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Vasco Ariston de Carvalho Azevedohttp://lattes.cnpq.br/1020477751003832Pamela Del Carmen Mancha AgrestiRodrigo Dias de Oliveira CarvalhoÁlvaro Cantini NunesFlaviano dos Santos Martinshttp://lattes.cnpq.br/8315573039724612Nina Dias Coelho Rocha2021-02-24T17:00:59Z2021-02-24T17:00:59Z2020-12-18http://hdl.handle.net/1843/35057Os benefícios exercidos pelos probióticos têm impulsionado uma série de pesquisas em busca de novas linhagens com este potencial. Nesse contexto, este estudo visou identificar e caracterizar 21 novas linhagens provenientes de ambientes diversos baseando-se em testes para avaliação de probiose sugeridos por diretrizes de agências reguladoras para posterior verificação do potencial protetor em modelo murino de mucosite intestinal induzida por 5-fluourouracil (5-FU). A identificação por MALDI-TOF revelou que todas as linhagens pertencem ao grupo das bactérias láticas (BLs), sendo relatados os gêneros Lactobacillus, Lactiplantibacillus, Lacticaseibacillus, Pediococcus e Enterococcus. Foram realizados testes in vitro de segurança por meio da análise da resistência à antibióticos, degradação da mucina e potencial hemolítico e de eficácia e funcionalidade, por meio da avaliação da resistência das linhagens aos sais biliares, tolerância ao pH ácido, produção de substâncias antimicrobianas, adesão e imunoestimulação em células Caco-2. Nenhum destes critérios foram excludentes quanto ao enquadramento das linhagens como probióticos, uma vez que todas apresentaram bons parâmetros na prospecção. Quatro delas pertencentes a espécies diferentes foram inicialmente direcionadas a estudos in vivo, sendo elas Lacticaseibacillus paracasei Lpa1 (Lactobacillus paracasei Lpa1), Enterococcus faecalis Efc15, Lactiplantibacillus plantarum Lpl19 (Lactobacillus plantarum Lpl19) e Lactobacillus crispatus Lc21. Ao serem administradas aos animais sem a indução da mucosite, todas as linhagens apresentaram parâmetros semelhantes aos animais controle, sugerindo não toxicidade. Todavia, as quatro não foram capazes de prevenir significativamente a perda de peso, redução do consumo hídrico e alimentar, redução das vilosidades e inflamação intestinais, característicos da doença. Contudo, mesmo não apresentando eficácia no tratamento da mucosite intestinal quando administradas, não se pode excluir o potencial destas linhagens em outros modelos de doenças e aplicações. Desta forma, o presente estudo abre uma gama de linhas de pesquisa para explorar o papel não somente das quatro linhagens testadas in vivo, mas também de todas as outras que apresentaram características sugestivas de probiose na prospecção.The benefits exerted by probiotics have driven a series of studies in search for new strains with this potential. In this context, this study aimed to identify and characterize 21 new strains provided from different environments based on tests for evaluation of probiosis suggested by guidelines of regulatory agencies for further verification of the protective effect in murine model of mucositis induced by 5-fluourouracil (5-FU). Identification by MALDI-TOF revealed that all strains belong to the group of lactic bacteria (BLs), belonging to the genera Lactobacillus, Lactiplantibacillus, Lacticaseibacillus, Pediococcus and Enterococcus. In vitro safety tests were performed through the analysis of antibiotic resistance, mucin degradation and hemolytic potential and efficacy and functionality tests, through the resistance of the strains to bile salts, tolerance to acid pH, production of antimicrobial substances, adhesion and immune stimulation in Caco-2 cells. None of these criteria were excluding regarding the classification of the strains as probiotics, once all of them presented good parameters in prospection. Four of them belonging to different species were initially directed to in vivo studies, being Lacticaseibacillus paracasei Lpa1 (Lactobacillus paracasei Lpa1), Enterococcus faecalis Efc15, Lactiplantibacillus plantarum Lpl19 (Lactobacillus plantarum Lpl19) and Lactobacillus crispatus Lc21). When administered to the animals without the induction of mucositis, all strains showed similar parameters to the control animals, suggesting non-toxicity. However, the four were not able to significantly prevent weight loss, reduced water and food consumption, reduction of intestinal villi and inflammation, characteristic of the disease. However, even if they are not effective in the treatment of intestinal mucositis, the potential of these strains in other disease models and applications cannot be excluded. In this way, the present study opens up a range of lines of research to explore the role not only of the four strains tested in vivo, but also of all others that showed characteristics suggestive of probiosis in prospecting.CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorporUniversidade Federal de Minas GeraisPrograma de Pós-Graduação em GenéticaUFMGBrasilICB - INSTITUTO DE CIÊNCIAS BIOLOGICASGenéticaProbióticosMucositeFluoruracilaInfecções bacterianasProspecção de probióticosTestes de funcionalidadeBactérias lácticasMucosite intestinal5-FUProspecção de bactérias lácticas para uso como probióticos e avaliação do efeito protetor em modelo murino de mucosite intestinal induzida por 5-FUinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGORIGINALDISSERTAÇÃO NINA_VERSÃO FINAL.pdfDISSERTAÇÃO NINA_VERSÃO FINAL.pdfapplication/pdf4540827https://repositorio.ufmg.br/bitstream/1843/35057/1/DISSERTA%c3%87%c3%83O%20NINA_VERS%c3%83O%20FINAL.pdfa11c1bf8e2a28a9c465259567c8ce5d8MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-82119https://repositorio.ufmg.br/bitstream/1843/35057/2/license.txt34badce4be7e31e3adb4575ae96af679MD521843/350572021-02-24 14:00:59.384oai:repositorio.ufmg.br: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Repositório InstitucionalPUBhttps://repositorio.ufmg.br/oaiopendoar:2021-02-24T17:00:59Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
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