Isolation and characterization of phage AHP-1 and its combined effect with chloramphenicol to control Aeromonas hydrophila
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To develop an alternative bio-control measure for multi-drug resistant pathogenic Aeromonas hydrophila, which causes motile Aeromonas septicemia in fish, novel virulent phage (AHP-1) was isolated from carp tissues. Morphological analysis by transmission electron microscopy revealed that AHP-1 belongs to Myoviridae family. AHP-1 displayed 81% of moderate adsorption by 25 min, and latent period of 40 min with burst size of 97 PFU mL−1 at an optimal multiplicity of infection (MOI) 0.1. AHP-1 was stable over a broad range of pH (4–11), temperature (4–50 °C), and salinity (0.1–3.5%). Both time and MOI dependent in vitro A. hydrophila growth inhibition was observed with AHP-1. AHP-1 (10 MOI) showed higher growth inhibition against A. hydrophila than chloramphenicol (5 μg mL−1), and combined treatment was more promising than individuals. Immune gene expression analysis of zebrafish upon continuous bath exposure to AHP-1 resulted significantly higher (il-6 and sod-1) or slight induction (tnf-α, il1-β, il-10, and cxcl-8a) than controls at beginning of the phage exposure, but those lowered to basal level by day 12 post-phage exposure. It suggests no adverse immune responses have occurred for the AHP-1 dose that used, and have potential for the phage therapy. Further detailed in vivo studies are needed to confirm the protective efficacy of newly isolated AHP-1 against A. hydrophila infection.
KeywordsA. hydrophila Chloramphenicol Fish pathogen Immune responses Phage AHP-1 Multi-drug resistance
This work was supported by the National Research Foundation of Korea (NRF) grant funded by the government of Korea (MSIT) (2017010990).
Compliance with ethical standards
Zebrafish experiments were conducted in accordance with the institutional animal care guidelines and supervision of committees of Chungnam National University (CNU-00866).
Conflict of interest
The authors declare that they have no conflict of interest.
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