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Chromosomal Polymorphism in the Sporothrix schenckii Complex  [PDF]
Alexandre A. Sasaki, Geisa F. Fernandes, Anderson M. Rodrigues, Fábio M. Lima, Marjorie M. Marini, Luciano dos S. Feitosa, Marcus de Melo Teixeira, Maria Sueli Soares Felipe, José Franco da Silveira, Zoilo P. de Camargo
PLOS ONE , 2014, DOI: 10.1371/journal.pone.0086819
Abstract: Sporotrichosis is a polymorphic disease caused by a complex of thermodimorphic fungi including S. brasiliensis, S. schenckii sensu stricto (s. str.), S. globosa and S. luriei. Humans and animals can acquire the disease through traumatic inoculation of propagules into the subcutaneous tissue. Despite the importance of sporotrichosis as a disease that can take epidemic proportions there are just a few studies dealing with genetic polymorphisms and genomic architecture of these pathogens. The main objective of this study was to investigate chromosomal polymorphisms and genomic organization among different isolates in the S. schenckii complex. We used pulsed field gel electrophoresis (PFGE) to separate chromosomal fragments of isolated DNA, followed by probe hybridization. Nine loci (β-tubulin, calmodulin, catalase, chitin synthase 1, Internal Transcribed Spacer, Pho85 cyclin-dependent kinase, protein kinase C Ss-2, G protein α subunit and topoisomerase II) were mapped onto chromosomal bands of Brazilian isolates of S. schenckii s. str. and S. brasiliensis. Our results revealed the presence of intra and interspecies polymorphisms in chromosome number and size. The gene hybridization analysis showed that closely related species in phylogenetic analysis had similar genetic organizations, mostly due to identification of synteny groups in chromosomal bands of similar sizes. Our results bring new insights into the genetic diversity and genome organization among pathogenic species in the Sporothrix schenckii complex.
Adhesion of the human pathogen Sporothrix schenckii to several extracellular matrix proteins
Lima, O.C.;Figueiredo, C.C.;Pereira, B.A.S.;Coelho, M.G.P.;Morandi, V.;Lopes-Bezerra, L.M.;
Brazilian Journal of Medical and Biological Research , 1999, DOI: 10.1590/S0100-879X1999000500020
Abstract: the pathogenic fungus sporothrix schenckii is the causative agent of sporotrichosis. this subcutaneous mycosis may disseminate in immunocompromised individuals and also affect several internal organs and tissues, most commonly the bone, joints and lung. since adhesion is the first step involved with the dissemination of pathogens in the host, we have studied the interaction between s. schenckii and several extracellular matrix (ecm) proteins. the binding of two morphological phases of s. schenckii, yeast cells and conidia, to immobilized type ii collagen, laminin, fibronectin, fibrinogen and thrombospondin was investigated. poly (2-hydroxyethyl methacrylate) (poly-hema) was used as the negative control. cell adhesion was assessed by elisa with a rabbit anti-s. schenckii antiserum. the results indicate that both morphological phases of this fungus can bind significantly to type ii collagen, fibronectin and laminin in comparison to the binding observed with bsa (used as blocking agent). the adhesion rate observed with the ecm proteins (type ii collagen, fibronectin and laminin) was statistically significant (p<0.05) when compared to the adhesion obtained with bsa. no significant binding of conidia was observed to either fibrinogen or thrombospondin, but yeast cells did bind to the fibrinogen. our results indicate that s. schenckii can bind to fibronectin, laminin and type ii collagen and also show differences in binding capacity according to the morphological form of the fungus.
Adhesion of the human pathogen Sporothrix schenckii to several extracellular matrix proteins  [cached]
Lima O.C.,Figueiredo C.C.,Pereira B.A.S.,Coelho M.G.P.
Brazilian Journal of Medical and Biological Research , 1999,
Abstract: The pathogenic fungus Sporothrix schenckii is the causative agent of sporotrichosis. This subcutaneous mycosis may disseminate in immunocompromised individuals and also affect several internal organs and tissues, most commonly the bone, joints and lung. Since adhesion is the first step involved with the dissemination of pathogens in the host, we have studied the interaction between S. schenckii and several extracellular matrix (ECM) proteins. The binding of two morphological phases of S. schenckii, yeast cells and conidia, to immobilized type II collagen, laminin, fibronectin, fibrinogen and thrombospondin was investigated. Poly (2-hydroxyethyl methacrylate) (poly-HEMA) was used as the negative control. Cell adhesion was assessed by ELISA with a rabbit anti-S. schenckii antiserum. The results indicate that both morphological phases of this fungus can bind significantly to type II collagen, fibronectin and laminin in comparison to the binding observed with BSA (used as blocking agent). The adhesion rate observed with the ECM proteins (type II collagen, fibronectin and laminin) was statistically significant (P<0.05) when compared to the adhesion obtained with BSA. No significant binding of conidia was observed to either fibrinogen or thrombospondin, but yeast cells did bind to the fibrinogen. Our results indicate that S. schenckii can bind to fibronectin, laminin and type II collagen and also show differences in binding capacity according to the morphological form of the fungus.
Phenotypic polomorphism in indigenous strains of sporothrix schenckii Polimorfismo fenotípico de cepas autóctonas de Sporothrix schenckii
Herta Vélez Arango,Lucía Santamaría de Uribe,María Elena Vargas de Betancur,Fernando Montoya Maya
Iatreia , 1996,
Abstract: We studied macroscopic colony findings, sugar assimilation patterns and virulence of 55 Sporothrix schenckij strains obtained from patients with cutaneous sporothrichosis. They were collected during a 10-year period at the Mycology Laboratory, University of Antioquia, School of Medicine, Medellín, Colombia. Pigmentation types and macroscopic morphological characteristics’ were studied on mycosel agar and malt extract. In most cases 2 or 3 colony colors were present In both media. In mycosel agar only 5 strains (9.1%) were monochromatic. Pigmentation was very stable in that medium. Eighty five percent of the mycosel agar colonies were beige, brown, pleated or pleated and umbilicated. All strains assimilated D-glucosa, glycerol and D-xylosa. We established 9 patterns of assimilation (blotypes), from A to I In 25 strains. Both pigmented and albino strains were virulent for mice. We emphasize the diversity of our Indigenous strains, and the importance of genotypic characterization and of the correlation studies of phenotypic and genotypic variation with the clinical and geographical patterns of the disease. Se presentan los resultados macroscópicos, de asimilación de azúcares y de virulencia de 55 cepas de Sporothrix schenckii aisladas a partir de lesiones de pacientes con esporotricosis cutánea, que consultaron al laboratorio de micología de la Facultad de Medicina de la Universidad de Antioquia en Medellín, Colombia, y que fueron coleccionadas en el transcurso de 10 a os. La morfología macroscópica de las colonias y su pigmentación se evaluaron tanto en agar mycosel como en extracto de malta. La mayoría de las cepas en los dos medios presentaban colonias con 2 ó 3 colores diferentes. En mycosel 5 cepas (9.1%) fueron monocromáticas y éste fue el medio más estable para definir las características de pigmentación. El 85% de las cepas en mycosel fueron café claras, café oscuras, plisadas o plisadas y umbilicadas. Todas las cepas asimilaron D-glucosa, glicerol y D-xilosa en el sistema Api 20C y 25 cepas se clasificaron en 9 biotipos de asimilación de la A a la I. La mayoría de las cepas tanto pigmentadas como albinas, resultaron virulentas para ratones. En éstos predominaron los cuerpos en cigarro en forma de naveta y no se visual izaron cuerpos asteroides en los exudados testiculares. Se demuestra así la gran heterogeneidad fenotípica de las cepas autóctonas de S. schenckii, se plantea la importancia de correlacionar estos hallazgos con los patrones de heterogeneidad gen ética informados por investigadores Japoneses y quizás explicar por esta diversidad fenot
2D-immunoblotting analysis of Sporothrix schenckii cell wall
Ruiz-Baca, Estela;Mora-Montes, Héctor M;López-Romero, Everardo;Toriello, Conchita;Mojica-Marín, Virgilio;Urtiz-Estrada, Norma;
Memórias do Instituto Oswaldo Cruz , 2011, DOI: 10.1590/S0074-02762011000200021
Abstract: we utilized two-dimensional gel electrophoresis and immunoblotting (2d-immunoblotting) with anti-sporothrix schenckii antibodies to identify antigenic proteins in cell wall preparations obtained from the mycelial and yeast-like morphologies of the fungus. results showed that a 70-kda glycoprotein (gp70) was the major antigen detected in the cell wall of both morphologies and that a 60-kda glycoprotein was present only in yeast-like cells. in addition to the gp70, the wall from filament cells showed four proteins with molecular weights of 48, 55, 66 and 67 kda, some of which exhibited several isoforms. to our knowledge, this is the first 2d-immunoblotting analysis of the s. schenckii cell wall.
Ultrastructural study of the mycelial phase of clinical isolates of Sporothrix schenckii obtained from feline, canine and human cases of sporotrichosis
Madrid, Isabel Martins;Mattei, Antonella Souza;Soares, Mauro Pereira;Nobre, Márcia de Oliveira;Meireles, Mário Carlos Araújo;
Brazilian Journal of Microbiology , 2011, DOI: 10.1590/S1517-83822011000300037
Abstract: using transmission electron microscopy, we studied the presence of melanin and cell wall thickness of clinical isolates of sporothrix schenckii obtained from cats, dogs and humans as compared to reference strains. we detected differences regarding presence of the melanin among the clinical isolates of s. schenckii and a correlation between presence of melanin and cell wall thickness.
Atividade in vitro do óleo essencial de Origanum vulgare frente à Sporothrix Schenckii
Cleff, M.B.;Meinerz, A.R.M.;Schuch, L.F.D.;Rodrigues, M.R.A.;Meireles, M.C.A.;Mello, J.R.B.;
Arquivo Brasileiro de Medicina Veterinária e Zootecnia , 2008, DOI: 10.1590/S0102-09352008000200039
Abstract: in vitro activity of the essential oil origanum vulgare against sporothrix schenckii was determined by the mic (minimum inhibitory concentration). for this, seven samples of s. schenckii were studied, two isolated from two cases of human sporotrichosis and five isolated from cats. analysis of the essential oil was carried out in a gas chromatograph (gc/fid) for the identification and quantification of thymol and carvacrol (antifungal agents). mic was obtained based on the microdilution method according to the adapted document nccls-m 27a2 for fitopharmacy. all the isolates presented sensibility to the essential oil. s. schenckii was inhibited in a concentration of 0.25% (250m l/ml). chromatographic analysis showed that thymol concentration was bigger than carvacrol. the antifungal activity demonstrated by the essential oil of o. vulgare against s. schenckii stimulates the accomplishment of more studies, including in vivo studies.
Utilidade do coágulo sangüíneo para o isolamento de Sporothrix schenckii de gatos naturalmente infectados
Schubach, Tania Maria Pacheco;Schubach, Armando de Oliveira;Okamoto, Thais;Figueiredo, Fabiano Borges;Pereira, Sandro Antonio;Leme, Luiz Rodrigo Paes;Santos, Isabele Barbiere dos;Reis, Rosani Santos dos;Paes, Rodrigo de Almeida;Perez, Mauricio de Andrade;Marzochi, Mauro Célio de Almeida;Francesconi-do-Valle, Ant?nio Carlos;Wanke, Bodo;
Brazilian Journal of Veterinary Research and Animal Science , 2004, DOI: 10.1590/S1413-95962004000600008
Abstract: the diagnosis of disseminated sporotrichosis is usually obtained by necropsy and the isolation of sporothrix schenckii from blood is rare. fungemia was shown in vivo through the isolation of s. schenckii from peripheral blood of 13 (n=38, 34.2%) cats with naturally acquired sporotrichosis. the coinfection with fiv and with felv found, respectively, in 6 (n=34, 17.6%) cases and 1 (n=34, 2.9%), apparently did not alter the frequency of the isolation of s. schenckii from peripheral blood. there was agreement of 84.2% comparing these results to the blood culture results simultaneously achieved. in this way, we propose the clot culture as a practical alternative method, efficient and cheap for the diagnosis of disseminated sporotrichosis in cats in vivo.
Comportamiento experimental del Sporothrix schenckii y la Leishmania mexicana en el hamster
González de Polanía, Luz Angela;Alzate, Alberto;Saravia, Nancy;
Revista do Instituto de Medicina Tropical de S?o Paulo , 1990, DOI: 10.1590/S0036-46651990000500002
Abstract: the macroscopic description of the pathogenic process of sporothrix schenckii and leishmania mexicana spp in hamsters inoculated subcutaneously in the nose provided bases for the differentiation of the behavior of these two microorganisms in a model frequently utilized for their study. sequential observations over 150 days demonstrated that infections caused by these pathogens results initially in edema and erythema followed by loss of hair, necrosis and ulceration. the pus production was a characteristic presented only by s. schenckii. these clinical signs were observed earlier in sporotrichosis than in l. mexicana infection. differences in the mean day of appearance were statistically significant. the lesions produced by s. schenckii and l. mexicana in this experimental model share clinical signs, but their incubated period and relative frequency allow us to differentiate them. the circumstances of inoculation such as strain, dose of inoculum, place and route of inoculation are related to the experimental behavior of them.
Molecular characterisation of Sporothrix schenckii isolates from humans and cats involved in the sporotrichosis epidemic in Rio de Janeiro, Brazil
Reis, Rosani Santos;Almeida-Paes, Rodrigo;Muniz, Mauro de Medeiros;Tavares, Patrícia Morais e Silva;Monteiro, Paulo Cezar Fialho;Schubach, Tania Maria Pacheco;Gutierrez-Galhardo, Maria Clara;Zancopé-Oliveira, Rosely Maria;
Memórias do Instituto Oswaldo Cruz , 2009, DOI: 10.1590/S0074-02762009000500018
Abstract: an epidemic of sporotrichosis, a subcutaneous mycosis caused by the fungus sporothrix schenckii, is ongoing in rio de janeiro, brazil, in which cases of human infection are related to exposure to cats. in an attempt to demonstrate the zoonotic character of this epidemic using molecular methodology, we characterised by dna-based typing methods 19 human and 25 animal s. schenckii isolates from the epidemic, as well as two control strains. to analyse the isolates, the random amplified polymorphic dna (rapd) technique was performed using three different primers, together with dna fingerprinting using the minisatellite derived from the wild-type phage m13 core-sequence. the analyses generated amplicons with considerable polymorphism. although isolates exhibited high levels of genetic relatedness, they could be clustered into 5-10 genotypes. the rapd profiles of epidemic s. schenckii isolates could be distinguished from that of the united states isolate, displaying 20% similarity to each primer and 60% when amplified with the m13 primer. dna fingerprinting of s. schenckii isolated from the nails (42.8%) and the oral cavities (66%) of cats were identical to related human samples, suggesting that there is a common infection source for animals and humans in this epidemic. it is clear that cats act as a vehicle for dissemination of s. schenckii.
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