Bioterapia com Lucilia cuprina: atividade antimicrobiana e eficácia em feridas infectadas experimentalmente

Detalhes bibliográficos
Autor(a) principal: Dillmann, Janaína Brand
Data de Publicação: 2022
Tipo de documento: Tese
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
Texto Completo: http://repositorio.ufsm.br/handle/1/25785
Resumo: Skin wounds are frequent in animals due to behavioral and management factors, and a subject of constant study due to their clinical, scientific, and economic interest. Although bacteria are part of the normal microbiota of the skin and wounds, the onset of an infection by their exacerbated proliferation and potential biofilm formation slows the healing process. In addition, inappropriate use of antibiotic therapy is a major cause of the spread of resistant microorganisms. Larval therapy emerges as an alternative for wound treatment, since necrophagous larvae such as the species Lucilia cuprina have present debridement potential, promote lesion cleansing by consuming local dead tissue and decrease the degree of infection, since bacteria are ingested and destroyed in the gut of the larvae. In addition to these benefits, pH change, stimulation of neovascularization, and tissue washing by induced local exudation accelerate the healing process. Thus, the objectives of this study were: (i) to evaluate the in vitro antimicrobial potential of L. cuprina larvae challenged by bacteria and (ii) to evaluate the efficacy and therapeutic safety of L. cuprina larvae on induced infected wounds in Wistar rats. Objective (i) was achieved by collecting whole body extracts from larvae incubated for 24 hours with a bacterial suspension of Staphylococcus aureus, Pseudomonas aeruginosa or methicillin-resistant S. aureus (MRSA). An antimicrobial analysis by Colony Forming Unit (CFU) assay was performed at two incubation times (0 and 6 hours) with pretreated and untreated (PBS) groups of larval extracts compared to a control group of bacterial growth. The results of experiment (i) showed significant activity against P. aeruginosa, a slight decrease in bacterial growth for S. aureus and MRSA. The results of objective (ii) were achieved through clinical, microbiological, histopathological analyses of the wounds and hematological and biochemical analyses of the animals. A significant shrinkage of the wound area (>95%) in the larvae-treated group was achieved at 9th day compared to 15th day in the antibiotic-treated group. There was complete elimination of MRSA colonies after the second treatment (6th day), compared to an increase in bacterial proliferation in the control group. On histopathology, a clean wound was observed in the larval treated group, as also adequate collagen formation and re-epithelialization on 15th day, and an increase in blood platelet levels in the hematological evaluation. It is then concluded that L. cuprina larvae are safe and effective in accelerating wound treatment and eliminating MRSA, and may represent an alternative for use in wound healing resistant to conventional treatments.
id UFSM_1d40ca8f89244b0b5218badcd8671a8e
oai_identifier_str oai:repositorio.ufsm.br:1/25785
network_acronym_str UFSM
network_name_str Manancial - Repositório Digital da UFSM
repository_id_str
spelling Bioterapia com Lucilia cuprina: atividade antimicrobiana e eficácia em feridas infectadas experimentalmenteBiotherapy with Lucilia cuprina: antimicrobial activity and effectiveness in experimentally infected woundsCalliphoridaeFeridasStaphylococcus aureus resistente a meticilinaTerapia larvalCalliphoridaeMaggot debridement therapyMethicillin-resistant Staphylococcus aureusWoundCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIASkin wounds are frequent in animals due to behavioral and management factors, and a subject of constant study due to their clinical, scientific, and economic interest. Although bacteria are part of the normal microbiota of the skin and wounds, the onset of an infection by their exacerbated proliferation and potential biofilm formation slows the healing process. In addition, inappropriate use of antibiotic therapy is a major cause of the spread of resistant microorganisms. Larval therapy emerges as an alternative for wound treatment, since necrophagous larvae such as the species Lucilia cuprina have present debridement potential, promote lesion cleansing by consuming local dead tissue and decrease the degree of infection, since bacteria are ingested and destroyed in the gut of the larvae. In addition to these benefits, pH change, stimulation of neovascularization, and tissue washing by induced local exudation accelerate the healing process. Thus, the objectives of this study were: (i) to evaluate the in vitro antimicrobial potential of L. cuprina larvae challenged by bacteria and (ii) to evaluate the efficacy and therapeutic safety of L. cuprina larvae on induced infected wounds in Wistar rats. Objective (i) was achieved by collecting whole body extracts from larvae incubated for 24 hours with a bacterial suspension of Staphylococcus aureus, Pseudomonas aeruginosa or methicillin-resistant S. aureus (MRSA). An antimicrobial analysis by Colony Forming Unit (CFU) assay was performed at two incubation times (0 and 6 hours) with pretreated and untreated (PBS) groups of larval extracts compared to a control group of bacterial growth. The results of experiment (i) showed significant activity against P. aeruginosa, a slight decrease in bacterial growth for S. aureus and MRSA. The results of objective (ii) were achieved through clinical, microbiological, histopathological analyses of the wounds and hematological and biochemical analyses of the animals. A significant shrinkage of the wound area (>95%) in the larvae-treated group was achieved at 9th day compared to 15th day in the antibiotic-treated group. There was complete elimination of MRSA colonies after the second treatment (6th day), compared to an increase in bacterial proliferation in the control group. On histopathology, a clean wound was observed in the larval treated group, as also adequate collagen formation and re-epithelialization on 15th day, and an increase in blood platelet levels in the hematological evaluation. It is then concluded that L. cuprina larvae are safe and effective in accelerating wound treatment and eliminating MRSA, and may represent an alternative for use in wound healing resistant to conventional treatments.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESAs feridas cutâneas são frequentes em animais devido a fatores comportamentais e de manejo, tema de estudo constante devido seu interesse clínico, científico e econômico. Embora bactérias façam parte da microbiota da pele e de feridas, sua proliferação exacerbada e potencial formação de biofilme retarda o processo de cicatrização. Além disso, o uso inapropriado de terapia antibiótica é a principal causa da disseminação de microrganismos resistentes. Com isso, a terapia larval surge como uma alternativa no tratamento de feridas, uma vez que larvas necrobiontófagas como da espécie Lucilia cuprina possuem potencial de desbridamento, promovem a limpeza das lesões através do consumo do tecido morto local e diminuem o grau de infecção, já que as bactérias são ingeridas e destruídas no trato digestório das larvas. Além desses benefícios, a mudança de pH, estímulo da neovascularização e lavagem tecidual pela exsudação local induzida aceleram o processo cicatricial. Dessa forma, os objetivos desse estudo foram: (i) avaliar o potencial antimicrobiano in vitro de larvas de L. cuprina estimuladas por bactérias e (ii) avaliar a eficácia e segurança terapêutica de larvas de L. cuprina em feridas infectadas induzidas em ratos Wistar. O objetivo (i) foi alcançado através de coleta de extratos do corpo inteiro das larvas incubadas durante 24 horas com uma suspensão bacteriana de Staphylococcus aureus, Pseudomonas aeruginosa ou S. aureus resistente à meticilina (SARM); e análise antimicrobiana através de ensaio de Unidade Formadora de Colônia (UFC) em dois tempos de incubação (0 e 6 horas) com grupos de extratos larvais pré-tratados e não tratados (PBS), em comparação com um grupo de controle do crescimento bacteriano. Os resultados do experimento (i) mostraram significativa atividade contra P. aeruginosa, uma ligeira diminuição do crescimento bacteriano para S. aureus e SARM. Já os resultados do objetivo (ii) foram alcançados através de análises clínicas, microbiológicas, histopatológicas das feridas, e hematológicas e bioquímicas dos animais. Uma contração significativa da área da ferida (>95%) no grupo tratado com larvas foi alcançado ao 9º dia, em comparação com o 15º dia no grupo tratado com antibiótico. Houve eliminação completa das colônias bacterianas de SARM após o segundo tratamento (6º dia) em comparação com um aumento na proliferação bacteriana do grupo controle. Na histopatologia, observou-se uma ferida limpa no grupo tratado com larvas, formação adequada de colágeno e reepitelização no 15º dia, além de aumento nos níveis de plaquetas na avaliação hematológica. Concluiu-se então, que as larvas de L. cuprina são seguras e eficazes na cicatrização da lesão e na eliminação de SARM e que podem representar uma alternativa para a utilização em cicatrização de feridas resistentes aos tratamentos convencionais.Universidade Federal de Santa MariaBrasilMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaCentro de Ciências RuraisMonteiro, Silvia Gonzalezhttp://lattes.cnpq.br/3762606653182779Silva Júnior, José Valter JoaquimCargnelutti, Juliana FelipettoDalla Rosa, LucianaBassuino, Daniele MariathDillmann, Janaína Brand2022-08-04T11:16:10Z2022-08-04T11:16:10Z2022-02-23info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/25785porAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2022-08-04T11:16:10Zoai:repositorio.ufsm.br:1/25785Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2022-08-04T11:16:10Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Bioterapia com Lucilia cuprina: atividade antimicrobiana e eficácia em feridas infectadas experimentalmente
Biotherapy with Lucilia cuprina: antimicrobial activity and effectiveness in experimentally infected wounds
title Bioterapia com Lucilia cuprina: atividade antimicrobiana e eficácia em feridas infectadas experimentalmente
spellingShingle Bioterapia com Lucilia cuprina: atividade antimicrobiana e eficácia em feridas infectadas experimentalmente
Dillmann, Janaína Brand
Calliphoridae
Feridas
Staphylococcus aureus resistente a meticilina
Terapia larval
Calliphoridae
Maggot debridement therapy
Methicillin-resistant Staphylococcus aureus
Wound
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Bioterapia com Lucilia cuprina: atividade antimicrobiana e eficácia em feridas infectadas experimentalmente
title_full Bioterapia com Lucilia cuprina: atividade antimicrobiana e eficácia em feridas infectadas experimentalmente
title_fullStr Bioterapia com Lucilia cuprina: atividade antimicrobiana e eficácia em feridas infectadas experimentalmente
title_full_unstemmed Bioterapia com Lucilia cuprina: atividade antimicrobiana e eficácia em feridas infectadas experimentalmente
title_sort Bioterapia com Lucilia cuprina: atividade antimicrobiana e eficácia em feridas infectadas experimentalmente
author Dillmann, Janaína Brand
author_facet Dillmann, Janaína Brand
author_role author
dc.contributor.none.fl_str_mv Monteiro, Silvia Gonzalez
http://lattes.cnpq.br/3762606653182779
Silva Júnior, José Valter Joaquim
Cargnelutti, Juliana Felipetto
Dalla Rosa, Luciana
Bassuino, Daniele Mariath
dc.contributor.author.fl_str_mv Dillmann, Janaína Brand
dc.subject.por.fl_str_mv Calliphoridae
Feridas
Staphylococcus aureus resistente a meticilina
Terapia larval
Calliphoridae
Maggot debridement therapy
Methicillin-resistant Staphylococcus aureus
Wound
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
topic Calliphoridae
Feridas
Staphylococcus aureus resistente a meticilina
Terapia larval
Calliphoridae
Maggot debridement therapy
Methicillin-resistant Staphylococcus aureus
Wound
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description Skin wounds are frequent in animals due to behavioral and management factors, and a subject of constant study due to their clinical, scientific, and economic interest. Although bacteria are part of the normal microbiota of the skin and wounds, the onset of an infection by their exacerbated proliferation and potential biofilm formation slows the healing process. In addition, inappropriate use of antibiotic therapy is a major cause of the spread of resistant microorganisms. Larval therapy emerges as an alternative for wound treatment, since necrophagous larvae such as the species Lucilia cuprina have present debridement potential, promote lesion cleansing by consuming local dead tissue and decrease the degree of infection, since bacteria are ingested and destroyed in the gut of the larvae. In addition to these benefits, pH change, stimulation of neovascularization, and tissue washing by induced local exudation accelerate the healing process. Thus, the objectives of this study were: (i) to evaluate the in vitro antimicrobial potential of L. cuprina larvae challenged by bacteria and (ii) to evaluate the efficacy and therapeutic safety of L. cuprina larvae on induced infected wounds in Wistar rats. Objective (i) was achieved by collecting whole body extracts from larvae incubated for 24 hours with a bacterial suspension of Staphylococcus aureus, Pseudomonas aeruginosa or methicillin-resistant S. aureus (MRSA). An antimicrobial analysis by Colony Forming Unit (CFU) assay was performed at two incubation times (0 and 6 hours) with pretreated and untreated (PBS) groups of larval extracts compared to a control group of bacterial growth. The results of experiment (i) showed significant activity against P. aeruginosa, a slight decrease in bacterial growth for S. aureus and MRSA. The results of objective (ii) were achieved through clinical, microbiological, histopathological analyses of the wounds and hematological and biochemical analyses of the animals. A significant shrinkage of the wound area (>95%) in the larvae-treated group was achieved at 9th day compared to 15th day in the antibiotic-treated group. There was complete elimination of MRSA colonies after the second treatment (6th day), compared to an increase in bacterial proliferation in the control group. On histopathology, a clean wound was observed in the larval treated group, as also adequate collagen formation and re-epithelialization on 15th day, and an increase in blood platelet levels in the hematological evaluation. It is then concluded that L. cuprina larvae are safe and effective in accelerating wound treatment and eliminating MRSA, and may represent an alternative for use in wound healing resistant to conventional treatments.
publishDate 2022
dc.date.none.fl_str_mv 2022-08-04T11:16:10Z
2022-08-04T11:16:10Z
2022-02-23
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://repositorio.ufsm.br/handle/1/25785
url http://repositorio.ufsm.br/handle/1/25785
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
Centro de Ciências Rurais
publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
Centro de Ciências Rurais
dc.source.none.fl_str_mv reponame:Manancial - Repositório Digital da UFSM
instname:Universidade Federal de Santa Maria (UFSM)
instacron:UFSM
instname_str Universidade Federal de Santa Maria (UFSM)
instacron_str UFSM
institution UFSM
reponame_str Manancial - Repositório Digital da UFSM
collection Manancial - Repositório Digital da UFSM
repository.name.fl_str_mv Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)
repository.mail.fl_str_mv atendimento.sib@ufsm.br||tedebc@gmail.com
_version_ 1805922092202328064