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Acinetobacter baumannii em meio hospitalar

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Universidade de Aveiro Ano 2010/2011

Departamento de Biologia

Maria José da Costa

Pinho Silveira

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Universidade de Aveiro Ano 2010/2011

Departamento de Biologia

Maria José da Costa

Pinho Silveira

Acinetobacter baumannii em meio hospitalar

Dissertação apresentada à Universidade de Aveiro para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Microbiologia, realizada sob a orientação científica da Doutora Sónia Alexandra Leite Velho Mendo Barroso, Professora auxiliar do Departamento de biologia da

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o júri

presidente Professora Doutora Maria Ângela Sousa Dias Alves Cunha Professora auxiliar do departamento de biologia da Universidade de Aveiro

Doutora Cláudia Sofia Soares de Oliveira Investigadora em Pós-Doutoramento do CESAM

Professora Doutora Sónia Alexandra Leite Velho Mendo Barroso Professora auxiliar do departamento de biologia da Universidade de Aveiro

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agradecimentos Agradeço à minha orientadora, professora Sónia Mendo, um exemplo singular de profissionalismo, por toda a dedicação, companheirismo e ajuda prestada durante a realização deste trabalho.

Ao hospital Infante D. Pedro, em especial ao director do serviço de patologia clínica, Dr. Elmano Ramalheira, pela cedência das estirpes, tornando possível a realização deste estudo. Neste contexto prolongo os meus agradecimentos à coordenadora dos técnicos de análises clínicas Manuela Luís e à técnica Raquel Diaz por todo a amizade, incentivo, ajuda e dedicação.

À Cátia, que além de ser uma pessoa fantástica, me ajudou em todos os momentos, incentivou e ensinou. Por toda a dedicação, compreensão, amizade e companheirismo e porque sem ti nada teria sido igual.

E por fim, mas não menos importante, a todas as pessoas do LBM, que me ajudaram em todos os instantes e fizeram-me passar horas maravilhosas e fantásticas que jamais esquecerei.

Muitissimo obrigada a todos. !

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palavras-chave Acinetobacter baumannii; Multirresistência; β-lactamases; Integrões classe 1.

resumo A espécie Acinetobacter baumannii é um microrganismo multirresistente,

frequentemente associado a surtos e infecções nos cuidados de saúde. A presença de uma grande variedade de factores determinantes na resistência aos antibióticos, juntamente com a sua capacidade de regular esses mecanismos e se adaptar sob condições ambientais adversas, faz com que esta espécie seja centro de preocupação para a Saúde pública.

A sua disseminação na maior parte das vezes clonal, dentro das instituições de saúde, faz com que este patogéneo nosocomial, esteja associado a elevadas taxas de mortalidade e morbilidade.

Os 47 isolados de A. baumannii foram isolados a partir de diferentes amostras biológicas provenientes de pacientes com exame bacteriológico positivo. Estas amostras deram entrada no Serviço de Patologia Clínica do Hospital Infante D. Pedro, e a faixa etária em que A. baumannii foi isolada com maior frequência está compreendida entre 81 e 90 anos de idade, facto provavelmente devido a um sistema imunitário mais fragilizado. Relativamente ao serviço onde surgiu um maior número de A. baumannii foi o Serviço de Medicina Intensiva (SMI), sendo a prevalência por produto biológico verificado em expectorações e urinas, facto justificado pela capacidade deste microrganismo aderir facilmente a superfícies, como algálias e tubos de ventilação.

No que refere aos Testes de Sensibilidade aos Antimicrobianos (TSA), A. baumannii é mais sensível à colistina, uma polimixina E, usada actualmente como último recurso para o tratamento de infecções por este patogéneo. Sendo este um microrganismo multirresistente, isso implica resistência a praticamente todas as classes de antibióticos testados. No entanto, os mecanismos responsáveis por estas resistências devem ser estudados, uma vez que apesar do fenótipo observado, a pesquisa de alguns dos genes frequentemente associados a essas resistências foi negativa. Foram encontrados genes de resistência associados a integrões de classe 1, e foi detectada a presença de elementos genéticos móveis, nomeadamente a presença de plasmídeos.

O estudo mais aprofundado dos mecanismos de resistência destes patogéneos é fundamental, de forma a melhor conhecer as opções terapêuticas existentes. Por outro lado torna-se importante perceber os mecanismos de disseminação deste patogéneo em ambiente hospitalar.

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keywords Acinetobacter baumannii; Multidrug resistant; β-lactamases enzymes; Class 1 integron

abstract Acinetobacter baumannii is a multiresistant and pathogenic organism,

commonly associated to outbreaks, clinical infections and hospital environment. These species present different antibiotic resistant determinants and different resistance mechanisms, but they have the ability to adapt to adverse environment conditions, and are therefore of concern to public health of the society.

The dissemination of these nosocomial pathogenic species, frequently clonal, is the main problem associated to the high mortality and morbidity rate, within the hospital environment.

To perform this work 47 A. baumannii isolates were collected from different biological samples from patients with a positive bacteriologic test. The statistic analysis was performed by the Clinical Pathology Service of the Hospital Infante D. Pedro, Aveiro. A. baumannii were frequently isolated from the age group between 81 and 90 years old. Probably, these results are due to a weak immune system. The isolates were collected mainly from intensive care unit (SMI) and from the following biological samples: sputum and urines, maybe because these isolates have the ability to adhere to the surfaces, such as catheters and ventilation tubes.

Antibiotics susceptibility tests were performed, showing that A. baumannii isolates are only susceptible to colistin, a polimixin E, that are used as the last resource to threat A. baumannii infections. These tests also showed that A. baumannii isolates are multidrug resistant, however the presence of β-lactamases genes was negative. Nevertheless some antibiotic resistance genes were associated to class 1 integrons. The presence of mobile genetic elements, namely plasmids, was found in some isolates.

Further studies exploring the resistance mechanisms of these pathogenic isolates are still required in order to increase the therapeutic options to threat nosocomial infections caused by this organism. Also, identify the source of dissemination of this pathogen within the hospital environment is of utmost importance.

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