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3. Paracoccidioides brasiliensis AND Paracoccidioides lutzii, A SECRET LOVE AFFAIR

Author

ARANTES, Thales Domingos, BAGAGLI, Eduardo, NIÑO-VEGA, Gustavo, SAN-BLAS, Gioconda, THEODORO, Raquel Cordeiro, Universidade Estadual Paulista (Unesp), Laboratório de Virologia e Micologia, Centro de Microbiología y Biología Celular, and Depto. De Biologia Celular e Genética

Subjects
Ecology, Paracoccidioides lutzii, Molecular phylogeny, Paracoccidioides brasiliensis, Biology, Dimorphism
Abstract

Made available in DSpace on 2018-12-11T16:58:48Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-09-01. Added 1 bitstream(s) on 2019-10-09T18:30:00Z : No. of bitstreams: 1 S0036-46652015000800025.pdf: 126533 bytes, checksum: 9236dc3d6017b1691b85e36acfe43ab8 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) To commemorate Prof. Carlos da Silva Lacaz’s centennial anniversary, the authors have written a brief account of a few, out of hundreds, biological, ecological, molecular and phylogenetic studies that led to the arrival of Paracoccidioides lutzii, hidden for more than a century within Paracoccidioides brasiliensis. Lacaz’s permanent interest in this fungus, and particularly his conviction on the benefits that research on paracoccidioidomycosis would bring to patients, were pivotal in the development of the field. Universidade Estadual Paulista Instituto de Biociências/UNESP Depto. Microbiologia e Imunologia Instituto de Medicina Tropical/IMT Laboratório de Virologia e Micologia Campus Universitário Lagoa Nova Instituto Venezolano de Investigaciones Científicas (IVIC) Laboratorio de Micología Centro de Microbiología y Biología Celular Universidade Federal do Rio Grande do Norte Depto. De Biologia Celular e Genética Campus Universitário Lagoa Nova Universidade Estadual Paulista Instituto de Biociências/UNESP Depto. Microbiologia e Imunologia FAPESP: 2012/03233-3 FAPESP: 2012/14047-6

Published
2015

4. Detection of Paracoccidioides spp. in environmental aerosol samples.

Author

Arantes, Thales Domingos, Theodoro, Raquel Cordeiro, Da Graça Macoris, Severino Assis, and Bagagli, Eduardo

Abstract

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]

Published
2013
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5. Ecological study of Paracoccidioides brasiliensis in soil: growth ability, conidia production and molecular detection.

Author

Terçarioli, Gisela Ramos, Bagagli, Eduardo, Reis, Gabriela Martins, Theodoro, Raquel Cordeiro, Bosco, Sandra De Moraes Gimenes, Macoris, Severino Assis da Graça, and Richini-Pereira, Virgínia Bodelão

Subjects
*ENVIRONMENTAL sciences, *ANIMAL behavior, *ALUMINUM, *ECOLOGY, *CULTURE
Abstract

Background: Paracoccidioides brasiliensis ecology is not completely understood, although several pieces of evidence point to the soil as its most probable habitat. The present study aimed to investigate the fungal growth, conidia production and molecular pathogen detection in different soil conditions. Methods: Soils samples of clayey, sandy and medium textures were collected from ground surface and the interior of armadillo burrows in a hyperendemic area of Paracoccidioidomycosis. P. brasiliensis was inoculated in soil with controlled humidity and in culture medium containing soil extracts. The molecular detection was carried out by Nested PCR, using panfungal and species specific primers from the ITS-5.8S rDNA region. Results: The soil texture does not affect fungus development and the growth is more abundant on/in soil saturated with water. Some soil samples inhibited the development of P. brasiliensis, especially those that contain high values of Exchangeable Aluminum (H+Al) in their composition. Some isolates produced a large number of conidia, mainly in soil-extract agar medium. The molecular detection was positive only in samples collected from armadillo burrows, both in sandy and clayey soil. Conclusion: P. brasiliensis may grow and produce the infectious conidia in sandy and clayey soil, containing high water content, mainly in wild animal burrows, but without high values of H+Al. [ABSTRACT FROM AUTHOR]

Published
2007
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6. Phylogenetic and evolutionary aspects of Paracoccidioides brasiliensis reveal a long coexistence with animal hosts that explain several biological features of the pathogen

Author

Bagagli, Eduardo, Bosco, Sandra M.G., Theodoro, Raquel Cordeiro, and Franco, Marcello

Subjects
*PARACOCCIDIOIDES brasiliensis, *PATHOGENIC fungi, *MYCOSES, *ARMADILLOS, *PHYLOGENY
Abstract

Abstract: The habitat of the mycelial saprobic form of Paracoccidioides brasiliensis, which produces the infectious propagula, has not been determined and has proven difficult for mycologists to describe. The fungus has been rarely isolated from the environment, the disease has a prolonged latency period and no outbreaks have been reported. These facts have precluded the adoption of preventive measures to avoid infection. The confirmation of natural infections in nine-banded armadillos (Dasypus novemcinctus) with P. brasiliensis, in high frequency and wide geographic distribution, has opened new avenues for the study and understanding of its ecology. Armadillos belong to the order Xenarthra, which has existed in South America ever since the Paleocene Era (65 million years ago), when the South American subcontinent was still a detached land, before the consolidation of what is now known as the American continent. On the other hand, strong molecular evidence suggests that P. brasiliensis and other dimorphic pathogenic fungi – such as Blastomyces dermatitidis, Coccidioides immitis and Histoplasma capsulatum – belong to the family Onygenaceae sensu lato (order Onygenales, Ascomycota), which appeared around 150 million years ago. P. brasiliensis ecology and relation to its human host are probably linked to the fungal evolutionary past, especially its long coexistence with and adaptation to animal hosts other than Homo sapiens, of earlier origin. Instead of being a blind alley, the meaning of parasitism for dimorphic pathogenic fungi should be considered as an open two-way avenue, in which the fungus may return to the environment, therefore contributing to preserve its teleomorphic (sexual) and anamorphic (asexual) forms in a defined and protected natural habitat. [Copyright &y& Elsevier]

Published
2006
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7. 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

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

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