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Rodrigo Otávio Silveira Silvahttp://lattes.cnpq.br/1439472527834751Carlos Augusto de Oliveira JúniorFrancisco Carlos Faria LobatoCecília Leite Costahttp://lattes.cnpq.br/8987695103593496Diogo Soares Gonçalves Cruz2021-05-19T11:14:47Z2021-05-19T11:14:47Z2021-01-25http://hdl.handle.net/1843/35990https://orcid.org/0000-0002-8577-6849Clostridioides difficile é um enteropatógeno bacteriano que infecta animais e seres humanos, capaz de promover distúrbios intestinais que podem levar o seu hospedeiro ao óbito. A infecção por C. difficile (ICD) representa atualmente uma das principais causas de infecção hospitalar, sendo a antibioticoterapia o principal fator predisponente. Na década de 2000, surtos ocasionados por C. difficile estiveram intimamente relacionados ao uso indiscriminado de antibióticos da classe das fluoroquinolonas. No Brasil, o tratamento da ICD é baseado no uso do metronidazol e da vancomicina. O uso de antimicrobianos parece ter relação direta com a seleção de determinadas estirpes de C. difficile. Nessa perspectiva, estudos epidemiológicos relacionados à resistência antimicrobiana desse patógeno são fundamentais para evitar a emergência de estirpes que podem representar um problema de saúde pública. Entretanto, o teste de sensibilidade antimicrobiana “padrão-ouro” recomendado para C. difficile é laborioso e de custo elevado. Dessa forma, o teste de disco difusão torna-se uma opção mais atrativa, uma vez que o método é simples de executar, mais barato e eficaz. O estudo teve como objetivo avaliar o perfil de sensibilidade ao metronidazol, à vancomicina e à moxifloxacina através do método de disco difusão de 120 estirpes de C. difficile isoladas de animais e seres humanos no Brasil. De acordo com pontos de corte já estabelecidos em estudos anteriores, as estirpes consideradas resistentes nesse teste foram submetidas ao método E-test para confirmação do fenótipo através da concentração inibitória mínima (CIM), mediante os critérios preconizados pelos órgãos de referência. Adicionalmente, três isolados classificados como resistentes à moxifloxacina foram sequenciados (gene gyrA) para detectar mutações que poderiam indicar o mecanismo genotípico de resistência à droga. Nenhum isolado foi resistente à vancomicina e somente uma estirpe (0,8%), considerada não toxigênica (RT010, clado 1), apresentou resistência ao metronidazol (CIM > 256 µg/mL). Em relação à moxifloxacina, 14% (n = 17) das estirpes avaliadas foram resistentes, as quais obtiveram CIM > 32 µg/mL. O ribotipo RT126 (clado 5) foi o único a apresentar associação estatística positiva de resistência à moxifloxacina (P < 0,0001), bem como os isolados das espécies equina e suína (P = 0,0139 e P = 0,0228, respectivamente). A mutação do tipo Thr82Ile foi encontrada nos três isolados sequenciados. O estudo revelou uma baixa frequência de resistência ao metronidazol e à vancomicina, enquanto a resistência à moxifloxacina foi superior a trabalhos anteriores realizados no Brasil.Clostridioides difficile is a bacterial enteropathogen that infects animals and humans, able to promote intestinal disorders that can lead to its host's death. Infection with C. difficile (CDI) currently represents one of the main causes of nosocomial infection, with antibiotic therapy being the main predisposing factor. In the 2000s, outbreaks caused by C. difficile were closely related to the indiscriminate use of antibiotics of the fluoroquinolone class. In Brazil, the treatment of CDI is based on the use of metronidazole and vancomycin. The use of antimicrobials seems to have a direct relationship with the selection of certain strains of C. difficile. In this perspective, epidemiological studies related to the antimicrobial resistance of this pathogen are essential to avoid the emergence of strains that may represent a public health problem. However, the “gold standard” antimicrobial susceptibility test recommended for C. difficile is laborious and expensive. Thus, the diffusion disk test becomes a more attractive option, since the method is simple to perform, cheaper and more effective. The study aimed to evaluate the sensitivity profile to metronidazole, vancomycin and moxifloxacin using the disk diffusion method from 120 strains of C. difficile from animals and humans isolated in Brazil. According to cutoff points already established in previous studies, strains considered resistant in this test were submitted to the E-test method to confirm the phenotype through the minimum inhibitory concentration (MIC), according to the criteria recommended by the reference institution. In addition, three isolates classified as resistant to moxifloxacin were sequenced (gyrA gene) to detect mutations that could indicate the genotypic mechanism of drug resistance. There was no vancomycin-resistant isolate and one strain (0.8%), non toxigenic (RT010, clade 1), was resistant to metronidazole (MIC > 256 µg/mL). Regarding moxifloxacin, 14% (n = 17) of the strains evaluated were resistant, which obtained MIC > 32 µg/mL. The ribotype RT126 (clade 5) was the only one to present a positive statistical association of resistance to moxifloxacin (P < 0.0001), as well as isolates of the equine and swine species (P = 0.0139 and P = 0.0228, respectively). The Thr82Ile mutation was found in the three sequenced isolates. The study revealed a low frequency of resistance to metronidazole and vancomycin, while resistance to moxifloxacin was superior to previous studies carried out in Brazil.CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoCAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorporUniversidade Federal de Minas GeraisPrograma de Pós-Graduação em Ciência AnimalUFMGBrasilVET - DEPARTAMENTO DE MEDICINA VETERINÁRIA PREVENTIVAClostridium difficileMedicina veterináriaDiarreia em animaisClostrídiosesPerfil de sensibilidade antimicrobiana de Clostridioides (Clostridium) difficile ao metronidazol, vancomicina e moxifloxacina isolados de animais e seres humanos em Minas Gerais, Brasilinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGORIGINALDissertação - Diogo Soares Gonçalves Cruz.pdfDissertação - Diogo Soares Gonçalves Cruz.pdfapplication/pdf1524169https://repositorio.ufmg.br/bitstream/1843/35990/1/Disserta%c3%a7%c3%a3o%20-%20Diogo%20Soares%20Gon%c3%a7alves%20Cruz.pdf1742d7e2cb09edc96ae66cba1b4d1b66MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-82119https://repositorio.ufmg.br/bitstream/1843/35990/2/license.txt34badce4be7e31e3adb4575ae96af679MD521843/359902021-05-19 08:14:47.236oai:repositorio.ufmg.br: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Repositório InstitucionalPUBhttps://repositorio.ufmg.br/oaiopendoar:2021-05-19T11:14:47Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
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