Preservation methods in isolates of Sporothrix characterized by polyphasic approach
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Outros Autores: | , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | https://hdl.handle.net/1822/81912 |
Resumo: | Sporotrichosis is a subcutaneous mycosis with worldwide distribution and caused by eight pathogenic species of the Sporothrix genus. Different ex situ preservation methods are used around the world to maintain the survival, morphophysiological and genetic traits of fungal strains isolated from patients with sporotrichosis for long terms. The main aim of the present study was to evaluate the survival, phenotypic and genotypic stability of Sporothrix strains after preservation on PDA slant stored at 4 °C, sterile water and cryopreservation at −80 °C, for a period of time of 6, 12, 18 and 24 months of storage. Eight clinical Sporothrix isolates were identified based on a polyphasic approach consisting of classical macro- and micro-morphological traits, biochemical assays, proteomic profiles by MALDI-TOF MS and molecular biology. According to the final identification, one strain was identified as S. schenckii (CMRVS 40428) and seven strains were re-identified as S. brasiliensis (CMRVS 40421, CMRVS 40423, CMRVS 40424, CMRVS 40425, CMRVS 40426, CMRVS 40427 and CMRVS 40433). In addition, it was observed that the isolates survived after the different time points of storage in distilled water, PDA slant and cryopreservation at −80 °C. For fungi preserved in water, low polymorphisms were detected by the partial sequencing of β-tubulin. Cryopreservation at −80 °C induced morphological changes in one single isolate. The proteomic profiles obtained by MALDI-TOF MS after preservation showed differences among the methods. In conclusion, preservation on agar slant stored at 4 °C was the most effective method to preserve the eight clinical Sporothrix strains. This method produced less change in the phenotypic traits and kept the genetic integrity of all strains. Agar slant stored at 4 °C is a simple and inexpensive method and can be especially used in culture collections of limited funding and resources. |
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Preservation methods in isolates of Sporothrix characterized by polyphasic approachpreservationSporothrixphenotypegenotypeproteomicstabilityEngenharia e Tecnologia::Outras Engenharias e TecnologiasScience & TechnologySporotrichosis is a subcutaneous mycosis with worldwide distribution and caused by eight pathogenic species of the Sporothrix genus. Different ex situ preservation methods are used around the world to maintain the survival, morphophysiological and genetic traits of fungal strains isolated from patients with sporotrichosis for long terms. The main aim of the present study was to evaluate the survival, phenotypic and genotypic stability of Sporothrix strains after preservation on PDA slant stored at 4 °C, sterile water and cryopreservation at −80 °C, for a period of time of 6, 12, 18 and 24 months of storage. Eight clinical Sporothrix isolates were identified based on a polyphasic approach consisting of classical macro- and micro-morphological traits, biochemical assays, proteomic profiles by MALDI-TOF MS and molecular biology. According to the final identification, one strain was identified as S. schenckii (CMRVS 40428) and seven strains were re-identified as S. brasiliensis (CMRVS 40421, CMRVS 40423, CMRVS 40424, CMRVS 40425, CMRVS 40426, CMRVS 40427 and CMRVS 40433). In addition, it was observed that the isolates survived after the different time points of storage in distilled water, PDA slant and cryopreservation at −80 °C. For fungi preserved in water, low polymorphisms were detected by the partial sequencing of β-tubulin. Cryopreservation at −80 °C induced morphological changes in one single isolate. The proteomic profiles obtained by MALDI-TOF MS after preservation showed differences among the methods. In conclusion, preservation on agar slant stored at 4 °C was the most effective method to preserve the eight clinical Sporothrix strains. This method produced less change in the phenotypic traits and kept the genetic integrity of all strains. Agar slant stored at 4 °C is a simple and inexpensive method and can be especially used in culture collections of limited funding and resources.The authors are grateful to the Carlos Chagas Filho State Funding Agency (FAPERJ-Grants: JCNE E297 26/203.301/2017; JCNE E-26/201.433/2021–MMEO; E-26/202.737/2019), CAPES (Process 001; D.C.-M fellowship 88882.317297/2019-01), and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq-Grant Proc. 409227/2016-1). C. Santos thanks the funding from the Universidad de La Frontera through the research project FAPESP-UFRO 2020/07546-2. This study was also partially supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of the UID/BIO/04469/2019 unit, the BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020-Programa Operacional Regional do Norte and it also had the partial support of the European Union’s Horizon 2020 research and innovation programme under grant agreement No 871129 - IS_MIRRI21 Project.info:eu-repo/semantics/publishedVersionMDPIUniversidade do Minhode Souza Rabello, Vanessa BritoCorrêa-Moreira, DaniellySantos, CledirAbreu Pinto, Tatiana CastoProcopio-Azevedo, Anna CarolinaBoechat, JéssicaCoelho, Rowena AlvesAlmeida-Paes, RodrigoCosta, GiselaLima, NelsonZancopé-Oliveira, Rosely MariaMarques Evangelista Oliveira, Manoel20232023-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/1822/81912engde Souza Rabello, V.B.; Corrêa-Moreira, D.; Santos, C.; Abreu Pinto, T.C.; Procopio-Azevedo, A.C.; Boechat, J.; Coelho, R.A.; Almeida-Paes, R.; Costa, G.; Lima, N.; Zancopé-Oliveira, R.M.; Marques Evangelista Oliveira, M. Preservation Methods in Isolates of Sporothrix Characterized by Polyphasic Approach. J. Fungi 2023, 9, 34. https://doi.org/10.3390/jof90100342309-608X10.3390/jof9010034https://www.mdpi.com/2309-608X/9/1/34info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:14:27Zoai:repositorium.sdum.uminho.pt:1822/81912Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:06:46.029084Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Preservation methods in isolates of Sporothrix characterized by polyphasic approach |
title |
Preservation methods in isolates of Sporothrix characterized by polyphasic approach |
spellingShingle |
Preservation methods in isolates of Sporothrix characterized by polyphasic approach de Souza Rabello, Vanessa Brito preservation Sporothrix phenotype genotype proteomic stability Engenharia e Tecnologia::Outras Engenharias e Tecnologias Science & Technology |
title_short |
Preservation methods in isolates of Sporothrix characterized by polyphasic approach |
title_full |
Preservation methods in isolates of Sporothrix characterized by polyphasic approach |
title_fullStr |
Preservation methods in isolates of Sporothrix characterized by polyphasic approach |
title_full_unstemmed |
Preservation methods in isolates of Sporothrix characterized by polyphasic approach |
title_sort |
Preservation methods in isolates of Sporothrix characterized by polyphasic approach |
author |
de Souza Rabello, Vanessa Brito |
author_facet |
de Souza Rabello, Vanessa Brito Corrêa-Moreira, Danielly Santos, Cledir Abreu Pinto, Tatiana Casto Procopio-Azevedo, Anna Carolina Boechat, Jéssica Coelho, Rowena Alves Almeida-Paes, Rodrigo Costa, Gisela Lima, Nelson Zancopé-Oliveira, Rosely Maria Marques Evangelista Oliveira, Manoel |
author_role |
author |
author2 |
Corrêa-Moreira, Danielly Santos, Cledir Abreu Pinto, Tatiana Casto Procopio-Azevedo, Anna Carolina Boechat, Jéssica Coelho, Rowena Alves Almeida-Paes, Rodrigo Costa, Gisela Lima, Nelson Zancopé-Oliveira, Rosely Maria Marques Evangelista Oliveira, Manoel |
author2_role |
author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
de Souza Rabello, Vanessa Brito Corrêa-Moreira, Danielly Santos, Cledir Abreu Pinto, Tatiana Casto Procopio-Azevedo, Anna Carolina Boechat, Jéssica Coelho, Rowena Alves Almeida-Paes, Rodrigo Costa, Gisela Lima, Nelson Zancopé-Oliveira, Rosely Maria Marques Evangelista Oliveira, Manoel |
dc.subject.por.fl_str_mv |
preservation Sporothrix phenotype genotype proteomic stability Engenharia e Tecnologia::Outras Engenharias e Tecnologias Science & Technology |
topic |
preservation Sporothrix phenotype genotype proteomic stability Engenharia e Tecnologia::Outras Engenharias e Tecnologias Science & Technology |
description |
Sporotrichosis is a subcutaneous mycosis with worldwide distribution and caused by eight pathogenic species of the Sporothrix genus. Different ex situ preservation methods are used around the world to maintain the survival, morphophysiological and genetic traits of fungal strains isolated from patients with sporotrichosis for long terms. The main aim of the present study was to evaluate the survival, phenotypic and genotypic stability of Sporothrix strains after preservation on PDA slant stored at 4 °C, sterile water and cryopreservation at −80 °C, for a period of time of 6, 12, 18 and 24 months of storage. Eight clinical Sporothrix isolates were identified based on a polyphasic approach consisting of classical macro- and micro-morphological traits, biochemical assays, proteomic profiles by MALDI-TOF MS and molecular biology. According to the final identification, one strain was identified as S. schenckii (CMRVS 40428) and seven strains were re-identified as S. brasiliensis (CMRVS 40421, CMRVS 40423, CMRVS 40424, CMRVS 40425, CMRVS 40426, CMRVS 40427 and CMRVS 40433). In addition, it was observed that the isolates survived after the different time points of storage in distilled water, PDA slant and cryopreservation at −80 °C. For fungi preserved in water, low polymorphisms were detected by the partial sequencing of β-tubulin. Cryopreservation at −80 °C induced morphological changes in one single isolate. The proteomic profiles obtained by MALDI-TOF MS after preservation showed differences among the methods. In conclusion, preservation on agar slant stored at 4 °C was the most effective method to preserve the eight clinical Sporothrix strains. This method produced less change in the phenotypic traits and kept the genetic integrity of all strains. Agar slant stored at 4 °C is a simple and inexpensive method and can be especially used in culture collections of limited funding and resources. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023 2023-01-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://hdl.handle.net/1822/81912 |
url |
https://hdl.handle.net/1822/81912 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
de Souza Rabello, V.B.; Corrêa-Moreira, D.; Santos, C.; Abreu Pinto, T.C.; Procopio-Azevedo, A.C.; Boechat, J.; Coelho, R.A.; Almeida-Paes, R.; Costa, G.; Lima, N.; Zancopé-Oliveira, R.M.; Marques Evangelista Oliveira, M. Preservation Methods in Isolates of Sporothrix Characterized by Polyphasic Approach. J. Fungi 2023, 9, 34. https://doi.org/10.3390/jof9010034 2309-608X 10.3390/jof9010034 https://www.mdpi.com/2309-608X/9/1/34 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
MDPI |
publisher.none.fl_str_mv |
MDPI |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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