Microbiota de ninhos de espuma no combate à quitridiomicose: estratégias para conservação de anfíbios

Detalhes bibliográficos
Ano de defesa: 2024
Autor(a) principal: Sousa, Paulo Ricardo Santos de
Orientador(a): Hissa, Denise Cavalcante
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Não Informado pela instituição
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Área do conhecimento CNPq:
Link de acesso: http://repositorio.ufc.br/handle/riufc/77408
Resumo: Anurans are the vertebrates experiencing the highest population decline globally, with chytridiomycosis caused by the pathogenic fungus Batrachochytrium dendrobatidis (Bd) reported as the primary cause of this decline. It has been reported that the skin microbiota of anurans plays a crucial role in controlling Bd. These bacteria are considered a promising strategy for the development of probiotics that strengthen the host's defenses, preventing chytridiomycosis. Foam nests are biofoams composed of ranaspumin proteins and carbohydrates that provide shelter for eggs until they develop into tadpoles. Similar to anuran skin, foam nests have a unique microbiota, yet unexplored for the presence of anti-Bd bacteria. Thus, this research aimed to prospect anti-Bd bacteria in foam nests of the frog species Adenomera hylaedactyla, Leptodactylus vastus, and Physalaemus cuvieri, aiming to identify potential isolates that can act in controlling the pathogen during the early stages of tadpole development. The study evaluated 182 bacterial isolates from nests of the three frog species. Anti-Bd activity was assessed directly by inoculating the test bacteria at equidistant points on the surface of 1% Tryptone Agar plates containing a standardized Bd layer. Results were analyzed based on the presence or absence of inhibition halos around the isolates. To assess if the activity was due to an extracellular product, each test bacterium's culture was co-cultivated with the fungus, and the co-culture supernatant was cultured in liquid medium inoculated with a standardized Bd suspension. The result was expressed by the number of viable total Bd. Based on the results of the two tests and the compatibility test among the isolates, 11 candidates were selected to form an anti-Bd probiotic consortium from the 55 with hability to inhibit Bd growth. The isolates were combined in18 different consortia. As a result, 30.77% of the isolates were capable of directly inhibiting Bd growth, while only 4.95% of the culture supernatants showed significant Bd inhibition. Additionally, 64.84% of the culture supernatants stimulated Bd growth. In both tests, the most promising isolates with anti-Bd activity were belonging to the genus Pseudomonas. Thirteen of the eighteen consortia showed moderate inhibition. Five consortia stood out, composed of bacteria belonging to the genera Aeromonas, Acinetobacter, Chryseobacterium, Microbacterium, Pseudomonas, and Serratia. The results of this study demonstrate that components of the microbiota in frog foam nests may be the key to Bd control in the early developmental stages. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001.
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spelling Sousa, Paulo Ricardo Santos deMelo, Vânia Maria MacielHissa, Denise Cavalcante2024-07-30T13:54:35Z2024-07-30T13:54:35Z2024SOUSA, Paulo Ricardo Santos de. Microbiota de ninhos de espuma no combate à quitridiomicose: estratégias para conservação de anfíbios. 2024. 74 f. Dissertação (Mestrado em Sistemática, Uso e Conservação da Biodiversidade) - Universidade Federal do Ceará, Fortaleza, 2024.http://repositorio.ufc.br/handle/riufc/77408Anurans are the vertebrates experiencing the highest population decline globally, with chytridiomycosis caused by the pathogenic fungus Batrachochytrium dendrobatidis (Bd) reported as the primary cause of this decline. It has been reported that the skin microbiota of anurans plays a crucial role in controlling Bd. These bacteria are considered a promising strategy for the development of probiotics that strengthen the host's defenses, preventing chytridiomycosis. Foam nests are biofoams composed of ranaspumin proteins and carbohydrates that provide shelter for eggs until they develop into tadpoles. Similar to anuran skin, foam nests have a unique microbiota, yet unexplored for the presence of anti-Bd bacteria. Thus, this research aimed to prospect anti-Bd bacteria in foam nests of the frog species Adenomera hylaedactyla, Leptodactylus vastus, and Physalaemus cuvieri, aiming to identify potential isolates that can act in controlling the pathogen during the early stages of tadpole development. The study evaluated 182 bacterial isolates from nests of the three frog species. Anti-Bd activity was assessed directly by inoculating the test bacteria at equidistant points on the surface of 1% Tryptone Agar plates containing a standardized Bd layer. Results were analyzed based on the presence or absence of inhibition halos around the isolates. To assess if the activity was due to an extracellular product, each test bacterium's culture was co-cultivated with the fungus, and the co-culture supernatant was cultured in liquid medium inoculated with a standardized Bd suspension. The result was expressed by the number of viable total Bd. Based on the results of the two tests and the compatibility test among the isolates, 11 candidates were selected to form an anti-Bd probiotic consortium from the 55 with hability to inhibit Bd growth. The isolates were combined in18 different consortia. As a result, 30.77% of the isolates were capable of directly inhibiting Bd growth, while only 4.95% of the culture supernatants showed significant Bd inhibition. Additionally, 64.84% of the culture supernatants stimulated Bd growth. In both tests, the most promising isolates with anti-Bd activity were belonging to the genus Pseudomonas. Thirteen of the eighteen consortia showed moderate inhibition. Five consortia stood out, composed of bacteria belonging to the genera Aeromonas, Acinetobacter, Chryseobacterium, Microbacterium, Pseudomonas, and Serratia. The results of this study demonstrate that components of the microbiota in frog foam nests may be the key to Bd control in the early developmental stages. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001.Os anuros são os vertebrados que mais sofrem declínio populacional no mundo, sendo a quitridiomicose, causada pelo fungo patogênico Batrachochytrium dendrobatidis (Bd), relatada como a principal causa desse declínio. Já foi reportado que a microbiota da pele dos anuros tem um papel importante no controle do Bd. Essas bactérias são apontadas como uma estratégia promissora para o desenvolvimento de probióticos que atuam fortalecendo as defesas do hospedeiro, prevenindo a quitridiomicose. Os ninhos de espuma, constituem bioespumas compostas de proteínas ranaspuminas e carboidratos que permitem abrigar os ovos até seu desenvolvimento em girinos. Assim como a pele dos anuros, os ninhos de espuma possuem uma microbiota exclusiva, porém ainda não explorada quanto a presença de bactérias anti-Bd. Dessa forma, esta pesquisa teve por objetivo a prospecção de bactérias anti-Bd em ninhos de espuma das espécies de rãs Adenomera hylaedactyla, Leptodactylus vastus e Physalaemus cuvieri visando identificar potenciais isolados que possam atuar no controle do patógeno nas fases iniciais do desenvolvimento do girino. No estudo foram avaliados 182 isolados bacterianos provenientes de ninhos das três espécies de rãs. A atividade anti-Bd foi avaliada de forma direta, inoculando-se as bactérias-testes em pontos equidistantes da superfície de placas de Ágar Triptona 1% contendo uma camada padronizada de Bd. Os resultados foram analisados de acordo com a presença ou ausência de halos de inibição ao redor dos isolados. Para avaliar se a atividade era devida a um produto extracelular, a cultura de cada bactéria-teste foi co- cultivada com a cultura do fungo, e o sobrenadante do co-cultivo foi subsequentemente cultivado em meio líquido inoculado com uma suspensão padronizada de Bd. O resultado foi expresso pelo número de Bd viáveis totais. Com base nos resultados dos dois testes e no teste de compatibilidade entre os isolados foram selecionados 11 isolados candidatos a compor um consórcio probiótico anti-Bd, dos 55 que apresentaram resultados positivos. Os isolados foram combinados gerando 18 consórcios diferentes. Como resultados tem-se que 30,77% dos isolados foram capazes de inibir diretamente o crescimento do Bd, enquanto apenas 4,95% dos sobrenadantes das culturas apresentaram inibição significativa do Bd. Além disso, 64,84% dos sobrenadantes das culturas estimularam o crescimento do Bd. Em ambos os testes, os isolados mais promissores com atividade anti-Bd foram identificados como pertencentes ao gênero Pseudomonas. Dos 18 consórcios testados, 13 apresentaram inibição moderada. Cinco consórcios se destacaram, sendo compostos por bactérias pertencentes aos gêneros Aeromonas, Acinetobacter, Chryseobacterium, Microbacterium, Pseudomonas e Serratia. Os resultados desse estudo mostram que componentes da microbiota dos ninhos de espuma de rãs podem ser a chave para o controle do Bd nas fases iniciais do desenvolvimento. O presente trabalho foi realizado com apoio da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Código de Financiamento 001.Microbiota de ninhos de espuma no combate à quitridiomicose: estratégias para conservação de anfíbiosinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisBatrachochytrium dendrobatidisAnurosProbióticosBioaumentaçãoBatrachochytrium dendrobatidisAnuransProbioticsBioaugmentationCNPQ::CIENCIAS BIOLOGICAS::ECOLOGIAinfo:eu-repo/semantics/openAccessporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFChttp://lattes.cnpq.br/1872305912279121http://lattes.cnpq.br/4154200299160135http://lattes.cnpq.br/15725046509306052024-07-30LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/77408/4/license.txt8a4605be74aa9ea9d79846c1fba20a33MD54ORIGINAL2024_dis_prssousa.pdf2024_dis_prssousa.pdfapplication/pdf2369378http://repositorio.ufc.br/bitstream/riufc/77408/3/2024_dis_prssousa.pdf26fede9538cd31a92ae5bab4d0211ba2MD53riufc/774082024-07-30 10:55:47.327oai:repositorio.ufc.br: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Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-07-30T13:55:47Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.pt_BR.fl_str_mv Microbiota de ninhos de espuma no combate à quitridiomicose: estratégias para conservação de anfíbios
title Microbiota de ninhos de espuma no combate à quitridiomicose: estratégias para conservação de anfíbios
spellingShingle Microbiota de ninhos de espuma no combate à quitridiomicose: estratégias para conservação de anfíbios
Sousa, Paulo Ricardo Santos de
CNPQ::CIENCIAS BIOLOGICAS::ECOLOGIA
Batrachochytrium dendrobatidis
Anuros
Probióticos
Bioaumentação
Batrachochytrium dendrobatidis
Anurans
Probiotics
Bioaugmentation
title_short Microbiota de ninhos de espuma no combate à quitridiomicose: estratégias para conservação de anfíbios
title_full Microbiota de ninhos de espuma no combate à quitridiomicose: estratégias para conservação de anfíbios
title_fullStr Microbiota de ninhos de espuma no combate à quitridiomicose: estratégias para conservação de anfíbios
title_full_unstemmed Microbiota de ninhos de espuma no combate à quitridiomicose: estratégias para conservação de anfíbios
title_sort Microbiota de ninhos de espuma no combate à quitridiomicose: estratégias para conservação de anfíbios
author Sousa, Paulo Ricardo Santos de
author_facet Sousa, Paulo Ricardo Santos de
author_role author
dc.contributor.co-advisor.none.fl_str_mv Melo, Vânia Maria Maciel
dc.contributor.author.fl_str_mv Sousa, Paulo Ricardo Santos de
dc.contributor.advisor1.fl_str_mv Hissa, Denise Cavalcante
contributor_str_mv Hissa, Denise Cavalcante
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::ECOLOGIA
topic CNPQ::CIENCIAS BIOLOGICAS::ECOLOGIA
Batrachochytrium dendrobatidis
Anuros
Probióticos
Bioaumentação
Batrachochytrium dendrobatidis
Anurans
Probiotics
Bioaugmentation
dc.subject.ptbr.pt_BR.fl_str_mv Batrachochytrium dendrobatidis
Anuros
Probióticos
Bioaumentação
dc.subject.en.pt_BR.fl_str_mv Batrachochytrium dendrobatidis
Anurans
Probiotics
Bioaugmentation
description Anurans are the vertebrates experiencing the highest population decline globally, with chytridiomycosis caused by the pathogenic fungus Batrachochytrium dendrobatidis (Bd) reported as the primary cause of this decline. It has been reported that the skin microbiota of anurans plays a crucial role in controlling Bd. These bacteria are considered a promising strategy for the development of probiotics that strengthen the host's defenses, preventing chytridiomycosis. Foam nests are biofoams composed of ranaspumin proteins and carbohydrates that provide shelter for eggs until they develop into tadpoles. Similar to anuran skin, foam nests have a unique microbiota, yet unexplored for the presence of anti-Bd bacteria. Thus, this research aimed to prospect anti-Bd bacteria in foam nests of the frog species Adenomera hylaedactyla, Leptodactylus vastus, and Physalaemus cuvieri, aiming to identify potential isolates that can act in controlling the pathogen during the early stages of tadpole development. The study evaluated 182 bacterial isolates from nests of the three frog species. Anti-Bd activity was assessed directly by inoculating the test bacteria at equidistant points on the surface of 1% Tryptone Agar plates containing a standardized Bd layer. Results were analyzed based on the presence or absence of inhibition halos around the isolates. To assess if the activity was due to an extracellular product, each test bacterium's culture was co-cultivated with the fungus, and the co-culture supernatant was cultured in liquid medium inoculated with a standardized Bd suspension. The result was expressed by the number of viable total Bd. Based on the results of the two tests and the compatibility test among the isolates, 11 candidates were selected to form an anti-Bd probiotic consortium from the 55 with hability to inhibit Bd growth. The isolates were combined in18 different consortia. As a result, 30.77% of the isolates were capable of directly inhibiting Bd growth, while only 4.95% of the culture supernatants showed significant Bd inhibition. Additionally, 64.84% of the culture supernatants stimulated Bd growth. In both tests, the most promising isolates with anti-Bd activity were belonging to the genus Pseudomonas. Thirteen of the eighteen consortia showed moderate inhibition. Five consortia stood out, composed of bacteria belonging to the genera Aeromonas, Acinetobacter, Chryseobacterium, Microbacterium, Pseudomonas, and Serratia. The results of this study demonstrate that components of the microbiota in frog foam nests may be the key to Bd control in the early developmental stages. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001.
publishDate 2024
dc.date.accessioned.fl_str_mv 2024-07-30T13:54:35Z
dc.date.available.fl_str_mv 2024-07-30T13:54:35Z
dc.date.issued.fl_str_mv 2024
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.citation.fl_str_mv SOUSA, Paulo Ricardo Santos de. Microbiota de ninhos de espuma no combate à quitridiomicose: estratégias para conservação de anfíbios. 2024. 74 f. Dissertação (Mestrado em Sistemática, Uso e Conservação da Biodiversidade) - Universidade Federal do Ceará, Fortaleza, 2024.
dc.identifier.uri.fl_str_mv http://repositorio.ufc.br/handle/riufc/77408
identifier_str_mv SOUSA, Paulo Ricardo Santos de. Microbiota de ninhos de espuma no combate à quitridiomicose: estratégias para conservação de anfíbios. 2024. 74 f. Dissertação (Mestrado em Sistemática, Uso e Conservação da Biodiversidade) - Universidade Federal do Ceará, Fortaleza, 2024.
url http://repositorio.ufc.br/handle/riufc/77408
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