Estudo in vitro e in vivo da atividade Leishmanicida do extrato acetato de etila de Plectranthus amboinicus (Lour.) Spreng. frente à Leishmania (Viannia) braziliensis

Detalhes bibliográficos
Ano de defesa: 2017
Autor(a) principal: Gonçalves, Thially Braga
Orientador(a): Nagao-Dias, Aparecida Tiemi
Banca de defesa: Não Informado pela instituição
Tipo de documento: Tese
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
Palavras-chave em Português:
Link de acesso: http://www.repositorio.ufc.br/handle/riufc/40249
Resumo: Cutaneous leishmaniasis are anthropozoonoses caused by protozoa of the Leishmania genus. Leishmania braziliensis is most popular specie in Brazil. Plectranthus amboinicus, popularly known in Brazil as “malvarisco”, belongs to the family Lamiaceae. It has a great economic importance because of its widely medicinal use. In this work, the in vitro and in vivo effect of the ethyl acetate extracted of Plectranthus amboinicus (EAEPA) on L. braziliensis infection has been evaluated. The extracts of hexane, dichloromethane and ethyl acetate of P. amboinicus were initially tested in vitro for activity against Leishmania promastigotes. Since the EAEPA has shown the inhibition of 98.5% promastigotes at the concentration 4mg/mL, it was selected for the in vivo assays. To evaluate the effect against amastigote forms, murine intraperitoneal macrophages were infected or not with L. braziliensis and treated with different concentrations of EAEPA for 24h. Cytotoxicity, parasite load, and cytokine production have been analyzed. For the in vivo assay, Hamsters (Mesocricetus auratus; n = 20) were infected with 106 promastigotes and BALB/c mices (n = 15) with 107 of L. braziliensis on the right paw cushion and on the right ear, respectively. After the appearance of the lesions, the Hamsters were divided in 4 groups that received the following treatments: DMS0 1%; Antimony (20mg / kg / day, IM, 30 days); EAEPA (4mg / kg I.P. and I.L., 30 days). The BALB/c mice were divided into three groups: DMSO 1%, Antimony (20mg / kg / day, IM, 15 days), EAEPA (250mg / kg / day, PI, 15 days). Afterwards, the lesion thickness and parasite load have been evaluated and histopathological analyses have been done. Phytochemical analysis of EAEPA revealed the presence of flavonoids, digitalis glycosides, tannins and steroids. In vitro results have presented that EAEPA exhibited cytotoxicity from the 0.125 mg / mL concentration with 43% inhibition of cell growth. Treatment with EAEPA in vitro has presented a significant reduction of parasite load from the 0.03 mg/mL concentration. In cells treated with EAEPA, increased levels of TNF-α and IL-10, maintenance of TGF-β levels, and decreased IL-4 production have been observed. In vivo experiments, EAEPA administered intraperitoneally efficiently reduced lesion thickness, parasite load, and inflammatory parameters in Hamsters and BALB/c mice, compared to the untreated control. In summary, EAEPA was able to induce control of the lesion in the animals infected with L. braziliensis. It suggests that this may occur because the reduction of parasitic load, such as through immunoregulation of the inflammatory process. The leishmanicidal action of Plectranthus amboinicus elucidated in this work through the treatment in two animal models of infection by Leishmania braziliensis opens possibilities for the development of a new herbal medicine capable of reducing parasitemia or even cure, with low toxicity and few adverse effects.
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spelling Gonçalves, Thially BragaNagao-Dias, Aparecida Tiemi2019-03-11T22:15:25Z2019-03-11T22:15:25Z2017GONÇALVES, Thially Braga. Estudo in vitro e in vivo da atividade Leishmanicida do extrato acetato de etila de Plectranthus amboinicus (Lour.) Spreng. frente à Leishmania (Viannia) braziliensis. 2017. 139 f. Tese (Doutorado em Biotecnologia - Rede Nordeste de Biotecnologia) - Universidade Federal do Ceará, Fortaleza, 2017.http://www.repositorio.ufc.br/handle/riufc/40249Cutaneous leishmaniasis are anthropozoonoses caused by protozoa of the Leishmania genus. Leishmania braziliensis is most popular specie in Brazil. Plectranthus amboinicus, popularly known in Brazil as “malvarisco”, belongs to the family Lamiaceae. It has a great economic importance because of its widely medicinal use. In this work, the in vitro and in vivo effect of the ethyl acetate extracted of Plectranthus amboinicus (EAEPA) on L. braziliensis infection has been evaluated. The extracts of hexane, dichloromethane and ethyl acetate of P. amboinicus were initially tested in vitro for activity against Leishmania promastigotes. Since the EAEPA has shown the inhibition of 98.5% promastigotes at the concentration 4mg/mL, it was selected for the in vivo assays. To evaluate the effect against amastigote forms, murine intraperitoneal macrophages were infected or not with L. braziliensis and treated with different concentrations of EAEPA for 24h. Cytotoxicity, parasite load, and cytokine production have been analyzed. For the in vivo assay, Hamsters (Mesocricetus auratus; n = 20) were infected with 106 promastigotes and BALB/c mices (n = 15) with 107 of L. braziliensis on the right paw cushion and on the right ear, respectively. After the appearance of the lesions, the Hamsters were divided in 4 groups that received the following treatments: DMS0 1%; Antimony (20mg / kg / day, IM, 30 days); EAEPA (4mg / kg I.P. and I.L., 30 days). The BALB/c mice were divided into three groups: DMSO 1%, Antimony (20mg / kg / day, IM, 15 days), EAEPA (250mg / kg / day, PI, 15 days). Afterwards, the lesion thickness and parasite load have been evaluated and histopathological analyses have been done. Phytochemical analysis of EAEPA revealed the presence of flavonoids, digitalis glycosides, tannins and steroids. In vitro results have presented that EAEPA exhibited cytotoxicity from the 0.125 mg / mL concentration with 43% inhibition of cell growth. Treatment with EAEPA in vitro has presented a significant reduction of parasite load from the 0.03 mg/mL concentration. In cells treated with EAEPA, increased levels of TNF-α and IL-10, maintenance of TGF-β levels, and decreased IL-4 production have been observed. In vivo experiments, EAEPA administered intraperitoneally efficiently reduced lesion thickness, parasite load, and inflammatory parameters in Hamsters and BALB/c mice, compared to the untreated control. In summary, EAEPA was able to induce control of the lesion in the animals infected with L. braziliensis. It suggests that this may occur because the reduction of parasitic load, such as through immunoregulation of the inflammatory process. The leishmanicidal action of Plectranthus amboinicus elucidated in this work through the treatment in two animal models of infection by Leishmania braziliensis opens possibilities for the development of a new herbal medicine capable of reducing parasitemia or even cure, with low toxicity and few adverse effects.Leishmanioses cutâneas são antropozoonoses causadas por protozoários do gênero Leishmania no qual Leishmania braziliensis é a principal espécie no Brasil. Plectranthus amboinicus, conhecida popularmente como malvarisco, pertence à família Lamiaceae, com grande importância econômica por possuir amplo uso medicinal. Neste trabalho, avaliou-se o efeito in vitro e in vivo do extrato acetato de etila de Plectranthus amboinicus (EAEPA) na infecção por L. braziliensis. Os extratos hexano, diclorometano e acetato de etila de P. amboinicus foram inicialmente testados in vitro quanto à atividade contra as formas promastigotas de Leishmania. Uma vez que o EAEPA mostrou a inibição de 98,5% de promastigotas na concentração 4mg/mL, foi selecionado para os demais ensaios in vitro e in vivo. A composição química de EAEPA foi analisada por UPLC–QTOF–MS. Para avaliar o efeito contra as formas amastigotas, macrófagos intraperitoneais murinos foram infectados ou não com L. braziliensis e tratados com diferentes concentrações de EAEPA, por 24h. Foram analisadas a citotoxicidade, carga parasitária e a produção de citocinas. Para o ensaio in vivo, Hamsters (Mesocricetus auratus; n=20) foram infectados com 106 promastigotas e camundongos BALB/c (n=15) com 107 de L. braziliensis no coxim da pata direita e na orelha direita, respectivamente. Após o aparecimento das lesões, os Hamsters foram divididos em 4 grupos que receberam os seguintes tratamentos: DMS0 1%; Antimônio (20mg/kg/dia, I.M.,30 dias); EAEPA (4mg/kg I.P. e I.L.,30 dias), Os camundongos BALB/c foram divididos em três grupos: DMSO 1%; Antimônio (20mg/kg/dia, I.M.,15 dias); EAEPA (250mg/kg/dia, I.P.,15 dias). Em seguida, avaliou-se espessura da lesão, a carga parasitária e realizou-se análises histopatológicas. A análise fitoquímica de EAEPA revelou a presença de flavonoides, glicosídeos digitálicos, taninos e esteroides. A identificação dos compostos químicos de EAEPA revelou a presença de ácido quinico, ácido protocatecuico, ácido cafeico, cumaroilmalato, timoquinona, ácido siringico, ácido rosmarinico, luteolina, apigenina, tricine e cirsimaritina. Os resultados in vitro mostraram que EAEPA apresentou citotoxicidade a partir da concentração 0,125mg/mL com 43% de inibição do crescimento celular. O tratamento com EAEPA in vitro mostrou redução significativa da carga parasitária a partir da concentração 0,03mg/mL. Nas células tratadas com EAEPA, observou-se aumento dos níveis de TNFα e IL-10, manutenção dos níveis de TGFβ; e diminuição da produção de IL-4. Nos experimentos in vivo, EAEPA administrado via intraperitoneal reduziu eficientemente a espessura da lesão, a carga parasitária e os parâmetros inflamatórios em Hamsters e em camundongos BALB/c, comparados ao controle não tratado. Em suma, os dados mostraram que, em animais infectados com L. braziliensis, EAEPA foi capaz de induzir o controle da lesão, sugerindo que isso possa ocorrer em função da diminuição da carga parasitária, bem como, através de uma imunorregulação do processo inflamatório. A ação leishmanicida de Plectranthus amboinicus elucidada nesse trabalho através do tratamento em dois modelos animais de infecção por Leishmania braziliensis abre possibilidades ao desenvolvimento de um novo fitoterápico capaz de diminuir a parasitemia ou até mesmo a cura, com baixa toxicidade e poucos efeitos adversos.Leishmania braziliensisPlectranthus amboinicusLeishmaniose cutâneaEstudo in vitro e in vivo da atividade Leishmanicida do extrato acetato de etila de Plectranthus amboinicus (Lour.) Spreng. frente à Leishmania (Viannia) braziliensisinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccessLICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/40249/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52ORIGINAL2017_tese_tbgoncalves.pdf2017_tese_tbgoncalves.pdfapplication/pdf2834845http://repositorio.ufc.br/bitstream/riufc/40249/1/2017_tese_tbgoncalves.pdfa7931de42bfd918ab181b54e038b4cc5MD51riufc/402492019-03-11 19:15:25.703oai:repositorio.ufc.br:riufc/40249Tk9URTogUExBQ0UgWU9VUiBPV04gTElDRU5TRSBIRVJFClRoaXMgc2FtcGxlIGxpY2Vuc2UgaXMgcHJvdmlkZWQgZm9yIGluZm9ybWF0aW9uYWwgcHVycG9zZXMgb25seS4KCk5PTi1FWENMVVNJVkUgRElTVFJJQlVUSU9OIExJQ0VOU0UKCkJ5IHNpZ25pbmcgYW5kIHN1Ym1pdHRpbmcgdGhpcyBsaWNlbnNlLCB5b3UgKHRoZSBhdXRob3Iocykgb3IgY29weXJpZ2h0Cm93bmVyKSBncmFudHMgdG8gRFNwYWNlIFVuaXZlcnNpdHkgKERTVSkgdGhlIG5vbi1leGNsdXNpdmUgcmlnaHQgdG8gcmVwcm9kdWNlLAp0cmFuc2xhdGUgKGFzIGRlZmluZWQgYmVsb3cpLCBhbmQvb3IgZGlzdHJpYnV0ZSB5b3VyIHN1Ym1pc3Npb24gKGluY2x1ZGluZwp0aGUgYWJzdHJhY3QpIHdvcmxkd2lkZSBpbiBwcmludCBhbmQgZWxlY3Ryb25pYyBmb3JtYXQgYW5kIGluIGFueSBtZWRpdW0sCmluY2x1ZGluZyBidXQgbm90IGxpbWl0ZWQgdG8gYXVkaW8gb3IgdmlkZW8uCgpZb3UgYWdyZWUgdGhhdCBEU1UgbWF5LCB3aXRob3V0IGNoYW5naW5nIHRoZSBjb250ZW50LCB0cmFuc2xhdGUgdGhlCnN1Ym1pc3Npb24gdG8gYW55IG1lZGl1bSBvciBmb3JtYXQgZm9yIHRoZSBwdXJwb3NlIG9mIHByZXNlcnZhdGlvbi4KCllvdSBhbHNvIGFncmVlIHRoYXQgRFNVIG1heSBrZWVwIG1vcmUgdGhhbiBvbmUgY29weSBvZiB0aGlzIHN1Ym1pc3Npb24gZm9yCnB1cnBvc2VzIG9mIHNlY3VyaXR5LCBiYWNrLXVwIGFuZCBwcmVzZXJ2YXRpb24uCgpZb3UgcmVwcmVzZW50IHRoYXQgdGhlIHN1Ym1pc3Npb24gaXMgeW91ciBvcmlnaW5hbCB3b3JrLCBhbmQgdGhhdCB5b3UgaGF2ZQp0aGUgcmlnaHQgdG8gZ3JhbnQgdGhlIHJpZ2h0cyBjb250YWluZWQgaW4gdGhpcyBsaWNlbnNlLiBZb3UgYWxzbyByZXByZXNlbnQKdGhhdCB5b3VyIHN1Ym1pc3Npb24gZG9lcyBub3QsIHRvIHRoZSBiZXN0IG9mIHlvdXIga25vd2xlZGdlLCBpbmZyaW5nZSB1cG9uCmFueW9uZSdzIGNvcHlyaWdodC4KCklmIHRoZSBzdWJtaXNzaW9uIGNvbnRhaW5zIG1hdGVyaWFsIGZvciB3aGljaCB5b3UgZG8gbm90IGhvbGQgY29weXJpZ2h0LAp5b3UgcmVwcmVzZW50IHRoYXQgeW91IGhhdmUgb2J0YWluZWQgdGhlIHVucmVzdHJpY3RlZCBwZXJtaXNzaW9uIG9mIHRoZQpjb3B5cmlnaHQgb3duZXIgdG8gZ3JhbnQgRFNVIHRoZSByaWdodHMgcmVxdWlyZWQgYnkgdGhpcyBsaWNlbnNlLCBhbmQgdGhhdApzdWNoIHRoaXJkLXBhcnR5IG93bmVkIG1hdGVyaWFsIGlzIGNsZWFybHkgaWRlbnRpZmllZCBhbmQgYWNrbm93bGVkZ2VkCndpdGhpbiB0aGUgdGV4dCBvciBjb250ZW50IG9mIHRoZSBzdWJtaXNzaW9uLgoKSUYgVEhFIFNVQk1JU1NJT04gSVMgQkFTRUQgVVBPTiBXT1JLIFRIQVQgSEFTIEJFRU4gU1BPTlNPUkVEIE9SIFNVUFBPUlRFRApCWSBBTiBBR0VOQ1kgT1IgT1JHQU5JWkFUSU9OIE9USEVSIFRIQU4gRFNVLCBZT1UgUkVQUkVTRU5UIFRIQVQgWU9VIEhBVkUKRlVMRklMTEVEIEFOWSBSSUdIVCBPRiBSRVZJRVcgT1IgT1RIRVIgT0JMSUdBVElPTlMgUkVRVUlSRUQgQlkgU1VDSApDT05UUkFDVCBPUiBBR1JFRU1FTlQuCgpEU1Ugd2lsbCBjbGVhcmx5IGlkZW50aWZ5IHlvdXIgbmFtZShzKSBhcyB0aGUgYXV0aG9yKHMpIG9yIG93bmVyKHMpIG9mIHRoZQpzdWJtaXNzaW9uLCBhbmQgd2lsbCBub3QgbWFrZSBhbnkgYWx0ZXJhdGlvbiwgb3RoZXIgdGhhbiBhcyBhbGxvd2VkIGJ5IHRoaXMKbGljZW5zZSwgdG8geW91ciBzdWJtaXNzaW9uLgo=Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2019-03-11T22:15:25Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.pt_BR.fl_str_mv Estudo in vitro e in vivo da atividade Leishmanicida do extrato acetato de etila de Plectranthus amboinicus (Lour.) Spreng. frente à Leishmania (Viannia) braziliensis
title Estudo in vitro e in vivo da atividade Leishmanicida do extrato acetato de etila de Plectranthus amboinicus (Lour.) Spreng. frente à Leishmania (Viannia) braziliensis
spellingShingle Estudo in vitro e in vivo da atividade Leishmanicida do extrato acetato de etila de Plectranthus amboinicus (Lour.) Spreng. frente à Leishmania (Viannia) braziliensis
Gonçalves, Thially Braga
Leishmania braziliensis
Plectranthus amboinicus
Leishmaniose cutânea
title_short Estudo in vitro e in vivo da atividade Leishmanicida do extrato acetato de etila de Plectranthus amboinicus (Lour.) Spreng. frente à Leishmania (Viannia) braziliensis
title_full Estudo in vitro e in vivo da atividade Leishmanicida do extrato acetato de etila de Plectranthus amboinicus (Lour.) Spreng. frente à Leishmania (Viannia) braziliensis
title_fullStr Estudo in vitro e in vivo da atividade Leishmanicida do extrato acetato de etila de Plectranthus amboinicus (Lour.) Spreng. frente à Leishmania (Viannia) braziliensis
title_full_unstemmed Estudo in vitro e in vivo da atividade Leishmanicida do extrato acetato de etila de Plectranthus amboinicus (Lour.) Spreng. frente à Leishmania (Viannia) braziliensis
title_sort Estudo in vitro e in vivo da atividade Leishmanicida do extrato acetato de etila de Plectranthus amboinicus (Lour.) Spreng. frente à Leishmania (Viannia) braziliensis
author Gonçalves, Thially Braga
author_facet Gonçalves, Thially Braga
author_role author
dc.contributor.author.fl_str_mv Gonçalves, Thially Braga
dc.contributor.advisor1.fl_str_mv Nagao-Dias, Aparecida Tiemi
contributor_str_mv Nagao-Dias, Aparecida Tiemi
dc.subject.por.fl_str_mv Leishmania braziliensis
Plectranthus amboinicus
Leishmaniose cutânea
topic Leishmania braziliensis
Plectranthus amboinicus
Leishmaniose cutânea
description Cutaneous leishmaniasis are anthropozoonoses caused by protozoa of the Leishmania genus. Leishmania braziliensis is most popular specie in Brazil. Plectranthus amboinicus, popularly known in Brazil as “malvarisco”, belongs to the family Lamiaceae. It has a great economic importance because of its widely medicinal use. In this work, the in vitro and in vivo effect of the ethyl acetate extracted of Plectranthus amboinicus (EAEPA) on L. braziliensis infection has been evaluated. The extracts of hexane, dichloromethane and ethyl acetate of P. amboinicus were initially tested in vitro for activity against Leishmania promastigotes. Since the EAEPA has shown the inhibition of 98.5% promastigotes at the concentration 4mg/mL, it was selected for the in vivo assays. To evaluate the effect against amastigote forms, murine intraperitoneal macrophages were infected or not with L. braziliensis and treated with different concentrations of EAEPA for 24h. Cytotoxicity, parasite load, and cytokine production have been analyzed. For the in vivo assay, Hamsters (Mesocricetus auratus; n = 20) were infected with 106 promastigotes and BALB/c mices (n = 15) with 107 of L. braziliensis on the right paw cushion and on the right ear, respectively. After the appearance of the lesions, the Hamsters were divided in 4 groups that received the following treatments: DMS0 1%; Antimony (20mg / kg / day, IM, 30 days); EAEPA (4mg / kg I.P. and I.L., 30 days). The BALB/c mice were divided into three groups: DMSO 1%, Antimony (20mg / kg / day, IM, 15 days), EAEPA (250mg / kg / day, PI, 15 days). Afterwards, the lesion thickness and parasite load have been evaluated and histopathological analyses have been done. Phytochemical analysis of EAEPA revealed the presence of flavonoids, digitalis glycosides, tannins and steroids. In vitro results have presented that EAEPA exhibited cytotoxicity from the 0.125 mg / mL concentration with 43% inhibition of cell growth. Treatment with EAEPA in vitro has presented a significant reduction of parasite load from the 0.03 mg/mL concentration. In cells treated with EAEPA, increased levels of TNF-α and IL-10, maintenance of TGF-β levels, and decreased IL-4 production have been observed. In vivo experiments, EAEPA administered intraperitoneally efficiently reduced lesion thickness, parasite load, and inflammatory parameters in Hamsters and BALB/c mice, compared to the untreated control. In summary, EAEPA was able to induce control of the lesion in the animals infected with L. braziliensis. It suggests that this may occur because the reduction of parasitic load, such as through immunoregulation of the inflammatory process. The leishmanicidal action of Plectranthus amboinicus elucidated in this work through the treatment in two animal models of infection by Leishmania braziliensis opens possibilities for the development of a new herbal medicine capable of reducing parasitemia or even cure, with low toxicity and few adverse effects.
publishDate 2017
dc.date.issued.fl_str_mv 2017
dc.date.accessioned.fl_str_mv 2019-03-11T22:15:25Z
dc.date.available.fl_str_mv 2019-03-11T22:15:25Z
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dc.identifier.citation.fl_str_mv GONÇALVES, Thially Braga. Estudo in vitro e in vivo da atividade Leishmanicida do extrato acetato de etila de Plectranthus amboinicus (Lour.) Spreng. frente à Leishmania (Viannia) braziliensis. 2017. 139 f. Tese (Doutorado em Biotecnologia - Rede Nordeste de Biotecnologia) - Universidade Federal do Ceará, Fortaleza, 2017.
dc.identifier.uri.fl_str_mv http://www.repositorio.ufc.br/handle/riufc/40249
identifier_str_mv GONÇALVES, Thially Braga. Estudo in vitro e in vivo da atividade Leishmanicida do extrato acetato de etila de Plectranthus amboinicus (Lour.) Spreng. frente à Leishmania (Viannia) braziliensis. 2017. 139 f. Tese (Doutorado em Biotecnologia - Rede Nordeste de Biotecnologia) - Universidade Federal do Ceará, Fortaleza, 2017.
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