Development of alginate microparticles coated with HPMCAS containing the antimicrobial peptide Ctx(Ile²¹)-Ha for application as a natural oral additive in laying hens against systemic Salmonella Enteritidis infection

Detalhes bibliográficos
Autor(a) principal: Roque Borda, Cesar Augusto
Data de Publicação: 2021
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://hdl.handle.net/11449/204500
Resumo: The search for new compounds that can increase and improve performance or reduce chemical residues in products of animal origin is becoming relevant and in demand. At the same time, the constant use of synthetic antibiotics as growth promoters causes great bacterial resistance in laying birds, as well as traces and residues in their products, which can affect animal health and, consequently, human health. In this sense, the antimicrobial peptide (AMP) Ctx(Ile21)-Ha, extracted and isolated from the Brazilian Cerrado frog Hypsiboas albopunctatus, is a very promising molecule in the control of pathogenic bacteria. However, AMPs applied in an in vivo system can undergo degradation or denaturation due to the presence of proteases or hydrolysis by the acidic environment. Thus, this study aimed at the synthesis and purification of the Ctx(Ile21)-Ha peptide, and the coating of alginate microparticles with hypromellose acetate/succinate (HPMCAS), in addition to its physicochemical characterization. Hemolytic activity and simulated in vitro gastrointestinal release of the coated microparticles were also evaluated. In vivo study was carried out with newly hatched laying hens, challenged with Salmonella Enteritidis and treated with microparticles in the feed. Bacterial counts were performed in liver, spleen, and cecum to verify the effects of peptide. The results obtained showed thermostable microparticles, with encapsulation efficiency greater than 70%, slight gastric release, excellent intestinal release, the hemolytic activity of peptide decreased by 95%, and in the in vivo experiment a significant difference was obtained in bacterial count, mainly in liver, followed by spleen. Therefore, this study demonstrated that the coated microparticles containing AMP Ctx(Ile21)-Ha have a high antimicrobial activity and serve as a tool for the prevention of systemic infections, without contaminating residues compared to conventional drugs.
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spelling Development of alginate microparticles coated with HPMCAS containing the antimicrobial peptide Ctx(Ile²¹)-Ha for application as a natural oral additive in laying hens against systemic Salmonella Enteritidis infectionDesenvolvimento de micropartículas de alginato revestidas com HPMCAS contendo o peptídeo antimicrobiano Ctx(Ile²¹)-Ha para aplicação como aditivo oral natural em galinhas de postura contra infecção sistêmica por Salmonella EnteritidisAdditivesEnteric coatingLaying hensSalmonellaPoultry healthAditivosAves PoedeirasMicropartículasRevestimento entéricoSanidadeThe search for new compounds that can increase and improve performance or reduce chemical residues in products of animal origin is becoming relevant and in demand. At the same time, the constant use of synthetic antibiotics as growth promoters causes great bacterial resistance in laying birds, as well as traces and residues in their products, which can affect animal health and, consequently, human health. In this sense, the antimicrobial peptide (AMP) Ctx(Ile21)-Ha, extracted and isolated from the Brazilian Cerrado frog Hypsiboas albopunctatus, is a very promising molecule in the control of pathogenic bacteria. However, AMPs applied in an in vivo system can undergo degradation or denaturation due to the presence of proteases or hydrolysis by the acidic environment. Thus, this study aimed at the synthesis and purification of the Ctx(Ile21)-Ha peptide, and the coating of alginate microparticles with hypromellose acetate/succinate (HPMCAS), in addition to its physicochemical characterization. Hemolytic activity and simulated in vitro gastrointestinal release of the coated microparticles were also evaluated. In vivo study was carried out with newly hatched laying hens, challenged with Salmonella Enteritidis and treated with microparticles in the feed. Bacterial counts were performed in liver, spleen, and cecum to verify the effects of peptide. The results obtained showed thermostable microparticles, with encapsulation efficiency greater than 70%, slight gastric release, excellent intestinal release, the hemolytic activity of peptide decreased by 95%, and in the in vivo experiment a significant difference was obtained in bacterial count, mainly in liver, followed by spleen. Therefore, this study demonstrated that the coated microparticles containing AMP Ctx(Ile21)-Ha have a high antimicrobial activity and serve as a tool for the prevention of systemic infections, without contaminating residues compared to conventional drugs.A pesquisa por novos compostos que possam aumentar e melhorar o desempenho ou diminuir resíduos químicos em produtos de origem animal torna-se relevante e demandada. Paralelamente, o uso constante de antibióticos sintéticos como promotores de crescimento causa uma grande resistência bacteriana em aves poedeiras, além de rastros e resíduos em seus produtos, os quais podem afetar a sanidade animal e, consequentemente, a saúde humana. Neste sentido, o peptídeo antimicrobiano (PAM) Ctx(Ile21)-Ha, extraído e isolado da rã do cerrado brasileiro Hypsiboas albopunctatus, é uma molécula muito promissora no controle de bactérias patogênicas. No entanto, os PAMs aplicados em sistemas in vivo, podem sofrer degradação ou desnaturação pela presença de proteases ou hidrolise pelo entorno ácido. Assim, este estudo objetivou a síntese e purificação peptídeo Ctx(Ile21)-Ha, e o revestimento de micropartículas de alginato com acetato/succinato de hipromelose (HPMCAS), além de sua caracterização fisioquímica. Foi também avaliada a atividade hemolítica e liberação in vitro gastrointestinal simulada das micropartículas revestidas. O estudo in vivo foi realizado com galinhas de postura recém-nascidas, desafiadas com Salmonella Enteritidis, e tratadas com as micropartículas na ração. Foi realizado a contagem bacteriana em fígado, baço e ceco para verificação dos efeitos do peptídeo. Os resultados obtidos mostraram micropartículas termoestáveis, com eficiência de encapsulação acima do 70%, leve liberação gástrica, ótima liberação intestinal, atividade hemolítica do peptídeo diminuída em 95%, além no experimento in vivo obteve-se diferença significativa na contagem principalmente no fígado, seguida do baço. Portanto, este estudo demonstrou que as micropartículas revestidas contendo o PAM Ctx(Ile21)-Ha possuem alta atividade antimicrobiana e serve de ferramenta para prevenção de infecção sistêmica, sem resíduos contaminantes em comparação com os fármacos convencionais.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP: 18/25707-3Universidade Estadual Paulista (Unesp)Vicente, Eduardo FestozoUniversidade Estadual Paulista (Unesp)Roque Borda, Cesar Augusto2021-04-26T18:38:23Z2021-04-26T18:38:23Z2021-04-06info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfhttp://hdl.handle.net/11449/20450033004102002P0enginfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESP2024-06-05T14:25:18Zoai:repositorio.unesp.br:11449/204500Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T16:00:28.482373Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Development of alginate microparticles coated with HPMCAS containing the antimicrobial peptide Ctx(Ile²¹)-Ha for application as a natural oral additive in laying hens against systemic Salmonella Enteritidis infection
Desenvolvimento de micropartículas de alginato revestidas com HPMCAS contendo o peptídeo antimicrobiano Ctx(Ile²¹)-Ha para aplicação como aditivo oral natural em galinhas de postura contra infecção sistêmica por Salmonella Enteritidis
title Development of alginate microparticles coated with HPMCAS containing the antimicrobial peptide Ctx(Ile²¹)-Ha for application as a natural oral additive in laying hens against systemic Salmonella Enteritidis infection
spellingShingle Development of alginate microparticles coated with HPMCAS containing the antimicrobial peptide Ctx(Ile²¹)-Ha for application as a natural oral additive in laying hens against systemic Salmonella Enteritidis infection
Roque Borda, Cesar Augusto
Additives
Enteric coating
Laying hens
Salmonella
Poultry health
Aditivos
Aves Poedeiras
Micropartículas
Revestimento entérico
Sanidade
title_short Development of alginate microparticles coated with HPMCAS containing the antimicrobial peptide Ctx(Ile²¹)-Ha for application as a natural oral additive in laying hens against systemic Salmonella Enteritidis infection
title_full Development of alginate microparticles coated with HPMCAS containing the antimicrobial peptide Ctx(Ile²¹)-Ha for application as a natural oral additive in laying hens against systemic Salmonella Enteritidis infection
title_fullStr Development of alginate microparticles coated with HPMCAS containing the antimicrobial peptide Ctx(Ile²¹)-Ha for application as a natural oral additive in laying hens against systemic Salmonella Enteritidis infection
title_full_unstemmed Development of alginate microparticles coated with HPMCAS containing the antimicrobial peptide Ctx(Ile²¹)-Ha for application as a natural oral additive in laying hens against systemic Salmonella Enteritidis infection
title_sort Development of alginate microparticles coated with HPMCAS containing the antimicrobial peptide Ctx(Ile²¹)-Ha for application as a natural oral additive in laying hens against systemic Salmonella Enteritidis infection
author Roque Borda, Cesar Augusto
author_facet Roque Borda, Cesar Augusto
author_role author
dc.contributor.none.fl_str_mv Vicente, Eduardo Festozo
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Roque Borda, Cesar Augusto
dc.subject.por.fl_str_mv Additives
Enteric coating
Laying hens
Salmonella
Poultry health
Aditivos
Aves Poedeiras
Micropartículas
Revestimento entérico
Sanidade
topic Additives
Enteric coating
Laying hens
Salmonella
Poultry health
Aditivos
Aves Poedeiras
Micropartículas
Revestimento entérico
Sanidade
description The search for new compounds that can increase and improve performance or reduce chemical residues in products of animal origin is becoming relevant and in demand. At the same time, the constant use of synthetic antibiotics as growth promoters causes great bacterial resistance in laying birds, as well as traces and residues in their products, which can affect animal health and, consequently, human health. In this sense, the antimicrobial peptide (AMP) Ctx(Ile21)-Ha, extracted and isolated from the Brazilian Cerrado frog Hypsiboas albopunctatus, is a very promising molecule in the control of pathogenic bacteria. However, AMPs applied in an in vivo system can undergo degradation or denaturation due to the presence of proteases or hydrolysis by the acidic environment. Thus, this study aimed at the synthesis and purification of the Ctx(Ile21)-Ha peptide, and the coating of alginate microparticles with hypromellose acetate/succinate (HPMCAS), in addition to its physicochemical characterization. Hemolytic activity and simulated in vitro gastrointestinal release of the coated microparticles were also evaluated. In vivo study was carried out with newly hatched laying hens, challenged with Salmonella Enteritidis and treated with microparticles in the feed. Bacterial counts were performed in liver, spleen, and cecum to verify the effects of peptide. The results obtained showed thermostable microparticles, with encapsulation efficiency greater than 70%, slight gastric release, excellent intestinal release, the hemolytic activity of peptide decreased by 95%, and in the in vivo experiment a significant difference was obtained in bacterial count, mainly in liver, followed by spleen. Therefore, this study demonstrated that the coated microparticles containing AMP Ctx(Ile21)-Ha have a high antimicrobial activity and serve as a tool for the prevention of systemic infections, without contaminating residues compared to conventional drugs.
publishDate 2021
dc.date.none.fl_str_mv 2021-04-26T18:38:23Z
2021-04-26T18:38:23Z
2021-04-06
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/11449/204500
33004102002P0
url http://hdl.handle.net/11449/204500
identifier_str_mv 33004102002P0
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.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Universidade Estadual Paulista (Unesp)
publisher.none.fl_str_mv Universidade Estadual Paulista (Unesp)
dc.source.none.fl_str_mv reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
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