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2.
Biomédica (Bogotá) ; 43(Supl. 1): 216-228, ago. 2023.
Article in English | LILACS | ID: biblio-1533885

ABSTRACT

Introduction. For over a century, Sporothrix schenckii was considered the sole species responsible for sporotrichosis. In 2007, scientific community confirmed the disease could be caused by various Sporothrix species. These species differed in their virulence factors and their antifungal sensitivity. Objective. This study aims to characterize 42 Colombian clinical isolates of Sporothrix spp. phenotypically and genotypically. Materials and methods. Forty-two clinical isolates were characterized using phenotypic methods. It involved various culture media to determine their growth range at different temperatures and to assess the type and distribution of pigment and colony texture. Microscopic morphology was evaluated through microcultures, as well as the conidia diameter, type of sporulation, and morphology. Additionally, the assimilation of carbohydrates was selected as a physiological trait for species identification. Genotyping of 40 isolates was performed through partial amplification of the calmodulin gene, followed by sequence analysis. Results. Molecular studies enabled the identification of 32 isolates of S. schenckii and 8 isolates of S. globosa. The combination of phenotypic and genotypic methods eased these species characterizations and the recognition keys development based on parameters such as growth diameter at 25 and 30 °C, colony texture (membranous or velvety) on potato dextrose agar, and microscopic morphology with predominance of pigmented triangular, elongated oval globose, or subglobose conidia. Conclusions. Confirmation of the phenotypic characteristics and molecular analysis is crucial for identifying Sporothrix species and determining adequate treatment. This study represents the first phenotypical and genotypical characterization of clinical isolates of Sporothrix spp. reported in Colombia.


Introducción. Por más de un siglo se creyó que Sporothrix schenckii era la única especie responsable de la esporotricosis. Sin embargo, en el 2007, se consideró que podría ser causada por diferentes especies de Sporothrix, que difieren en sus factores de virulencia y su sensibilidad a los antifúngicos. Objetivo. Caracterizar fenotípica y genotípicamente 42 aislamientos clínicos colombianos de Sporothrix spp. Materiales y métodos. Se caracterizaron 42 aislamientos clínicos mediante métodos fenotípicos. Se usaron varios medios de cultivo para determinar el rango de crecimiento a diferentes temperaturas, el tipo y la distribución del pigmento, y la textura de las colonias. Se evaluó la morfología microscópica por microcultivos mediante la determinación del diámetro, el tipo de esporulación y la morfología de las conidias. La asimilación de carbohidratos se usó como una característica fisiológica para identificar las especies. La genotipificación de los 40 aislamientos se llevó a cabo mediante la amplificación parcial del gen que codifica para la calmodulina y se confirmó por secuenciación. Resultados. Mediante estudios moleculares, se identificaron 32 aislamientos de S. schenckii y ocho de S. globosa. La combinación de métodos fenotípicos y genotípicos permitió caracterizar las especies y construir claves para su reconocimiento, con base en parámetros como el diámetro de crecimiento a 25 y 30 °C, la textura de las colonias (membranosa, aterciopelada) en agar papa dextrosa y la morfología microscópica con predominio de conidias (triangulares pigmentadas, ovales globosas elongadas, subglobosas). Conclusiones. La caracterización fenotípica y los análisis moleculares son necesarios para identificar las especies de Sporothrix y, de esta forma, elegir el tratamiento indicado. Esta es la primera caracterización fenotípica y genotípica reportada de aislamientos clínicos colombianos de Sporothrix spp.


Subject(s)
Sporotrichosis , Phenotype , Sporothrix , Genotype
3.
Biomédica (Bogotá) ; 43(Supl. 1): 109-119, ago. 2023. tab, graf
Article in Spanish | LILACS | ID: biblio-1533882

ABSTRACT

Introducción. La esporotricosis es una micosis de implantación causada por Sporothrix spp. Este se encuentra distribuido mundialmente y se puede encontrar en la vegetación y en el suelo. La ruta más frecuente de adquisición de la infección es por traumatismos con elementos contaminados con propágulos del hongo. Los gatos domésticos son los animales más afectados y pueden transmitirla a los humanos, por lo que es considerada una zoonosis. Las formas clínicas incluyen: la linfangítica nodular, la cutánea fija, la pulmonar (poco habitual) y la diseminada (excepcional). Objetivo. Analizar la epidemiología de la esporotricosis en Argentina entre los años 2010 y 2022. Describir la presentación clínica, los métodos de diagnóstico y el tratamiento de los casos diagnosticados en este período. Conocer los genotipos circulantes y observar su relación con el lugar geográfico de adquisición de la infección. Materiales y métodos. Se llevó a cabo un estudio analítico, retrospectivo y observacional, en el que se analizaron las historias clínicas de los pacientes con esporotricosis de 12 instituciones de salud de Argentina, entre los años 2010 y 2022. Resultados. Se presentan 54 casos en los que la forma clínica más frecuente fue la linfangítica nodular y el tratamiento de elección fue el itraconazol. En todos los casos se realizó diagnóstico convencional. El cultivo de las muestras clínicas resultó más sensible que el examen directo, ya que permitió el desarrollo de Sporothrix spp. en los 54 casos. En 22 casos se hizo identificación molecular y Sporothrix schenkii sensu stricto fue la especie más frecuentemente aislada. Conclusiones. Este estudio permitió conocer la epidemiología de esta micosis en Argentina, así como la disponibilidad de métodos diagnósticos y el tratamiento de elección.


Introduction. Sporotrichosis is an implantation mycosis caused by Sporothrix spp. It is distributed worldwide and can be found in vegetation and soil. The most frequent route of infection is by trauma with elements contaminated with fungal propagules. Since domestic cats are the most affected animals and can transmit this infection to humans, sporotrichosis is considered a zoonosis. Clinical presentations include nodular lymphangitis, fixed cutaneous, pulmonary (rare), and disseminated (exceptional). Objective. To analyze the epidemiology of sporotrichosis in Argentina during 2010 and 2022. To describe the clinical presentation, diagnostic methods, and treatment of cases diagnosed during this period. To know the circulating genotypes and to observe possible associations with the geographic location where the infection was acquired. Materials and methods. Analytical, retrospective, and observational study. We analyzed the medical records of patients with sporotrichosis from 12 health institutions in Argentina, between 2010 and 2022. Results. We present 54 cases in which the most frequent clinical form was nodular lymphangitis, and the treatment of choice was itraconazole. Conventional diagnosis was made in all cases. Culture of clinical samples was more sensitive than direct examination because it allowed the isolation of Sporothrix spp. in all 54 cases. Molecular identification was performed in 22 cases, with Sporothrix schenkii sensu stricto being the most frequently isolated species. Conclusions. This study allowed to know the epidemiology of this mycosis in Argentina, as well as the availability of diagnostic methods and the treatment of choice.


Subject(s)
Sporotrichosis , Argentina , Sporothrix , Mycoses
4.
An. bras. dermatol ; 97(1): 1-13, Jan.-Feb. 2022. graf
Article in English | LILACS | ID: biblio-1360090

ABSTRACT

Abstract In recent decades, an alarming increase in the number of sporotrichosis cases has been reported in southern and southeastern Brazil, especially in the state of Rio de Janeiro, has been considered a long-term hyperendemic condition associated with feline transmission. According to phenotypic classifications, the Sporothrix species recovered from cats were classified as S. brasiliensis in 96.5% of the studied cases. This finding has also been demonstrated in humans, which confirms the zoonotic transmission associated with this predominant species in Brazil. The zoonotic transmission of the fungus and its important virulence in the context of the hyperendemic situation in Rio de Janeiro have changed the approach to the disease, which in its classic form was restricted to certain professional groups and very specific regions in the Brazilian territory, into a public health challenge of scientific interest. Its atypical manifestations and hypersensitivity reactions are increasingly frequent, constituting a new sporotrichosis aspect, which deserves attention from the medical community, as well as from other health professionals.


Subject(s)
Humans , Animals , Cats , Sporotrichosis/epidemiology , Sporothrix , Cat Diseases/epidemiology , Hypersensitivity , Brazil/epidemiology
5.
Braz. J. Pharm. Sci. (Online) ; 58: e20152, 2022. tab, graf
Article in English | LILACS | ID: biblio-1403756

ABSTRACT

Abstract Sporothrix spp. are the major dimorphic fungus associated with a type of subcutaneous mycosis, sporotrichosis. The limitation of antifungal availability and the past reports of in vitro resistance of Sporothrix spp. clinical isolates makes it important to search for new compounds with antifungal activities. In this study, we therefore evaluate the in vitro activities of complexes coordinated with Co(II) and cobalt chloride hexahydrate against clinical isolates of Sporothrix spp. Broth microdilution test was performed as per M38-A2 from CLSI (2008) in duplicate for 31 clinical isolates of Sporothrix spp. (27 S. brasiliensis e 04 S. schenckii stricto sensu). The antifungal activities of the complexes coordinated with Co(II) and cobalt chloride hexahydrate were detected at a concentration range of 32-128 µg/mL for all isolates. None of the compounds demonstrated any cytotoxicity (to macrophage cells) at the concentration of 200 µg/mL. The activity against Sporothrix spp. recorded in this study instigate the continuity of experimental studies with Co(II) to search for the mechanisms of antifungal action as well as to evaluate its interaction with the commercial antifungal drugs.


Subject(s)
In Vitro Techniques/instrumentation , Macrophages/classification , Sporotrichosis/drug therapy , Sporothrix/classification , Pharmaceutical Preparations/administration & dosage , Chlorides/agonists , Fungi
6.
Rev. chil. infectol ; 38(6): 811-815, dic. 2021. ilus
Article in Spanish | LILACS | ID: biblio-1388310

ABSTRACT

Resumen La esporotricosis es una infección fúngica de evolución subagudacrónica causada por hongos dimórficos del Complejo Sporothrix schenckii. Es más frecuente en zonas tropicales. La prevalencia en Argentina se estima entre 0,01 y 0,02%. En la mitad de los pacientes se manifiesta como una esporotricosis linfocutánea, la cual se produce tras la inoculación del hongo en la piel luego de un trauma menor. La lesión inicial es una pápula o nódulo que se sucede con la aparición de una cadena ascendente de nódulos subcutáneos móviles, indoloros y eritematosos. El diagnóstico se realiza a partir del cultivo micológico. El antifúngico de elección es itraconazol y el pronóstico es usualmente favorable. Se presenta el caso de una niña de 4 años, previamente sana, que consultó por adenopatías axilares de evolución subaguda sin respuesta a múltiples esquemas antimicrobianos, confirmándose el diagnóstico de una esporotricosis linfocutánea por el cultivo de una biopsia ganglionar.


Abstract Sporotrichosis is a subacute-chronic fungal infection caused by dimorphic fungi of the Sporothrix schenckii Complex. It is more common in tropical areas. The prevalence in Argentina is estimated between 0.01 and 0.02%. In half of the patients it manifests as lymphocutaneous sporotrichosis, which occurs after inoculation of the fungus into the skin after minor trauma. The initial lesion is a papule or nodule that occurs with the appearance of an ascending chain of mobile, painless and erythematous subcutaneous nodules. The diagnosis is made from mycological culture. The antifungal of choice is itraconazole and the prognosis is usually favorable. We present the case of a healthy 4-year-old girl who consulted for subacute axillary lymphadenopathy without response to multiple antimicrobial regimens, arriving at the diagnosis of lymphocutaneous sporotrichosis from the culture of a lymph node biopsy sample.


Subject(s)
Humans , Male , Child, Preschool , Sporotrichosis/diagnosis , Sporotrichosis/microbiology , Sporotrichosis/drug therapy , Skin/pathology , Sporothrix , Itraconazole/therapeutic use , Antifungal Agents/therapeutic use
7.
An. bras. dermatol ; 96(3): 349-351, May-June 2021. graf
Article in English | LILACS | ID: biblio-1285054

ABSTRACT

Abstract Sporotrichosis is a subcutaneous mycosis that affects humans and animals, with a typically subacute or chronic evolution, caused by Sporothrix spp., a dimorphic fungus. Although the cutaneous form is the most frequent presentation, the ocular involvement has been more frequently diagnosed in endemic areas, affecting mainly children and the elderly. Approximately 80% of affected patients have the lymphocutaneous form, while only 2.3% have conjunctival lesions, with 0.7% showing primary ocular involvement. We describe two cases of sporotrichosis with ocular involvement in children through inoculation by felines, with a good response to antifungal treatment.


Subject(s)
Humans , Animals , Child , Aged , Cats , Sporotrichosis/diagnosis , Sporotrichosis/drug therapy , Sporothrix , Dermatomycoses/drug therapy , Eye , Antifungal Agents/therapeutic use
8.
Rev. Méd. Clín. Condes ; 32(2): 240-245, mar.-abr. 2021. tab, ilus
Article in Spanish | LILACS | ID: biblio-1518400

ABSTRACT

OBJETIVO: Reportar un caso de esporotricosis linfocutánea contraída en Chile y realizar una revisión de la literatura con énfasis en los hallazgos dermatoscópicos de las lesiones y en los casos reportados en el país. CASO CLÍNICO: Paciente adulto mayor con lesiones nodulares ulceradas siguiendo trayecto linfático en extremidad superior derecha. RESULTADOS: Estudio micológico confirmó la presencia de Sporothrix schenckii. Se inició tratamiento con antifúngico oral con buena respuesta terapéutica. CONCLUSIONES: Existe un escaso número de reporte de esporotricosis linfocutánea contraída en Chile con confirmación micológica. Este es el único caso chileno con descripción dermatoscópica de las lesiones.


OBJETIVE: To report a case of lymphocutaneous sporotrichosis adquired in Chile and provide a review of the literature with emphasis on dermoscopic findings and previous cases reported in the country. CLINICAL CASE: An elderly patient presented with ulcerated nodular lesions following lymphatic vessels in his right upper limb.RESULTS: Mycological work-up confirmed the presence of Sporothrix schenkii. Oral antifungal treatment was initiated with good therapeutic response. CONCLUSIONS: Few case reports of lymphocutaneous sporotricosis adquired in Chile can be found in literature. Until now this is the only chilean case with dermoscopic description of the lesions.


Subject(s)
Humans , Male , Aged , Sporotrichosis/diagnosis , Sporotrichosis/drug therapy , Sporotrichosis/microbiology , Sporothrix/isolation & purification , Dermoscopy , Antifungal Agents/therapeutic use
9.
An. bras. dermatol ; 96(2): 231-233, Mar.-Apr. 2021. graf
Article in English | LILACS | ID: biblio-1248728

ABSTRACT

Abstract Sporotrichosis is a subcutaneous mycosis caused by dimorphic fungi of the genus Sporothrix. The authors report a case of fixed cutaneous sporotrichosis with therapeutic failure after 18 months of itraconazole and terbinafine associated with cryosurgery. The patient was cured after the introduction of saturated potassium iodide solution. Sporothrix brasiliensis was the identified species, presenting a susceptibility profile to itraconazole and terbinafine. This fact suggests that therapeutic failure is probably related to the host-fungus interaction rather than drug resistance. It is possible that the immunomodulatory action of the saturated potassium iodide solution may have played an important role in curing this patient.


Subject(s)
Sporotrichosis/drug therapy , Sporothrix , Potassium Iodide/therapeutic use , Itraconazole , Antifungal Agents/therapeutic use
11.
An. bras. dermatol ; 95(6): 737-739, Nov.-Dec. 2020. graf
Article in English | LILACS, ColecionaSUS | ID: biblio-1142122

ABSTRACT

Abstract Sporothrix spp. infection can occur through the inoculation of the organism in the skin through direct contact with the soil (sapronotic infection), through contact with animals, such as infected cats and dogs (zoonotic infection), or less frequently via inhalation. With a subacute or chronic evolution, approximately 80% of patients affected by the disease present with the lymphocutaneous form; episodes associated with a hypersensitivity reaction are rare. The authors report the case of a 12-year-old child with immunoreactive sporotrichosis manifested clinically as erythema nodosum lesions in the lower limbs, associated with an ulcerated lesion in the left arm.


Subject(s)
Humans , Animals , Male , Child , Cats , Dogs , Sporotrichosis/diagnosis , Erythema Nodosum , Hypersensitivity , Sporothrix , Zoonoses
12.
Rev. bras. ciênc. vet ; 27(4): 183-187, out./dez. 2020. il.
Article in Portuguese | LILACS, VETINDEX | ID: biblio-1369656

ABSTRACT

A esporotricose é uma zoonose causada por fungos do gênero Sporothrix. Os gatos doentes têm importante potencial zoonótico e frequentemente revelam uma apresentação disseminada da doença. O objetivo deste relato de caso é descrever as características clínicas e anatomopatológicas de um caso de esporotricose felina refratária ao tratamento com apresentação clínica cutânea disseminada e sistêmica. Um felino macho de 3 anos de idade foi tratado para esporotricose com itraconazol e obteve resolução completa das lesões cutâneas. Porém, um ano após alta clínica, o animal apresentou aumento e ulceração da região nasal. Após cultura fúngica positiva para Sporothrix spp foi instituído tratamento com itraconazol 100mg associado ao iodeto de potássio 20mg por via oral diariamente e transcorrido 3 meses de tratamento, foi observada piora progressiva do sinais clínicos com disseminação de lesões para outras regiões do corpo. O animal foi submetido à eutanásia e encaminhado para necrópsia no Serviço de Anatomia Patológica da Faculdade de Veterinária da Universidade Federal Fluminense. Amostras da língua, pulmão, fígado, baço, rim, cérebro, linfonodo e pele foram coletadas para evidência de disseminação do agente fúngico e avaliação de alterações microscópicas. A coloração de metenamina de prata de Grocott foi utilizada para facilitar a visualização de leveduras sugestivas de Sporothrix spp nos cortes histológicos. Leveduras foram visualizadas em amostras de pele, língua, linfonodo, rim, fígado, baço e cérebro. No presente relato a disseminação do Sporothrixspp para diversos órgãos sinaliza a necessidade de uma minuciosa investigação dos casos graves de esporotricose felina para o tratamento adequado.


Sporotrichosis is a zoonosis caused by fungi of the genus Sporothrix. Sick cats have an significant zoonotic potential and often show a widespread presentation of the disease. The purpose of this case report is to describe the clinical and anatomopathological characteristics of a case of feline sporotrichosis refractory to treatment with disseminated and systemic cutaneous clinical presentation. A 3-year-old male feline was treated for sporotrichosis with itraconazole and achieved complete resolution of the skin lesions. However, one year after clinical discharge, the animal presented an increase and ulceration of the nasal region. After a positive fungal culture for Sporothrix spp, treatment with itraconazole 100mg associated with potassium iodide 20mg was instituted orally daily and after 3 months of treatment, a progressive worsening of the symptoms was observed with the spread of lesions to other regions of the body. The animal was euthanized and sent for necropsy at the Pathological Anatomy Service of the Veterinary Faculty of Universidade Federal Fluminense. Samples of the tongue, lung, liver, spleen, kidney, brain, lymph node, and skin were collected for evidence of spread of the fungal agent and evaluation of microscopic changes. Grocott silver methenamine staining was used to facilitate the visualization of yeasts suggestive of Sporothrix spp in histological sections. Yeasts were visualized in samples of skin, tongue, lymph node, kidney, liver, spleen, and brain. In the present report, the spread of Sporothrix spp to different organs signals the need for a thorough investigation of severe cases of feline sporotrichosis for the proper treatment.


Subject(s)
Animals , Cats , Autopsy/veterinary , Sporotrichosis/veterinary , Sporothrix/pathogenicity , Cats/physiology , Itraconazole/therapeutic use , Immunodeficiency Virus, Feline , Leukemia Virus, Feline
13.
Arq. bras. med. vet. zootec. (Online) ; 72(5): 2002-2006, Sept.-Oct. 2020. tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1131562

ABSTRACT

A esporotricose é uma doença emergente e a incidência de esporotricose zoonótica tem aumentado, principalmente no Brasil; a maioria dos casos está relacionada à transmissão de gatos infectados. O diagnóstico definitivo da esporotricose felina é feito por cultura fúngica; no entanto, aguardar o longo período de cultura pode atrasar o início do tratamento. O objetivo deste estudo foi detectar e determinar as espécies de Sporothrix por PCR realizado diretamente das lesões de gatos, provenientes de área endêmica, nos quais a esporotricose fazia parte do diagnóstico diferencial. Um total de 87,1% dos casos foi positivo por PCR ou cultura fúngica para Sporothrix; 81,4% foram confirmados como S. brasiliensis por PCR, 71,4% por isolamento e 65,7% pelos dois métodos. Em conclusão, a análise direta por PCR de lesões sugestivas de esporotricose em gatos é um bom método para confirmar a infecção e determinar as espécies de Sporothrix, garantindo um diagnóstico rápido; esse método tem uma boa concordância com o isolamento fúngico.(AU)


Subject(s)
Animals , Cats , Skin/injuries , Sporotrichosis/veterinary , Sporothrix/isolation & purification , Polymerase Chain Reaction/veterinary
14.
An. bras. dermatol ; 95(3): 376-378, May-June 2020. graf
Article in English | LILACS, ColecionaSUS | ID: biblio-1130870

ABSTRACT

Abstract A 44-year-old male patient presented with nodules that evolved with inflammation, following drainage of seropurulent secretion and ulceration. The patient had a 6 year-history of alcohol addiction and reported contact with cats. At the physical examination, the patient had skin-colored and erythematous nodules, and ulcers covered with thick, blackened crusts on the face, trunk and limbs. A culture of a nodule fluid revealed growth of Sporotrix sp. He also had pulmonary involvement and therefore the disease was classified as systemic sporotrichosis, a rare form that usually affect patients infected with HIV. Chronic alcohol abuse was considered the factor of immunosuppression for the patient.


Subject(s)
Humans , Male , Adult , Sporotrichosis/immunology , Sporotrichosis/pathology , Immunocompromised Host , Alcoholism/complications , Alcoholism/immunology , Immunocompetence , Sporotrichosis/chemically induced , Sporothrix/isolation & purification , Erythema/immunology , Erythema/pathology
15.
Arq. bras. med. vet. zootec. (Online) ; 71(3): 819-827, May-June 2019. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1011330

ABSTRACT

In this study, we described the antifungal activity of three Brazilian propolis extracts: brown, green and from jataí bees against Sporothrix brasiliensis. The extracts were obtained from ethanolic extraction and their chemical composition was determined by high-performance liquid chromatography coupled to mass spectrometry. The cellular toxicity was measured in MDBK (Madin-Darby Bovine Kidney) cells and quantified by the MTT assay (3- (4,5 dimethylthiazol-2yl -2,5-diphenyl-2H bromine tetrazolato). For antifungal activity, the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were determined by broth microdilution. The results showed that cell toxicity was not observed at lower concentrations (0.097 to 0.39μg/ml) for all extracts in comparison to cell control. Among the chemical compounds identified, caffeic acid, p-coumaric acid, chlorogenic acid, ferulic acid and rutin were quantified. In antifungal activity, green and jataí did not exhibit activity against the isolates (MIC and MFC greater than 0.78mg/ml). However, all isolates of S. brasiliensis were sensitive to brown propolis (MIC of 0.09 to 0.78mg/ml), including the standard strain (P<0.001). Among the Brazilian propolis studied, the brown propolis showed activity against the S. brasiliensis isolates and more studies should be undertaken in order to evaluate its promising use in the treatment of sporotrichosis.(AU)


Neste estudo, descreveu-se a atividade antifúngica de três extratos de própolis brasileiras: marrom, verde e de abelhas jataí (Tetragonisca angustula), contra Sporothrix brasiliensis. Os extratos foram obtidos de extração etanólica, e a sua composição química foi determinada por cromatografia líquida de alta eficiência, acoplada à espectrometria de massa. A toxicidade celular foi medida em células MDBK (Madin-Darby Bovine Kidney), avaliada por observação microscópica e quantificada pelo ensaio MTT (3- (4,5-dimetiltiazol-2-ilo -2,5-difenil-2H bromo tetrazolato). Para a atividade antifúngica, determinou-se a concentração inibitória mínima (CIM) e a concentração fungicida mínima (CFM) por meio de microdiluição em caldo. Os resultados mostraram que a toxicidade celular não foi observada em concentrações menores (0,097 a 0,39μg/ml). Entre os compostos químicos identificados, foram quantificados o ácido cafeico, ácido p-cumárico, ácido clorogênico, ácido ferúlico e a rutina. Na atividade antifúngica, as própolis verde e jataí não apresentaram atividade contra os isolados (CIM e CFM maior que 0,78mg/ml), porém todos os isolados de S. brasiliensis foram sensíveis à própolis marrom (CIM de 0,09 a 0,78mg/ml), incluindo a cepa padrão (P<0,001). Entre as própolis brasileiras estudadas, a marrom mostrou atividade contra S. brasiliensis, e mais estudos devem ser realizados para avaliar seu uso promissor no tratamento da esporotricose.(AU)


Subject(s)
Humans , Animals , Propolis/analysis , Propolis/therapeutic use , Sporothrix/isolation & purification , Itraconazole/therapeutic use , Drug Resistance, Fungal , Apitherapy/veterinary , Antifungal Agents/analysis
16.
Mem. Inst. Oswaldo Cruz ; 114: e190260, 2019. tab, graf
Article in English | LILACS | ID: biblio-1040612

ABSTRACT

BACKGROUND Sporotrichosis is a subcutaneous mycosis caused by dimorphic pathogenic fungi belonging to the Sporothrix genus. Pathogenic Sporothrix species typically produce melanin, which is known to be a virulence factor. OBJECTIVES The aim of this study was to perform phenotypic, genotypic, and virulence analyses of two distinct Sporothrix brasiliensis strains isolated from the same lesion on a patient from Rio de Janeiro. METHODS AND FINDINGS Genotypic analyses by partial sequencing of the calmodulin, β-tubulin, and chitin synthase genes, as well as polymerase chain reaction (PCR)-fingerprinting by T3B, M13, and GACA, showed that the isolates were very similar but not identical. Both isolates had similar phenotypic characteristics and effectively produced melanin in their yeast forms, accounting for their ability of causing disease in a murine sporotrichosis model. Remarkably, isolate B was albino in its environmental form but caused more severe disease than the pigmented A isolate. CONCLUSIONS These findings indicate that the patient was infected by two genetically and biologically distinct S. brasiliensis that vary in their production of melanin in their environmental forms. The results underscore the importance of characterizing phenotypically different isolates found in the same clinical specimen or patient.


Subject(s)
Humans , Animals , Mice , Sporotrichosis/pathology , Sporotrichosis/virology , Sporothrix/pathogenicity , Antifungal Agents/pharmacology , Phenotype , Sporothrix/drug effects , Sporothrix/genetics , Virulence , Microbial Sensitivity Tests , Polymerase Chain Reaction , DNA Fingerprinting , Disease Models, Animal , Genotype , Mice, Inbred BALB C
17.
Mem. Inst. Oswaldo Cruz ; 113(3): 185-196, Mar. 2018. tab, graf
Article in English | LILACS | ID: biblio-894908

ABSTRACT

BACKGROUND Sporotrichosis is caused by species of the genus Sporothrix. From 1998 to 2015, 4,703 cats were diagnosed at the Fundação Oswaldo Cruz (Fiocruz), Rio de Janeiro, Brazil. Even after the description of the Sporothrix species, the characterisation of feline isolates is not performed routinely. OBJECTIVES To characterise the clinical isolates from cats at the species level and correlate them with the clinical and epidemiological characteristics of the cats. METHODS Forty seven Sporothrix spp. isolates from cats assisted at Fiocruz from 2010 to 2011 were included. Medical records were consulted to obtain the clinical and epidemiological data. The isolates were identified through their morphological and physiological characteristics. T3B polymerase chain reaction (PCR) fingerprinting was used for molecular identification of the species. FINDINGS In phenotypic tests, 34 isolates were characterised as S. brasiliensis, one as S. schenckii and 12 as Sporothrix spp. PCR identified all isolates as S. brasiliensis. MAIN CONCLUSIONS S. brasiliensis is the only etiological agent of feline sporotrichosis in Rio de Janeiro to date. None association was found between the isolates and the clinical and epidemiological data. In addition, we strongly recommend the use of molecular techniques for the identification of isolates of Sporothrix spp.


Subject(s)
Sporothrix/classification , Cat Diseases/microbiology , Cat Diseases/epidemiology , Sporothrix/genetics , Brazil/epidemiology , DNA Fingerprinting
18.
Mem. Inst. Oswaldo Cruz ; 113(1): 68-70, Jan. 2018. graf
Article in English | LILACS | ID: biblio-1040580

ABSTRACT

There is a paucity of studies on the cell biology of Sporothrix luriei, the less common of the pathogenic Sporothrix species worldwide. The production of DHN-melanin, eumelanin, and pyomelanin were evaluated on the mycelial and yeast forms of the S. luriei ATCC 18616 strain. The mycelial form of this species produced only pyomelanin, which protected the fungus against environmental stressors such as ultraviolet light, heat, and cold. The yeast form was unable to produce any of the tested melanin types. The lack of melanin in the parasitic form of S. luriei may be an explanation for its low frequency in human infections.


Subject(s)
Sporothrix/metabolism , Melanins/biosynthesis
19.
Rio de Janeiro; s.n; 2018. 97 p. ilus.
Thesis in Portuguese | LILACS | ID: biblio-1046368

ABSTRACT

Os métodos de preservação empregados em micologia buscam manter a viabilidade e a estabilidade das características fenotípicas e genéticas. Entretanto, alterações podem ocorrer dependendo da metodologia empregada. Atualmente são escassos os dados da estabilidade dos isolados de Sporothrix spp., incluindo perfis fenotípicos, genotípicos e proteômicos após o armazenamento. Com a recente descrição das novas espécies patogênicas, para o homem e outros mamíferos, torna-se necessário o armazenamento desses fungos em coleções de culturas para estudos futuros, possibilitando maior conhecimento sobre a taxonomia, fisiologia e virulência de Sporothrix spp. Portanto, o objetivo desse estudo foi avaliar métodos de preservação que permitam manter a estabilidade dos perfis fenotípicos, genotípicos e proteômicos dos isolados de Sporothrix spp.. Com esse propósito foram utilizados 8 isolados de Sporotrix spp., armazenados por três diferentes métodos. Preservação em água destilada, sob refrigeração a 4ºC e criopreservação a -80°C, por períodos de 6, 12, 18 e 24 meses de armazenamento. Os isolados foram identificados pelo sequenciamento parcial dos genes calmodulina e ß-tubulina e pela PCR fingerprinting. A estabilidade do fenótipo foi realizada, por meio das provas fenotípicas sugeridas na chave de identificação das espécies de Sporothrix spp. que incluem: morfologia dos conídios, diâmetro da colônia a 30°C, teste de termotolerância a 37°C e assimilação de carboidratos, glicose sacarose e rafinose. O dimorfismo também foi avaliado após a preservação. A estabilidade genotípica foi realizada por meio do sequenciamento parcial do gene codificador da ß-tubulina


A estabilidade proteômica foi realizada por espectrometria de massa (MALDI-TOF MS), após 24 meses de preservação. Todos os isolados foram recuperados dos três métodos em todos os períodos avaliados. No entanto, os isolados preservados em água destilada apresentaram contaminações. Alterações morfológicas foram observadas após preservação a -80°C, um isolado deixou de apresentar colônia pigmentada e conídios demáceos. O diâmetro das colônias assim como a termotolerância permaneceram estáveis nos três métodos. A principal alteração ocorreu com o perfil de assimilação de carboidratos e essa variação modificou a identificação das espécies pela chave taxonômica. A estabilidade genotípica para o gene codificador da ß-tubulina foi preservada após 24 meses em ambos os métodos de armazenamento. Entretanto, baixo polimorfismo foi detectado após preservação em água destilada. Os isolados foram identificados corretamente por espectometria de massa após o armazenamento, porém alterações no perfil protéico foram observadas. O método de preservação a 4°C foi o mais efetivo para preservar os isolados de Sporothrix spp., manteve estável a maioria das características fenotípicas, como também a estabilidade do genótipo por até 24 meses de armazenamento. Esse é um método simples e de baixo custo que pode seu usado em laboratórios de recursos limitados como, um método alternativo de preservação. (AU)


Subject(s)
Animals , Phenotype , Preservation, Biological , Sporothrix , Preservation of Water Samples , Proteomics , Mitosporic Fungi , Genotype
20.
São Paulo; s.n; s.n; 2018. 93 p. tab, graf, ilus.
Thesis in Portuguese | LILACS | ID: biblio-881085

ABSTRACT

A esporotricose é uma doença crônica que envolve o tecido subcutâneo afetando seres humanos e animais, causada pelo fungo termodimórfico Sporothrix spp.. A esporotricose é endêmica na América latina, principalmente no Brasil que teve o maior surto zoonótico já registrado, ocorrendo na cidade do Rio de Janeiro. A espécie Sporothrix brasiliensis é a mais diagnosticada no surto e a mais virulenta entre as especies de Sporothrix spp., causando formas mais graves da doença. A esporotricose em gatos é endêmica, fatal e um dos principais fatores pelo alto número de casos no Rio de Janeiro. O tratamento é longo e não vem sendo o suficiente para conter o número de casos da doença. Uma vacina contra a esporotricose poderia mudar esse paradigma no Brasil. O presente trabalho obteve o proteoma da cepa S. brasiliensis 5110 por meio de uma eletroforese 2D, e caracterizou e identificou as possíveis proteínas imunogênicas do fungo por espectrometria de massa. Por meio de programas de predição, foi avaliado e sintetizado 7 sequências de aminoácidos,das proteínas identificadas com maiores chances de se acoplar a molécula MHC de classe II. Apenas 3 foram capazes de induzir proliferação in vitro, os peptídeos ZR3, ZR4 e ZR8, que foram utilizados como vacina na esporotricose subcutânea e avaliados sua eficácia por meio da carga fúngica, diâmetro das lesões, perfil celular e níveis de citocinas. Neste trabalho concluímos que o peptídeo ZR8 foi o melhor candidato à vacina na esporotricose, pois foi capaz de diminuir o diâmetro das lesões, aumentar os níveis de citocinas protetoras (IFN-γ, IL-17A e IL-1ß) e aumentar o número de células TCD4+ e CD3-/CD19+, sendo assim induzindo uma resposta imunológica protetora na esporotricose subcutânea


Sporotrichosis is a chronic disease, which involves the subcutaneous tissue affecting humans and animals caused by the thermodymorphic fungus Sporothrix spp. Sporotrichosis is endemic in Latin America, mainly in Brazil that had the largest zoonotic outbreak ever recorded, occurring in the city of Rio de Janeiro. The Sporothrix brasiliensis is the species more diagnosed in the outbreak and most virulent, causing severe forms of the disease. Sporotrichosis in cats is endemic, fatal and the main factors due to the high number of cases of the disease in Rio de Janeiro. The treatment is long, and has not been enough to contain the number of cases of sporotrichosis. A vaccine against sporotrichosis could change this paradigm in Brazil. The present work obtained the proteome of S. brasiliensis 5110 strain by 2D electrophoresis, and characterized and identified possible immunogenic proteins by mass spectrometry. By prediction programs were evaluated and synthesized 7 peptide sequence from antigenic proteins that have the highest chances of coupling to the MHC class II molecule. From these 7 peptides only 3 were able to induce proliferation in vitro, called ZR3, ZR4 and ZR8 peptides, that were used as a vaccine in subcutaneous sporotrichosis and evaluated their efficacy through fungal load, lesion diameter, cell profile and cytokine levels. We conclude that ZR8 peptide was the best candidate for sporotrichosis vaccine, since it was able to decrease the lesion diameter, increase the levels of protective cytokines (IFN-γ, IL-17A and IL-1ß) and increase the number of CD4+ T cells and CD3-/CD19+ inducing a protective immune response in subcutaneous sporotrichosis


Subject(s)
Animals , Male , Female , Mice , Sporotrichosis/complications , Sporothrix/classification , Vaccines/analysis , Mass Spectrometry/methods , Blotting, Western/methods , Proteome/pharmacology , Forecasting/methods
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