abstract
The occurrence of infections by pathogenic bacteria is one of the main sources of financial loss for the aquaculture industry. This problem often cannot be solved with antibiotic treatment or vaccination. Phage therapy seems to be an alternative environmentally-friendly strategy to control infections. Recognizing the cellular modifications that bacteriophage therapy may cause to the host is essential in order to confirm microbial inactivation, while understanding the mechanisms that drive the development of phage-resistant strains. The aim of this work was to detect cellular modifications that occur after phage AS-A treatment in A. salmonicida, an important fish pathogen. Phage-resistant and susceptible cells were subjected to five successive streak-plating steps and analysed with infrared spectroscopy, a fast and powerful tool for cell study. The spectral differences of both populations were investigated and compared with a phage sensitivity profile, obtained through the spot test and efficiency of plating. Changes in protein associated peaks were found, and these results were corroborated by 1-D electrophoresis of intracellular proteins analysis and by phage sensitivity profiles. Phage AS-A treatment before the first streaking-plate step clearly affected the intracellular proteins expression levels of phage-resistant clones, altering the expression of distinct proteins during the subsequent five successive streak-plating steps, making these clones recover and be phenotypically more similar to the sensitive cells.
keywords
ESCHERICHIA-COLI; SHOCK-PROTEIN; INFRARED-SPECTROSCOPY; IR SPECTROSCOPY; IN-VITRO; BACTERIOPHAGES; THERAPY; IDENTIFICATION; BIOCONTROL; INFECTION
subject category
Pharmacology & Pharmacy
authors
Moreirinha, C; Osorio, N; Pereira, C; Simoes, S; Delgadillo, I; Almeida, A
our authors
Catarina Moreirinha
Post-Doc Fellowshipacknowledgements
This work was supported by FEDER through COMPETE-Programa Operacional Factores de Competitividade, and by National funding through Fundacao para a Ciencia e Tecnologia (FCT), within the research projects FCOMP-01-0124-FEDER-013934 and ENV/ES/001048. Financial support was provided to Catarina Moreirinha in the form of a Postdoctoral grant (ENV/ES/001048), and Carla Pereira in the form of a PhD grant (SFRH/BD/76414/2011).