plectranthus barbatus Andrews (Lamiaceae) é amplamente distribuída no mundo e com uma série de indicações terapêuticas populares. Este trabalho teve como objetivo avaliar a caracterização fitoquímica de dois extratos da folha de P. barbatus e seu potencial antimicrobiano, antineoplásico e imunomodulador. Após coleta, herborização e obtenção do extrato aquoso (PBA) e acetona: água 7: 3 (orgânico) (PBO) de P. barbatus, a caracterização fitoquímica foi realizada por cromatografia líquida de alta eficiência (CLAE). A atividade antimicrobiana foi realizada para determinar a concentração inibitória mínima (CIM) contra oito cepas bacterianas usando o teste de microdiluição e o fungo Trichophyton rubrum por ensaio de difusão em disco e teste de microdiluição. A citotoxicidade foi avaliada por métodos MTT e azul de tripan em células normais mononucleares do sangue periférico (CMSP) em concentrações variadas entre 0,1 a 100 µg.mL-1 e nas linhagens celulares neoplásicas Toledo, K562, DU-145 e PANC-1 em 1, 10 e 100 µg.mL-1 . A atividade imunomoduladora foi avaliada por ELISA sanduíche de citocinas pró-inflamatórias em sobrenadante de culturas de esplenócitos de camundongos BALB/c. Ambos os extratos apresentaram flavonoides, derivados cinâmicos, esteróides e ácido elágico. O PBO mostrou atividade bacteriostática contra Acinetobacter baumannii (CIM = 250 µg.mL-1) e atividade fungistática do PBA contra Trichophyton rubrum (CIM = 800 µg.mL-1). Os extratos não apresentaram toxicidade para CMSP e células neoplásicas (IC50 > 100 µg.mL-1). Além disso, o PBO a 100 µg.mL-1 inibiu significativamente as citocinas IFN-γ e IL-17A (p = 0,03). Plectranthus barbatus é um candidato potencial para uso terapêutico devido à sua baixa toxicidade em células humanas saudáveis e exibe atividade de interesse médico como bacteriostática, fungistática e imunomoduladora.

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.

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

Micosis en los pies: descripción clínico-epidemiológica en un centro de referencia de Bogotá, Colombia / Foot mycosis: clinical and epidemiological characterization in one referal center in Bogota, Colombia

Objetivo: Describir las características clínico-epidemiológicas de una población con diagnóstico de infección cutánea micótica en los pies confirmada por examen directo con KOH y cultivo en un centro de referencia de Bogotá, Colombia. Material y método: Estudio observacional descriptivo en el que se incluyeron todos los pacientes con lesiones en los pies que fueron atendidos en el servicio de micología entre el año 2011 y el 2016. Se analizaron las características sociodemográficas, clínicas, etiologías y el perfil de tratamiento por medio de un análisis bivariado. Resultados: Se incluyeron 305 pacientes, de los cuales el 82% residía en zona urbana de la ciudad de Bogotá. El hábito más frecuente fue bañarse descalzo, la forma clínica que predominó fue la interdigital y el 35% de los casos presentó de forma simultánea tiña del pie y onicomicosis. Los agentes etiológicos más comunes fueron los dermatofitos con el 95,2% de los casos. Discusión: La presentación clínica sugestiva de micosis, además del resultado positivo del examen directo y del cultivo, permiten hacer el diagnóstico de estas in fecciones. Las características sociodemográficas de quienes sufren este tipo de micosis en Colombia se relacionan con su contagiosidad y tendencia a la cronicidad. La intervención de tales aspectos debe hacer parte de las estrategias para su prevención.

Objective: To describe the clinical and epidemiological characteristics of the population with cutaneous mycosis in the feet confirmed by positive mycological studies diagnosed in a reference center in Bogota, Colombia. Methods: Descriptive observational study in which all patients with lesions in the feet that were treated in the mycology service between 2011 and 2016 were included. In all cases KOH examination and fungal culture were performed. The sociodemographic and clinical characteristics, etiologies and the treatment profile were assessed using a bivariate analysis. Results: A total of 305 patients were included, of which 61% were men and 82% lived in an urban area of the city of Bogota. The most common behavior was to take a barefoot bath, the most important comorbidities were venous insufficiency and psoriasis, the main clinical form was interdigital and 35% of the cases presented simultaneously tinea pedis and onychomycosis. The group of dermatophytes was the most frequently isolated (95.2%). Discussion: Clinical examination corresponding with mycosis in the feet, direct examination, and positive culture, allow the physician accurate diagnosis and guide the most appropriate treatment of these infections according to their etiology. The sociodemographic characteristics of those who suffer from this mycosis in Co lombia are related to their contagiousness and tendency to chronicity, therefore, the intervention of such aspects must be part of the strategies for their prevention.

Ocorrência de dermatófitos em tegumento de bovinos e ovinos hígidos / Occurrence of dermatophytes in healthy cattle and sheep tegument / Ocurrencia de dermatofitos en tegumento de bovinos y ovinos hígidos

O objetivo desta pesquisa foi verificar a prevalência de dermatófitos em tegumento de bovinos e ovinos hígidos e a sua capacidade de transmissão e desenvolvimento da doença. Para isso, foram coletadas amostras de pelo e descamações de 90 bovinos e 90 ovinos hígidos. Essas amostras foram semeadas em meio DTM e ágar Sabouraud Dextrose enriquecido com extrato de levedura, tiamina, antibióticos (estreptomicina e cloranfenicol) e suplementado de cicloheximida, sendo essas incubadas a 35 oC por 10 dias. As culturas positivas foram avaliadas macro e microscopicamente e, os fungos foram identificados por métodos bioquímicos. Verificou-se que as espécies isoladas com maior frequência, nos bovinos, foram T. mentagrophytes e M. gypseum. Nos ovinos, foi constatada maior ocorrência do agente T. verrucosum, seguido por T. mentagrophytes e M. gypseum. Houve também isolamento de M. canis, porém, em um número reduzido de amostras. Assim, concluiu-se que o tegumento de bovinos e de ovinos hígidos apresentou incidência elevada de dermatófitos de diferentes espécies, em amostras coletadas durante o período chuvoso do ano. Por isso, em animais jovens ou naqueles submetidos a elevados níveis de estresse e, consequente queda da resposta imunológica, o risco de desenvolvimento da dermatofitose e da transmissão dos dermatófitos neste período são eminentes.(AU)

The purpose of this study was to determine the prevalence of dermatophytes in healthy cattle and sheep teguments, and their ability to transmit and develop diseases. Samples of fur and flaking from 90 healthy cattle and 90 healthy sheep were collected. These samples were plated on DTM medium and Sabouraud Dextrose agar supplemented with yeast extract, thiamine, antibiotics (streptomycin and chloramphenicol) enhanced with cycloheximide. They were then incubated at 35 °C for 10 days. Positive cultures were macroscopically and microscopically evaluated, and fungi were identified by biochemical methods. It was found that the most frequent species isolated in cattle were T. mentagrophytes and M. gypseum. In sheep, a higher occurrence of the agent T. verrucosum could be observed, followed by T. mentagrophytes and M. gypseum. There was also the isolation of M. canis, but in a small number of samples. Thus, it could be concluded that the tegument of healthy cattle and sheep showed high incidence of dermatophytes from different species in samples collected during the rainy season of the year. Therefore, in young animals, in those subjected to high levels of stress and consequent drop in immune response, there is eminent risk of developing dermatophytosis and transmission of dermatophytes in this period.(AU)

El objetivo de esta investigación ha sido verificar la prevalencia de dermatofitos en tegumento de bovinos y ovinos hígidos y su capacidad de transmisión y desarrollo de la enfermedad. Para ello, se recolectó muestras de pelo y descamaciones de 90 bovinos y 90 ovinos hígidos. Esas muestras fueron sembradas en medio DTM y agar Sabouraud Dextrosis enriquecida con extracto de levadura, tiamina, antibióticos (estreptomicina y cloranfenicol) y suplementado de cicloheximida, siendo esas incubadas a 35ºC durante 10 días. Los cultivos positivos fueron evaluados macro y microscópicamente y los hongos fueron identificados por métodos bioquímicos. Se verificó que las especies aisladas con mayor frecuencia, en los bovinos, fueron T. mentagrophytes y M. gypseum. En los ovinos, se constató mayor ocurrencia del agente T. verrucosum, seguido por T. mentagrophytes y M. gypseum. También hubo aislamiento de M. canis, sin embargo, en un número reducido de muestras. Así, se concluyó que el tegumento de bovinos y de ovinos hígidos presentó una elevada incidencia de dermatofitos de diferentes especies, en muestras recogidas durante el período lluvioso del año. Por lo tanto, en animales jóvenes o en aquellos sometidos a altos niveles de estrés y, consecuente caída de la respuesta inmunológica, el riesgo de desarrollo de la dermatofitosis y de la transmisión de los dermatofitos en este período son eminentes.(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.

The presence of dermatophytes in infected pets and their household environment / Presença de dermatófitos em pets infectados e em seu ambiente domiciliar

The aim of this study was to diagnose dermatophytosis in pets and investigate the presence of dermatophytes in their home environment. Samples from hair coat were collected from 70 pets: 47 dogs, 19 cats, three guinea pigs and one rabbit. After mycological culture, 188 samples were collected from the household environments in 26 homes: 78 from places were of predominantly used by the tutors, 66 from places used by the animals, 44 from flooring, and 24 samples from contactees. Samples were seeded on Mycosel agar, incubated at 25°C, and the colonies were identified by their macro-and-microscopic characteristics. Dermatophytes were found in 37.1% of the samples originating from the sick animals. Microsporum canis was the most prevalent species, isolated in 12 dogs and eight cats; Trichophyton quinckeanum in three guinea pigs, Microsporum gypseum in two dogs and Trichophyton mentagrophytes in one cat. Dermatophytes were found in 69.2% of the surveyed homes; 29.5% of the places/objects predominantly used by the tutors, 42.4% mainly used by the animals, 31.8% from floors, and 50% from contactees. The meeting of dermatophytes in animals and in the household environment confirms the possibility of transmission by direct or indirect contact and their importance in public health.(AU)

O objetivo deste trabalho foi diagnosticar dermatofitose em pets e pesquisar dermatófitos em seu ambiente domiciliar. Colheram-se amostras de 70 pets, 47 cães, 19 gatos, três cobaias e um coelho. Visitaram-se 26 residências dos animais positivos em cultura micológica para a doença, colhendo-se 188 amostras do ambiente: 78 de locais de uso predominante dos tutores, 66 de uso dos animais e 44 de pisos; também foram colhidas 24 amostras de animais contactantes. As amostras clínicas foram semeadas em ágar Mycosel, incubadas a 25°C, e as colônias identificadas por suas características macro e microscópicas. Dermatófitos foram isolados em 37,1% dos animais suspeitos. Microsporum canis foi o mais frequente, sendo isolado de 12 cães e oito gatos, Trichophyton quinckeanum de três cobaias, Microsporum gypseum de dois cães e Trichophyton mentagrophytes de um gato. Foram encontrados dermatófitos em 69,2% das casas pesquisadas, isolando-se esses fungos em 29,5% dos locais/objetos de uso predominante dos tutores, 42,4% de uso predominante dos animais, 31,8% de pisos e 50% dos animais contactantes. O encontro de dermatófitos nos animais e em superfícies inanimadas nas residências confirma a possibilidade de transmissão de dermatofitose por contato direto e indireto e sua importância em saúde pública.(AU)

Dermatófitos como agentes de onicomicoses em população adulta de serviço ambulatorial do Hospital de Aeronáutica de São Paulo, na cidade de São Paulo

A onicomicose é uma infecção micótica das unhas, com interesse emSaúde Pública pela alta morbidade e ocorrência de surtos, em especial, ematletas e militares. Algumas condições mórbidas estão associadas a essaforma de micose o que pode acentuar seu caráter recidivante e crônico. Aidentificação correta dos agentes etiológicos das onicomicoses, sejamleveduras ou fungos filamentosos, pode ser subsídio para tratamentoadequado da infecção, além de fornecer indicação sobre possíveis fontes deinfecção. Este estudo teve como objetivo caracterizar e determinar asuscetibilidade a antifúngicos de fungos filamentosos agentes deonicomicoses em população militar da aeronáutica. Amostras ungueais delesões de 50 pacientes, com suspeita de onicomicose em pés ou mãos,foram coletadas para exame micológico direto, cultura, isolamento,identificação fenotípica e molecular dos agentes etiológicos. O perfil desuscetibilidade dos agentes etiológicos, frente a 3 fármacos antifúngicosusados no tratamento das onicomicoses, foi determinado pelo método dereferência americano (CLSI ). Fungos filamentosos foram implicados em 50(62,5%) casos, prevalecendo dermatófitos (43; 86%), como T. rubrum (22;44%), T. mentagrophytes var. interdigitale (19; 38%) e M. gypseum (3; 6%) enão dermatófitos (6; 12%), incluindo feo-hifomicetos como Chaetomiumglobosum (4; 8%) e hialo-hifomicetos como Fusarium solani e Scytalidiumspp. (1; 2% cada um). Em relação à suscetibilidade a antifúngicos, foiverificado que menores concentrações de terbinafina foram suficientes parainibir o crescimento fúngico do total de isolados, se comparadas às deitraconazol e fluconazol (MIC 50%/MIC 90%, respectivamente, de 0,015/0,06mg/L, 0,06/0,12 mg/L e 32/32 mg/L)...

Onychomycosis is a fungal infection of the nails, with interest in Public Healthdue to the high morbidity and occurrence of outbreaks, especially in athletesand soldiers. Some morbid conditions are associated with this form ofmycosis, which may accentuate its recurrent and chronic character. Thecorrect identification of the etiological agents of onychomycosis, whetheryeasts or filamentous fungi, can be a subsidy for adequate treatment of theinfection, as well as providing indication about possible sources of infection.This study aimed to characterize and determine de antifungal drugsusceptibility of filamentous fungi agents of onychomycosis in theaeronautical military population. Ungual samples of lesions of 80 patientswith suspicion of onychomycosis in feet or hands were collected for directmycological examination, culture, isolation, phenotypic and molecularidentification of the etiological agents. The susceptibility profile of theetiological agents to 3 antifungal drugs used in the treatment ofonychomycosis, was determined using the American reference method(CLSI). Filamentous fungi were implicated in 50 (62.5%) cases, withdermatophytes prevailing (43; 86%), such as T. rubrum (22; 44%), T.mentagrophytes var. interdigitale (19; 38%), and M. gypseum (3; 6%) ornon-dermatophytes (6; 12%), including feo-hyphomicetes such asChaetomium globosum (4; 8%), and hialo-hyphomycetes like Fusariumsolani and Scytalidium spp. (1; 2% each). Regarding susceptibility toantifungals, it was found that lower concentrations of terbinafine weresufficient to inhibit fungal growth in the total of isolates, compared toitraconazole and fluconazole (MIC 50%/90%, respectively: 0.015/0.06 mg/L,0.06/0.12 mg/L and 32/32 mg/L)...

In Vitro additive effect on griseofulvin and terbinafine combinations against multidrug-resistant dermatophytes

ABSTRACT Griseofulvin (GF) and terbinafine (TF) are commonly used drugs to treat dermatophytosis, a fungal infection of the skin. Today there is an increase in drug resistance to these antifungals which highlight the need for alternative synergistic therapies. Minimum Inhibitory Concentration (MIC) of GF and TF were determined against fungi clinical isolates from local hospitals with values ranging 0.03-2.0 µg mL-1 and 0.24-4.0 µg mL-1, respectively. A checkboard test was used to determine the combination of GF:TF which could induce an additive effect against the fungi isolates Multidrug-resistant isolates showed susceptibility after treatment with 16:2 µg mL-1 GF:TF. An MTT assay further verified that GF and TF combinations have greater additive effect against pathological and multidrug-resistant isolates than antifungals alone. Herein we disclose GF:TF combinations that could constitute as a possible new anti-dermatophyte therapy.

Dermatophyte colonization in rabbits kept in pet stores of Santiago of Chile / Colonización por dermatofitos en conejos mantenidos en tiendas de mascotas de Santiago de Chile

ABSTRACT Objective. The dermatophytes are keratinophilic fungi, of importance in public health because of their anthropozoophilic nature. Given the increasing acquisition of exotic animals as pets and the scarce studies on the state of colonization by dermatophytes on these animals; we raised the objective of determine the presence of dermatophytes in clinically healthy rabbits (Oryctolagus cuniculus) from pet stores in Santiago, Chile. Materials and Methods. 42 clinically healthy rabbits were studied. Clinical specimens were obtained from the hairy mantle and cultivated on Sabouraud glucose agar and dermatophyte test medium (DTM™); the identification of the fungal isolates was performed using classic mycological procedures that included direct microscopic examination and the analysis of micromorphological features on culture. Results. Of the total number of rabbits studied, three (7.1%) presented colonization by dermatophytes, being more frequent in males and in those animals that coexisted with Guinea Pigs. The species identified in all three cases was Trichophyton mentagrophytes. Conclusions. This study evidence colonization by dermatophytes in domestic rabbits, important information for veterinarians and owners of pets, at the time of being in contact with this animal species.

RESUMEN Objetivo. Los dermatofitos son hongos queratinofílicos, de importancia en salud pública por su rol antropozoofílico. Dado el aumento en la adquisición de animales exóticos como mascotas y los escasos estudios sobre el estado de colonización por dermatofitos en estos animales, nos planteamos el objetivo de determinar la presencia de dermatofitos en conejos (Oryctolagus cuniculus) clínicamente sanos procedentes de tiendas de mascotas en Santiago de Chile. Materiales y Métodos. Se estudiaron 42 conejos clínicamente sanos. Las muestras clínicas se obtuvieron desde el manto piloso y cultivadas en agar Sabouraud glucosado y dermatophyte test medium (DTM®). La identificación del agente aislado fue realizada por procedimientos micológicos clásicos que incluyeron examen microscópico directo y cultivo. Resultados. Del total de conejos estudiados, tres (7,1%) presentaron colonización por dermatofitos, siendo más frecuente en machos y en aquellos animales que convivían con cuyes. La especie identificada en los tres casos fue Trichophyton mentagrophytes. Conclusiones. Este estudio evidencia la colonización por dermatofitos en conejos domésticos, información importante para médicos veterinarios y propietarios de mascotas, al momento de estar en contacto con esta especie animal.

Anti-fungal potential of ozone against some dermatophytes

ABSTRACT Dermatophytes are classified in three genera, Epidermophyton, Microsporum and Trichophyton. They have the capacity to invade keratinized tissue to produce a cutaneous infection known as dermatophytoses. This investigation was performed to study the effect of gaseous ozone and ozonized oil on three specific properties of six different dermatophytes. These properties included sporulation, mycelia leakage of sugar and nutrients and the activity of their hydrolytic enzymes. Generally, ozonized oil was found to be more efficacious than gaseous ozone. Microsporum gypseum and Microsporum canis were the most susceptible, while Trichophyton interdigitale and T. mentagrophytes were relatively resistant. The study revealed a steady decline in spore production of M. gypseum and M. canis on application of ozonated oil. An increase in leakage of electrolytes and sugar was noticed after treatment with ozonized oil in the case of M. gypseum, M. canis, T. interdigitale, T. mentagrophytes and T. rubrum. The results also revealed loss in urease, amylase, alkaline phosphatase, lipase and keratinase enzyme producing capacity of the investigated fungi.

Photodynamic action of protoporphyrin IX derivatives on Trichophyton rubrum

Abstract: BACKGROUND: Dermatophytes are filamentous keratinophilic fungi. Trichophyton rubrum is a prevalent infectious agent in tineas and other skin diseases. Drug therapy is considered to be limited in the treatment of such infections, mainly due to low accessibility of the drug to the tissue attacked and development of antifungal resistance in these microorganisms. In this context, Photodynamic Therapy is presented as an alternative. OBJECTIVE: Evaluate, in vitro, the photodynamic activity of four derivatives of Protoporphyrin IX by irradiation with LED 400 nm in T. rubrum. METHOD: Assays were subjected to irradiation by twelve cycles of ten minutes at five minute intervals. RESULT: Photodynamic action appeared as effective with total elimination of UFCs from the second irradiation cycle. CONCLUSION: Studies show that the photodynamic activity on Trichophyton rubrum relates to a suitable embodiment of the photosensitizer, which can be maximized by functionalization of peripheral groups of the porphyrinic ring.

Detection of seasonal asymptomatic dermatophytes in Van cats

Abstract The Van cat is a domestic landrace found in the Van province of eastern Turkey. In this study, we aimed to determine the seasonal carriage of dermatophytes in Van cats without clinical lesions. A total of 264 hair specimens were collected from clinically healthy cats in and around the Van Province. Of these samples, 30.3% were obtained in spring, 30.6% in summer, 16.6% in autumn, and 22.3% in winter; 45.1% of samples were from male cats and the rest from female ones. Of the studied cats, 118 were younger than 1 year, 78 were 1–3 years old, and 68 were older than 3 years. The specimens were subjected to direct microscopic examination with 15% potassium hydroxide and cultured on Sabouraud dextrose agar and dermatophyte test medium supplemented with cycloheximide and chloramphenicol. Dermatophyte identification was carried out based on macroscopic and microscopic colony morphology, urease activities, in vitro hair perforation test, growth at 37 °C, and pigmentation on corn meal agar. Dermatophytes were isolated from 19 (7.1%) of the 264 specimens examined. The most frequently isolated fungi were Trichophyton terrestre (4.1%), followed by Microsporum gypseum (1.1%), M. nanum (1.1%), and T. mentagrophytes (0.7%), and these fungi may represent a health risk for humans in contact with clinically healthy Van cats. M. canis was not isolated from any of the specimens. Our results show no significant (p > 0.05) association between carriage of dermatophytes and the gender of cats. The carriage rate of dermatophytes was high in spring and winter, and the only possible risk factor for infection was age of the animal.

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.

Dermatofitoses: agentes etiológicos, formas clínicas, terapêutica e novas perspectivas de tratamento / Dermatophytoses: etiologic agents, clinical forms, therapy and new perspectives of treatment

As dermatofitoses têm ocorrência mundial, sendo mais prevalentes em países de clima tropical e subtropical. Dados epidemiológicos indicam que essas micoses estão entre as infecções fúngicas de maior ocorrência. O quadro clínico mais comum de dermatofitose inclui despigmentação, placas anulares, prurido e perda de cabelo, com lesões tipicamente conhecidas como tineas, ocasionadas por fungos filamentosos dermatofíticos de três gêneros anamórficos: Microsporum, Trichophyton e Epidermophyton. O tratamento das dermatofitoses, em geral, está relacionado ao uso de antifúngicos tópicos e/ou sistêmicos, apresentando como problemática o surgimento de espécies multirresistentes. Esta revisão aborda as dermatofitoses e seus agentes etiológicos de forma aprofundada em aspectos epidemiológicos, apresentando a importância clínica do tema, com ênfase na causa, prevenção, tratamento e prognóstico dessa micose cutânea (AU)

Dermatophytoses have worldwide occurrence with higher prevalence in tropical and subtropical countries. Epidemiological data show that these mycoses are among the most frequent fungal infections. The most common symptoms of dermatophytoses include depigmentation, annular plaques, itching and hair loss, with lesions such as tinea, caused by dermatophytic filamentous fungi of three anamorphic genera: Microsporum, Trichophyton and Epidermophyton. Topical and/or systemic antifungalmedications are used in the treatment of dermatophytoses in general, resulting in problems such as the emergence of multidrug-resistant species. This review discusses dermatophytoses and their etiological agents with a focus on epidemiological aspects, presenting the clinical importance of the issue, with emphasis on cause, prevention, treatment and prognosis of this skin mycosis (AU)