Determination of Chitin Content in Insects: An Alternate Method Based on Calcofluor Staining

Detalhes bibliográficos
Autor(a) principal: Henriques, Bianca Santos
Data de Publicação: 2020
Outros Autores: Garcia, Eloi Souza, Azambuja, Patricia, Genta, Fernando Ariel
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/40565
Resumo: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ, Brasil.
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spelling Henriques, Bianca SantosGarcia, Eloi SouzaAzambuja, PatriciaGenta, Fernando Ariel2020-03-30T20:00:53Z2020-03-30T20:00:53Z2020HENRIQUES. Bianca Santos et al. Determination of Chitin Content in Insects: An Alternate Method Based on Calcofluor Staining. Frontiers in Physiology, v. 11, Article 117, p. 1-10, Feb. 2020.1664-042Xhttps://www.arca.fiocruz.br/handle/icict/4056510.3389/fphys.2020.00117engFrontiers MediaQuitinaTécnicaPolissacarídeoInsetoQuantificaçãoChitinPolysaccharideCalcofluorInsectQuantitationTechniqueDetermination of Chitin Content in Insects: An Alternate Method Based on Calcofluor Staininginfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia para Entomologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia para Entomologia Molecular. Rio de Janeiro, RJ, Brasil.Chitin is an aminopolysaccharide present in yeast cells and arthropod cuticle and is one of the most abundant biopolymers. The conventional methods for the quantitation of chitin content in biological samples are based on its hydrolysis (acid or enzymatic), and the assessment of the byproduct, glucosamine. However, previously described methodologies are time-consuming, laborious, low throughput, and not applicable to insect samples in many cases. Here we describe a new approach to chitin content quantitation based on calcofluor fluorescent brightener staining of samples, followed by microplate fluorescence readings. Calcofluor is a specific chitin stain commonly used for topological localization of the polymer. The protocol was tested in three important disease vector species, namely Lutzomyia longipalpis, Aedes aegypti, and Rhodnius prolixus, and then compared to a classic colorimetric chitin assessment method. Results show that chitin content in the tested insects can vary largely in a range of 8-4600 micrograms of chitin per insect, depending on species, sex, and instar. Comparisons between measurements from the previous protocol and calcofluor method showed statistically significant differences in some samples. However, the difference might be due to interference in the classic method from non-chitin sources of glucosamine and reducing agents. Furthermore, chitinase hydrolysis reduces the total chitin mass estimated between 36 and 74%, consolidating the fluorescent measurements as actual stained chitin in the same extent that was observed with the standard protocol. Therefore, the calcofluor staining method revealed to be a fast and reliable technique for chitin quantitation in homogenized insect samples.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/40565/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALFernandoAGenta_BiancaHenriques_etal_IOC_2020.pdfFernandoAGenta_BiancaHenriques_etal_IOC_2020.pdfapplication/pdf1133140https://www.arca.fiocruz.br/bitstream/icict/40565/2/FernandoAGenta_BiancaHenriques_etal_IOC_2020.pdfbab1d40e90dc227986c9d47e7be294f6MD52TEXTFernandoAGenta_BiancaHenriques_etal_IOC_2020.pdf.txtFernandoAGenta_BiancaHenriques_etal_IOC_2020.pdf.txtExtracted texttext/plain48623https://www.arca.fiocruz.br/bitstream/icict/40565/3/FernandoAGenta_BiancaHenriques_etal_IOC_2020.pdf.txt69f946d5ce7e009365b70244b2a02c07MD53icict/405652020-03-31 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dc.title.pt_BR.fl_str_mv Determination of Chitin Content in Insects: An Alternate Method Based on Calcofluor Staining
title Determination of Chitin Content in Insects: An Alternate Method Based on Calcofluor Staining
spellingShingle Determination of Chitin Content in Insects: An Alternate Method Based on Calcofluor Staining
Henriques, Bianca Santos
Quitina
Técnica
Polissacarídeo
Inseto
Quantificação
Chitin
Polysaccharide
Calcofluor
Insect
Quantitation
Technique
title_short Determination of Chitin Content in Insects: An Alternate Method Based on Calcofluor Staining
title_full Determination of Chitin Content in Insects: An Alternate Method Based on Calcofluor Staining
title_fullStr Determination of Chitin Content in Insects: An Alternate Method Based on Calcofluor Staining
title_full_unstemmed Determination of Chitin Content in Insects: An Alternate Method Based on Calcofluor Staining
title_sort Determination of Chitin Content in Insects: An Alternate Method Based on Calcofluor Staining
author Henriques, Bianca Santos
author_facet Henriques, Bianca Santos
Garcia, Eloi Souza
Azambuja, Patricia
Genta, Fernando Ariel
author_role author
author2 Garcia, Eloi Souza
Azambuja, Patricia
Genta, Fernando Ariel
author2_role author
author
author
dc.contributor.author.fl_str_mv Henriques, Bianca Santos
Garcia, Eloi Souza
Azambuja, Patricia
Genta, Fernando Ariel
dc.subject.other.pt_BR.fl_str_mv Quitina
Técnica
Polissacarídeo
Inseto
Quantificação
topic Quitina
Técnica
Polissacarídeo
Inseto
Quantificação
Chitin
Polysaccharide
Calcofluor
Insect
Quantitation
Technique
dc.subject.en.pt_BR.fl_str_mv Chitin
Polysaccharide
Calcofluor
Insect
Quantitation
Technique
description Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ, Brasil.
publishDate 2020
dc.date.accessioned.fl_str_mv 2020-03-30T20:00:53Z
dc.date.available.fl_str_mv 2020-03-30T20:00:53Z
dc.date.issued.fl_str_mv 2020
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.citation.fl_str_mv HENRIQUES. Bianca Santos et al. Determination of Chitin Content in Insects: An Alternate Method Based on Calcofluor Staining. Frontiers in Physiology, v. 11, Article 117, p. 1-10, Feb. 2020.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/40565
dc.identifier.issn.pt_BR.fl_str_mv 1664-042X
dc.identifier.doi.none.fl_str_mv 10.3389/fphys.2020.00117
identifier_str_mv HENRIQUES. Bianca Santos et al. Determination of Chitin Content in Insects: An Alternate Method Based on Calcofluor Staining. Frontiers in Physiology, v. 11, Article 117, p. 1-10, Feb. 2020.
1664-042X
10.3389/fphys.2020.00117
url https://www.arca.fiocruz.br/handle/icict/40565
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Frontiers Media
publisher.none.fl_str_mv Frontiers Media
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
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collection Repositório Institucional da FIOCRUZ (ARCA)
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