RESUMO
Sporotrichosis is considered a neglected disease of humans and animals in many regions of the world and is the most frequent implantation mycosis in Latin America. OBJECTIVES: To illustrate the zoonotic importance of the disease, describing a case involving a veterinarian and an infant that acquired the disease from a domestic cat and to describe, genotype and characterize these new isolates. METHODS: Direct examination of tissue samples from the two patients and feline lesions revealed the presence of Sporothrix yeast-like organisms. Fungal cultures and molecular identification of the strains were performed. Since antifungal susceptibility data of animal-borne isolates are scarce, the in vitro susceptibility testing by a microdilution reference method was determined against azoles, amphotericin B and terbinafine. RESULTS: Fungal culture and sequence analysis of the ITS region of rDNA and calmodulin and ß-tubulin genes confirmed the diagnosis and the causative agent as Sporothrix brasiliensis. In all cases, terbinafine was the most active drug, followed by posaconazole, itraconazole and voriconazole; the least active drugs were amphotericine B and fluconazole. Lack of clinical response in the veterinarian and in the infant to itraconazole and potassium iodide, respectively was observed. CONCLUSIONS: This study contributed to the molecular epidemiology of Sporothrix species in Argentina and the characterization of the in vitro susceptibility pattern of S. brasiliensis isolates recovered from a cat and two humans involved in this case of zoonotic sporotrichosis. Bearing in mind the "One Health" concept, the experience described in the present study highlights the need for future strategies for sporotrichosis treatment, control and prevention.
Assuntos
Antifúngicos/uso terapêutico , Doenças do Gato , Esporotricose/diagnóstico , Esporotricose/tratamento farmacológico , Zoonoses/diagnóstico , Zoonoses/tratamento farmacológico , Adulto , Animais , Antifúngicos/farmacologia , Argentina , Doenças do Gato/diagnóstico , Doenças do Gato/tratamento farmacológico , Doenças do Gato/microbiologia , Doenças do Gato/transmissão , Gatos , Pré-Escolar , Feminino , Genótipo , Humanos , Testes de Sensibilidade Microbiana , Técnicas de Diagnóstico Molecular , Técnicas de Tipagem Micológica/métodos , Núcleo Familiar , Filogenia , Sporothrix/classificação , Sporothrix/efeitos dos fármacos , Sporothrix/genética , Sporothrix/isolamento & purificação , Esporotricose/microbiologia , Médicos VeterináriosRESUMO
Se describe un brote de histoplasmosis que afectó a 6 cadetes de la Fuerza Aérea Argentina, sin antecedentes patológicos previos. Todos consultaron por problemas respiratorios después de haber limpiado un hangar. En ese recinto se encontraron abundantes deyecciones de animales, presuntamente de palomas y murciélagos. Los pacientes sufrieron fiebre, mialgias, taquipnea y tos no productiva. Las radiografías y tomografías de tórax mostraron imágenes pulmonares micronodulares, engrosamiento de los tabiques interalveolares y adenopatías hiliares. Todos tuvieron una evolución favorable y no requirieron tratamiento antifúngico. Las pruebas de inmunodifusión y contrainmunoelectroforesis con antígenos de Histoplasma capsulatum fueron positivas, al igual que las intradermorreacciones con histoplasmina. Se recogieron 5 muestras de tierra del lugar, las que fueron inoculadas por vía intraperitoneal a 20 hámsteres. De los cultivos de hígado y bazo de dichos animales se consiguió aislar la fase micelial de H. capsulatum. La cepa aislada se comparó con las obtenidas de 12 pacientes argentinos utilizando perfiles genéticos y se observó un clado único con más de 96% de similitud, lo que confirma la homogeneidad de las cepas argentinas. Si bien la histoplasmosis es endémica en la Pampa húmeda, este es el primer brote totalmente documentado al sur del paralelo 34°.
An histoplasmosis outbreak affecting 6 previously healthy Air Force cadets is herein presented. The patients suffered from fever and respiratory symptoms after having cleaned an abandoned hangar soiled with pigeons and bat droppings. They all presented fever, myalgia, tachypnea, and nonproductive cough. Chest X-ray and CT scan studies showed disseminated reticulonodular images affecting both lungs. Hiliar adenomegalies were also observed. All patients achieved a favourable outcome without antifungal treatment. Both serologic tests searching for specificic antibodies (immunodiffusion and counterimmunoelectrophoresis) and histoplasmin skin tests were positive in all cases. Five soil samples mixed with pigeons and bat droppings were collected from the hangar. Suspensions of these samples were inoculated into 20 hamsters by intraperitoneal injection; mycelial phase of H. capsulatum was isolated from liver and spleen cultures. The genetic profile of this strain was compared with 12 isolates obtained from Argentinean patients, and a great degree of homogeneity was observed (> 96% similarity). Although histoplasmosis is endemic in the wet Pampas, this is the first epidemic outbreak reported south of the 34th parallel.
Assuntos
Adulto , Animais , Cricetinae , Humanos , Masculino , Adulto Jovem , Surtos de Doenças , Histoplasmose/epidemiologia , Militares , Argentina/epidemiologia , Quirópteros/microbiologia , Columbidae/microbiologia , DNA Fúngico/análise , Plumas/microbiologia , Fezes/microbiologia , Histoplasma/classificação , Histoplasma/genética , Histoplasma/crescimento & desenvolvimento , Histoplasma/isolamento & purificação , Histoplasmina , Histoplasmose/diagnóstico , Histoplasmose/transmissão , Mesocricetus , Exposição Ocupacional , Testes CutâneosRESUMO
Cryptococcus is an encapsulated yeast of class Basidiomycetes, etiologic agent of cryptococcosis. Cryptococcosis is one of the most common opportunistic infections in immunosuppressed patients, although it can affect immunocompetent individuals. In recent years, the identification of medically important fungal species has been achieved through the amplification of specific regions or genes of fungal DNA by polymerase chain reaction (PCR). The CAP59 gene is involved in the synthesis of the capsule in Cryptococcus neoformans and Cryptococcus gattii, and is useful in the molecular identification of serotypes. In this research, we use yeasts of different serotypes from collection strains and C. neoformans isolates recovered from patients with cryptococcosis. A standardized yeast suspension from different Cryptococcus isolates as template allowed CAP59 gene amplification. This procedure was quick, simple, and inexpensive and required no PCR steps. This is important for taxonomic studies in laboratories with implemented molecular biology tools.
Assuntos
Cryptococcus gattii/genética , Cryptococcus neoformans/genética , DNA Fúngico/genética , Proteínas Fúngicas/genética , Micologia/métodos , Técnicas de Amplificação de Ácido Nucleico , Criptococose/microbiologia , Cryptococcus gattii/classificação , Cryptococcus gattii/crescimento & desenvolvimento , Cryptococcus neoformans/classificação , Cryptococcus neoformans/crescimento & desenvolvimento , DNA Fúngico/isolamento & purificação , Humanos , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Sensibilidade e Especificidade , Sorotipagem/métodos , SuspensõesRESUMO
A histoplasmosis outbreak affecting 6 previously healthy Air Force cadets is herein presented. The patients suffered from fever and respiratory symptoms after having cleaned an abandoned hangar soiled with pigeons and bat droppings. They all presented fever, myalgia, tachypnea, and nonproductive cough. Chest X-ray and CT scan studies showed disseminated reticulonodular images affecting both lungs. Hiliar adenomegalies were also observed. All patients achieved a favourable outcome without antifungal treatment. Both serologic tests searching for specificic antibodies (immunodiffusion and counterimmunoelectrophoresis) and histoplasmin skin tests were positive in all cases. Five soil samples mixed with pigeons and bat droppings were collected from the hangar. Suspensions of these samples were inoculated into 20 hamsters by intraperitoneal injection; mycelial phase of H. capsulatum was isolated from liver and spleen cultures. The genetic profile of this strain was compared with 12 isolates obtained from Argentinean patients, and a great degree of homogeneity was observed (> 96% similarity). Although histoplasmosis is endemic in the wet Pampas, this is the first epidemic outbreak reported south of the 34th parallel.
Assuntos
Surtos de Doenças , Histoplasmose/epidemiologia , Militares , Adulto , Animais , Argentina/epidemiologia , Quirópteros/microbiologia , Columbidae/microbiologia , Cricetinae , DNA Fúngico/análise , Plumas/microbiologia , Fezes/microbiologia , Histoplasma/classificação , Histoplasma/genética , Histoplasma/crescimento & desenvolvimento , Histoplasma/isolamento & purificação , Histoplasmina , Histoplasmose/diagnóstico , Histoplasmose/transmissão , Humanos , Masculino , Mesocricetus , Exposição Ocupacional , Testes Cutâneos , Adulto JovemRESUMO
INTRODUCTION: It is generally recognized that Candida dubliniensis is commonly found in immunocompromised patients, such as those with advanced human immunodeficiency virus infection, at sites of periodontal disease. Since there are no data available for Argentina, the aim of this study was to determine the prevalence of and to identify C. dubliniensis in periodontal pockets from immunocompetent subjects living in Buenos Aires, Argentina, through a comparison of phenotypic and molecular assays. METHODS: Yeasts recovered from subgingival plaque samples were studied for 180 immunocompetent non-smoking patients with periodontal disease. Yeasts were identified by conventional mycological methods and by specific polymerase chain reaction (PCR) assay. Fluconazole and voriconazole susceptibility studies were performed in keeping with the Clinical and Laboratory Standards Institute. RESULTS: Among 76 yeasts isolated, C. dubliniensis comprised 10.5% (n = 8; 95% confidence interval 4.7-19.7), which corresponded to 4.4% of patients studied (8/180). C. albicans was the most frequently isolated species of yeast. A great majority of C. dubliniensis isolates was susceptible with only one isolate resistant to both antifungals. CONCLUSION: Micromorphology on Staib agar was the phenotypic method that was most concordant with PCR and it was useful for selecting presumptive C. dubliniensis. This is the first report to use PCR to identify C. dubliniensis in subgingival fluid from immunocompetent individuals with periodontal disease in Argentina. On the basis of the findings presented here, we confirm that C. dubliniensis can colonize periodontal pockets of immunocompetent patients with periodontal disease.
Assuntos
Candida/genética , Candidíase Bucal/microbiologia , Placa Dentária/microbiologia , Bolsa Periodontal/microbiologia , Adolescente , Adulto , Idoso , Argentina/epidemiologia , Candida/classificação , Candida/efeitos dos fármacos , Candidíase Bucal/epidemiologia , DNA Fúngico/genética , Farmacorresistência Fúngica , Feminino , Fluconazol/farmacologia , Genótipo , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Técnicas de Tipagem Micológica , Fenótipo , Reação em Cadeia da Polimerase , Pirimidinas/farmacologia , Triazóis/farmacologia , Voriconazol , Adulto JovemRESUMO
Comparison of different methods of DNA extraction from blood to detect fungal DNA by PCR. Invasive fungal infections (IFI) are associated with high mortality by reaching levels of 50%, and also with a significant failure in antifungical treatments. This fact mostly obeys to difficulties in obtaining a fast and accurate mycologic diagnosis due to the low sensitivity of conventional methods, mainly in neutropenic and AIDS patients. Various methods based on fungal DNA study are currently being used for the diagnosis of mycotic infections. We herein evaluated two procedures of extraction and purification of fungal DNA in blood for their use in PCR detection. Both of them showed equal efficiency in obtaining high performance DNA with universal primers ITS land ITS 4 as target.
Assuntos
DNA Fúngico/sangue , DNA Fúngico/isolamento & purificação , Reação em Cadeia da Polimerase , Análise Química do Sangue/métodos , HumanosRESUMO
La infección fúngica invasora (IFI) está asociada a un alto índice de mortalidad, que alcanza el 50% debido a la frecuente falla en el tratamiento antifúngico. Existen dificultades para realizar un diagnóstico micológico rápido y certero dada la baja sensibilidad de los métodos convencionales, especialmente en pacientes neutropénicos y con SIDA. Numerosos métodos para diagnosticar infecciones micóticas basados en el estudio del ADN fúngico están actualmente en desarrollo. Nosotros evaluamos la utilidad de dos procedimientos de extracción y purificación del ADN fúngico presente en sangre para su posterior detección por PCR. Ambos métodos resultaron igualmente eficientes para obtener ADNs de óptima calidad y para realizar la técnica de PCR con los iniciadores universales para hongos ITS 1 e ITS 4.
Invasive fungal infections (IFI) are associated with high mortality by reaching levels of 50%, and also with a significant failure in antifungical treatments. This fact mostly obeys to difficulties in obtaining a fast and accurate mycologic diagnosis due to the low sensitivity of conventional methods, mainly in neutropenic and AIDS patients. Various methods based on fungal DNA study are currently being used for the diagnosis of mycotic infections. We herein evaluated two procedures of extraction and purification of fungal DNA in blood for their use in PCR detection. Both of them showed equal efficiency in obtaining high performance DNA with universal primers ITS 1and ITS 4 as target.
Assuntos
Humanos , DNA Fúngico/sangue , DNA Fúngico/isolamento & purificação , Reação em Cadeia da Polimerase , Análise Química do Sangue/métodosRESUMO
The importance of epidemiological monitoring of yeasts involved in pathologic processes is unquestionable due to the increase of these infections over the last decade, the changes observed in species causing candidiasis, and empirical antifungal treatment. At the Mycology Center, 1006 isolates from a wide range of clinical samples were studied during 1999-2001. Candida albicans (40.3%) was the most isolated species, although, the Candida no albicans species with 54.9% showed the major prevalence. In blood cultures Candida parapsilosis (34.9%), C. albicans (30.2%) and C. tropicalis (25.6%) were recovered most frequently while C. glabrata represented only 2.3%. C. albicans with 60%-80% was the predominant specie in mucosal surface. We also detected Candida mediastinistis, which alert us over the importance at this location. Urinary tract infections caused by yeasts were more frequent in hospitalized patients, being C. albicans (47.7%), the most commonly isolated, followed by C. glabrata (24.8%) and C. tropicalis (20.0%). In the candidal onychomycoses, C. parapsilosis (37.7%) outplaced C. albicans (22.0%). Fluconazole susceptibility studies of Candida species allowed us to conclude that the majority of C. albicans islolates are susceptible, and that the highest resistance averages were observed in C. glabrata (21.41%) and C. krusei (69.23%).
Assuntos
Candida albicans/isolamento & purificação , Candida/isolamento & purificação , Candidíase/epidemiologia , Antifúngicos/farmacologia , Argentina/epidemiologia , Líquidos Corporais/microbiologia , Candida/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Candida glabrata/efeitos dos fármacos , Candida glabrata/isolamento & purificação , Candida tropicalis/efeitos dos fármacos , Candida tropicalis/isolamento & purificação , Candidíase/microbiologia , Candidíase Cutânea/microbiologia , Candidíase Vulvovaginal/microbiologia , Cateterismo Periférico , Farmacorresistência Fúngica , Feminino , Fluconazol/farmacologia , Fungemia/microbiologia , Humanos , Masculino , Mediastinite/microbiologia , Mucosa/microbiologia , Onicomicose/microbiologia , Especificidade de Órgãos , Prevalência , Estudos Retrospectivos , Especificidade da Espécie , Infecções Urinárias/microbiologiaRESUMO
Las levaduras implicadas en procesos patológicos son de indiscutible importancia debido al incremento experimentado por estas infecciones en las últimas décadas, a los cambios observados en las especies causales y al uso empírico de antifúngicos. En el Centro de Micología se estudiaron 1006 aislamientos provenientes de una amplia gama de muestras clínicas durante el periodo 1999-2001. Candida albicans con 40,3% resultó la especie de mayor frecuencia de aislamiento, pero las especies de Candida no albicans con 54,9% resultaron de mayor prevalencia y el 4,8% fueron otras levaduras. En los hemocultivos Candida parapsilosis con 34,9%, C. albicans con 30,2% y C. tropicalis con 25,6% resultaron las más recuperadas, mientras que C. glabrata se presentó con un 2,3%. En las secreciones mucosas C.albicans con 60%-80% fue la especie preponderante. Hemos detectado especies de Candida causantes de mediastinitis, lo que nos alerta sobre su importancia en estos procesos. Las infecciones del tracto urinario por levaduras se detectaron en mayor frecuencia en individuos hospitalizados, resultando C. albicans con 47,7% la especie más aislada, y dentro de Candida no albicans, C. glabrata con 24,8% y C. tropicalis con 20,0%. En las onixis candidiásicas C. parapsilosis con 37,7% desplazó a C.albicans con 22,0% de este lugar anatómico. Los estudios de sensiblidad al fluconazol de las especies de Candida nos permiten concluir que C.albicans es una especie sensible y que los mayores porcentajes de resistencia se observaron en C. glabrata (21,41%) y and C. krusei (69,23%).
The importance of epidemiological monitoring of yeasts involved in pathologic processes is unquestionable due to the increase of these infections over the last decade, the changes observed in species causing candidiasis, and empirical antifungal treatment. At the Mycology Center, 1006 isolates from a wide range of clinical samples were studied during 1999-2001. Candida albicans (40.3%) was the most isolated species, although, the Candida no albicans species with 54.9% showed the major prevalence. In blood cultures Candida parapsilosis (34.9%), C. albicans (30.2%) and C. tropicalis (25.6%) were recovered most frequently while C. glabrata represented only 2.3%. C. albicans with 60%-80% was the predominant specie in mucosal surface. We also detected Candida mediastinistis, which alert us over the importance at this location. Urinary tract infections caused by yeasts were more frequent in hospitalized patients, being C. albicans (47.7%), the most commonly isolated, followed by C. glabrata (24.8%) and C. tropicalis (20.0%). In the candidal onychomycoses, C. parapsilosis (37.7%) outplaced C. albicans (22.0%). Fluconazole susceptibility studies of Candida species allowed us to conclude that the majority of C. albicans islolates are susceptible, and that the highest resistance averages were observed in C. glabrata (21.41%) and C. krusei (69.23%).
Assuntos
Feminino , Humanos , Masculino , Candida albicans/isolamento & purificação , Candida/isolamento & purificação , Candidíase/epidemiologia , Antifúngicos/farmacologia , Argentina/epidemiologia , Líquidos Corporais/microbiologia , Cateterismo Periférico , Candida albicans/efeitos dos fármacos , Candida glabrata/efeitos dos fármacos , Candida glabrata/isolamento & purificação , Candida tropicalis/efeitos dos fármacos , Candida tropicalis/isolamento & purificação , Candida/efeitos dos fármacos , Candidíase Cutânea/microbiologia , Candidíase Vulvovaginal/microbiologia , Candidíase/microbiologia , Farmacorresistência Fúngica , Fluconazol/farmacologia , Fungemia/microbiologia , Mediastinite/microbiologia , Mucosa/microbiologia , Especificidade de Órgãos , Onicomicose/microbiologia , Prevalência , Estudos Retrospectivos , Especificidade da Espécie , Infecções Urinárias/microbiologiaRESUMO
The importance of epidemiological monitoring of yeasts involved in pathologic processes is unquestionable due to the increase of these infections over the last decade, the changes observed in species causing candidiasis, and empirical antifungal treatment. At the Mycology Center, 1006 isolates from a wide range of clinical samples were studied during 1999-2001. Candida albicans (40.3
) was the most isolated species, although, the Candida no albicans species with 54.9
showed the major prevalence. In blood cultures Candida parapsilosis (34.9
), C. albicans (30.2
) and C. tropicalis (25.6
) were recovered most frequently while C. glabrata represented only 2.3
. C. albicans with 60
-80
was the predominant specie in mucosal surface. We also detected Candida mediastinistis, which alert us over the importance at this location. Urinary tract infections caused by yeasts were more frequent in hospitalized patients, being C. albicans (47.7
), the most commonly isolated, followed by C. glabrata (24.8
) and C. tropicalis (20.0
). In the candidal onychomycoses, C. parapsilosis (37.7
) outplaced C. albicans (22.0
). Fluconazole susceptibility studies of Candida species allowed us to conclude that the majority of C. albicans islolates are susceptible, and that the highest resistance averages were observed in C. glabrata (21.41
) and C. krusei (69.23
).