325 results on '"Bagagli, Eduardo"'
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102. Ecological study of Paracoccidioides brasiliensis in soil: growth ability, conidia production and molecular detection
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Terçarioli, Gisela Ramos, primary, Bagagli, Eduardo, additional, Reis, Gabriela Martins, additional, Theodoro, Raquel Cordeiro, additional, Bosco, Sandra De Moraes Gimenes, additional, Macoris, Severino Assis da Graça, additional, and Richini-Pereira, Virgínia Bodelão, additional
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- 2007
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103. Pythium insidiosum: relato do primeiro caso de infecção humana no Brasil
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Marques, Silvio Alencar, primary, Bagagli, Eduardo, additional, Bosco, Sandra M. G., additional, Camargo, Rosangela M. P., additional, and Marques, Mariangela E. A., additional
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- 2006
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104. Phylogenetic and evolutionary aspects of Paracoccidioides brasiliensis reveal a long coexistence with animal hosts that explain several biological features of the pathogen
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Bagagli, Eduardo, primary, Bosco, Sandra M.G., additional, Theodoro, Raquel Cordeiro, additional, and Franco, Marcello, additional
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- 2006
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105. Study of Toxoplasma infection in Brazilian wild mammals: Serological evidence in Dasypus novemcinctus Linnaeus, 1758 and Euphractus sexcinctus Wagler, 1830
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da Silva, Aristeu Vieira, primary, de Moraes Gimenes Bosco, Sandra, additional, Langoni, Helio, additional, and Bagagli, Eduardo, additional
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- 2006
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106. Reply to Dr Conti-Diaz
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Bagagli, Eduardo, primary
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- 2006
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107. Cryptic Speciation and Recombination in the Fungus Paracoccidioides brasiliensis as Revealed by Gene Genealogies
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Matute, Daniel R., primary, McEwen, Juan G., additional, Puccia, Rosana, additional, Montes, Beatriz A., additional, San-Blas, Gioconda, additional, Bagagli, Eduardo, additional, Rauscher, Jason T., additional, Restrepo, Angela, additional, Morais, Favia, additional, Niño-Vega, Gustavo, additional, and Taylor, John W., additional
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- 2005
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108. Primers for Clinical Detection of Paracoccidioides brasiliensis
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San-Blas, Gioconda, primary, Niño-Vega, Gustavo, additional, Barreto, Laura, additional, Hebeler-Barbosa, Flavia, additional, Bagagli, Eduardo, additional, Olivero de Briceño, Rosa, additional, and Mendes, Rinaldo Poncio, additional
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- 2005
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109. Molecular detection ofParacoccidioides brasiliensisin soil
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Theodoro, Raquel Cordeiro, primary, Candeias, João Manuel Grisi, additional, Araújo, João Pessoa, additional, Bosco, Sandra De Moraes Gimenes, additional, Macoris, Severino Assis Da Graça, additional, Junior, Luis Otavio Padula, additional, Franco, Marcello, additional, and Bagagli, Eduardo, additional
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- 2005
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110. Distribution of paracoccidioidomycosis: determination of ecologic correlates through spatial analyses
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Simões, Ligia Barrozo, primary, Marques, Sílvio Alencar, additional, and Bagagli, Eduardo, additional
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- 2004
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111. Comparison of the Sequences of the Internal Transcribed Spacer Regions and PbGP43 Genes of Paracoccidioides brasiliensis from Patients and Armadillos ( Dasypus novemcinctus )
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Hebeler-Barbosa, Flavia, primary, Morais, Flavia V., additional, Montenegro, Mario R., additional, Kuramae, Eiko E., additional, Montes, Beatriz, additional, McEwen, Juan G., additional, Bagagli, Eduardo, additional, and Puccia, Rosana, additional
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- 2003
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112. Analysis of the synaptonemal complex of the nine-banded armadillo, Dasypus novemcinctus
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Scavone, Márcia Denise da Paixão, primary, Oliveira, Claudio, additional, Bagagli, Eduardo, additional, and Foresti, Fausto, additional
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- 2000
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113. Paracoccidioides lutzii sp. nov.: biological and clinical implications.
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Teixeira, Marcus de Melo, Theodoro, Raquel Cordeiro, Oliveira, Fabiana Freire Mendes de, Machado, Gabriel Capella, Hahn, Rosane Christine, Bagagli, Eduardo, San-Blas, Gioconda, and Soares Felipe, Maria Sueli
- Abstract
Paracoccidioides lutzii, formerly known as ‘Pb01-like’ strains in the P. brasiliensis complex, is proposed as a new species based on phylogenetic and comparative genomics data, recombination analysis, and morphological characteristics. Conidia of P. lutzii are elongated, different from those of P. brasiliensis. P. lutzii occurs in the central and northern regions of Brazil. Studies comparing P. brasiliensis and P. lutzii may have significant clinical consequences for the diagnosis and treatment of paracoccidioidomycosis. [ABSTRACT FROM PUBLISHER]
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- 2014
114. Genetic recombinants inTrichoderma pseudokoningii(Rifai) without typical parasexuality
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Bagagli, Eduardo, primary, Furlaneto, Márcia Cristina, additional, Pizzirani-Kleiner, Aline, additional, and Azevedo, João Lúcio, additional
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- 1995
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115. Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Differentiation of the Dimorphic Fungal Species Paracoccidioides brasiliensisand Paracoccidioides lutzii
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Nobrega de Almeida, João, Del Negro, Gilda M. B., Grenfell, Rafaella C., Vidal, Monica S. M., Thomaz, Danilo Y., de Figueiredo, Dulce S. Y., Bagagli, Eduardo, Juliano, Luiz, and Benard, Gil
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ABSTRACTIsolates of Paracoccidioides brasiliensisand Paracoccidioides lutzii, previously characterized by molecular techniques, were identified for the first time by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS). All isolates were correctly identified, with log score values of >2.0. Thus, MALDI-TOF MS is a new tool for differentiating species of the genus Paracoccidioides.
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- 2015
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116. Detection of Paracoccidioides spp. in environmental aerosol samples.
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Arantes, Thales Domingos, Theodoro, Raquel Cordeiro, Da Graça Macoris, Severino Assis, and Bagagli, Eduardo
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Taking into account that paracoccidioidomycosis infection occurs by inhalation of the asexual conidia produced by Paracoccidioides spp. in its saprobic phase, this work presents the collection of aerosol samples as an option for environmental detection of this pathogen, by positioning a cyclonic air sampler at the entrance of armadillo burrows. Methods included direct culture, extinction technique culture and Nested PCR of the rRNA coding sequence, comprising the ITS1-5.8S-ITS2 region. In addition, we evaluated one armadillo ( Dasypus novemcinctus) as a positive control for the studied area. Although the pathogen could not be isolated by the culturing strategies, the aerosol sampling associated with molecular detection through Nested PCR proved the best method for discovering Paracoccidioides spp. in the environment. Most of the ITS sequences obtained in this investigation proved to be highly similar with the homologous sequences of Paracoccidioides lutzii from the GenBank database, suggesting that this Paracoccidioides species may not be exclusive to mid-western Brazil as proposed so far. [ABSTRACT FROM AUTHOR]
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- 2013
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117. Paracoccidioidomycosis in a Dog: Case Report of Generalized Lymphadenomegaly.
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de Farias, Marconi Rodrigues, Condas, Larissa Anuska Zeni, Ribeiro, Márcio Garcia, de Moraes Gimenes Bosco, Sandra, Muro, Marisol Dominguez, Werner, Juliana, Theodoro, Raquel Cordeiro, Bagagli, Eduardo, Marques, Sílvio Alencar, and Franco, Marcello
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Paracoccidioidomycosis (PCM) is a severe systemic mycosis, endemic in Latin America and highly prevalent in Brazil, where it ranks eighth as a mortality cause among infectious and parasitic diseases in humans. The disease in animals has been little explored. It is observed that armadillos can harbor the fungus at high frequencies, although the active disease has not been well documented in this wild mammal. Dogs are susceptible to experimental infection, and the naturally acquired PCM-disease was reported only recently in a dog from Brazil. The present work reports the second case of naturally acquired PCM in a 6-year-old female dog that presented emaciation, lymphadenomegaly, and hepatosplenomegaly. Biochemical and pulmonary radiographic evaluation did not reveal any abnormalities. PCM was diagnosed by clinical findings, culturing, immunohistochemistry, and histopathology of popliteal lymph node. The fungus was recovered from popliteal lymph node, and the molecular analysis showed respective sequencing similarities of 99 and 100% for 803 nucleotides of the Gp43 gene and 592 nucleotides from the ITS- 5.8S region of Paracoccidioides brasiliensis. Immunohistochemistry revealed severe lymphadenitis and presented numerous yeasts, which reacted against the gp43 antibody. Histopathology revealed a severe granulomatous lymphadenitis associated with numerous single or multiple budding yeasts. After diagnosis, the dog was successfully treated with itraconazol for 2 years. Veterinarians should be aware of the importance of considering PCM for differential diagnosis, especially in dogs from PCM-endemic areas, whose monophagocytic system involvement is evident. [ABSTRACT FROM AUTHOR]
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- 2011
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118. Morphological and molecular characterization of an equine isolate of Pythium insidiosum and comparison with the first human isolate from the same geographic region.
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De Moraes Gimenes Bosco, Sandra, Martins Reis, Gabriela, Cordeiro Theodoro, Raquel, Assis Da Graca Macoris, Severino, Alencar Marques, Silvio, Da Graca Macoris, Delphim, and Bagagli, Eduardo
- Abstract
Pythium insidiosum causes pythiosis, a life-threatening disease that occurs in tropical regions and affects man and animals. Although pythiosis in Brazil had been described in various animal species, the first human case was only recently reported. The present study aimed to characterize the morphologic and molecular characteristics of a new equine isolate of P. insidiosum and compare them with those of the first Brazilian human isolate. Both isolates were recovered from the same region of the country. Macroscopic and microscopic features were evaluated in two culture media. Sporangia formation and zoospore release were obtained after culturing the isolates with fragments of grasses and crops in an appropriate liquid induction medium. The molecular analysis of the isolates consisted of the complete sequencing of the ITS-5.8S rDNA region and sequences of both showed identical composition of 836 bp and 99% similarity with the isolates M16, 65, M12, 339 and 394 deposited at GenBank. Simple mycological procedures such as the production of sporangia and zoospores may distinguish P. insidiosum from zygomycetes. The rDNA sequencing indicates that, in Brazil, both humans and animals might be infected by a common genotype of the pathogen. [ABSTRACT FROM AUTHOR]
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- 2008
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119. Molecular detection of Paracoccidioides brasiliensis in road-killed wild animals.
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Richini-Pereira, Virgínia Bodelão, Bosco, Sandra De Moraes Gimenes, Griese, Juliana, Theodoro, Raquel Cordeiro, Macoris, Severino Assis Da Graça, Da Silva, Reinaldo José, Barrozo, Lígia, Tavares, Patrícia Morais E Silva, Zancopé-Oliveira, Rosely Maria, and Bagagli, Eduardo
- Abstract
Paracoccidioides brasiliensis infections have been little studied in wild and/or domestic animals, which may represent an important indicator of the presence of the pathogen in nature. Road-killed wild animals have been used for surveillance of vectors of zoonotic pathogens and may offer new opportunities for eco-epidemiological studies of paracoccidiodomycosis (PCM). The presence of P. brasiliensis infection was evaluated by Nested-PCR in tissue samples collected from 19 road-killed animals; 3 Cavia aperea (guinea pig), 5 Cerdocyon thous (crab-eating-fox), 1 Dasypus novemcinctus (nine-banded armadillo), 1 Dasypus septemcinctus (seven-banded armadillo), 2 Didelphis albiventris (white-eared opossum), 1 Eira barbara (tayra), 2 Gallictis vittata (grison), 2 Procyon cancrivorus (raccoon) and 2 Sphiggurus spinosus (porcupine). Specific P. brasiliensis amplicons were detected in (a) several organs of the two armadillos and one guinea pig, (b) the lung and liver of the porcupine, and (c) the lungs of raccoons and grisons. P. brasiliensis infection in wild animals from endemic areas might be more common than initially postulated. Molecular techniques can be used for detecting new hosts and mapping 'hot spot' areas of PCM. [ABSTRACT FROM AUTHOR]
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- 2008
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120. Molecular detection of Paracoccidioides brasiliensis in soil.
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Theodoro, Raquel Cordeiro, Candeias, João Manuel Grisi, Araújo Jr., João Pessoa, Bosco, Sandra De Moraes, Macoris, Severino Assis Da Graça, Junior, Luis Otavio Padula, Franco, Marcello, and Bagagli, Eduardo
- Subjects
RECOMBINANT DNA ,PARACOCCIDIOIDES brasiliensis ,DNA ,GENES ,SOILS - Abstract
The objective of the present study was to evaluate different techniques for the detection of Paracoccidioides brasiliensis in soil, e.g., culture, animal inoculation and specific DNA amplification by Nested PCR. We designed species-specific inner primers derived from rDNA regions (ITS, 5.8S gene) and found their sensitivity to be higher than culture and animal inoculation. In addition, the sensitivity of these primers was higher than p27-gene primers developed for detection of P. brasiliensis in soil in a previous study. DNA from P. brasiliensis was detected in soil artificially seeded with the fungus (positive soil control) and from environmental samples collected in an armadillo burrow. [ABSTRACT FROM AUTHOR]
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- 2005
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121. Human Pythiosis, Brazil.
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De Moraes Gimenes Bosco, Sandra, Bagagli, Eduardo, Araújo Jr., João Pessoa, Candeias, João Manuel Grisi, De Franco, Marcello Fabiano, Marques, Mariangela Esther Alencar, Mendoza, Leonel, De Camargo, Rosangela Pires, and Marques, Silvio Alencar
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MEDICAL research , *WATERBORNE infection , *AQUATIC organisms , *PATHOGENIC microorganisms - Abstract
Pythiosis. caused by Pythium insidiosum, occurs in humans and animals and is acquired from aquatic environments that harbor the emerging pathogen. Diagnosis is difficult because clinical and histopathologic features are not pathognomonic. We report the first human case of pythiosis from Brazil, diagnosed by using culture and rDNA sequencing. [ABSTRACT FROM AUTHOR]
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- 2005
122. Analysis of Inteins in the Candida parapsilosisComplex for Simple and Accurate Species Identification
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Prandini, Tâmara Heloísa Rocha, Theodoro, Raquel Cordeiro, Bruder-Nascimento, Ariane C. M. O., Scheel, Christina M., and Bagagli, Eduardo
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ABSTRACTInteins are coding sequences that are transcribed and translated with flanking sequences and then are excised by an autocatalytic process. There are two types of inteins in fungi, mini-inteins and full-length inteins, both of which present a splicing domain containing well-conserved amino acid sequences. Full-length inteins also present a homing endonuclease domain that makes the intein a mobile genetic element. These parasitic genetic elements are located in highly conserved genes and may allow for the differentiation of closely related species of the Candida parapsilosis(psilosis) complex. The correct identification of the three psilosis complex species C. parapsilosis, Candida metapsilosis, and Candida orthopsilosisis very important in the clinical setting for improving antifungal therapy and patient care. In this work, we analyzed inteins that are present in the vacuolar ATPase gene VMAand in the threonyl-tRNA synthetase gene ThrRSin 85 strains of the Candidapsilosis complex (46 C. parapsilosis, 17 C. metapsilosis, and 22 C. orthopsilosis). Here, we describe an accessible and accurate technique based on a single PCR that is able to differentiate the psilosis complex based on the VMA intein. Although the ThrRS intein does not distinguish the three species of the psilosis complex by PCR product size, it can differentiate them by sequencing and phylogenetic analysis. Furthermore, this intein is unusually present as both mini- and full-length forms in C. orthopsilosis. Additional population studies should be performed to address whether this represents a common intraspecific variability or the presence of subspecies within C. orthopsilosis.
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- 2013
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123. Molecular and Morphological Data Support the Existence of a Sexual Cycle in Species of the Genus Paracoccidioides
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Teixeira, Marcus de Melo, Theodoro, Raquel Cordeiro, Derengowski, Lorena da Silveira, Nicola, André Moraes, Bagagli, Eduardo, and Felipe, Maria Sueli
- Abstract
ABSTRACTThe genus Paracoccidioidesincludes the thermodimorphic species Paracoccidioides brasiliensisand P. lutzii, both of which are etiologic agents of paracoccidioidomycosis, a systemic mycosis that affects humans in Latin America. Despite the common occurrence of a sexual stage among closely related fungi, this has not been observed with Paracoccidioidesspecies, which have thus been considered asexual. Molecular evolutionary studies revealed recombination events within isolated populations of the genus Paracoccidioides, suggesting the possible existence of a sexual cycle. Comparative genomic analysis of all dimorphic fungi and Saccharomyces cerevisiaedemonstrated the presence of conserved genes involved in sexual reproduction, including those encoding mating regulators such as MAT, pheromone receptors, pheromone-processing enzymes, and mating signaling regulators. The expression of sex-related genes in the yeast and mycelial phases of both Paracoccidioidesspecies was also detected by real-time PCR, with nearly all of these genes being expressed preferentially in the filamentous form of the pathogens. In addition, the expression of sex-related genes was responsive to the putative presence of pheromone in the supernatants obtained from previous cocultures of strains of two different mating types. In vitrocrossing of isolates of different mating types, discriminated by phylogenetic analysis of the a-box (MAT1-1) and the high-mobility-group (HMG) domain (MAT1-2), led to the identification of the formation of young ascocarps with constricted coiled hyphae related to the initial stage of mating. These genomic and morphological analyses strongly support the existence of a sexual cycle in species of the genus Paracoccidioides.
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- 2013
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124. Comparison of the Sequences of the Internal Transcribed Spacer Regions and PbGP43Genes of Paracoccidioides brasiliensisfrom Patients and Armadillos (Dasypus novemcinctus)
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Hebeler-Barbosa, Flavia, Morais, Flavia V., Montenegro, Mario R., Kuramae, Eiko E., Montes, Beatriz, McEwen, Juan G., Bagagli, Eduardo, and Puccia, Rosana
- Abstract
ABSTRACTParacoccidioides brasiliensisisolates from 10 nine-banded armadillos (Dasypus novemcinctus) were comparable with 19 clinical isolates by sequence analysis of the PbGP43gene and ribosomal internal transcribed spacer 1 (ITS1) and ITS2 and by random amplified polymorphic DNA. In this original ITS study, eight isolates differed by one or three sites among five total substitution sites.
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- 2003
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125. Soil mycobiome associated with subtropical region of Brazil
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Yamauchi, Danielle Hamae, Universidade Estadual Paulista (Unesp), Bagagli, Eduardo [UNESP], and Trovão, Flávia Hebeler Barbosa
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Solo ,Fungos patogênicos ,NGS ,Micobioma ,Metagenômica - Abstract
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Mas, nos agradecimentos de sua dissertação, você cita também a Fapesp. E a FAPESP faz algumas exigências na inclusão dos agradecimentos pelo financiamento recebido, uma delas é a obrigatoriedade de se ter o número do processo também no agradecimento. Vi que você agradeceu, faltou incluir o número do processo. Assim que tiver efetuado a correção submeta o arquivo, em formato PDF, novamente. Agradecemos a compreensão. on 2020-06-16T14:51:35Z (GMT) Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Made available in DSpace on 2020-06-18T12:40:54Z (GMT). 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No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Rejected by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo as orientações abaixo: Problema 1: Financiamento Fapesp No formulário de submissão consta somente a Capes como agência financiadora do seu projeto. Mas, nos agradecimentos de sua dissertação, você cita também a Fapesp. E a FAPESP faz algumas exigências na inclusão dos agradecimentos pelo financiamento recebido, uma delas é a obrigatoriedade de se ter o número do processo também no agradecimento. Vi que você agradeceu, faltou incluir o número do processo. Assim que tiver efetuado a correção submeta o arquivo, em formato PDF, novamente. Agradecemos a compreensão. on 2020-06-16T14:51:35Z (GMT) Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2020-06-18T12:40:54Z (GMT) No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Submitted by Danielle Hamae Yamauchi (danielle.yamauchi@unesp.br) on 2020-06-15T15:59:31Z No. of bitstreams: 1 dissertação.pdf: 1574303 bytes, checksum: 3d7add800f75aa926d6ced2f9aaff9dc (MD5) Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2020-06-18T12:40:54Z (GMT) No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2020-06-18T12:40:54Z (GMT) No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Submitted by Danielle Hamae Yamauchi (danielle.yamauchi@unesp.br) on 2020-06-16T17:53:23Z No. of bitstreams: 1 dissertação_danielle.yamauchi.pdf: 1578364 bytes, checksum: 9d1139590da89e8401add4159ba3d074 (MD5) Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2020-06-18T12:40:54Z (GMT) No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Submitted by Danielle Hamae Yamauchi (danielle.yamauchi@unesp.br) on 2020-06-16T17:53:23Z No. of bitstreams: 1 dissertação_danielle.yamauchi.pdf: 1578364 bytes, checksum: 9d1139590da89e8401add4159ba3d074 (MD5) Submitted by Danielle Hamae Yamauchi (danielle.yamauchi@unesp.br) on 2020-06-16T17:53:23Z No. of bitstreams: 1 dissertação_danielle.yamauchi.pdf: 1578364 bytes, checksum: 9d1139590da89e8401add4159ba3d074 (MD5) Submitted by Danielle Hamae Yamauchi (danielle.yamauchi@unesp.br) on 2020-06-16T17:53:23Z No. of bitstreams: 1 dissertação_danielle.yamauchi.pdf: 1578364 bytes, checksum: 9d1139590da89e8401add4159ba3d074 (MD5) Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2020-06-18T12:40:54Z (GMT) No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Rejected by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo as orientações abaixo: Problema 1: Financiamento Fapesp No formulário de submissão consta somente a Capes como agência financiadora do seu projeto. Mas, nos agradecimentos de sua dissertação, você cita também a Fapesp. E a FAPESP faz algumas exigências na inclusão dos agradecimentos pelo financiamento recebido, uma delas é a obrigatoriedade de se ter o número do processo também no agradecimento. Vi que você agradeceu, faltou incluir o número do processo. Assim que tiver efetuado a correção submeta o arquivo, em formato PDF, novamente. Agradecemos a compreensão. on 2020-06-16T14:51:35Z (GMT) Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2020-06-18T12:40:54Z (GMT) No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2020-06-18T12:40:54Z (GMT) No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Submitted by Danielle Hamae Yamauchi (danielle.yamauchi@unesp.br) on 2020-06-16T17:53:23Z No. of bitstreams: 1 dissertação_danielle.yamauchi.pdf: 1578364 bytes, checksum: 9d1139590da89e8401add4159ba3d074 (MD5) Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2020-06-18T12:40:54Z (GMT) No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Submitted by Danielle Hamae Yamauchi (danielle.yamauchi@unesp.br) on 2020-06-16T17:53:23Z No. of bitstreams: 1 dissertação_danielle.yamauchi.pdf: 1578364 bytes, checksum: 9d1139590da89e8401add4159ba3d074 (MD5) Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2020-06-18T12:40:54Z (GMT) No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2020-06-18T12:40:54Z (GMT) No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Submitted by Danielle Hamae Yamauchi (danielle.yamauchi@unesp.br) on 2020-06-16T17:53:23Z No. of bitstreams: 1 dissertação_danielle.yamauchi.pdf: 1578364 bytes, checksum: 9d1139590da89e8401add4159ba3d074 (MD5) Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Submitted by Danielle Hamae Yamauchi (danielle.yamauchi@unesp.br) on 2020-06-16T17:53:23Z No. of bitstreams: 1 dissertação_danielle.yamauchi.pdf: 1578364 bytes, checksum: 9d1139590da89e8401add4159ba3d074 (MD5) Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2020-06-18T12:40:54Z (GMT) No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2020-06-18T12:40:54Z (GMT) No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Made available in DSpace on 2020-06-18T12:40:54Z (GMT). No. of bitstreams: 1 yamauchi_dh_me_bot_par.pdf: 701723 bytes, checksum: 817bf08d96ac36874c257954d9d8240a (MD5) Previous issue date: 2020-02-28 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) O reino fungi é constituído por amplos grupos de organismos com reconhecida importância ecológica, industrial ou médica. O solo é um importante reservatório para microrganismos, especialmente fungos, que atuam na decomposição e reciclagem de nutrientes ou se associam com animais e plantas em relações simbióticas ou causando patogenias. Ainda são poucos os estudos relacionados a estrutura do micobioma, sendo rasa a compreensão da diversidade e ecologia de fungos. A compreensão dos padrões de distribuição da diversidade fúngica é essencial para mensurar as mudanças naturais e antrópicas relacionadas a estas comunidades, tal como entender a ecologia e epidemiologia dos fungos considerados patogênicos. Neste trabalho caracterizamos a diversidade fúngica das amostras de solo em diferentes ambientes no município de Botucatu (Brasil), descrevendo a estrutura da comunidade fúngica e mensurando a prevalência dos fungos patogênicos, mediante a técnica de Sequenciamento de Nova Geração (NGS). No sequenciamento mediante plataforma Illumina, analisamos 44 amostras provenientes de 8 ambientes diferentes, em dois períodos (outono e primavera), incluindo tocas de tatus, corujas e outros ambientes relacionados. A riqueza e diversidade variaram entre os ambientes, sendo favorecido nas amostras ambientais da floresta sazonal semidecidual e das tocas de tatus, os quais possuem uma maior cobertura vegetal. Ainda, ambientes mais antropizados, com menores efeitos de cobertura vegetal, como edifício abandonado e toca de coruja, proporcionaram as menores diversidades, apesar de favorecer crescimento de alguns fungos patogênicos. Os fungos patogênicos dimórficos, quando comparada com os fungos oportunistas, foram encontrados em baixas frequências, como foi o caso de Paracoccidioides spp., endêmico da região, que foi encontrado em diversos ambientes em baixa abundância. A estrutura da comunidade se mostrou mais similar nas amostras coletadas no mesmo ambiente, com fatores ambientais semelhantes. A pesquisa evidenciou a influência do ambiente nas comunidades fúngicas, mesmo em locais geograficamente próximos. CAPES: 88882.183597/2018-01 FAPESP: 2018/06390-9
- Published
- 2020
126. Ecology of Paracoccidioides brasiliensis and Paracoccidioides lutzii and its association with the Dasypus novemcinctus armadillo in the states of São Paulo and Mato Grosso, Brazil
- Author
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Hrycyk, Marluce Francisca, Universidade Estadual Paulista (Unesp), and Bagagli, Eduardo [UNESP]
- Subjects
filogenia ,tatus ,armadillos ,paracoccidioidomycosis ,ecologia ,Paracoccidioides spp ,paracoccidioidomicose ,ecology ,phylogeny - Abstract
Submitted by MARLUCE FRANCISCA HRYCYK null (marluce@unemat.br) on 2018-03-23T15:42:38Z No. of bitstreams: 1 Texto final 23 03 18.pdf: 2572888 bytes, checksum: 00a5717448a63a2138d8dbcd62ca9c06 (MD5) Approved for entry into archive by Luciana Pizzani null (luciana@btu.unesp.br) on 2018-03-26T17:45:23Z (GMT) No. of bitstreams: 1 hrycyk_mf_dr_bot.pdf: 2473289 bytes, checksum: 4273a8f7803f0e2dda1dadd73582c9a2 (MD5) Made available in DSpace on 2018-03-26T17:45:23Z (GMT). No. of bitstreams: 1 hrycyk_mf_dr_bot.pdf: 2473289 bytes, checksum: 4273a8f7803f0e2dda1dadd73582c9a2 (MD5) Previous issue date: 2018-03-01 Fundação de Amparo à Pesquisa do Estado de Mato Grosso (FAPEMAT) Paracoccidioides spp. são fungos que vivem no ambiente (solo) e em associação com tecidos de hospedeiros mamíferos. Apresentam como característica de patogenicidade o termo-dimorfismo, em que, a 25º C apresenta a fase micelial e produz suas partículas infectantes (conídios) e a 35-37º C está sob forma de leveduras (forma parasitária). As características morfológicas e micromorfológicas são importantes, mas insuficientes para a diferenciação entre os genótipos. Contudo, o avanço no conhecimento da Biologia Molecular nos últimos anos permitiu uma grande compreensão da biologia e das relações filogenéticas de Paracoccidioides spp. A região de ITS1-5.8S-ITS2 (rDNA) tem sido largamente usada na identificação de diversos fungos, sendo muito útil para discriminar entre P. brasiliensis e P. lutzii, no entanto, ela não separa entre as genótipos do complexo P. brasiliensis (S1, PS2, PS3 e PS4), no qual, recentemente foi proposto um rearranjo taxonômico, em que foram descritas como novas espécies de Paracoccidioides: P. brasiliensis para S1, P. americana para PS2, P. restrepiensis para PS3 e P. venezuelensis para PS4. Além do seqüenciamento do ITS, é necessário incluir outras regiões gênicas mais ou menos polimórficas para estudos filogenéticos que visam identificar Paracoccidioides spp. como nesse estudo. Este trabalho teve como objetivo mapear áreas de ocorrência e isolar Paracoccidioides a partir de tatus (Dasypus novemcinctus), bem com detectar molecularmente por Nested-PCR o DNA do fungo em amostras de solo e nos órgãos (baço, fígado e linfonodos mesentéricos) de tatus. Nós isolamos o fungo de tatus do Sudeste (Botucatu/SP) e Centro-Oeste (Alta Floresta/MT). Todos os isolados foram caracterizados molecularmente pelo seqüenciamento ITS-rDNA e gp43 exon 2 e por PCR-RFLP de alfa tubulina (tub1) como P. brasiliensis e P. americana. A reação de Nested-PCR foi sensível e permitiu detectar o DNA de Paracoccidioides nas amostras de tecido (baço, fígado e linfonodos mesentéricos) de D. novemcinctus e também no solo dos Estados de São Paulo e Mato Grosso. A construção filogenética com os amplicons ambientais de solo mostrou que P. lutzii está amplamente presente nessas regiões (Sudeste e Centro-Oeste), além do que, amplicons de P. lutzii de Alta Floresta/MT clusterizaram em um clado separadamente, sugerindo que espécies crípticas podem existir, dentro de P. lutzii. Paracoccidioides spp. are fungi that live in the environment (soil) and in association with tissues of mammalian hosts. They present as a characteristic of pathogenicity the thermo - dimorphism, in which, at 25º C it presents the m ycelial phase and produces its infective particles (conidia) and at 35 - 37º C it is in the form of yeasts (parasitic form). The morphological and micromorphological characteristics are important, but insufficient for the differentiation between the genotype s. However, the advance in the knowledge of Molecular Biology in recent years has allowed a great understanding of the biology and phylogenetic relationships of Paracoccidioides spp. The region of ITS1 - 5.8S - ITS2 (rDNA) has been widely used in the identific ation of several fungi, in which it is very useful in the discrimination between P. brasiliensis and P. lutzii , however, it does not separate between genotypes of the complex P. brasiliensis (S1, PS2, PS3 and PS4), in which recently a taxonomic rearrangeme nt was proposed, described as new species of Paracoccidioides : P. brasiliensis for S1, P. americana for PS2, P. restrepiensis for PS3 and P. venezuelensis for PS4 . In addition to the sequencing of ITS, it is necessary to include other more or less polymorp hic gene regions for phylogenetic studies aimed at identifying Paracoccidioides spp. as in this study . The objective of this work was to map out occurrence areas and isolate Paracoccidioides from armadillos ( Dasypus novemcinctus ), as well as molecularly de tect the DNA of the fungus in soil samples and organs (spleen, liver and mesenteric lymph nodes) by Nested - PCR . We isolated the armadillo fungus from the Southeast (Botucatu / SP) and Central West (Alta Floresta / MT). All isolates were molecularly charac terized by ITS - rDNA and gp43 exon 2 sequencing and by alpha tubulin (tub1) PCR - RFLP as P. brasiliensis and P. americana . The Nested - PCR reaction was sensitive and allowed to detect the DNA of Paracoccidioides in tissue samples (spleen, liver and mesenteric lymph nodes) of D. novemcinctus and also in the soil of the States of São Paulo and Mato Grosso. Phylogenetic construction with environmental soil amplicons showed that P. lutzii is widely present in these regions (Southeast and Center - West), in addition, P. lutzii amplicons from Alta Floresta / MT cluster in a clade separately, suggesting that species may exist within P. lutzii .
- Published
- 2018
127. Microbiological and environmental aspects of candidemias in the tertiary hospital (HC/FMB/UNESP/Botucatu) located in the south center region of the state of São Paulo, Brazil
- Author
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Giacobino, Juliana, Universidade Estadual Paulista (Unesp), and Bagagli, Eduardo Luiz [UNESP]
- Subjects
complexo Candida parapsilosis ,fatores de virulência ,virulence factors ,molecular identification ,exposição ambiental ,antifungal agents ,environmental exposures ,agentes antifúngicos ,Candida parapsilosis complex ,identificação molecular - Abstract
Submitted by Juliana Giacobino (julianagiacobino@gmail.com) on 2018-04-02T13:36:42Z No. of bitstreams: 2 Tese Doutorado Final 19 mar 2018.pdf: 943288 bytes, checksum: d33104aa54aeddf7c9607be3604dbdb2 (MD5) Tese Doutorado Final 19 mar 2018.pdf: 943288 bytes, checksum: d33104aa54aeddf7c9607be3604dbdb2 (MD5) Approved for entry into archive by Luciana Pizzani null (luciana@btu.unesp.br) on 2018-04-03T15:05:50Z (GMT) No. of bitstreams: 1 giacobino_j_dr_bot_par.pdf: 380724 bytes, checksum: f5256c73eb690c57f0d5ed0c91edd6cf (MD5) Made available in DSpace on 2018-04-03T15:05:50Z (GMT). No. of bitstreams: 1 giacobino_j_dr_bot_par.pdf: 380724 bytes, checksum: f5256c73eb690c57f0d5ed0c91edd6cf (MD5) Previous issue date: 2018-02-23 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Infecções fúngicas causadas por leveduras constituem um dos maiores problemas em pacientes hospitalizados em todo o mundo e têm se tornado uma importante causa de morbidade e mortalidade. Embora Candida albicans seja a espécie mais frequentemente isolada e sua forma de infecção ocorra geralmente por translocação endógena, tem-se observado um aumento das espécies não-albicans, com destaque para o complexo C. parapsilosis, cuja principal forma de infecção é exógena, provavelmente pelas mãos dos profissionais de saúde. Este trabalho propôs estudar os aspectos microbiológicos e ambientais das candidemias, com especial atenção para o complexo Candida parapsilosis, no hospital terciário HC/FMB/UNESP, Botucatu, utilizando-se de métodos moleculares, busca ativa dos agentes no ambiente hospitalar (ar, superfícies e mãos de profissionais de saúde), bem como determinar os fatores de virulência e susceptibilidade aos antifúngicos destas espécies e associação com o desfecho clínico. Os isolados clínicos (obtidos de hemoculturas, período 2007-2015) e ambientais (período 2014-2015) de C. parapsilosis sensu lato foram identificados pelo meio CHROMagar Candida, Vitek-2, sequenciamento do rDNA e perfis dos inteins VMA e ThrRS. Fatores de virulência (produção de proteinase, fosfolipase e biofilme) e perfis de susceptibilidade aos antifúngicos anfotericina B, fluconazol, voriconazol, caspofungina e micafungina foram estimados, e dados clínicos obtidos junto aos prontuários dos pacientes. Dentre os isolados clínicos (n=45), C. parapsilosis sensu stricto (s.s.) representou 84%, C. orthopsilosis 16% e ausência de C. metapsilosis. Os perfis de inteins confirmaram o padrão híbrido incomum da espécie C. orthopsilosis. Dentre os ambientais (n=14), todos pertenceram à C. parapsilosis s.s. A produção de proteinase foi positiva ou fortemente positiva em 55% dos isolados clínicos de C. parapsilosis s.s., e negativa nos isolados de C. orthopsilosis. Todos os isolados ambientais de C. parapsilosis s.s. foram produtores de proteinase (64% fortemente positivos). A produção de fosfolipase foi positiva para apenas um isolado ambiental de C. parapsilosis s.s. A produção de biofilme foi significativamente maior em C. orthopsilosis. Todos os isolados clínicos de C. parapsilosis s.s. foram sensíveis à anfotericina B e voriconazol, cinco foram sensíveis dose-dependentes e dois resistentes ao fluconazol, três foram simultaneamente sensíveis dose-dependentes à caspofungina e micafungina. Os isolados de C. orthopsilosis foram sensíveis a todos os antifúngicos. A mortalidade foi alta (51%) e as mortes ocorreram principalmente em recém-nascidos e idosos. A melhor compreensão das leveduras do complexo C. parapsilosis terá impactos positivos no tratamento e prevenção de novas infecções. Fungal infections caused by yeasts are serious problems in hospitalized patients around the globe and have become a major cause of morbidity and mortality. Although Candida albicans is the most frequently isolated species and its infection usually occurs by endogenous translocation, an increase of non-albicans species has been observed, with emphasys for the C. parapsilosis complex, whose main route of infection is exogenous, probably by the hands of health professionals. This work proposes to study the microbiological and environmental aspects of candidemia, with special attention to the C. parapsilosis complex, in a public tertiary hospital HC/FMB/UNESP, in Botucatu, using the molecular methods, active search of agents in the hospital environment (air, surfaces and hands of health professionals), as well as to determine the virulence factors and susceptibility to the antifungals of these species and correlate with the patient clinical outcomes. The clinical isolates (obtained of the blood cultures, 2007-2015 period) and environmental (2014-2015 period) of the C. parapsilosis sensu lato were identified by CHROMagar Candida medium, Vitek-2, rDNA sequencing and VMA and ThrRS inteins profiles. Virulence factors (production of proteinase, phospholypase and biofilm) and profiles of susceptibility to antifungals amphotericin B, fluconazole, voriconazole, caspofungin and micafungin were estimated, and the clinical data were obtained from patient’s records. Among the clinical isolates (n=45), C. parasilosis sensu stricto (s.s.) represented 84%, C. orthopsilosis 16% and absence of C. metapsilosis. The inteins profiles confirmed the unusual hybrid pattern of the C. orthopsilosis species. Among the environmental isolates (n=14), all belong to C. parapsilosis s.s. Proteinase production was positive or strongly positive in 55% of clinical isolates of C. parapsilosis s.s., and negative in the C. orthopsilosis isolates. All the environment isolates of C. parapsilosis s.s. were proteinase producers (64% of them were strongly positive). The phospholipase production was positive for only one environmental isolate of C. parapsilosis s.s. The biofilm production was significantly higher in C. orthopsilosis. All the clinical isolates of C. parapsilosis s.s. were sensitive to amphoterycin B and voriconazole, five were dose-dependent sensitivity and two resistant to fluconazole, three were simultaneously dose-dependent sensitivity to caspofungin and micafungin. The isolates of C. orthopsilosis were sensitive to all antifungals. The mortality was high (51%) and deaths occurred mainly in newborns and elderly patients. A better understanding of C. parapsilosis complex yeasts will have positive impacts on the treatment and prevention of new infections in our hospitals.
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- 2018
128. Prp8 intein in dermatophytes: molecular identification and evolution
- Author
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Garces, Hans Garcia, Universidade Estadual Paulista (Unesp), and Bagagli, Eduardo [UNESP]
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filogenia ,Dermatophytes ,Intein PRP8 ,molecular identification ,PRP8 intein ,Dermatófitos ,Identificação molecular ,phylogeny - Abstract
Submitted by HANS GARCIA GARCES null (atiweb@gmail.com) on 2018-02-15T14:17:44Z No. of bitstreams: 1 tese final hans garcia.pdf: 5207018 bytes, checksum: dc8235a547fe1b9abb0de219b802a19f (MD5) Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2018-02-20T17:09:18Z (GMT) No. of bitstreams: 1 garces_hg_dr_bot_par.pdf: 414383 bytes, checksum: a87b367ff5d75b8f43af0ee14876ad21 (MD5) Made available in DSpace on 2018-02-20T17:09:18Z (GMT). No. of bitstreams: 1 garces_hg_dr_bot_par.pdf: 414383 bytes, checksum: a87b367ff5d75b8f43af0ee14876ad21 (MD5) Previous issue date: 2018-02-08 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Os dermatófitos são um grupo de fungos constituídos pelos gêneros Trichophyton, Epidermophyton e Microsporum que têm a habilidade de degradar a queratina. É por essa razão que podem colonizar a pele do homem e dos animais, embora também possam crescer no ambiente, geralmente em solos com restos de queratina. As características morfológicas permitem a identificação e classificação taxonômica das espécies, mas realizar um diagnóstico baseado nestas características pode levar a erros. As técnicas moleculares ajudam a realizar diagnósticos mais rápidos baseados em uma identificação molecular e também têm possibilitado importantes avanços quanto aos estudos filogenéticos, sendo as sequências ITS1-5.8S-ITS2 as mais utilizadas. No entanto, estes marcadores não são suficientes para identificar e elucidar todas as relações filogenéticas e taxonômicas dos dermatófitos devido à ampla variedade de espécies existentes, -sendo necessário novos marcadores genéticos, como o intein PRP8. Os inteins são elementos genéticos parasitas, de natureza proteica, presentes em alguns fungos e estão associados a importantes genes altamente conservados. Ointein PRP8 está localizado nogenePRP8 que codifica para a proteína PRP8 associada ao complexo do spliciosoma. O presente estudo visa caracterizar o intein PRP8 em fungos dermatófitos, de forma a empregar estes elementos genéticos como marcadores moleculares para uma correta identificação destas espécies e elucidações das relações filogenéticas do grupo. Para realizar este objetivo, foram caracterizadasmolecularmente 45 cepas usando as regiões nucleares ribossomais ITS1-5.8S-ITS2 e D1/D2 e posteriormente a região do intein PRP8 de 40 cepas foi sequenciada para determinar as características do intein. A presença do intein foi comprovada em todas as espécies avaliadas, apresentando um full-intein com uma Homing Endonuclease presumivelmente ativa. As construções filogenéticas obtidas usando as sequências do intein são correspondentes as construídas a partir de genes nucleares ribossomais, demonstrando a validade do intein para ser incluído em futuros estudos filogenético associados a taxonomia e determinação de espécies. O polimorfismo na região da Homing Endonuclease permitiu a identificação de algumas espécies de Microsporum mediante um ensaio de PCR-Eletroforese. Dermatophytes are a fungal group composed by the genera Trichophyton, Epidermophyton and Microsporum with the ability to degrade keratin. That is why they can colonize the skin of man and animals, although they can also grow in the environment, usually in soils with remains of keratin. The morphological characteristics allow the identification and taxonomic classification of the species, but a diagnosis based on these characteristics is difficult. Molecular techniques made diagnoses based on molecular identification faster and made possible important advances in phylogenetic studies, with ITS1-5.8s-ITS2 sequences being the most used. However, these markers are not enough to elucidate all the phylogenetic relationships and taxonomic of dermatophytes due to the wide variety of existing species, and new genetic markers may be needed for correct identification and phylogenetic studies, such as the PRP8 intein. Inteins are parasitic genetic elements of a protein nature present in some fungi and are associated with important highly conserved genes. The PRP8 intein is located within the PRP8 gene coding for the PRP8 protein associated with the spliciosome complex. The present study aims to characterize the PRP8 intein in dermatophyte fungi, in order to use these genetic elements as molecular markers for a correct identification of these species and elucidations of the phylogenetic relationships of the group. To accomplish this goal, 45 strains were molecularly characterized using the ITS1-5.8S-ITS2 and D1 / D2 ribosomal nuclear regions and subsequently the PRP8 intein region of 40 strains was sequenced to determine the intein characteristics. The presence of intein was confirmed in all evaluated species, presenting a full-intein with a presumably active Homing Endonuclease. The phylogenetic construction obtained using the intein sequences correspond to those constructed from ribosomal nuclear genes, demonstrating the intein validity to be included in future phylogenetic studies associated with taxonomy and species determination. Polymorphism in the Homing Endonuclease region allowed the identification of some Microsporum species by a PCR-Electrophoresis assay. 12481-13-0
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- 2018
129. Comparação entre o processo de virulência em Paracoccidioides brasiliensis e P. lutzii com utilização de modelo alternativo de bioensaio e knockdown gênico
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Machado, Gabriel Capella [UNESP], Universidade Estadual Paulista (Unesp), and Bagagli, Eduardo [UNESP]
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Genetica molecular ,Antigenos ,Virulencia (Microbiologia) ,Paracoccidioides brasiliensis ,Paracoccidioidomicose ,Paracoccidioidomycosis - Abstract
Made available in DSpace on 2016-09-27T13:40:04Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-07-31. Added 1 bitstream(s) on 2016-09-27T13:45:14Z : No. of bitstreams: 1 000868835.pdf: 1467491 bytes, checksum: 8ee2f2f1a6a55757ee87dffc67df02b2 (MD5) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) A paracoccidioidomicose, uma das mais importantes micoses sistêmicas da América Latina, é causada pelas espécies crípticas Paracoccidioides brasiliensis e P. lutzii. Ainda não é claro como o processo de especiação dentro do gênero Paracoccidioides influi em aspectos como virulência e patogenia da doença. O objetivo deste trabalho é caracterizar algumas dessas diferenças entre as duas espécies crípticas, utilizando-se de modelo experimental alternativo, as larvas da traça de cera Galleria mellonella. Este modelo tem sido utilizado em estudos envolvendo outros patógenos, porém ainda é pouco explorado em Paracoccidioides spp. No presente estudo, G. mellonella refletiu diferenças entre a virulência de cargas fúngicas distintas, bem como dentre isolados variados e destes com os grupos controle. Além disso, foi possível a recuperação do patógeno a partir de larvas experimentalmente infectadas. Isolados P. lutzii apresentaram níveis de virulência variáveis, sendo pertencentes a esta espécie os isolados com maior e menor virulência observadas neste estudo (8334 e Pb01, respectivamente). Já os isolados pertencentes a espécie P. brasiliensis (Pb339, Pb192 e T15LN1) apresentaram taxas de mortalidade mais homogêneas e intermediárias. Tais resultados credenciam as larvas de G. mellonella como modelo de experimentação adequado para o estudo do processo de virulência em Paracoccidioides spp. Ainda, foi realizado knockdown do gene codificador da gp43 (PbGP43), o antígeno imunodominante em P. brasiliensis e do seu homólogo em P. lutzii (PlP43) utilizando-se a estratégia de RNA antisense combinada com transformação mediada por Agrobacterium tumefaciens. Foram obtidos três transformantes P. brasiliensis com expressão de PbGP43 de cerca de 8%, 14% e 36% quando comparados com a expressão do isolado selvagem original Pb192. Além disso foram obtidos dois transformante P. lutzii com expressão de PlP43 por volta de 23% e... Paracoccidioidomycosis (PCM), an important systemic mycosis in Latin America, is caused by the cryptic species Paracoccidioides brasiliensis and P. lutzii. How these species differ in their virulence factors is still unknown and needs to be properly evaluated. While the role of PbGP43, which codes gp43, as a virulence factor is relatively well established for P. brasiliensis, the same is not true for its ortholog in P. lutzii, namely PlP43. PbGP43 and PlP43 present differences in their nucleotide sequences, expression levels, epitope occurrence and influences on PCM diagnosis. Herein, we obtained PbGP43 and PlP43 knockdown strains by antisense RNA technology combined with ATMT, in order to comparatively evaluate their effects on virulence, employing the alternative experimental host Galleria mellonella larvae. Our results suggest that p43 is important in the P. lutzii infectious process, as previously demonstrated by gp43 in P. brasiliensis. We confirmed G. mellonella as a useful model for studying P. brasiliensis and P. lutzii virulence, since it reflects variation between inoculum size and different strains, as well as between wild-type and respective knockdown transformant strains. Virulence levels may vary among isolates of the same species, probably reflecting that the physiological condition is more important than the species effect. Re-isolation of Paracoccidioides from experimentally infected G. mellonella larvae is possible and might be useful for epigenetic studies related to infection CNPq: 143371/2011-8
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- 2015
130. Detecção do Paracoccidioides spp. em amostras ambientais e diferenciação do complexo P. brasiliensis da espécie P. lutzii por Nested PCR e hibridização in situ
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Arantes, Thales Domingos [UNESP], Universidade Estadual Paulista (Unesp), Bagagli, Eduardo [UNESP], and Theodoro, Raquel Cordeiro [UNESP]
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Reação em cadeia da polimerase ,Hibridização in situ ,Fungos do solo ,Paracoccidioides brasiliensis ,In situ hybridization - Abstract
Made available in DSpace on 2016-01-13T13:27:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-02-27. Added 1 bitstream(s) on 2016-01-13T13:33:22Z : No. of bitstreams: 1 000855701.pdf: 1569730 bytes, checksum: 3d2a8da3b6cbd65de62a52de0c54630d (MD5) Made available in DSpace on 2016-01-13T13:27:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-02-27. Added 1 bitstream(s) on 2016-01-13T13:33:22Z : No. of bitstreams: 1 000855701.pdf: 1569730 bytes, checksum: 3d2a8da3b6cbd65de62a52de0c54630d (MD5) Made available in DSpace on 2016-01-13T13:27:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-02-27. Added 1 bitstream(s) on 2016-01-13T13:33:22Z : No. of bitstreams: 1 000855701.pdf: 1569730 bytes, checksum: 3d2a8da3b6cbd65de62a52de0c54630d (MD5) Made available in DSpace on 2016-01-13T13:27:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-02-27. Added 1 bitstream(s) on 2016-01-13T13:33:22Z : No. of bitstreams: 1 000855701.pdf: 1569730 bytes, checksum: 3d2a8da3b6cbd65de62a52de0c54630d (MD5) Made available in DSpace on 2016-01-13T13:27:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-02-27. Added 1 bitstream(s) on 2016-01-13T13:33:22Z : No. of bitstreams: 1 000855701.pdf: 1569730 bytes, checksum: 3d2a8da3b6cbd65de62a52de0c54630d (MD5) Made available in DSpace on 2016-01-13T13:27:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-02-27. Added 1 bitstream(s) on 2016-01-13T13:33:22Z : No. of bitstreams: 1 000855701.pdf: 1569730 bytes, checksum: 3d2a8da3b6cbd65de62a52de0c54630d (MD5) Made available in DSpace on 2016-01-13T13:27:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-02-27. Added 1 bitstream(s) on 2016-01-13T13:33:22Z : No. of bitstreams: 1 000855701.pdf: 1569730 bytes, checksum: 3d2a8da3b6cbd65de62a52de0c54630d (MD5) Made available in DSpace on 2016-01-13T13:27:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-02-27. Added 1 bitstream(s) on 2016-01-13T13:33:22Z : No. of bitstreams: 1 000855701.pdf: 1569730 bytes, checksum: 3d2a8da3b6cbd65de62a52de0c54630d (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) O solo é o provável habitat do fungo patogênico Paracoccidioides spp., devido à detecção molecular deste patógeno neste tipo de amostra, associada à frequente infecção de trabalhadores rurais e isolamento em animais silvestres (Dasypus novemcinctus e Cabassous centralis). O presente estudo visou detectar e diferenciar as espécies Paracoccidioides brasiliensis (complexo) e Paracoccidioides lutzii no ambiente, pelas técnicas de Nested PCR, FISH, respectivamente, em amostras ambientais aerossóis e solo de tocas de tatus provenientes de áreas endêmicas para a Paracoccidioidomicose nas regiões Sudeste, Centro-Oeste e Norte brasileiras. Além da detecção ambiental de Paracoccidioides sp. por métodos moleculares, foi estudada a ocorrência de tatus infectados com P. lutzii no centro-oeste brasileiro, região de maior prevalência desta espécie, onde nenhum animal havia sido avaliado até o presente momento. Obtivemos a detecção positiva para ambas as espécies de Paracoccidioides por ambas as técnicas de detecção (Nested PCR e hibridização in situ), além da diferenciação por ITS (Internal Transcribed Spacer) pudemos visualizar os espécimes fúngicos em algumas amostras aerossóis. Acreditamos que os dados refletem a real ocorrência dos fungos em seu nicho, no entanto, o isolamento animal em tatus demonstrou-se negativo para 7 animais avaliados no trabalho, o que pode indicar que a relação da espécie P. lutzii com os tatus pode não ser a mesma com os casos de tatus infectados com P. brasiliensis em outras regiões brasileiras Soil is probably the habitat of pathogenic fungi Paracoccidioides spp., due to the molecular detection of this pathogen in these samples, associated with the frequent of infection in rural workers and the isolation in wild animals (Dasypus novemcinctus and Cabassous centralis). This project aimed to detect and differentiate the species P. brasiliensis (complex) and P. lutzii in the environment, by the techniques of Nested PCR and FISH, respectively, in environmental aerosol samples and soil from armadillo's burrows, from endemic and non-endemic areas to Paracoccidioidomycosis in the Southeast, Midwest and North regions of Brazil. Besides the environmental detection of Paracoccidioides spp. by molecular methods, the occurrence of armadillos infected with P. lutzii the Brazilian center-west region with the highest prevalence of this kind, where no animals were evaluated at the present time will be studied. We achieved positive detection of both species of Paracoccidioides by both detection techniques (PCR and in situ hybridization), and further the differentiation by the ITS (Internal Transcribed Spacer) we could visualize fungal species in some aerosol samples also differentiating the two species. We believe of the data reflect the actual occurrence of fungi in their niche, however the animal isolation in armadillo's showed up negative for 7 animals evaluated at work, which indicated that the ratio of the species P. lutzii with armadillo may not be the same with cases of armadillo's infected with P. brasiliensis in other regions of Brazil FAPESP: 2012/03233-3
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- 2015
131. Produção de biofilme em isolados de Candida spp do Hospital das Clínicas de Botucatu, UNESP
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Nascimento, Ariane Cristina Mendes de Oliveira Bruder [UNESP], Universidade Estadual Paulista (Unesp), and Bagagli, Eduardo [UNESP]
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Candida albicans ,Antimicóticos ,Biofilme ,Infeccão hospitalar - Abstract
Made available in DSpace on 2014-08-13T14:50:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-08-16Bitstream added on 2014-08-13T18:01:10Z : No. of bitstreams: 1 000736352_20150816.pdf: 622518 bytes, checksum: 9cd0a00890850158d786a74933e00102 (MD5) Bitstreams deleted on 2015-08-17T11:07:08Z: 000736352_20150816.pdf,. Added 1 bitstream(s) on 2015-08-17T11:07:54Z : No. of bitstreams: 1 000736352.pdf: 2845178 bytes, checksum: cd1b9d03cabd50c9aedba9d2463f1aa2 (MD5) Made available in DSpace on 2014-08-13T14:50:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-08-16Bitstream added on 2014-08-13T18:01:10Z : No. of bitstreams: 1 000736352_20150816.pdf: 622518 bytes, checksum: 9cd0a00890850158d786a74933e00102 (MD5) Bitstreams deleted on 2015-08-17T11:07:08Z: 000736352_20150816.pdf,. Added 1 bitstream(s) on 2015-08-17T11:07:54Z : No. of bitstreams: 1 000736352.pdf: 2845178 bytes, checksum: cd1b9d03cabd50c9aedba9d2463f1aa2 (MD5) Made available in DSpace on 2014-08-13T14:50:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-08-16Bitstream added on 2014-08-13T18:01:10Z : No. of bitstreams: 1 000736352_20150816.pdf: 622518 bytes, checksum: 9cd0a00890850158d786a74933e00102 (MD5) Bitstreams deleted on 2015-08-17T11:07:08Z: 000736352_20150816.pdf,. Added 1 bitstream(s) on 2015-08-17T11:07:54Z : No. of bitstreams: 1 000736352.pdf: 2845178 bytes, checksum: cd1b9d03cabd50c9aedba9d2463f1aa2 (MD5) Made available in DSpace on 2014-08-13T14:50:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-08-16Bitstream added on 2014-08-13T18:01:10Z : No. of bitstreams: 1 000736352_20150816.pdf: 622518 bytes, checksum: 9cd0a00890850158d786a74933e00102 (MD5) Bitstreams deleted on 2015-08-17T11:07:08Z: 000736352_20150816.pdf,. Added 1 bitstream(s) on 2015-08-17T11:07:54Z : No. of bitstreams: 1 000736352.pdf: 2845178 bytes, checksum: cd1b9d03cabd50c9aedba9d2463f1aa2 (MD5) Made available in DSpace on 2014-08-13T14:50:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-08-16Bitstream added on 2014-08-13T18:01:10Z : No. of bitstreams: 1 000736352_20150816.pdf: 622518 bytes, checksum: 9cd0a00890850158d786a74933e00102 (MD5) Bitstreams deleted on 2015-08-17T11:07:08Z: 000736352_20150816.pdf,. Added 1 bitstream(s) on 2015-08-17T11:07:54Z : No. of bitstreams: 1 000736352.pdf: 2845178 bytes, checksum: cd1b9d03cabd50c9aedba9d2463f1aa2 (MD5) Nas últimas décadas, a epidemiologia das infecções por Candida spp. e o perfil de susceptibilidade a drogas vêm sofrendo importantes alterações. A emergência dessas espécies, como agente de infecções invasivas, tem se destacado principalmente entre pacientes imunocomprometidos. O principal objetivo desse estudo foi avaliar a epidemiologia das infecções por Candida spp. no Hospital das Clínicas da Faculdade de Medicina de Botucatu, UNESP, e auxiliar no suprimento de dados para o melhor controle da infecção hospitalar. Os objetivos específicos incluíram a comparação entre as metodologias tradicional, automatizada e molecular de identificação em nível de espécie atualmente disponíveis nesta instituição, avaliar o perfil de susceptibilidade aos antifúngicos, bem como a produção de biofilme por estes isolados e caracterizar, utilizando ferramentas de biologia molecular, genes envolvidos na produção de biofilme de C. albicans e C. parapsilosis e o emprego dos introns do grupo I para genotipagem de C. albicans. Um total de 196 isolados de Candida spp. provenientes de hemocultura de pacientes foram avaliados. As espécies mais frequentes foram C. albicans (39,3 44,4%) e C. parapsilosis (33,2-42,3%). Foi observada boa correlação (>80%) entre os métodos de identificação avaliados – fenotípico tradicional, automatizado e molecular; e a caracterização dos introns do grupo I detectou predominância de C. albicans genótipo A (74%). O método tradicional é o mais simples e de baixo- custo, e o método automatizado mais rápido na liberação do resultado, quando comparados às técnicas moleculares, as quais apresentam maior poder discriminatório. Sugere-se a utilização do meio cromogênico, em paralelo ao método de identificação selecionado, para auxiliar na interpretação dos resultados, principalmente nas espécies mais frequentes. O método empregado em laboratórios de rotina deve ser considerado quando ... Over the last decade, there have been important changes in the epidemiology of Candida infections and antifungal agents used to treat these infections. In recent years, Candida species have emerged as important causes of invasive infections mainly among immunocompromised patients. The principal aim of this study was to evaluate the epidemiology of infectious Candida species isolated in our hospital and accordingly supply data for hospital infection control The other aims were to compare the traditional, automated and molecular methods used to identify Candida isolates to the species level current available in the Clinical Hospital of the Boucatu Medical School, UNESP, to evaluate the drug susceptibility as well as to biofilm production by this yeasts and to characterize, by molecular tools, several genes involved in biofilm production of C. albicans and C. parapsilosis and the use of group I introns to genotype the species C. albicans. A total of 196 Candida strains that were isolated from blood specimens of hospitalized patients were included in the study. The most common Candida species identified were C. albicans (39,3-44,4%) and C. parapsilosis (33,2-42,3%). According to the insertion of group I introns, the genotype A of C. albicans were the most frequent (74%). The evaluation of traditional phenotypic methods, the automated and some molecular revealed good agreement (>80%) and since both phenotypic methods were simple and cost effective and automated were few time-consuming when compared to molecular techniques, they should be considered in the routine lab identification of Candida species. The chromogenic media must be employed to avoid misinterpretations, independently of the method used. Antifungal susceptibility testing was performed by the reference microdilution method (CLSI) and all strains showed good susceptible against the drugs evaluated (fluconazole, voriconazole, anidulafungin and ...
- Published
- 2013
132. Caracterização fenotípica e molecular das espécies fúngicas causadoras de peritonites em pacientes submetidos à diálise peritoneal ambulatorial continua do Hospital das Clínicas da UNESP, Botucatu
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Giacobino, Juliana [UNESP], Universidade Estadual Paulista (Unesp), and Bagagli, Eduardo [UNESP]
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Dialise peritoneal ,Biofilme ,Peritonite ,Atividade antifúngica ,Peritonitis ,Candida - Abstract
Made available in DSpace on 2014-08-13T14:50:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-07-19Bitstream added on 2014-08-13T18:00:53Z : No. of bitstreams: 1 000750764.pdf: 553502 bytes, checksum: d8b9b96b06bbcfe3fb747913ca8ae054 (MD5) Made available in DSpace on 2014-08-13T14:50:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-07-19Bitstream added on 2014-08-13T18:00:53Z : No. of bitstreams: 1 000750764.pdf: 553502 bytes, checksum: d8b9b96b06bbcfe3fb747913ca8ae054 (MD5) Made available in DSpace on 2014-08-13T14:50:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-07-19Bitstream added on 2014-08-13T18:00:53Z : No. of bitstreams: 1 000750764.pdf: 553502 bytes, checksum: d8b9b96b06bbcfe3fb747913ca8ae054 (MD5) Made available in DSpace on 2014-08-13T14:50:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-07-19Bitstream added on 2014-08-13T18:00:53Z : No. of bitstreams: 1 000750764.pdf: 553502 bytes, checksum: d8b9b96b06bbcfe3fb747913ca8ae054 (MD5) Made available in DSpace on 2014-08-13T14:50:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-07-19Bitstream added on 2014-08-13T18:00:53Z : No. of bitstreams: 1 000750764.pdf: 553502 bytes, checksum: d8b9b96b06bbcfe3fb747913ca8ae054 (MD5) As peritonites são sérias e frequentes complicações em pacientes submetidos à terapia de reposição renal, principalmente na diálise peritoneal ambulatorial contínua (CAPD, “continuous ambulatory peritoneal dialysis”). As infecções fúngicas, embora menos frequentes que as bacterianas, estão associadas a uma maior taxa de morbidade e mortalidade, e normalmente tornam impossíveis a CAPD, devido à necessidade de remoção do cateter. Este trabalho visou caracterizar e identificar as peritonites fúngicas com o uso de métodos morfofisiológicos e moleculares, além de avaliar seus perfis de sensibilidade e produção de biofilme in vitro em pacientes submetidos à CAPD do Hospital das Clínicas da Faculdade de Medicina de Botucatu. Foram avaliados trinta isolados de leveduras obtidos de episódios distintos de peritonites, que foram identificados por métodos morfofisiológicos (CHROMagar e auxanograma) e moleculares, tais como o sequenciamento de rDNA, perfil de restrição do gene SADH e padrão do intron do grupo I. Foram também realizados o perfil de sensibilidade antifúngica para as drogas anfotericina B, fluconazol, voriconazol e caspofungina pela técnica de microdiluição em caldo, seguindo o protocolo do CLSI M27-A3 e a produção de biofilme utilizando-se discos de silicone em reação com XTT (sal de tetrazol). Dos 30 isolados de leveduras, observou-se que as espécies de Candida parapsilosis foram as predominantes (n=10), seguida de Candida albicans (n=7), Candida tropicalis (n=6), Candida orthopsilosis (n=5), Candida guilliermondii (n=1) e Kodamaea ohmeri (n=1). Dentre os isolados de C. albicans, caracterizados pelo perfil de intron do grupo I, observou-se predomínio de genótipos do grupo A (n=5) e também a ocorrência de genótipos do grupo B (n=2). Todos os isolados foram sensíveis ao voriconazol e caspofungina, amostras de C.orthopsilosis (n=5), K.ohmeri (n=1), C.guilliermondii (n=1) e C. albicans (n=1) ...
- Published
- 2013
133. Pesquisa dos inteins VMA e THRRS em leveduras do gênero Candida isoladas de hemocultura
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Prandini, Tâmara Heloísa Rocha [UNESP], Universidade Estadual Paulista (Unesp), Bagagli, Eduardo [UNESP], and Theodoro, Raquel Cordeiro [UNESP]
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Micoses ,Biologia molecular ,Candida - Abstract
Made available in DSpace on 2014-06-11T19:23:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-29Bitstream added on 2014-06-13T20:29:34Z : No. of bitstreams: 1 prandini_thr_me_botib.pdf: 557611 bytes, checksum: d3fc30980a3b1907390cb1a9dcc2047b (MD5) Made available in DSpace on 2014-06-11T19:23:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-29Bitstream added on 2014-06-13T20:29:34Z : No. of bitstreams: 1 prandini_thr_me_botib.pdf: 557611 bytes, checksum: d3fc30980a3b1907390cb1a9dcc2047b (MD5) Made available in DSpace on 2014-06-11T19:23:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-29Bitstream added on 2014-06-13T20:29:34Z : No. of bitstreams: 1 prandini_thr_me_botib.pdf: 557611 bytes, checksum: d3fc30980a3b1907390cb1a9dcc2047b (MD5) Made available in DSpace on 2014-06-11T19:23:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-29Bitstream added on 2014-06-13T20:29:34Z : No. of bitstreams: 1 prandini_thr_me_botib.pdf: 557611 bytes, checksum: d3fc30980a3b1907390cb1a9dcc2047b (MD5) Made available in DSpace on 2014-06-11T19:23:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-29Bitstream added on 2014-06-13T20:29:34Z : No. of bitstreams: 1 prandini_thr_me_botib.pdf: 557611 bytes, checksum: d3fc30980a3b1907390cb1a9dcc2047b (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Leveduras do gênero Candida são consideradas os principais agentes de micoses sistêmicas oportunistas, que aumentaram nas duas últimas décadas, devido ao número cada vez maior de hospedeiros com algum grau de comprometimento do sistema imune. Diferentes espécies podem apresentar diferentes perfis clínico-epidemiológicos, assim, um maior conhecimento do perfil molecular e uma correta identificação dos isolados faz-se necessária. Com o sequenciamento de genomas completos de algumas espécies de Candida, tornou-se possível a descoberta e melhor compreensão de algumas regiões gênicas peculiares com importantes implicações evolutivas, biotecnológicas e de desenvolvimento de novas opções terapêuticas. Dentre estas regiões gênicas, destacam-se alguns elementos genéticos parasitas, dentre eles os inteins ou internal proteins, descobertos recentemente e ainda pouco estudados em populações de Candida. Este trabalho avaliou a ocorrência dos inteins nos genes VMA (ATPase vacuolar) e ThrRS (treonil RNAt-sintetase) na população de Candida de interesse médico (C. albicans, C. parapsilosis, C. tropicalis e C. glabrata), possibilitando um maior conhecimento dos perfis moleculares e filogenéticos dos isolados causadores de infecção em nossa região. Foi comprovado também que ambos os inteins aqui pesquisados apresentam um potencial como marcador molecular para a identificação destas espécies, bem como as espécies crípticas do complexo psilosis Not available
- Published
- 2012
134. Espécies crípticas em Paracoccidioides brasiliensis: diferenciação antigênica e de resposta a fatores estressantes
- Author
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Machado, Gabriel Capella [UNESP], Universidade Estadual Paulista (Unesp), and Bagagli, Eduardo [UNESP]
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Serology ,Sorologia ,Genetics ,Paracoccidioides brasiliensis ,Genética - Abstract
Made available in DSpace on 2014-06-11T19:26:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-04-29Bitstream added on 2014-06-13T18:53:57Z : No. of bitstreams: 1 machado_gc_me_botib.pdf: 359425 bytes, checksum: 96a3b6e269405927664fd4f77072b888 (MD5) Made available in DSpace on 2014-06-11T19:26:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-04-29Bitstream added on 2014-06-13T18:53:57Z : No. of bitstreams: 1 machado_gc_me_botib.pdf: 359425 bytes, checksum: 96a3b6e269405927664fd4f77072b888 (MD5) Made available in DSpace on 2014-06-11T19:26:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-04-29Bitstream added on 2014-06-13T18:53:57Z : No. of bitstreams: 1 machado_gc_me_botib.pdf: 359425 bytes, checksum: 96a3b6e269405927664fd4f77072b888 (MD5) Made available in DSpace on 2014-06-11T19:26:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-04-29Bitstream added on 2014-06-13T18:53:57Z : No. of bitstreams: 1 machado_gc_me_botib.pdf: 359425 bytes, checksum: 96a3b6e269405927664fd4f77072b888 (MD5) Made available in DSpace on 2014-06-11T19:26:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-04-29Bitstream added on 2014-06-13T18:53:57Z : No. of bitstreams: 1 machado_gc_me_botib.pdf: 359425 bytes, checksum: 96a3b6e269405927664fd4f77072b888 (MD5) Made available in DSpace on 2014-06-11T19:26:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-04-29Bitstream added on 2014-06-13T18:53:57Z : No. of bitstreams: 1 machado_gc_me_botib.pdf: 359425 bytes, checksum: 96a3b6e269405927664fd4f77072b888 (MD5) Made available in DSpace on 2014-06-11T19:26:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-04-29Bitstream added on 2014-06-13T18:53:57Z : No. of bitstreams: 1 machado_gc_me_botib.pdf: 359425 bytes, checksum: 96a3b6e269405927664fd4f77072b888 (MD5) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) O fungo termodimórfico Paraccocidiodes brasiliensis, em sua fase leveduriforme, é o agente etiológico de uma das mais importantes micoses sistêmicas da América Latina, a paracoccidioidomicose. A doença pode ser fatal em sua forma aguda, além de gerar, na forma crônica, lesões principalmente de ordem dermatológica e pulmonar. O fato de P. brasiliensis ser considerado uma única entidade biológica vem sendo questionado pelos recentes achados de filogenia molecular. Atualmente acredita-se que P. brasiliensis seja um complexo de quatro espécies crípticas, S1, PS2, PS3 e P. lutzii, sendo esta última a mais distante filogeneticamente. É possível que o fato esteja interferindo em aspectos relacionados à doença, como, por exemplo, a positividade do diagnóstico sorológico. Além disso, tal especiação pode significar diferenças na biologia do fungo, em aspectos como sua morfologia, fisiologia e ocupação de nichos ecológicos.A primeira frente deste trabalho visou produzir e caracterizar antígenos a partir de cepas pertencentes às diferentes espécies de P. brasiliensis, e avaliar o desempenho das mesmas em testes de imunodifusão (ID) contra pacientes das diferentes regiões do Brasil. Melhores resultados foram obtidos com o antígeno produzido por filtrado de cultura de um isolado pertencente ao grupo PS3, o qual apresentou altas taxas do antígeno imunodominante, a glicoproteína gp43, detectável por SDS-PAGE, e apresentou boa positividade nas IDs quando cruzados com soros de pacientes da região de Botucatu, reagindo inclusive com alguns soros falso-negativos para o antígeno considerado padrão, produzido pela cepa B-339. Entretanto, este antígeno, bem como outros aqui avaliados, não foi capaz de apresentar desempenho satisfatório frente a soros de pacientes da Região Centro-Oeste, justamente os que apresentam maiores taxas de negatividade, provavelmente por serem... The thermo-dimorphic fungus Paraccocidiodes brasiliensis is the causative agent of paracoccidioidomycosis, one of the most important systemic mycosis in Latin America. The disease can be fatal in its acute form, besides causing important lesions especially in lungs and skin when occurs in its chronic form. The fact that P. brasiliensis is considered a single biological entity has been challenged by the recent findings in molecular phylogenetics. Currently, it has been observed that P. brasiliensis is a complex of four cryptic species, S1, PS2, PS3 and Pb01-like, which is the most distantly related, being proposed as new formal species, P. lutzii. It is possible that these facts are interfering with the host/pathogen interactions, as well as with the results of serological diagnosis. Furthermore, this speciation could reflect differences in the biology of the fungus, especially on their morphology, physiology and occupation of ecological niches.The first approach of this work was to produce and characterize antigens from strains belonging to different species of P. brasiliensis, and evaluate their performance in immunodiffusion tests (ID) against sera of pacients from different regions of Brazil. The better results were obtained with the antigen produced by culture filtrate of an isolate belonging to the PS3, AgEpm83, which showed high positivity rates in IDs, against sera of patients from Botucatu. This antigen, in addition, presented high levels of the glycoprotein gp43, the immunodominant component, detectable by SDS-PAGE. However, this antigen, as well as other here evaluated, was unable to provide satisfactory performance against the sera of patients from the Brazilian Midwest Region, probably because they are infected with P. lutzii isolates. The in silico analysis of exon2 of the gene that encodes gp43 indicated that this gene is under purifying selection both... (Complete abstract click electronic access below)
- Published
- 2011
135. Estudo da prevalência de fungos em travesseiros de crianças com rinite e, ou, asma
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Olbrich, Sandra Regina Leite Rosa [UNESP], Universidade Estadual Paulista (Unesp), and Bagagli, Eduardo [UNESP]
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Asma em crianças ,Rinite ,Alergia em crianças ,Fungos - Abstract
Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-10-15Bitstream added on 2014-06-13T20:01:11Z : No. of bitstreams: 1 olbrich_srlr_dr_botib.pdf: 1124820 bytes, checksum: d6bdfea755aadfb3c11171a7283ac8f6 (MD5) Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-10-15Bitstream added on 2014-06-13T20:01:11Z : No. of bitstreams: 1 olbrich_srlr_dr_botib.pdf: 1124820 bytes, checksum: d6bdfea755aadfb3c11171a7283ac8f6 (MD5) Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-10-15Bitstream added on 2014-06-13T20:01:11Z : No. of bitstreams: 1 olbrich_srlr_dr_botib.pdf: 1124820 bytes, checksum: d6bdfea755aadfb3c11171a7283ac8f6 (MD5) Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-10-15Bitstream added on 2014-06-13T20:01:11Z : No. of bitstreams: 1 olbrich_srlr_dr_botib.pdf: 1124820 bytes, checksum: d6bdfea755aadfb3c11171a7283ac8f6 (MD5) Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-10-15Bitstream added on 2014-06-13T20:01:11Z : No. of bitstreams: 1 olbrich_srlr_dr_botib.pdf: 1124820 bytes, checksum: d6bdfea755aadfb3c11171a7283ac8f6 (MD5) Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-10-15Bitstream added on 2014-06-13T20:01:11Z : No. of bitstreams: 1 olbrich_srlr_dr_botib.pdf: 1124820 bytes, checksum: d6bdfea755aadfb3c11171a7283ac8f6 (MD5) Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-10-15Bitstream added on 2014-06-13T20:01:11Z : No. of bitstreams: 1 olbrich_srlr_dr_botib.pdf: 1124820 bytes, checksum: d6bdfea755aadfb3c11171a7283ac8f6 (MD5) Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-10-15Bitstream added on 2014-06-13T20:01:11Z : No. of bitstreams: 1 olbrich_srlr_dr_botib.pdf: 1124820 bytes, checksum: d6bdfea755aadfb3c11171a7283ac8f6 (MD5) O crescente interesse por micro-organismos alergênicos, a procura por novos reservatórios, além do interesse em fungos anemófilos, a grande frequência e diversidade, bem como a associação com doenças alérgicas, motivaram o estudo. A identificação e quantificação dos fungos em travesseiros poderão proporcionar avanços no diagnóstico e nas medidas ambientais. Avaliar ocorrência de fungos em travesseiros de crianças alérgicas, o ambiente e os aspectos envolvidos.Pacientes do Ambulatório de Imunologia e Alergia do Hospital das Clínicas de Botucatu preencheram questionário com dados epidemiológicos e clínicos e os travesseiros utilizados pelas crianças foram coletados, na residência, colocados em saco plástico estéril. Para o controle, foram adquiridos três travesseiros novos. Todos foram trazidos ao laboratório de Micologia do Instituto de Biociências de Botucatu - UNESP e foram aspirados, dentro da câmara de fluxo, com amostrador de ar MAS- 100NT® por cinco minutos em cada área- externa e interna. Utilizadas três placas para cada aspirado com três diferentes meios de cultura. As identificações foram feitas pelos métodos micológicos tradicionais e, para alguns, foram realizadas extrações de DNA e sequenciamento. Todos procedentes de diferentes regiões de Botucatu , 4,46% dos pacientes do Ambulatório de Alergia e Imunologia, que realizaram teste de sensibilização, eram positivo para os fungos testados; a maioria dos pacientes era sensibilizada a mais de um alérgeno, os mais frequentes foram: Dermatophagoides Farinae , Dermatophagoides Pteronyssinus , Blomia tropicalis e fungos; 80% dos pacientes tinham rinite e 50% também tinham asma; nenhum ambiente, avaliado no domicílio dos pacientes, revelou-se adequado para pacientes alérgicos; somente dois travesseiros tinham capa adequada e se encontravam visivelmente limpos; todos os travesseiros dos... The increasing interest in allergenic microorganisms, the search for new reservoirs in addition to the interest in anemophilous fungi, their high frequency and diversity as well their association with allergic diseases motivated this study. Fungal identification and quantification in pillows can lead to advancement in diagnosis and in environmental measures. To evaluate the occurrence of fungi in pillows of allergic children, the environment and the issues involved. Patients from the Immunology and allergy outpatient unit of the Botucatu University Hospital completed a questionnaire with epidemiological and clinical data, and the pillows used by the children were collected from their homes and placed in a sterile plastic bag; three new pillows were acquired for control. All the pillows were brought to the Mycology Laboratory of the Biosciences Institute and aspirated within a flow chamber by an MAS-100NT® air sampler for five minutes on each area – external and internal. Three plates were used for each aspirate with three different culture media. Fungi were identified by conventional mycological methods, and DNA extraction and sequencing were also performed. All the participants were from different regions of Botucatu; 4.46% of the patients from the Immunology and Allergy Outpatient Unit who underwent the sensitization test were positive for the tested fungi; most of the patients were sensitized to more than one allergen, of which the most frequent were: Dermatophagoides Farinae, Dermatophagoides Pteronyssinus, Blomia tropicalis and fungi; 80% of the patients had rhinitis, and 50% also had asthma. None of the environments evaluated at the patients’ homes showed to be adequate for allergic patients; only two pillows had an adequate cover and were visibly clean; all the patients’ pillows were contaminated with fungi and, in some of them, the presence of gram positive bacteria ... (Complete abstract click electronic access below)
- Published
- 2010
136. Susceptibilidade antifúngica, produção de biofilme e caracterização do gene ALS3 em isolados de Candida albicans e não-albicans do hospital das clínicas, UNESP, Botucatu
- Author
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Nascimento, Ariane Cristina Mendes de Oliveira Bruder [UNESP], Universidade Estadual Paulista (Unesp), and Bagagli, Eduardo [UNESP]
- Subjects
Biofilme (Microbiologia) ,Candida albicans ,Infeccão hospitalar ,Hospital infection - Abstract
Made available in DSpace on 2014-06-11T19:23:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-19Bitstream added on 2014-06-13T19:49:33Z : No. of bitstreams: 1 nascimento_acmob_me_botib.pdf: 365893 bytes, checksum: 7a173728f136f9b95dc827e38a28e865 (MD5) Made available in DSpace on 2014-06-11T19:23:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-19Bitstream added on 2014-06-13T19:49:33Z : No. of bitstreams: 1 nascimento_acmob_me_botib.pdf: 365893 bytes, checksum: 7a173728f136f9b95dc827e38a28e865 (MD5) Made available in DSpace on 2014-06-11T19:23:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-19Bitstream added on 2014-06-13T19:49:33Z : No. of bitstreams: 1 nascimento_acmob_me_botib.pdf: 365893 bytes, checksum: 7a173728f136f9b95dc827e38a28e865 (MD5) Made available in DSpace on 2014-06-11T19:23:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-19Bitstream added on 2014-06-13T19:49:33Z : No. of bitstreams: 1 nascimento_acmob_me_botib.pdf: 365893 bytes, checksum: 7a173728f136f9b95dc827e38a28e865 (MD5) Made available in DSpace on 2014-06-11T19:23:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-19Bitstream added on 2014-06-13T19:49:33Z : No. of bitstreams: 1 nascimento_acmob_me_botib.pdf: 365893 bytes, checksum: 7a173728f136f9b95dc827e38a28e865 (MD5) Made available in DSpace on 2014-06-11T19:23:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-19Bitstream added on 2014-06-13T19:49:33Z : No. of bitstreams: 1 nascimento_acmob_me_botib.pdf: 365893 bytes, checksum: 7a173728f136f9b95dc827e38a28e865 (MD5) Made available in DSpace on 2014-06-11T19:23:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-19Bitstream added on 2014-06-13T19:49:33Z : No. of bitstreams: 1 nascimento_acmob_me_botib.pdf: 365893 bytes, checksum: 7a173728f136f9b95dc827e38a28e865 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Leveduras oportunistas do gênero Candida são capazes de disseminar-se em hospedeiros susceptíveis, num processo crescente nos últimos anos. Um fator complicador destes quadros ocorre quando estas leveduras são capazes de produzir biofilme, principalmente quando associadas a cateteres ou outros dispositivos médicos, elevando o poder de penetração e invasão em órgãos do hospedeiro. Por também conferir maior resistência às drogas antifúngicas do que as células dispersas, o biofilme fúngico tornou-se um dos maiores problemas no combate a estas infecções. A base genética da produção de biofimes nestas leveduras é complexa, porém já foi determinado o envolvimento de genes da família ALS, codificadores de glicoproteínas de adesão. Dentre os oito genes desta família (ALS1 ao ALS7 e ALS9), destaca-se o papel de ALS3. O gene ALS3, assim como todos os outros genes da família, apresenta uma estrutura composta por 3 domínios. O domínio 5’, região bem conservada; um domínio central que apresenta motifs de 108pb repetidos em tandem, com variações de tamanho entre os genes da mesma família e entre o mesmo gene em diferentes espécies, em uma mesma espécie e até mesmo entre alelos de uma mesma cepa, e o domínio 3, menos conservado que o domínio 5’, que pode apresentar variações de tamanho e de algumas seqüências de aminoácidos. Tendo em vista a crescente incidência de infecções por esse microrganismo em todo o mundo, o presente estudo objetivou investigar a freqüência das diferentes espécies de Candida em nossa região e caracterizá-las quanto à susceptibilidade a drogas antifúngicas e produção de biofilme, e possível correlação da produção de biofilme com polimorfismos de tamanho do gene ALS3. Os resultados obtidos confirmam a crescente incidência de espécies não-albicans, principalmente isoladas de infecções invasivas como cultura... Opportunistic yeasts of the genus Candida are able to disseminate into the bloodstream in susceptible hosts, in an increasing course in the recent years. A complicating factor is when these yeasts are capable of producing biofilms, especially associated with catheters or other medical devices. Biofilm also confers greater resistance to antifungal drugs than dispersed cells, so the fungal biofilm has become one of the greatest problems in combating these infections. The genetic basis of the biofim production by yeasts is complex, but it has been know the involvement of ALS gene family, encoders of adhesion glycoproteins. Among the eight genes of this family (ALS1 to ALS7 and ALS9), the ALS3 are considered the most important. The ALS3 gene, such as the others members of the family, have three general domains: the 5’domain, conserved, with approximately 1300-pb; followed by a central domain consisting entirely of tandem-repeats of a 108-pb sequence, that are somewhat variable; and the 3’ domain, which is least conserved in length and sequence. Considering the increase incidence of these infections worldwide, the aims of this study were identify the frequency of Candida species in our region, to characterize the profile of antifungal susceptibility; to quantify the biofilm production and to correlate this production with the ALS3 gene length polymorphism. Our data confirm the increase incidence of non-albicans species, mainly when obtained from invasive infections, such as blood and peritoneal fluid, in which C. parapsilosis was the most frequent isolated species. The same was also observed to biofilm production, in which isolates obtained from invasive infections (blood and peritoneal fluid) are more biofilm producers than that obtained from vaginal secretion and urine. Among the different species, isolates of non-albicans also are more biofilm producers than C. albicans. Polimerase... (Complete abstract click electronic access below)
- Published
- 2009
137. Ecologia molecular de fungos patogênicos onygenales em animais silvestres do interior do estado de São Paulo
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Pereira, Vírginia Bodelão Richini [UNESP], Universidade Estadual Paulista (Unesp), Bagagli, Eduardo [UNESP], and Bosco, Sandra de Moraes Gimenes [UNESP]
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Molecular biology ,Fungos patogenicos ,Micologia ,Animais silvestres - Doenças ,Paracoccidioidomicose ,Dasypus novemcintus - Abstract
Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-13Bitstream added on 2014-06-13T20:21:29Z : No. of bitstreams: 1 pereira_vbr_dr_botib.pdf: 2768539 bytes, checksum: e0eceb3d0040712e43091635190b217c (MD5) Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-13Bitstream added on 2014-06-13T20:21:29Z : No. of bitstreams: 1 pereira_vbr_dr_botib.pdf: 2768539 bytes, checksum: e0eceb3d0040712e43091635190b217c (MD5) Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-13Bitstream added on 2014-06-13T20:21:29Z : No. of bitstreams: 1 pereira_vbr_dr_botib.pdf: 2768539 bytes, checksum: e0eceb3d0040712e43091635190b217c (MD5) Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-13Bitstream added on 2014-06-13T20:21:29Z : No. of bitstreams: 1 pereira_vbr_dr_botib.pdf: 2768539 bytes, checksum: e0eceb3d0040712e43091635190b217c (MD5) Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-13Bitstream added on 2014-06-13T20:21:29Z : No. of bitstreams: 1 pereira_vbr_dr_botib.pdf: 2768539 bytes, checksum: e0eceb3d0040712e43091635190b217c (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) A Paracoccidioidomicose (PCM) é uma micose sistêmica e a de maior ocorrência na América Latina, causada pelo fungo Paracoccidioides brasiliensis. Apesar dos esforços contínuos de diversos grupos de pesquisa principalmente do Brasil, Colômbia Venezuela e Argentina, a fase ambiental produtora de propágulos infectantes, seu nicho ecológico e outros aspectos fundamentais da biologia deste patógeno ainda representa um enigma. Sabe-se, no entanto, que há alguns indicadores biológicos onde se constata a infecção natural do P. brasiliensis em espécies de tatu Dasypus novemcinctus em áreas endêmicas. Assim, o isolamento sistemático deste patógeno nesta espécie animal tem despertado o interesse em avaliar outras espécies animais cuja distribuição geográfica seja coincidente com a da PCM. O conhecimento de reservatórios naturais de fungos patogênicos e o mapeamento de regiões habitadas pelos fungos são dados fundamentais para elucidar a eco-epidemiologia da PCM. O presente projeto focalizou a detecção ambiental do P. brasiliensis, bem como de outros fungos patogênicos geneticamente relacionados, em animais silvestres pelo emprego de métodos moleculares em amostras de tecido de animais atropelados em beiras de estradas. Sabe-se que esse material é muitas vezes negligenciado, sendo utilizado em estudos de conservação biológica e de parasitologia. Esta abordagem é inédita e útil no estudo da eco-epidemiologia da PCM. Foram avaliados também tatus (D. novemcintus) capturados em regiões geograficamente definidas, visando o isolamento fúngico do P. brasiliensis, bem como de outros fungos patogênicos, por cultura de órgãos, histopatologia e técnicas moleculares. A integração dos dados micológicos, moleculares e a aplicação de técnicas de geoprocessamento permitiu a caracterização da área geográfica dos animais avaliados e contribuiu para um melhor conhecimento sobre a ocorrência do patógeno no hospedeiro animal Paracoccidioidomycosis (PCM) is the most prevalent systemic mycosis of Latin America, caused by fungus Paracoccidioides brasiliensis. Despite the continuous efforts by several research groups mainly from Brazil, Colombia, Venezuela and Argentina, the saprobic form producing infective propagula, its exact niche in nature and other fundamental aspects of the biology of this pathogen still remains enigmatic. It is known however that some biological indicators where there is a natural infection of P. brasiliensis from the nine-banded armadillo Dasypus novemcinctus in an endemic area. Thus, the systematic isolation of this pathogen in this species has increased the necessity in evaluating several wild mammals whose geographical distribution is coincident of PCM. The knowledge about natural reservoirs of pathogenic fungi and mapping landscape of areas inhabited by fungi may be clarified the ecoepidemiology of PCM. This work focused on the environmental detection of P. brasiliensis, as well as other related pathogenic fungi in wild animals, the use of molecular tools in tissue samples from road-killed wild animals. While road-killed animals proved to be usefull both form parasitological and conservative studies, these materials have been negleted in the area of infections disease. This approach is new and has been useful to elucidate PCM ecoepidemiology. It was evaluated armadillos (D. novemcinctus) captured is some restricted areas, such as from Savanna and a County considered to be hyperendemic for PCM, by culture, histopathology and molecular techniques, in order to confirm if P. brasiliensis also occurs in such defined ecological conditions. The use the Geographical Information Systems (GIS), thus contributing to a better understanding about the occurrence of the pathogen in the host animal and the associated biotic and abiotic factors
- Published
- 2009
138. Detecção molecular de Paracoccidioides brasiliensis em amostras de tecido e sangue periférico de pacientes portadores de Pacoccidioidomicose: correlação com perfis histopatológico, sorológicos e produção de TNF-α
- Author
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Siqueira, Keila Zaniboni [UNESP], Universidade Estadual Paulista (Unesp), Marques, Silvio Alencar [UNESP], and Bagagli, Eduardo [UNESP]
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Imunologia ,Paracoccidioides brasiliensis ,Paraccoccidioidomicose - Patologia - Estudo de caso - Abstract
Made available in DSpace on 2014-06-11T19:33:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-07-25Bitstream added on 2014-06-13T20:44:53Z : No. of bitstreams: 1 siqueira_kz_dr_botfm.pdf: 354096 bytes, checksum: d00ae8c0bad924914a08763aaa47a5e3 (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) A paracoccidioidomicose (PCM) é uma doença granulomatosa sistêmica, causada pelo Paracoccidioides brasiliensis, um fungo imperfeito e termodimórfico. Embora o contágio ocorra principalmente pela via aérea, as vias transcutânea e mucosa não devem ser desconsideradas, devido à possibilidade de invasão através de pele ou mucosas. No presente estudo, a detecção molecular de P. brasiliensis por Nested- PCR foi realizada em fragmentos de biópsias de lesões cutâneo-mucosas de pacientes com PCM ativa e comparada a outras técnicas utilizadas como rotina no diagnóstico dessa micose. Para tanto, amostras de 29 pacientes atendidos no Serviço de Dermatologia, da Faculdade de Medicina de Botucatu/UNESP, foram coletadas no período de abril/2004 a abril/2007. Na análise histopatológica observou-se granulomas compactos bem organizados, algumas vezes ao lado de áreas contendo granulomas frouxos mal definidos. Tal técnica foi positiva em todas as amostras avaliadas, com presença de grande quantidade de células fúngicas em 65,5% dos casos, algumas com multi-brotamentos. Para as análises sorológicas, na técnica de ELISA, 74,1% dos casos apresentaram títulos de 1/12800, 11,1% apresentaram 1/3200, 3,7% apresentaram 1/800 e 11,1% foram negativos. Já para imunodifusão (ID), 3,5% apresentaram título de 1/512, 55,1% apresentaram títulos de 1/32, 3,5% apresentaram 1/16, 3,5% apresentaram 1/8, 10,3% apresentaram 1/4, 6,9% apresentaram 1/2 e 17,2% foram negativos. Foi observado um aumento nos níveis de TNF- , nos pacientes na forma grave, produzido por monócitos não estimulados, em comparação à forma leve e moderada, e ao controle. A produção desta citocina por monócitos estimulados com LPS também se mostrou gradativamente maior quanto maior a gravidade dos pacientes. Além disso, na análise do seguimento dos pacientes nota-se uma diminuição significativa... A paracoccidioidomicose (PCM) é uma doença granulomatosa sistêmica, causada pelo Paracoccidioides brasiliensis, fungo imperfeito e termodimórfico (Franco et al., 1987, 1989). Com distribuição endêmica no Brasil e outros países da América Latina (Restrepo et al., 1973; Restrepo, 1985; Wanke & Londero, 1994), esse agente etiológico apresenta-se sob a forma de levedura em tecidos do hospedeiro ou quando cultivado à temperatura de 35-37°C, e sob a forma de micélio à temperatura ambiente (4-28°C) (Restrepo, 1985; San- Blas & San-Blas, 1985), As lesões primárias da PCM ocorrem com mais freqüência nos pulmões, dando origem a infecção subclínica, que pode evoluir para cura ou tornar-se latente na presença de resposta imune eficiente, ou ainda, disseminar para outros órgãos, por via hematogênica ou linfática, causando doença grave (Giraldo et al., 1976; Londero & Severo, 1981; Restrepo, 1988; Montenegro & Franco, 1994; Bethlem et al., 1999). A PCM tem como apresentações clínicas as formas aguda-subaguda e crônica (Franco et al., 1987). A primeira, também chamada juvenil, ocorre predominantemente em crianças, adolescentes e adultos jovens. Geralmente se desenvolve a partir de lesão primária não detectada, que se dissemina rapidamente pelas vias linfática e hematogênica para órgãos linfóides e hematopoiéticos, constituindo quadro infeccioso grave (Mendes et al., 1988). A forma crônica da doença é mais freqüente e se desenvolve a partir do complexo primário ou do foco quiescente (reinfecção endógena). Esta geralmente afeta adultos do sexo masculino, com idade superior a 30 anos. Apresenta quadro clínico de duração prolongada, normalmente acima de seis meses, onde se pode dizer que o comprometimento pulmonar é constantemente observado e o tegumentar (cutâneo e/ou mucoso) é freqüente (Marques et al., 1994)... (Complete abstract click electronic access below)
- Published
- 2008
139. Evolução cromossômica na Ordem Xenarthra
- Author
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Pereira Júnior, Hélio Rubens Jacintho [UNESP], Universidade Estadual Paulista (Unesp), Jorge, Wilham [UNESP], and Bagagli, Eduardo [UNESP]
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Cromossomos ,Evolução (Biologia) ,Genética - Abstract
Made available in DSpace on 2014-06-11T19:32:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-03-01Bitstream added on 2014-06-13T21:03:47Z : No. of bitstreams: 1 pereirajunior_hrj_dr_botib.pdf: 1684166 bytes, checksum: 2f23cdb28a2397afd8b1cdb49d47d88e (MD5) Made available in DSpace on 2014-06-11T19:32:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-03-01Bitstream added on 2014-06-13T21:03:47Z : No. of bitstreams: 1 pereirajunior_hrj_dr_botib.pdf: 1684166 bytes, checksum: 2f23cdb28a2397afd8b1cdb49d47d88e (MD5) Made available in DSpace on 2014-06-11T19:32:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-03-01Bitstream added on 2014-06-13T21:03:47Z : No. of bitstreams: 1 pereirajunior_hrj_dr_botib.pdf: 1684166 bytes, checksum: 2f23cdb28a2397afd8b1cdb49d47d88e (MD5) Made available in DSpace on 2014-06-11T19:32:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-03-01Bitstream added on 2014-06-13T21:03:47Z : No. of bitstreams: 1 pereirajunior_hrj_dr_botib.pdf: 1684166 bytes, checksum: 2f23cdb28a2397afd8b1cdb49d47d88e (MD5) Made available in DSpace on 2014-06-11T19:32:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-03-01Bitstream added on 2014-06-13T21:03:47Z : No. of bitstreams: 1 pereirajunior_hrj_dr_botib.pdf: 1684166 bytes, checksum: 2f23cdb28a2397afd8b1cdb49d47d88e (MD5) Made available in DSpace on 2014-06-11T19:32:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-03-01Bitstream added on 2014-06-13T21:03:47Z : No. of bitstreams: 1 pereirajunior_hrj_dr_botib.pdf: 1684166 bytes, checksum: 2f23cdb28a2397afd8b1cdb49d47d88e (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) A Ordem Xenarthra é uma ordem de mamíferos placentários composta por três formas viventes: tamanduás, tatus e preguiças arborícolas. É originária da América Sul e tem como distribuição geográfica o centro sul da América do Norte até o extremo sul da América do Sul. A Ordem possui trinta espécies viventes divididas em quatro famílias: Dasypodidae formada pelos tatus, com vinte e uma espécies, Myrmecophagidae (tamanduás) com quatro espécies, Bradypodidae (preguiças de três dedos) com quatro espécies e Megalonychidae (preguiças de dois dedos) com duas espécies. As espécies Priodontes maximus (tatu-canastra), Tolypeutes tricinctus (tatu-bola da caatinga), Bradypus torquatus (preguiça de coleira) e Myrmecophaga tridactyla (tamanduá-bandeira) estão classificadas como vulneráveis pela IUCN (International Union of Conservation), sendo a preguiça de coleira e o tatubola da caatinga endêmicos do Brasil. A maioria dos xenartros possui uma constituição cariotípica que varia de 48 até 65 cromossomos, exceto a espécie Tolypeutes matacus com 2n=38 cromossomos. Das trinta espécies conhecidas atualmente, apenas dezenove tiveram seu cariótipo descrito ou relatado. Os xenartros apresentam uma ampla gama de eventos cromossômicos, como fusão / fissão entre as espécies do gênero Cabassous, Choloepus e Bradypus e na família Myrmecophagidae, eventos de rearranjos cromossômico no gênero Dasypus e a combinação de ambos no gênero Chaetophractus. A presente tese está dividida em quatro capítulos: 1 revisão bibliográfica sobre os dados cromossômicos na Ordem Xenarthra; 2 descrição do cariótipo da espécie Cabassous unicinctus; 3 descrição uma nova espécie do gênero Tamandua; 4 análise da evolução cromossômica da Ordem Xenarthra, utilizando abordagem filogenética. Not available.
- Published
- 2007
140. Caracterização da transição micélio-levedura em Paracoccidioides brasiliensis e sua relação com a expressão do gene do choque térmico 70(HSP70)
- Author
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Theodoro, Raquel Cordeiro [UNESP], Universidade Estadual Paulista (Unesp), Bagagli, Eduardo [UNESP], and Maia, Ivan de Godoy [UNESP]
- Subjects
Fungo ,Micose ,Fungus ,Paracoccidioidomicose - Aspectos genéticos ,Paracoccidioides brasilienses ,Mycosis - Abstract
Made available in DSpace on 2014-06-11T19:26:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-02-27Bitstream added on 2014-06-13T20:14:33Z : No. of bitstreams: 1 theodoro_rc_me_botib.pdf: 1627532 bytes, checksum: 84aa1424194b14fdd0953c57c4c5c2a6 (MD5) Made available in DSpace on 2014-06-11T19:26:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-02-27Bitstream added on 2014-06-13T20:14:33Z : No. of bitstreams: 1 theodoro_rc_me_botib.pdf: 1627532 bytes, checksum: 84aa1424194b14fdd0953c57c4c5c2a6 (MD5) Made available in DSpace on 2014-06-11T19:26:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-02-27Bitstream added on 2014-06-13T20:14:33Z : No. of bitstreams: 1 theodoro_rc_me_botib.pdf: 1627532 bytes, checksum: 84aa1424194b14fdd0953c57c4c5c2a6 (MD5) Made available in DSpace on 2014-06-11T19:26:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-02-27Bitstream added on 2014-06-13T20:14:33Z : No. of bitstreams: 1 theodoro_rc_me_botib.pdf: 1627532 bytes, checksum: 84aa1424194b14fdd0953c57c4c5c2a6 (MD5) Made available in DSpace on 2014-06-11T19:26:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-02-27Bitstream added on 2014-06-13T20:14:33Z : No. of bitstreams: 1 theodoro_rc_me_botib.pdf: 1627532 bytes, checksum: 84aa1424194b14fdd0953c57c4c5c2a6 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) O fungo termo dimórfico, Paracoccidioides brasiliensis é o agente etiológico da Paracoccidioidomicose (PCM), a micose sistêmica mais prevalente da América Latina. Este fungo vem sendo frequentemente isolado de amostras clínicas, tecidos de tatu (Dasypus novemcinctus) e recentemente foi também isolado de cão. Este trabalho avaliou a transição de micélio para levedura (M-L), a termo tolerância e o perfil de virulência em nove isolados de P. brasiliensis (quatro de pacientes humanos, quatro de tatus e um de cão), bem como a sua relação com a seqüência parcial e expressão do gene hsp70 (Heat Shock Protein 70) através de Real Time RT-PCR. Tanto os dados morfológicos como moleculares se mostraram variáveis dentre os diferentes isolados. Alguns destes dados, como sequenciamento e morfologia leveduriforme corroboram com a divisão de nossos isolados nas duas espécies crípticas simpátricas previamente propostas por Matute et al (2006). Nossos resultados confirmam que a HSP70 pode ser um importante fator de virulência por estar associado à termo tolerância, mas sua expressão parece não ser diretamente associada a altos padrões de virulência. The thermo dimorphic fungus Paracoccidioides brasiliensis is the etiological agent of Paracoccidioidomycosis (PCM), the most prevalent systemic mycosis in Latin America. The previous phylogenetic species recognition proved the existence of, at least, three cryptic species in this pathogen. In this work we evaluated the mycelia to yeast (M-Y) transition, thermo tolerance and virulence profiles of nine isolates of P. brasiliensis, (including members of two of the three species) as well as its relation to the partial sequence and expression of hsp70 gene. It was observed a large phenotypic variability concerning the M-Y transition. The isolates Bt84 and T10 took more time to convert to the yeast form. These same isolates presented stretched yeast cells at 36°C, instead of the typical round cells. It was also observed arthroconidia production during the M-Y transition for some of the nine isolates studied. The hsp70 expression showed to be variable among our isolates. The partial sequencing of hsp70 gene resulted in a Neighbour Joining tree that divided our isolates in two main groups. Our data confirm that hsp70 gene might be an important virulence factor, associated with the thermo tolerance, but its expression does not seem to be directly related to high virulence profiles. We also presented some preliminary results about mycological characters that could be important candidates for morphologic markers for species recognition, as well as the partial sequencing of one member of the hsp70 gene family that allowed the separation of our isolates in two clusters, that correspond to the two sympatric cryptic species that occur in our PCM hyper endemic area (Botucatu, SP, Brazil).
- Published
- 2007
141. Caracterização molecular de isolados de Paracoccidioides brasiliensis obtidos de Dasypus novemcinctus (tatu) e sua correlação com virulência e origem geográfica
- Author
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Barbosa, Flavia Hebeler [UNESP], Universidade Estadual Paulista (Unesp), and Bagagli, Eduardo [UNESP]
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Micose - Epidemiologia ,Mycosis - Epidemiology ,Paracoccidioides brasiliensis - Genética - Abstract
Made available in DSpace on 2014-06-11T19:27:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2002Bitstream added on 2014-06-13T19:15:40Z : No. of bitstreams: 1 barbosa_fh_me_botfm.pdf: 692507 bytes, checksum: a7f91088d5074d8d040c054c5989981c (MD5) O fungo dimórfico Paracoccidioides brasiliensis (Pb) é o agente etiológico da paracoccidioidomicose (PCM), a micose sistêmica mais importante da América Latina. A espécie de tatu Dasypus novemcinctus mostrou ser um importante reservatório do fungo e o estudo de isolados destes animais será útil para uma melhor compreensão da ecologia e epidemiologia desta doença. O objetivo deste trabalho foi comparar a virulência e variabilidade genética de isolados de tatus e de pacientes para melhor compreender a importância do tatu na PCM. Os isolados animais foram obtidos da área endêmica de Botucatu, São Paulo, Brasil, com exata informação dos locais de coleta. Os isolados clínicos, exceto a cepa padrão Pb265, foram obtidos de pacientes residentes nos municípios de Pratânia, Manduri e Botucatu. A virulência foi avaliada no modelo experimental do hamster através da contagem de UFC (Unidades Formadoras de Colônias), histopatologia e dosagem de anticorpos específicos e a caracterização molecular por meio da técnica de RAPD e sequenciamento do DNA ribossômico. Todos isolados de tatus foram virulentos, disseminando-se para vários órgãos. A cepa Pb265, isolada de paciente, apresentou baixa virulência. A contagem de UFC nos diferentes órgãos permitiu uma classificação do fungo em 4 categorias: virulência muito alta, virulência alta, virulência intermediária e baixa virulência. A análise molecular mostrou grande diversidade genética entre os isolados, porém não houve separação entre os fungos isolados de pacientes e de tatu. Estes resultados indicam que tanto o homem quanto o tatu podem estar sendo infectados por patógenos com mesmo genótipo, provavelmente localizados em foco ambiental comum. Portanto, os tatus como reservatórios do P. brasiliensis na natureza, representam importantes elos na cadeia da doença... Paracoccidioides brasiliensis is the etiologic agent of paracoccidioidomycosis (PCM), the most important systemic mycosis in Latin America. The armadillo Dasypus novemcinctus has been confirmed as the first natural reservoir of this fungus and its geographic distribution is similar to the distribution of human PCM. This work analysed the virulence profiles and genetic diversity of P. brasiliensis isolates from different armadillos and clinical isolates. The animal isolates were obtained from the Botucatu endemic area of PCM, with exact information of the collection sites. The clinical isolates, except Pb265, were obtained from patients that live in the same endemic area (Pratânia, Manduri and Botucatu county). Virulence was evaluated in an experimental hamster model by colony-forming units (CFU) and histopathological analysis in the testis, liver, spleen, lung and circulating specific antibodies measured by ELISA. Molecular characterisation was evaluated by RAPD methods and sequencing analyses of the internal transcribed spacer regions (ITS 1 and ITS 2) and 5.8 S ribosomal gene. All isolates from armadillos were virulent, with dissemination to many organs. The clinical isolate Pb 265 was less virulent. The isolates were classified into four categories according to number of CFU per gram of tissue: very high, high, intermediate and low virulence. Molecular analysis showed large genetic similarity between all isolates and did not separate armadillos from clinical isolates. This study confirms that armadillos harbor pathogenic genotypes of P. brasiliensis, probably the same ones that infect man. Thus, the armadillos are very important animals in the epidemiology this disease.
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- 2002
142. Epidemiological, clinical, and genomic landscape of coccidioidomycosis in northeastern Brazil.
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Eulálio KD, Kollath DR, Martins LMS, Filho AD, Cavalcanti MDAS, Moreira LM, Tenório BG, Alves LGB, Yamauchi D, Barrozo LV, Thompson Iii GR, Nacher M, Stajich JE, Benard G, Bagagli E, Felipe MSS, Barker BM, Trilles L, and Teixeira MM
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- Humans, Female, Male, Animals, Brazil epidemiology, Armadillos, Genomics, Genotype, Coccidioidomycosis epidemiology
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Coccidioidomycosis, listed as a priority mycosis by the WHO, is endemic in the United States but often overlooked in Central and South America. Employing a multi-institutional approach, we investigate how disease characteristics, pathogen genetic variation, and environmental factors impact coccidioidomycosis epidemiology and outcomes in South America. We identified 292 cases (1978-2021) and 42 outbreaks in Piauí and Maranhão states, Brazil, the largest series outside the US/Mexico epidemic zone. The male-to-female ratio was 57.4:1 and the most common activity was armadillo hunting (91.1%) 4 to 30 days before symptom onset. Most patients (92.8%) exhibited typical acute pulmonary disease, with cough (93%), fever (90%), and chest pain (77%) as predominant symptoms. The case fatality rate was 8%. Our negative binomial regression model indicates that reduced precipitation levels in the current (p = 0.015) and preceding year (p = 0.001) predict heightened incidence. Unlike other hotspots, acidic soil characterizes this region. Brazilian strains differ genomically from other C. posadasii lineages. Northeastern Brazil presents a distinctive coccidioidomycosis profile, with armadillo hunters facing elevated risks. Low annual rainfall emerges as a key factor in increasing cases. A unique C. posadasii lineage in Brazil suggests potential differences in environmental, virulence, and/or pathogenesis traits compared to other Coccidioides genotypes., (© 2024. The Author(s).)
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- 2024
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143. Molecular detection of Paracoccidioides spp. in environmental samples from the Southwestern Amazon.
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Ferreira IRS, de M Teixeira M, de Araújo AV, Bagagli E, Garces HG, Peters LP, and Carvalho CM
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- Animals, Humans, Soil Microbiology, Fungi, Soil, Brazil epidemiology, Paracoccidioides genetics, Paracoccidioidomycosis epidemiology, Paracoccidioidomycosis microbiology
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Paracoccidioidomycosis is an infection with the potential for environmental dissemination, especially in regions of hot and humid climate, where human cases have been recorded in the Southwestern Amazon of Brazil, specifically in the state of Acre. Despite studies providing information about the presence of these fungi in soil and animal samples, such as armadillos, further investigations are still needed to determine the epidemiological distribution of the genus Paracoccidioides. The aim of this study was to detect the occurrence of Paracoccidioides fungi in the Southwestern Amazon. To achieve this, 60 soil samples were collected from armadillo burrows on rural properties in the in the municipalities of Acrelândia, Bujari, Plácido de Castro, Rio Branco, Sena Madureira, and Senador Guiomard, located in the state of Acre, Brazil. Fungal DNA was extracted from these samples using the DNEASY® PowerSoil kit-Quiagen, followed by Nested PCR technique with ITS4 and ITS5 as external primers, and PBITS-E and PBITS-R as internal primers. DNA amplification products of about 380 bp compatible with Paracoccidioides spp. were detected in six samples (10%), being sequenced and identified as P. brasiliensis. These findings indicate that the soils of the Acre state could be considered a potential source for Paracoccidioides spp., suggesting that local infections are likely., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)
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- 2024
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144. A taxonomic review of the genus Paracoccidioides, with focus on the uncultivable species.
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Vilela R, de Hoog S, Bensch K, Bagagli E, and Mendoza L
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- Humans, Animals, Phylogeny, Saccharomyces cerevisiae, Paracoccidioides genetics, Paracoccidioidomycosis epidemiology, Paracoccidioidomycosis veterinary, Paracoccidioidomycosis microbiology, Dolphins
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Paracoccidioides species have always been surrounded by taxonomic uncertainties. The continuing nomenclatoral muddle was caused in part by the failure of Adolfo Lutz and Jorge Lôbo to name the etiologic agents of human paracoccidioidomycosis and Jorge Lôbo's diseases, respectively. Early in their history, it was postulated that the cultivable species causing systemic infections belonged in the genus Paracoccidioides, whereas the uncultivable species, causing skin disease, were not part of the genus. The taxonomy of these pathogens was further complicated when a similar skin disease with numerous yeast-like cells in infected dolphins was also reported. Due to its phenotypic similarities with that described by Jorge Lôbo in human and its uncultivable nature, it was assumed that the disease in dolphins was caused by the same fungus. Recent molecular and population genetic analysis, however, found the DNA extracted from the uncultivable yeast-like cells affecting dolphins shared common phylogenetic traits with cultivable Paracoccidioides species. The study revealed that the uncultivable pathogens comprised 2 different Paracoccidioides species, now known as P. ceti and P. loboi, correspondingly. To validate P. loboi binomial, a comprehensive historical critical review of Jorge Lôbo etiology was performed. This review showed the proposed binomial P. loboi was previously used, and, thus, a replacement name is introduced, Paracoccidioides lobogeorgii nom. nov. In addition, in this review, several cultivable human Paracoccidioides species are validated, and the generic type species, P. brasiliensis, is neotypified as the original material could not be traced., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Vilela et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
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- 2023
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145. Molecular Phylogenetic Analysis of Paracoccidioides Species Complex Present in Paracoccidioidomycosis Patient Tissue Samples.
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de Oliveira LBZ, Della Coletta AM, Gardizani TP, Garces HG, Bagagli E, Trilles L, Barrozo LV, Marques SA, Faveri J, and Dias-Melicio LA
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Paracoccidioidomycosis (PCM) is the main and most prevalent systemic mycosis in Latin America, that until recently, it was believed to be caused only by Paracoccidioides brasiliensis ( P. brasiliensis ). In 2006, researchers described three cryptic species: S1, PS2, PS3, and later, another one, PS4. In 2009, Paracoccidioides lutzii (Pb01-like) was described, and in 2017, a new nomenclature was proposed for the different agents: P. brasiliensis (S1), P. americana (PS2), P. restrepiensis (PS3), and P. venezuelensis (PS4). These species are not uniformly distributed throughout Latin America and, knowing that more than one cryptic species could coexist in some regions, we aimed to identify those species in patients' biopsy samples for a better understanding of the distribution and occurrence of these recently described species in Botucatu region. The Hospital of Medical School of Botucatu-UNESP, which is a PCM study pole, is located in São Paulo State mid-west region and is classified as a PCM endemic area. Genotyping analyses of clinical specimens from these patients that have been diagnosed and treated in our Hospital could favor a possible correlation between genetic groups and mycological and clinical characteristics. For this, molecular techniques to differentiate Paracoccidioides species in these biopsies, such as DNA extraction, PCR, and sequencing of three target genes (ITS, CHS2, and ARF) were conducted. All the sequences were analyzed at BLAST to testify the presence of P. brasiliensis . The phylogenetic trees were constructed using Mega 7.0 software and showed that 100% of our positive samples were from S1 cryptic species, therefore P. brasiliensis . This is important data, demonstrating the predominance of this species in the São Paulo State region.
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- 2023
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146. Cryptococcus neoformans Prp8 Intein: An In Vivo Target-Based Drug Screening System in Saccharomyces cerevisiae to Identify Protein Splicing Inhibitors and Explore Its Dynamics.
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Fernandes JAL, Zatti MDS, Arantes TD, Souza MFB, Santoni MM, Rossi D, Zanelli CF, Liu XQ, Bagagli E, and Theodoro RC
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Inteins are genetic mobile elements that are inserted within protein-coding genes, which are usually housekeeping genes. They are transcribed and translated along with the host gene, then catalyze their own splicing out of the host protein, which assumes its functional conformation thereafter. As Prp8 inteins are found in several important fungal pathogens and are absent in mammals, they are considered potential therapeutic targets since inhibiting their splicing would selectively block the maturation of fungal proteins. We developed a target-based drug screening system to evaluate the splicing of Prp8 intein from the yeast pathogen Cryptococcus neoformans (CnePrp8i) using Saccharomyces cerevisiae Ura3 as a non-native host protein. In our heterologous system, intein splicing preserved the full functionality of Ura3. To validate the system for drug screening, we examined cisplatin, which has been described as an intein splicing inhibitor. By using our system, new potential protein splicing inhibitors may be identified and used, in the future, as a new class of drugs for mycosis treatment. Our system also greatly facilitates the visualization of CnePrp8i splicing dynamics in vivo.
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- 2022
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147. Clinical and Eco-Epidemiological Aspects of a Novel Hyperendemic Area of Paracoccidioidomycosis in the Tocantins-Araguaia Basin (Northern Brazil), Caused by Paracoccidioides sp.
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Krakhecke-Teixeira AG, Yamauchi DH, Rossi A, de Sousa HR, Garces HG, Júnior JL, Júnior AOS, Felipe MSS, Bagagli E, de Andrade HF Jr, and Teixeira MM
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Paracoccidioidomycosis (PCM) is the most prevalent systemic mycosis in Brazil. The disease is caused by dimorphic fungi nested within the Paracoccidioides genus. We described 106 PCM cases (47.1 cases/year) at the Tropical Diseases Public Hospital of Tocantins State. PCM was prevalent in males and rural workers over 50 years; the chronic pulmonary form predominated in 67% of cases. The male-to-female ratio was 2.65:1, with more women affected than other endemic regions of Brazil. Urban or indoor activities were reported in women and are ascribed to disease urbanization. qPCR-based assays confirmed the identification of Paracoccidioides DNA in 37 biological specimens. Paracoccidioides sp. DNA was found in 53% of the environmental samples, suggesting autochthonous infections. Therefore, the Tocantins-Araguaia basin must be considered a novel hyperendemic area of PCM in Brazil, reinforcing the importance of including PCM as a notifiable disease, requiring specific diagnosis and health measures.
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- 2022
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148. Outbreak of equine pythiosis in a southeastern region of Brazil: Environmental isolation and phylogeny.
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Paz GSD, Camargo GG, Cury JE, Apolonio EVP, Garces HG, Prado ACD, Chechi JL, Oliveira AL, Watanabe MJ, Bagagli E, and Bosco SMG
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- Animals, Brazil epidemiology, Dogs, Horses, Phylogeny, Disease Outbreaks veterinary, Horse Diseases epidemiology, Horse Diseases microbiology, Pythiosis epidemiology, Pythium classification
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Pythiosis is a disease caused by the oomycete Pythium insidiosum, mainly reported in equines, dogs and humans and directly transmitted through contaminant zoospores in aquatic environments. We report the first outbreak of equine pythiosis in five equines. Wound samples were submitted for diagnostic testing including mycological culture and nested PCR. Treatment approaches consisted of conventional and alternative therapies. Microbiological analyses were performed using water samples from the riverbanks close to where the animals had grazed. All animals were positive for P. insidiosum cultures, and two animals responded successfully to alternative therapy (ozone therapy). After culture and molecular analysis of environmental samples, the presence of P. insidiosum in one section of the Tietê River was confirmed through a 99% sequence identity. Phylogenetic analyses using the cytochrome oxidase II gene showed that the animal isolates clustered in clade I and the environmental isolates clustered in clade III. Although the environmental and wound isolates belonged to different genetic clades, we concluded that the Tietê River is an important source of infection by P. insidiosum and that research concerning environmental isolation of P. insidiosum from rivers and lakes should be strongly facilitated in Brazil., (© 2021 Wiley-VCH GmbH.)
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- 2022
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149. Systemic Infection by Non- albicans Candida Species Affects the Development of a Murine Model of Multiple Sclerosis.
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Fraga-Silva TFC, Munhoz-Alves N, Mimura LAN, Oliveira LRC, Figueiredo-Godoi LMA, Garcia MT, Oliveira ES, Ishikawa LLW, Zorzella-Pezavento SFG, Bonato VLD, Junqueira JC, Bagagli E, and Sartori A
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Candidiasis may affect the central nervous system (CNS), and although Candida albicans is predominant, non- albicans Candida species can also be associated with CNS infections. Some studies have suggested that Candida infections could increase the odds of multiple sclerosis (MS) development. In this context, we investigated whether systemic infection by non- albicans Candida species would affect, clinically or immunologically, the severity of experimental autoimmune encephalomyelitis (EAE), which is an animal model used to study MS. For this, a strain of C. glabrata , C. krusei , and C. parapsilosis was selected and characterized using different in vitro and in vivo models. In these analysis, all the strains exhibited the ability to form biofilms, produce proteolytic enzymes, and cause systemic infections in Galleria mellonella , with C. glabrata being the most virulent species. Next, C57BL/6 mice were infected with strains of C. glabrata , C. krusei , or C. parapsilosis , and 3 days later were immunized with myelin oligodendrocyte glycoprotein to develop EAE. Mice from EAE groups previously infected with C. glabrata and C. krusei developed more severe and more prevalent paralysis, while mice from the EAE group infected with C. parapsilosis developed a disease comparable to non-infected EAE mice. Disease aggravation by C. glabrata and C. krusei strains was concomitant to increased IL-17 and IFN-γ production by splenic cells stimulated with fungi-derived antigens and with increased percentage of T lymphocytes and myeloid cells in the CNS. Analysis of interaction with BV-2 microglial cell line also revealed differences among these strains, in which C. krusei was the strongest activator of microglia concerning the expression of MHC II and CD40 and pro-inflammatory cytokine production. Altogether, these results indicated that the three non- albicans Candida strains were similarly able to reach the CNS but distinct in terms of their effect over EAE development. Whereas C. glabrata and C. Krusei aggravated the development of EAE, C. parapsilosis did not affect its severity. Disease worsening was partially associated to virulence factors in C. glabrata and to a strong activation of microglia in C. krusei infection. In conclusion, systemic infections by non- albicans Candida strains exerted influence on the experimental autoimmune encephalomyelitis in both immunological and clinical aspects, emphasizing their possible relevance in MS development.
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- 2022
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150. Candida tropicalis Systemic Infection Redirects Leukocyte Infiltration to the Kidneys Attenuating Encephalomyelitis.
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Munhoz-Alves N, Mimura LAN, Viero RM, Bagagli E, Peron JPS, Sartori A, and Fraga-Silva TFC
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Environmental factors, including infections, are strongly associated with the pathogenesis of multiple sclerosis (MS), which is an autoimmune and demyelinating disease of the central nervous system (CNS). Although classically associated with bacterial and viral agents, fungal species have also been suspected to affect the course of the disease. Candida tropicalis is an opportunistic fungus that affects immunocompromised individuals and is also able to spread to vital organs. As C. tropicalis has been increasingly isolated from systemic infections, we aimed to evaluate the effect of this fungus on experimental autoimmune encephalomyelitis (EAE), a murine model to study MS. For this, EAE was induced in female C57BL/6 mice 3 days after infection with 10
6 viable C. tropicalis yeasts. The infection decreased EAE prevalence and severity, confirmed by the less inflammatory infiltrate and less demyelization in the lumbar spinal cord. Despite this, C. tropicalis infection associated with EAE results in the death of some animals and increased urea and creatinine serum levels. The kidneys of EAE-infected mice showed higher fungal load associated with increased leukocyte infiltration (CD45+ cells) and higher expression of T-box transcription factor ( Tbx21 ) and forkhead box P3 ( Foxp3 ). Altogether, our results demonstrate that although C. tropicalis infection reduces the prevalence and severity of EAE, partially due to the sequestration of leukocytes by the inflamed renal tissue, this effect is associated with a poor disease outcome.- Published
- 2021
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