Abstract
Bordetella bronchiseptica is a respiratory animal pathogen that shows growing resistance to commonly used antibiotics, which has necessitated the examination of new antimicrobials, including bacteriophages. In this study, we examined the previously isolated and partially characterized B. bronchiseptica siphoviruses of the genus Vojvodinavirus (LK3, CN1, CN2, FP1 and MW2) for their ability to inhibit bacterial growth and biofilm, and we examined other therapeutically important properties through genomic analysis and lysogeny experiments. The phages inhibited bacterial growth at a low multiplicity of infection (MOI = 0.001) of up to 85% and at MOI = 1 for >99%. Similarly, depending on the phages and MOIs, biofilm formation inhibition ranged from 65 to 95%. The removal of biofilm by the phages was less efficient but still considerably high (40–75%). Complete genomic sequencing of Bordetella phage LK3 (59,831 bp; G + C 64.01%; 79 ORFs) showed integrase and repressor protein presence, indicating phage potential to lysogenize bacteria. Lysogeny experiments confirmed the presence of phage DNA in bacterial DNA upon infection using PCR, which showed that the LK3 phage forms more or less stable lysogens depending on the bacterial host. Bacterial infection with the LK3 phage enhanced biofilm production, sheep blood hemolysis, flagellar motility, and beta-lactam resistance. The examined phages showed considerable anti-B. bronchiseptica activity, but they are inappropriate for therapy because of their temperate nature and lysogenic conversion of the host bacterium.
Highlights
We aimed to determine if B. bronchiseptica-specific siphoviruses were appropriate for phage therapy. We examined their potential to inhibit bacterial growth and biofilm formation and reduce existing B. bronchiseptica biofilm
All bacteria were stored in Luria Bertani broth (LB) containing glycerol
Even at the lowest multiplicity of infection (MOI), inhibition of bacterial growth was considerable (85%), with maximal inhibition reached at MOI 1 (>99%) (Figure 4A)
Summary
Bordetella bronchiseptica is primarily a respiratory pathogen of domestic, wild, and laboratory animals [1] and is considered to be the evolutionary progenitor of all species of the Bordetella genus, including the well-known human pathogens B. pertussis and B. parapertussis [2]. B. bronchiseptica is one of the main causative agents of a few highly contagious and economically significant diseases, including kennel cough, atrophic rhinitis and upper respiratory tract disease (snuffles) [7,8,9]. It is generally considered a zoonotic agent, B. bronchiseptica rarely causes infection in humans, with the exception of immunocompromised individuals [10]
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