| Grant number: | 13/13358-0 |
| Support Opportunities: | Scholarships in Brazil - Doctorate (Direct) |
| Start date: | November 01, 2013 |
| End date: | August 31, 2016 |
| Field of knowledge: | Biological Sciences - Microbiology - Applied Microbiology |
| Principal Investigator: | Ana Lúcia da Costa Darini |
| Grantee: | Natália Candido Caçador |
| Host Institution: | Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil |
| Associated scholarship(s): | 14/19310-2 - Mutation, biofilm growth and phenotypic characteristics of Pseudomonas aeruginosa isolated from patients with cystic fibrosis, BE.EP.DD |
Abstract Pseudomonas aeruginosa is the major gram-negative bacterium associated with lung infections in Cystic Fibrosis (CF) patients, which contributes to the increased morbidity and mortality of such disease. P. aeruginosa has the ability to cause progressive damage to patients, mainly because of three factors: hypermutator phenotype; expression of virulence factors; and a remarkable and intrinsic resistance to antibiotics, associated with the ability to acquire new resistance determinants. In Brazil there are few studies related with bacteria isolated from CF patients, restricting the researches to few reference centers of FC. Within this context, the aim of this project is to evaluate strains of P. aeruginosa in CF patients treated at the Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto - Universidade de São Paulo (HCFMRP-USP), and relate all the main features of P. aeruginosa for obtaining of a complete overview of the isolates, namely: virulence factors, antimicrobial susceptibility of clinical-microbiological interest, genomic similarity and occurrence of hypermutable strains. Particularly, phenotypic and molecular methods will be used for: isolation and identification of P. aeruginosa, determination of antibiotic resistance and investigation of the occurrence of virulence factors. The occurrence of hypermutable strains will be determined by a modified disk diffusion method and the mutation frequency will be estimated from the rifampicin resistance displayed by the isolates. Sequencing assays will be realized for detection of: (i) mutations in genes belonging to the system that prevent or repair the mutations in the bacterial DNA; and (ii) mutations in the quorum sensing system genes and in the alginate production gene. The genetic variability of the isolates will be characterized considering the macrorestriction by pulsed-field gel electrophoresis. The isolates will be characterized as mucoid or non-mucoid according to colonial morphology displayed on the first culture, and determination/evaluation of patients with chronic and intermittent colonization will be held in conjunction with the medical team, with the aid of the results obtained by the techniques already described. It is expected that the results of this project contribute to a greater understanding of colonization/infection by P. aeruginosa and that they also play a role in the improvement of the quality of life of CF patients. (AU) | |
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