Avaliação da capacidade imunogênica de células leveduriformes de Sporothrix schenckii inativadas em modelo murino

Detalhes bibliográficos
Ano de defesa: 2010
Autor(a) principal: Antunes, Tatiana de Ávila
Orientador(a): Meireles, Mario Carlos Araújo
Banca de defesa: Não Informado pela instituição
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Pelotas
Programa de Pós-Graduação: Programa de Pós-Graduação em Veterinária
Departamento: Veterinária
País: BR
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: https://guaiaca.ufpel.edu.br/handle/123456789/2544
Resumo: Sporotrichosis is a subcutaneous mycosis of zoonotic character, cosmopolitan development subacute or chronic whose agent the dimorphic fungus Sporothrix schenckii affecting man and various species of animals, especially the domestic cat, being considered of interest to Public Healt. Considering the difficulties in the therapeutic treatment of ringworm in this animal species, including toxicity and the development of resistance to antifungal agents traditionally used to treat disease the study aimed evaluate the immunogenic capacity of yeast cells of S. schenckii inactivated both in immunoprophylaxis and immunotherapy of sporotrichosis. We used 160 Wistar albin rats (Rattus norvegicus) at 70 days (80 in immunoprophylaxis and 80 in immunotheraphy) were divided into four groups that were immunized three times every 15 days. The groups were divided as follows: G1 (control, mineral oil), G2 (Ag + incomplete freund adjuvant), G3 (Ag + Freund's complete adjuvant) and G4 (Ag + incomplete freund adjuvant + propolis). Preparation of the vaccine used was an isolate of S. schenckii from yeast in the form of a case of cutaneous sporotrichosis of domestic cat. The fungus was grown in medium liquid (Brain-Heart broth), incubated for 10 days at 370C and kept under constant agitation to obtain the yeast form. The culture was filtered through a double layer of sterile gauze, centrifuged, washed twice with buffered saline (PBS), homogenized and standardized in 108 cells of S. schenckii / ml. The cells were inactivated with 0.02% thimerosal and then emulsified with mineral oil, and vaccines packaged in sterilized sealed and kept at a temperature of 40C throughout the experimental period. Was used a dose of 0.1 ml/animal intramuscularly. Rats that received the vaccine as immunoprophylaxis after the three doses were challenged, and inoculated subcutaneously with 2X103 cells / ml of S. schenckii in the right footpad and evaluated for 10 days. Animals treated with immunotherapy were inoculated with the agent after 14 days and received three doses of immunogen. After the experimental period all were euthanized and necropsy to mycological examination, histopathology and counting colony forming units. The results were: immunoprophylaxis in the pathological changes showed statistically significant differences (P <0.05) in the vaccinated groups (G2, G3 and G4) compared to the control group (G1) to the point of inoculation, with no statistical difference the evaluation of internal organs. Immunotherapy clinical evaluation at the injection showed that there was no statistical difference among the four experimental groups, but at the end of the experiment the groups G1, G2, G3 and G4 had respectively 8.3%, 58.3%, 41.7 % and 50% of injuries in the process of regression and healing. In relation to injuries in other body areas only in the last week of the experiment, the G2 and G3 differ statistically (P <0.05) in the control group (G1). Pathological changes were found in the internal organs of the four experimental groups, but with a greater number of lesions in group CONT. Both in immunoprophylaxis and immunotherapy the retroisolation of the agent and count of colony forming units showed that there was growth of S. schenckii in the four experimental groups, but with less frequency and quantification of CFU at the point of inoculation and internal organs of the groups G2, G3 and G4. In histopathologic evaluation to determine the presence of granulomas and pyogranulomas focal and multifocal in the point of inoculation and internal organs of the four groups, but the animals that received immunoprophylactic changes were more restricted to the point of inoculation. The results indicate that the three vaccine formulations (AIF ACF and AIFP) used as immunoprophylactic and immunotherapy have not been effective for the remission of the lesions of experimental cutaneous sporotrichosis.
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spelling http://lattes.cnpq.br/7256629565934764http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4799478H9Schuch, Luiz Felipe Daméhttp://lattes.cnpq.br/7855228624176456Nascente, Patrícia da Silvahttp://lattes.cnpq.br/3560823637748071Meireles, Mario Carlos AraújoAntunes, Tatiana de Ávila2014-08-20T14:37:58Z2010-09-232014-08-20T14:37:58Z2010-03-10ANTUNES, Tatiana de Ávila. Avaliação da capacidade imunogênica de células leveduriformes de Sporothrix schenckii inativadas em modelo murino. 2010. 102 f. Tese (Doutorado em Veterinária) - Universidade Federal de Pelotas, Pelotas, 2010.https://guaiaca.ufpel.edu.br/handle/123456789/2544Sporotrichosis is a subcutaneous mycosis of zoonotic character, cosmopolitan development subacute or chronic whose agent the dimorphic fungus Sporothrix schenckii affecting man and various species of animals, especially the domestic cat, being considered of interest to Public Healt. Considering the difficulties in the therapeutic treatment of ringworm in this animal species, including toxicity and the development of resistance to antifungal agents traditionally used to treat disease the study aimed evaluate the immunogenic capacity of yeast cells of S. schenckii inactivated both in immunoprophylaxis and immunotherapy of sporotrichosis. We used 160 Wistar albin rats (Rattus norvegicus) at 70 days (80 in immunoprophylaxis and 80 in immunotheraphy) were divided into four groups that were immunized three times every 15 days. The groups were divided as follows: G1 (control, mineral oil), G2 (Ag + incomplete freund adjuvant), G3 (Ag + Freund's complete adjuvant) and G4 (Ag + incomplete freund adjuvant + propolis). Preparation of the vaccine used was an isolate of S. schenckii from yeast in the form of a case of cutaneous sporotrichosis of domestic cat. The fungus was grown in medium liquid (Brain-Heart broth), incubated for 10 days at 370C and kept under constant agitation to obtain the yeast form. The culture was filtered through a double layer of sterile gauze, centrifuged, washed twice with buffered saline (PBS), homogenized and standardized in 108 cells of S. schenckii / ml. The cells were inactivated with 0.02% thimerosal and then emulsified with mineral oil, and vaccines packaged in sterilized sealed and kept at a temperature of 40C throughout the experimental period. Was used a dose of 0.1 ml/animal intramuscularly. Rats that received the vaccine as immunoprophylaxis after the three doses were challenged, and inoculated subcutaneously with 2X103 cells / ml of S. schenckii in the right footpad and evaluated for 10 days. Animals treated with immunotherapy were inoculated with the agent after 14 days and received three doses of immunogen. After the experimental period all were euthanized and necropsy to mycological examination, histopathology and counting colony forming units. The results were: immunoprophylaxis in the pathological changes showed statistically significant differences (P <0.05) in the vaccinated groups (G2, G3 and G4) compared to the control group (G1) to the point of inoculation, with no statistical difference the evaluation of internal organs. Immunotherapy clinical evaluation at the injection showed that there was no statistical difference among the four experimental groups, but at the end of the experiment the groups G1, G2, G3 and G4 had respectively 8.3%, 58.3%, 41.7 % and 50% of injuries in the process of regression and healing. In relation to injuries in other body areas only in the last week of the experiment, the G2 and G3 differ statistically (P <0.05) in the control group (G1). Pathological changes were found in the internal organs of the four experimental groups, but with a greater number of lesions in group CONT. Both in immunoprophylaxis and immunotherapy the retroisolation of the agent and count of colony forming units showed that there was growth of S. schenckii in the four experimental groups, but with less frequency and quantification of CFU at the point of inoculation and internal organs of the groups G2, G3 and G4. In histopathologic evaluation to determine the presence of granulomas and pyogranulomas focal and multifocal in the point of inoculation and internal organs of the four groups, but the animals that received immunoprophylactic changes were more restricted to the point of inoculation. The results indicate that the three vaccine formulations (AIF ACF and AIFP) used as immunoprophylactic and immunotherapy have not been effective for the remission of the lesions of experimental cutaneous sporotrichosis.Esporotricose é uma micose subcutânea de caráter zoonótico, cosmopolita de evolução subaguda ou crônica que tem como agente etiológico o fungo dimórfico Sporothrix schenckii afetando o homem e várias espécies de animais, principalmente o felino doméstico, sendo considerada de interesse para a Saúde Pública. Considerando as dificuldades terapêuticas no tratamento da micose nessa espécie animal, incluindo toxicidade e o desenvolvimento de resistência aos antifúngicos tradicionalmente utilizados na terapia da enfermidade o estudo objetivou avaliar a capacidade imunogênica de células leveduriformes de S. schenckii inativadas tanto na imunoprofilaxia como na imunoterapia da esporotricose. Foram utilizados 160 ratos albinos wistar (Rattus norvergicus) com 70 dias (80 na imunoprofilaxia e 80 na imunoterapia) sendo divididos em quatro grupos que receberam três doses do imunógeno a cada 14 dias. Os grupos foram divididos em: G1 (controle- óleo mineral), G2 (Ag+ adjuvante incompleto de freund), G3 (Ag + adjuvante completo de freund) e G4 (Ag + adjuvante incompleto de freund + própolis). Para preparação da vacina foi utilizado um isolado de S. schenckii na forma leveduriforme proveniente de um caso de esporotricose cutânea de felino doméstico. O fungo foi cultivado em meio líquido (Brain-Heart broth), incubado durante 10 dias a 37oC e mantido sob agitação constante para obtenção da forma leveduriforme. A cultura foi filtrada em dupla camada de gaze estéril, centrifugada, lavada duas vezes com solução salina tamponada (PBS), homogeneizada e padronizada em 108 células de S. schenckii/ ml. As células foram inativadas com thimerosal a 0,02% e posteriormente emulsificadas com óleo mineral, sendo as vacinas acondicionadas em frascos estéreis fechados e mantidas a temperatura de 40C durante todo o período experimental. Foi utilizada uma dose de 0,1 ml/animal por via intramuscular. Os ratos que receberem a vacina como imunoprofilaxia após as três doses foram desafiados, sendo inoculados por via subcutânea com 2X103 células/ml de S. schenckii e avaliados durante 10 dias. Os animais tratados com imunoterápico foram inoculados com o agente e após 14 dias receberam três doses do imunógeno. Após o período experimental todos foram eutanasiados e feita necropsia para realização de análise micológica, histopatológica e contagem das Unidades Formadoras de Colônias. Os resultados obtidos foram: na imunoprofilaxia as alterações anatomopatológicas demonstraram diferença estatísticamente significativa (P<0,05) dos grupos vacinados (G2, G3 e G4) quando comparados ao grupo controle (G1) em relação ao ponto de inoculação, não havendo diferença estatística na avaliação dos órgãos internos. Na imunoterapia a avaliação clínica no ponto de inoculação evidenciou que não houve diferença estatística entre os quatro grupos experimentais, porém ao final do experimento os grupos G1, G2, G3 e G4 apresentavam respectivamente 8,3%, 58,3%, 41,7% e 50% das lesões em processo de regressão e cicatrização. Em relação a lesões em outras áreas corpóreas somente na última semana do experimento os grupos G2 e G3 diferiram estatisticamente (P<0,05) do grupo controle (G1). Alterações anatomopatológicas foram encontradas nos órgãos internos dos quatro grupos experimentais, porém com um maior numero de lesões nos animais do grupo CONT. Tanto na imunoprofilaxia quanto na imunoterapia o retroisolamento do agente e Contagem das Unidades Formadoras de colônias demonstraram que houve crescimento do S. schenckii nos quatro grupos experimentais, porém com menor freqüência e quantificação de UFCs no ponto de inoculação e órgãos internos dos grupos G2, G3 e G4. Na avaliação histopatológica foi verificada a presença de granulomas e piogranulomas focais e multifocais no ponto de inoculação e órgãos internos dos quatro grupos, porém os animais que receberam imunoprofilático as alterações ficaram mais restritas ao ponto de inoculação. Os resultados permitem concluir que as três formulações de vacinas (AIF, ACF e AIFP) utilizadas como imunoprofilático e imunoterápico não foram eficazes para a remissão completa das lesões de esporotricose cutânea experimental.application/pdfporUniversidade Federal de PelotasPrograma de Pós-Graduação em VeterináriaUFPelBRVeterináriaVeterináriaS. schenckiiEsporotricoseImunoprofilaxiaImunoterapiaVeterinaryS. schenckiiSporotrichosisImmunoprophylaxisImmunotherapyCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::PATOLOGIA ANIMALAvaliação da capacidade imunogênica de células leveduriformes de Sporothrix schenckii inativadas em modelo murinoEvaluation of the immunogenic capacity of yeast cells of Sporothrix schenckii inactivated in a murine modelinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFPel - Guaiacainstname:Universidade Federal de Pelotas (UFPEL)instacron:UFPELORIGINALtese_tatiana_antunes.pdfapplication/pdf3388546http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/2544/1/tese_tatiana_antunes.pdf6c26eaf4ab24c11f046c7e7b2f9f2566MD51open accessTEXTtese_tatiana_antunes.pdf.txttese_tatiana_antunes.pdf.txtExtracted Texttext/plain172797http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/2544/2/tese_tatiana_antunes.pdf.txtb58be652207d200650eb6f5251acfab4MD52open accessTHUMBNAILtese_tatiana_antunes.pdf.jpgtese_tatiana_antunes.pdf.jpgGenerated Thumbnailimage/jpeg1752http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/2544/3/tese_tatiana_antunes.pdf.jpg1934d29cb4b05b6a33a23e93cbe9a5feMD53open access123456789/25442022-01-27 09:46:39.096open accessoai:guaiaca.ufpel.edu.br:123456789/2544Repositório InstitucionalPUBhttp://repositorio.ufpel.edu.br/oai/requestrippel@ufpel.edu.br || repositorio@ufpel.edu.br || aline.batista@ufpel.edu.bropendoar:2022-01-27T12:46:39Repositório Institucional da UFPel - Guaiaca - Universidade Federal de Pelotas (UFPEL)false
dc.title.por.fl_str_mv Avaliação da capacidade imunogênica de células leveduriformes de Sporothrix schenckii inativadas em modelo murino
dc.title.alternative.eng.fl_str_mv Evaluation of the immunogenic capacity of yeast cells of Sporothrix schenckii inactivated in a murine model
title Avaliação da capacidade imunogênica de células leveduriformes de Sporothrix schenckii inativadas em modelo murino
spellingShingle Avaliação da capacidade imunogênica de células leveduriformes de Sporothrix schenckii inativadas em modelo murino
Antunes, Tatiana de Ávila
Veterinária
S. schenckii
Esporotricose
Imunoprofilaxia
Imunoterapia
Veterinary
S. schenckii
Sporotrichosis
Immunoprophylaxis
Immunotherapy
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::PATOLOGIA ANIMAL
title_short Avaliação da capacidade imunogênica de células leveduriformes de Sporothrix schenckii inativadas em modelo murino
title_full Avaliação da capacidade imunogênica de células leveduriformes de Sporothrix schenckii inativadas em modelo murino
title_fullStr Avaliação da capacidade imunogênica de células leveduriformes de Sporothrix schenckii inativadas em modelo murino
title_full_unstemmed Avaliação da capacidade imunogênica de células leveduriformes de Sporothrix schenckii inativadas em modelo murino
title_sort Avaliação da capacidade imunogênica de células leveduriformes de Sporothrix schenckii inativadas em modelo murino
author Antunes, Tatiana de Ávila
author_facet Antunes, Tatiana de Ávila
author_role author
dc.contributor.authorID.por.fl_str_mv
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/7256629565934764
dc.contributor.advisorID.por.fl_str_mv
dc.contributor.advisorLattes.por.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4799478H9
dc.contributor.referees1.pt_BR.fl_str_mv Nascente, Patrícia da Silva
dc.contributor.referees1ID.por.fl_str_mv
dc.contributor.referees1Lattes.por.fl_str_mv http://lattes.cnpq.br/3560823637748071
dc.contributor.advisor-co1.fl_str_mv Schuch, Luiz Felipe Damé
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/7855228624176456
dc.contributor.advisor1.fl_str_mv Meireles, Mario Carlos Araújo
dc.contributor.author.fl_str_mv Antunes, Tatiana de Ávila
contributor_str_mv Schuch, Luiz Felipe Damé
Meireles, Mario Carlos Araújo
dc.subject.por.fl_str_mv Veterinária
S. schenckii
Esporotricose
Imunoprofilaxia
Imunoterapia
topic Veterinária
S. schenckii
Esporotricose
Imunoprofilaxia
Imunoterapia
Veterinary
S. schenckii
Sporotrichosis
Immunoprophylaxis
Immunotherapy
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::PATOLOGIA ANIMAL
dc.subject.eng.fl_str_mv Veterinary
S. schenckii
Sporotrichosis
Immunoprophylaxis
Immunotherapy
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::PATOLOGIA ANIMAL
description Sporotrichosis is a subcutaneous mycosis of zoonotic character, cosmopolitan development subacute or chronic whose agent the dimorphic fungus Sporothrix schenckii affecting man and various species of animals, especially the domestic cat, being considered of interest to Public Healt. Considering the difficulties in the therapeutic treatment of ringworm in this animal species, including toxicity and the development of resistance to antifungal agents traditionally used to treat disease the study aimed evaluate the immunogenic capacity of yeast cells of S. schenckii inactivated both in immunoprophylaxis and immunotherapy of sporotrichosis. We used 160 Wistar albin rats (Rattus norvegicus) at 70 days (80 in immunoprophylaxis and 80 in immunotheraphy) were divided into four groups that were immunized three times every 15 days. The groups were divided as follows: G1 (control, mineral oil), G2 (Ag + incomplete freund adjuvant), G3 (Ag + Freund's complete adjuvant) and G4 (Ag + incomplete freund adjuvant + propolis). Preparation of the vaccine used was an isolate of S. schenckii from yeast in the form of a case of cutaneous sporotrichosis of domestic cat. The fungus was grown in medium liquid (Brain-Heart broth), incubated for 10 days at 370C and kept under constant agitation to obtain the yeast form. The culture was filtered through a double layer of sterile gauze, centrifuged, washed twice with buffered saline (PBS), homogenized and standardized in 108 cells of S. schenckii / ml. The cells were inactivated with 0.02% thimerosal and then emulsified with mineral oil, and vaccines packaged in sterilized sealed and kept at a temperature of 40C throughout the experimental period. Was used a dose of 0.1 ml/animal intramuscularly. Rats that received the vaccine as immunoprophylaxis after the three doses were challenged, and inoculated subcutaneously with 2X103 cells / ml of S. schenckii in the right footpad and evaluated for 10 days. Animals treated with immunotherapy were inoculated with the agent after 14 days and received three doses of immunogen. After the experimental period all were euthanized and necropsy to mycological examination, histopathology and counting colony forming units. The results were: immunoprophylaxis in the pathological changes showed statistically significant differences (P <0.05) in the vaccinated groups (G2, G3 and G4) compared to the control group (G1) to the point of inoculation, with no statistical difference the evaluation of internal organs. Immunotherapy clinical evaluation at the injection showed that there was no statistical difference among the four experimental groups, but at the end of the experiment the groups G1, G2, G3 and G4 had respectively 8.3%, 58.3%, 41.7 % and 50% of injuries in the process of regression and healing. In relation to injuries in other body areas only in the last week of the experiment, the G2 and G3 differ statistically (P <0.05) in the control group (G1). Pathological changes were found in the internal organs of the four experimental groups, but with a greater number of lesions in group CONT. Both in immunoprophylaxis and immunotherapy the retroisolation of the agent and count of colony forming units showed that there was growth of S. schenckii in the four experimental groups, but with less frequency and quantification of CFU at the point of inoculation and internal organs of the groups G2, G3 and G4. In histopathologic evaluation to determine the presence of granulomas and pyogranulomas focal and multifocal in the point of inoculation and internal organs of the four groups, but the animals that received immunoprophylactic changes were more restricted to the point of inoculation. The results indicate that the three vaccine formulations (AIF ACF and AIFP) used as immunoprophylactic and immunotherapy have not been effective for the remission of the lesions of experimental cutaneous sporotrichosis.
publishDate 2010
dc.date.available.fl_str_mv 2010-09-23
2014-08-20T14:37:58Z
dc.date.issued.fl_str_mv 2010-03-10
dc.date.accessioned.fl_str_mv 2014-08-20T14:37:58Z
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dc.identifier.citation.fl_str_mv ANTUNES, Tatiana de Ávila. Avaliação da capacidade imunogênica de células leveduriformes de Sporothrix schenckii inativadas em modelo murino. 2010. 102 f. Tese (Doutorado em Veterinária) - Universidade Federal de Pelotas, Pelotas, 2010.
dc.identifier.uri.fl_str_mv https://guaiaca.ufpel.edu.br/handle/123456789/2544
identifier_str_mv ANTUNES, Tatiana de Ávila. Avaliação da capacidade imunogênica de células leveduriformes de Sporothrix schenckii inativadas em modelo murino. 2010. 102 f. Tese (Doutorado em Veterinária) - Universidade Federal de Pelotas, Pelotas, 2010.
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