Resumo (PT):
Abstract (EN):
Bacteriophage–host interaction studies in biofilm structures are still challenging due to the technical
limitations of traditional methods. The aim of this study was to provide a direct fluorescence in situ
hybridization (FISH) method based on locked nucleic acid (LNA) probes, which targets the phage
replication phase, allowing the study of population dynamics during infection. Bacteriophages
specific for two biofilm-forming bacteria, Pseudomonas aeruginosa and Acinetobacter, were selected.
Four LNA probes were designed and optimized for phage-specific detection and for bacterial
counterstaining. To validate the method, LNA-FISH counts were compared with the traditional
plaque forming unit (PFU) technique. To visualize the progression of phage infection within a biofilm,
colony-biofilms were formed and infected with bacteriophages. A good correlation (r = 0.707) was
observed between LNA-FISH and PFU techniques. In biofilm structures, LNA-FISH provided a good
discrimination of the infected cells and also allowed the assessment of the spatial distribution of
infected and non-infected populations.
Language:
English
Type (Professor's evaluation):
Scientific
No. of pages:
12