Antimicrobial and antiparasitic potential of Dinoponera quadriceps venom
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UFC |
Texto Completo: | http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=11676 |
Resumo: | Animal toxins can be a source of molecular models for the design of new drugs. This study investigated the antimicrobial and trypanocidal potential from Dinoponera quadriceps ant venom (DqV) aiming to discover therapeutic value substances. We conducted the microdilution test, where it was determined the minimum inhibitory concentration (MIC) and Minimum Lethal Concentration (MLC) over Staphylococcus aureus ATCC 6538P , Escherichia coli ATCC 10536 , Pseudomonas aeruginosa ATCC 9027 , Salmonella subsp cholearaesuis choleraesuis serotype choleraesuis ATCC 10708 and Candida albicans ATCC 10231 strains and two microbial strains of Methicillin Resistant Staphylococcus aureus (MRSA), S. aureus ATCC 33591 and S. aureus CCBH 5330. MIC and MLC of DqV were respectively 6.25 Âg/mL and 12.5 Âg/mL for S. aureus ATCC 6538P, 3.12 Âg/mL and 3.12 Âg/mL for E. coli, 12.5 Âg/mL and 12.5 Âg/mL for P. aeruginosa, 12.5 Âg/mL and 25 Âg/mL for S. choleraesuis, 25 Âg/mL and 50 Âg/mL for C. albicans, 12.5 Âg/mL and 50 Âg/mL for S. aureus CCBH 5330 and 100 Âg/mL and 100 Âg/mL for S. aureus ATCC 33591. Mechanism of action experiments were performed for the strain of S. aureus ATCC 6538P methicillin-susceptible (MSSA), that changes in the permeability of the bacterial membrane of S. aureus treated with bacteriostatic and bactericidal concentrations of DqV was observed by the crystal violet assay and release of genetic material assay. A lowest MIC was observed when alkaline pH broth was used (7,5-9,0). Complete bacterial growth inhibition was observed after 4 h of incubation with the MLC of DqV. Bacterial morphology was analyzed by atomic force microscopy after exposure of bacteria to the CIM and CIM /2 of DqV for 4 hours, showing membrane damage. In antiparasitic assays, we determined the cytotoxic effects of the venom on epimastigote and trypomastigote forms of the Y strain of Trypanosoma cruzi. In epimastigotes, cytotoxicity was evaluated at 24 and 48 h, finding IC50 of 28.32 Âg/mL and 20.67 Âg/mL, respectively. The mechanism of cell death was assessed by flow cytometry and revealed the presence of necrotic and apoptotic involvement in the cytotoxic effect of DqV over epimastigote form, in addition, the appearance of double labeled cells with PI and Annexin V-FITC, indicating the occurrence of late apoptosis. Cytotoxicity was evaluated over trypomastigote finding an IC50 of 1.978 Âg/mL and over RAW 264.7 cells finding an IC50 of 32.44 Âg/mL. The venom showed antibacterial activity against S. aureus MSSA and MRSA, P. aeruginosa, S. choleraesuis, E. coli and C. albicans, suggesting membrane damage in S. aureus ATCC 6538P. Additionally, showed cytotoxic potential on epimastigote and tripomastigote forms of Y strain of T. cruzi. |
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info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisAntimicrobial and antiparasitic potential of Dinoponera quadriceps venomPotencial antimicrobiano e antiparasitÃrio do veneno da Dinoponera quadriceps2014-02-25Alice Maria Costa Martins43431690300http://lattes.cnpq.br/7532334620264577Renata de Sousa Alves74909126368http://lattes.cnpq.br/6730902567211867Nadia Accioly Pinto Nogueira19127057372http://lattes.cnpq.br/926132216566974102697539310http://lattes.cnpq.br/2010191025726363Danya Bandeira LimaUniversidade Federal do CearÃPrograma de PÃs-GraduaÃÃo em CiÃncias FarmacÃuticasUFCBRAnt venoms Anti-infectives AntiparasiticsFARMACIAAnimal toxins can be a source of molecular models for the design of new drugs. This study investigated the antimicrobial and trypanocidal potential from Dinoponera quadriceps ant venom (DqV) aiming to discover therapeutic value substances. We conducted the microdilution test, where it was determined the minimum inhibitory concentration (MIC) and Minimum Lethal Concentration (MLC) over Staphylococcus aureus ATCC 6538P , Escherichia coli ATCC 10536 , Pseudomonas aeruginosa ATCC 9027 , Salmonella subsp cholearaesuis choleraesuis serotype choleraesuis ATCC 10708 and Candida albicans ATCC 10231 strains and two microbial strains of Methicillin Resistant Staphylococcus aureus (MRSA), S. aureus ATCC 33591 and S. aureus CCBH 5330. MIC and MLC of DqV were respectively 6.25 Âg/mL and 12.5 Âg/mL for S. aureus ATCC 6538P, 3.12 Âg/mL and 3.12 Âg/mL for E. coli, 12.5 Âg/mL and 12.5 Âg/mL for P. aeruginosa, 12.5 Âg/mL and 25 Âg/mL for S. choleraesuis, 25 Âg/mL and 50 Âg/mL for C. albicans, 12.5 Âg/mL and 50 Âg/mL for S. aureus CCBH 5330 and 100 Âg/mL and 100 Âg/mL for S. aureus ATCC 33591. Mechanism of action experiments were performed for the strain of S. aureus ATCC 6538P methicillin-susceptible (MSSA), that changes in the permeability of the bacterial membrane of S. aureus treated with bacteriostatic and bactericidal concentrations of DqV was observed by the crystal violet assay and release of genetic material assay. A lowest MIC was observed when alkaline pH broth was used (7,5-9,0). Complete bacterial growth inhibition was observed after 4 h of incubation with the MLC of DqV. Bacterial morphology was analyzed by atomic force microscopy after exposure of bacteria to the CIM and CIM /2 of DqV for 4 hours, showing membrane damage. In antiparasitic assays, we determined the cytotoxic effects of the venom on epimastigote and trypomastigote forms of the Y strain of Trypanosoma cruzi. In epimastigotes, cytotoxicity was evaluated at 24 and 48 h, finding IC50 of 28.32 Âg/mL and 20.67 Âg/mL, respectively. The mechanism of cell death was assessed by flow cytometry and revealed the presence of necrotic and apoptotic involvement in the cytotoxic effect of DqV over epimastigote form, in addition, the appearance of double labeled cells with PI and Annexin V-FITC, indicating the occurrence of late apoptosis. Cytotoxicity was evaluated over trypomastigote finding an IC50 of 1.978 Âg/mL and over RAW 264.7 cells finding an IC50 of 32.44 Âg/mL. The venom showed antibacterial activity against S. aureus MSSA and MRSA, P. aeruginosa, S. choleraesuis, E. coli and C. albicans, suggesting membrane damage in S. aureus ATCC 6538P. Additionally, showed cytotoxic potential on epimastigote and tripomastigote forms of Y strain of T. cruzi.As toxinas animais podem ser fonte de modelos moleculares para o desenho de novos fÃrmacos. Este trabalho objetivou estudar o potencial antimicrobiano e tripanocida do veneno da formiga Dinoponera quadriceps (VDq) visando à descoberta de substÃncia de valor terapÃutico. Foi realizado o ensaio de microdiluiÃÃo em caldo, onde foi determinado a ConcentraÃÃo InibitÃria MÃnima (CIM) e ConcentraÃÃo Letal MÃnima (CLM) das cepas Staphylococcus aureus ATCC 6538P, Escherichia coli ATCC 10536, Pseudomonas aeruginosa ATCC 9027, Salmonella cholearaesuis subsp. choleraesuis sorotipo choleraesuis ATCC 10708 e Candida albicans ATCC 10231 e duas cepas Staphylococcus aureus Meticilina Resistente (MRSA), S. aureus ATCC 33591 e S. aureus CCBH 5330. CIM e CLM de VDq foram respectivamente 6,25 Âg/mL e 12,5 Âg/mL para S. aureus ATCC 6538P, 3,12 Âg/mL e 3,12 Âg/mL para E. coli, 12,5 Âg/mL e 12,5 Âg/mL para P. aeruginosa, 12,5 Âg/mL e 25 Âg/mL para S. choleraesuis, 25 Âg/mL e 50 Âg/mL para C. albicans, 12,5 Âg/mL e 50 Âg/mL para S. aureus CCBH 5330 e 100 Âg/mL e 100 Âg/mL para S. aureus ATCC 33591. Em seguida foram realizados experimentos de mecanismo de aÃÃo para a cepa de S. aureus ATCC 6538P SensÃvel à Meticilina (MSSA), onde se verificou alteraÃÃo na permeabilidade da membrana bacteriana de S. aureus tratado com concentraÃÃes bacteriostÃticas e bactericidas de VDq atravÃs do ensaio do cristal violeta e do ensaio de liberaÃÃo de material genÃtico. Uma menor CIM foi encontrada quando pHs alcalinos foram utilizados no teste (7,5-9,0). Uma completa inibiÃÃo de crescimento foi observada apÃs 4 h de incubaÃÃo com CLM de VDq. A morfologia bacteriana foi avaliada por microscopia de forÃa atÃmica apÃs exposiÃÃo da bactÃria à CIM e CIM/2 de VDq durante 4 h, mostrando o dano de membrana. Nos ensaios antiparasitÃrios, foram observados os efeitos citotÃxicos do veneno sobre formas epimastigotas e tripomastigotas da cepa Y de Trypanosoma cruzi. Nas formas epimastigotas, a citotoxicidade foi avaliada em 24 e 48 h, com IC50 de 28,32 Âg/mL e IC50= 20,67 Âg/mL, respectivamente. O mecanismo de morte celular foi avaliado por citometria de fluxo e revelou envolvimento necrÃtico e apoptÃtico no efeito do VDq sobre formas epimastigotas, alÃm do aparecimento de cÃlulas marcadas duplamente com PI e anexina V-FITC, indicando a ocorrÃncia de apoptose tardia. A citotoxicidade foi avaliada sobre formas tripomastigotas encontrando uma IC50 de 1,978 Âg/mL e sobre cÃlulas RAW 264.7 uma IC50 de 32,44 Âg/mL. O veneno apresentou atividade antimicrobiana sobre S. aureus MSSA e MRSA, P. aeruginosa, S. choleraesuis, E. coli e C. albicans, sugerindo lise de membrana em S. aureus ATCC 6538P. Adicionalmente, apresentou potencial citotÃxico sobre as formas epimastigota e tripomastigorta de cepa Y de T. cruzi.CoordenaÃÃo de AperfeiÃoamento de NÃvel Superiorhttp://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=11676application/pdfinfo:eu-repo/semantics/openAccessporreponame:Biblioteca Digital de Teses e Dissertações da UFCinstname:Universidade Federal do Cearáinstacron:UFC2019-01-21T11:24:51Zmail@mail.com - |
dc.title.en.fl_str_mv |
Antimicrobial and antiparasitic potential of Dinoponera quadriceps venom |
dc.title.alternative.pt.fl_str_mv |
Potencial antimicrobiano e antiparasitÃrio do veneno da Dinoponera quadriceps |
title |
Antimicrobial and antiparasitic potential of Dinoponera quadriceps venom |
spellingShingle |
Antimicrobial and antiparasitic potential of Dinoponera quadriceps venom Danya Bandeira Lima Ant venoms Anti-infectives Antiparasitics FARMACIA |
title_short |
Antimicrobial and antiparasitic potential of Dinoponera quadriceps venom |
title_full |
Antimicrobial and antiparasitic potential of Dinoponera quadriceps venom |
title_fullStr |
Antimicrobial and antiparasitic potential of Dinoponera quadriceps venom |
title_full_unstemmed |
Antimicrobial and antiparasitic potential of Dinoponera quadriceps venom |
title_sort |
Antimicrobial and antiparasitic potential of Dinoponera quadriceps venom |
author |
Danya Bandeira Lima |
author_facet |
Danya Bandeira Lima |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Alice Maria Costa Martins |
dc.contributor.advisor1ID.fl_str_mv |
43431690300 |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/7532334620264577 |
dc.contributor.referee1.fl_str_mv |
Renata de Sousa Alves |
dc.contributor.referee1ID.fl_str_mv |
74909126368 |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/6730902567211867 |
dc.contributor.referee2.fl_str_mv |
Nadia Accioly Pinto Nogueira |
dc.contributor.referee2ID.fl_str_mv |
19127057372 |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/9261322165669741 |
dc.contributor.authorID.fl_str_mv |
02697539310 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/2010191025726363 |
dc.contributor.author.fl_str_mv |
Danya Bandeira Lima |
contributor_str_mv |
Alice Maria Costa Martins Renata de Sousa Alves Nadia Accioly Pinto Nogueira |
dc.subject.eng.fl_str_mv |
Ant venoms Anti-infectives Antiparasitics |
topic |
Ant venoms Anti-infectives Antiparasitics FARMACIA |
dc.subject.cnpq.fl_str_mv |
FARMACIA |
dc.description.sponsorship.fl_txt_mv |
CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior |
dc.description.abstract.por.fl_txt_mv |
Animal toxins can be a source of molecular models for the design of new drugs. This study investigated the antimicrobial and trypanocidal potential from Dinoponera quadriceps ant venom (DqV) aiming to discover therapeutic value substances. We conducted the microdilution test, where it was determined the minimum inhibitory concentration (MIC) and Minimum Lethal Concentration (MLC) over Staphylococcus aureus ATCC 6538P , Escherichia coli ATCC 10536 , Pseudomonas aeruginosa ATCC 9027 , Salmonella subsp cholearaesuis choleraesuis serotype choleraesuis ATCC 10708 and Candida albicans ATCC 10231 strains and two microbial strains of Methicillin Resistant Staphylococcus aureus (MRSA), S. aureus ATCC 33591 and S. aureus CCBH 5330. MIC and MLC of DqV were respectively 6.25 Âg/mL and 12.5 Âg/mL for S. aureus ATCC 6538P, 3.12 Âg/mL and 3.12 Âg/mL for E. coli, 12.5 Âg/mL and 12.5 Âg/mL for P. aeruginosa, 12.5 Âg/mL and 25 Âg/mL for S. choleraesuis, 25 Âg/mL and 50 Âg/mL for C. albicans, 12.5 Âg/mL and 50 Âg/mL for S. aureus CCBH 5330 and 100 Âg/mL and 100 Âg/mL for S. aureus ATCC 33591. Mechanism of action experiments were performed for the strain of S. aureus ATCC 6538P methicillin-susceptible (MSSA), that changes in the permeability of the bacterial membrane of S. aureus treated with bacteriostatic and bactericidal concentrations of DqV was observed by the crystal violet assay and release of genetic material assay. A lowest MIC was observed when alkaline pH broth was used (7,5-9,0). Complete bacterial growth inhibition was observed after 4 h of incubation with the MLC of DqV. Bacterial morphology was analyzed by atomic force microscopy after exposure of bacteria to the CIM and CIM /2 of DqV for 4 hours, showing membrane damage. In antiparasitic assays, we determined the cytotoxic effects of the venom on epimastigote and trypomastigote forms of the Y strain of Trypanosoma cruzi. In epimastigotes, cytotoxicity was evaluated at 24 and 48 h, finding IC50 of 28.32 Âg/mL and 20.67 Âg/mL, respectively. The mechanism of cell death was assessed by flow cytometry and revealed the presence of necrotic and apoptotic involvement in the cytotoxic effect of DqV over epimastigote form, in addition, the appearance of double labeled cells with PI and Annexin V-FITC, indicating the occurrence of late apoptosis. Cytotoxicity was evaluated over trypomastigote finding an IC50 of 1.978 Âg/mL and over RAW 264.7 cells finding an IC50 of 32.44 Âg/mL. The venom showed antibacterial activity against S. aureus MSSA and MRSA, P. aeruginosa, S. choleraesuis, E. coli and C. albicans, suggesting membrane damage in S. aureus ATCC 6538P. Additionally, showed cytotoxic potential on epimastigote and tripomastigote forms of Y strain of T. cruzi. As toxinas animais podem ser fonte de modelos moleculares para o desenho de novos fÃrmacos. Este trabalho objetivou estudar o potencial antimicrobiano e tripanocida do veneno da formiga Dinoponera quadriceps (VDq) visando à descoberta de substÃncia de valor terapÃutico. Foi realizado o ensaio de microdiluiÃÃo em caldo, onde foi determinado a ConcentraÃÃo InibitÃria MÃnima (CIM) e ConcentraÃÃo Letal MÃnima (CLM) das cepas Staphylococcus aureus ATCC 6538P, Escherichia coli ATCC 10536, Pseudomonas aeruginosa ATCC 9027, Salmonella cholearaesuis subsp. choleraesuis sorotipo choleraesuis ATCC 10708 e Candida albicans ATCC 10231 e duas cepas Staphylococcus aureus Meticilina Resistente (MRSA), S. aureus ATCC 33591 e S. aureus CCBH 5330. CIM e CLM de VDq foram respectivamente 6,25 Âg/mL e 12,5 Âg/mL para S. aureus ATCC 6538P, 3,12 Âg/mL e 3,12 Âg/mL para E. coli, 12,5 Âg/mL e 12,5 Âg/mL para P. aeruginosa, 12,5 Âg/mL e 25 Âg/mL para S. choleraesuis, 25 Âg/mL e 50 Âg/mL para C. albicans, 12,5 Âg/mL e 50 Âg/mL para S. aureus CCBH 5330 e 100 Âg/mL e 100 Âg/mL para S. aureus ATCC 33591. Em seguida foram realizados experimentos de mecanismo de aÃÃo para a cepa de S. aureus ATCC 6538P SensÃvel à Meticilina (MSSA), onde se verificou alteraÃÃo na permeabilidade da membrana bacteriana de S. aureus tratado com concentraÃÃes bacteriostÃticas e bactericidas de VDq atravÃs do ensaio do cristal violeta e do ensaio de liberaÃÃo de material genÃtico. Uma menor CIM foi encontrada quando pHs alcalinos foram utilizados no teste (7,5-9,0). Uma completa inibiÃÃo de crescimento foi observada apÃs 4 h de incubaÃÃo com CLM de VDq. A morfologia bacteriana foi avaliada por microscopia de forÃa atÃmica apÃs exposiÃÃo da bactÃria à CIM e CIM/2 de VDq durante 4 h, mostrando o dano de membrana. Nos ensaios antiparasitÃrios, foram observados os efeitos citotÃxicos do veneno sobre formas epimastigotas e tripomastigotas da cepa Y de Trypanosoma cruzi. Nas formas epimastigotas, a citotoxicidade foi avaliada em 24 e 48 h, com IC50 de 28,32 Âg/mL e IC50= 20,67 Âg/mL, respectivamente. O mecanismo de morte celular foi avaliado por citometria de fluxo e revelou envolvimento necrÃtico e apoptÃtico no efeito do VDq sobre formas epimastigotas, alÃm do aparecimento de cÃlulas marcadas duplamente com PI e anexina V-FITC, indicando a ocorrÃncia de apoptose tardia. A citotoxicidade foi avaliada sobre formas tripomastigotas encontrando uma IC50 de 1,978 Âg/mL e sobre cÃlulas RAW 264.7 uma IC50 de 32,44 Âg/mL. O veneno apresentou atividade antimicrobiana sobre S. aureus MSSA e MRSA, P. aeruginosa, S. choleraesuis, E. coli e C. albicans, sugerindo lise de membrana em S. aureus ATCC 6538P. Adicionalmente, apresentou potencial citotÃxico sobre as formas epimastigota e tripomastigorta de cepa Y de T. cruzi. |
description |
Animal toxins can be a source of molecular models for the design of new drugs. This study investigated the antimicrobial and trypanocidal potential from Dinoponera quadriceps ant venom (DqV) aiming to discover therapeutic value substances. We conducted the microdilution test, where it was determined the minimum inhibitory concentration (MIC) and Minimum Lethal Concentration (MLC) over Staphylococcus aureus ATCC 6538P , Escherichia coli ATCC 10536 , Pseudomonas aeruginosa ATCC 9027 , Salmonella subsp cholearaesuis choleraesuis serotype choleraesuis ATCC 10708 and Candida albicans ATCC 10231 strains and two microbial strains of Methicillin Resistant Staphylococcus aureus (MRSA), S. aureus ATCC 33591 and S. aureus CCBH 5330. MIC and MLC of DqV were respectively 6.25 Âg/mL and 12.5 Âg/mL for S. aureus ATCC 6538P, 3.12 Âg/mL and 3.12 Âg/mL for E. coli, 12.5 Âg/mL and 12.5 Âg/mL for P. aeruginosa, 12.5 Âg/mL and 25 Âg/mL for S. choleraesuis, 25 Âg/mL and 50 Âg/mL for C. albicans, 12.5 Âg/mL and 50 Âg/mL for S. aureus CCBH 5330 and 100 Âg/mL and 100 Âg/mL for S. aureus ATCC 33591. Mechanism of action experiments were performed for the strain of S. aureus ATCC 6538P methicillin-susceptible (MSSA), that changes in the permeability of the bacterial membrane of S. aureus treated with bacteriostatic and bactericidal concentrations of DqV was observed by the crystal violet assay and release of genetic material assay. A lowest MIC was observed when alkaline pH broth was used (7,5-9,0). Complete bacterial growth inhibition was observed after 4 h of incubation with the MLC of DqV. Bacterial morphology was analyzed by atomic force microscopy after exposure of bacteria to the CIM and CIM /2 of DqV for 4 hours, showing membrane damage. In antiparasitic assays, we determined the cytotoxic effects of the venom on epimastigote and trypomastigote forms of the Y strain of Trypanosoma cruzi. In epimastigotes, cytotoxicity was evaluated at 24 and 48 h, finding IC50 of 28.32 Âg/mL and 20.67 Âg/mL, respectively. The mechanism of cell death was assessed by flow cytometry and revealed the presence of necrotic and apoptotic involvement in the cytotoxic effect of DqV over epimastigote form, in addition, the appearance of double labeled cells with PI and Annexin V-FITC, indicating the occurrence of late apoptosis. Cytotoxicity was evaluated over trypomastigote finding an IC50 of 1.978 Âg/mL and over RAW 264.7 cells finding an IC50 of 32.44 Âg/mL. The venom showed antibacterial activity against S. aureus MSSA and MRSA, P. aeruginosa, S. choleraesuis, E. coli and C. albicans, suggesting membrane damage in S. aureus ATCC 6538P. Additionally, showed cytotoxic potential on epimastigote and tripomastigote forms of Y strain of T. cruzi. |
publishDate |
2014 |
dc.date.issued.fl_str_mv |
2014-02-25 |
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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publishedVersion |
format |
masterThesis |
dc.identifier.uri.fl_str_mv |
http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=11676 |
url |
http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=11676 |
dc.language.iso.fl_str_mv |
por |
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por |
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info:eu-repo/semantics/openAccess |
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openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal do Cearà |
dc.publisher.program.fl_str_mv |
Programa de PÃs-GraduaÃÃo em CiÃncias FarmacÃuticas |
dc.publisher.initials.fl_str_mv |
UFC |
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BR |
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Universidade Federal do Cearà |
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reponame:Biblioteca Digital de Teses e Dissertações da UFC instname:Universidade Federal do Ceará instacron:UFC |
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Biblioteca Digital de Teses e Dissertações da UFC |
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Biblioteca Digital de Teses e Dissertações da UFC |
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Universidade Federal do Ceará |
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UFC |
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mail@mail.com |
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