Abstract
Aeromonas hydrophila is one of the major freshwater fish pathogens. In the current study, a cocktail of D6 and CF7 phages was given orally to Labeo rohita to assess phage survival in fish organs as well as to determine the therapeutic efficacy of phage treatment against fish mortality caused by A. hydrophila. In the phage-coated feed, prepared by simple spraying method, phage counts were quite stable for up to 2 months with a decline of ≤ 0.23 log10 and ≤ 1.66 log10 PFU/g feed during 4 oC and room temperature storage. Throughout the experimental period of 7 days, both phages could be detected in the gut of fish fed with phage-coated feed. Besides, both CF7 and D6 phages were also detected in fish kidneys indicating the ability of both the phage to cross the intestinal barrier. During challenge studies with LD50 dose of A. hydrophila, phage cocktail doses of 1 × 106 – 1 × 108 PFU/g feed prevented the mortality in L. rohita with relative percentage survival (RPS) of 8.7–65.2. When challenged with LD90 dose of A. hydrophila, an RPS value of 28.6 was obtained at a phage cocktail dose of 1 × 108 PFU/g feed. The RPS data showed that orally-fed phage cocktail protected the fish against the mortality caused by A. hydrophila in a dose-dependent manner. Simple practical approaches for phage cocktail development, medicated feed preparation and oral administration along with phage survival and protection data make the current study useful for farmer-level application.
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Data availability
The study data is available from the corresponding author on reasonable request.
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Acknowledgements
The authors are grateful to the Dean (College of Fisheries, Guru Angad Dev Veterinary & Animal Sciences University, Ludhiana, India) for all the necessary support during this study.
Funding
This work was supported by Rashtriya Krishi Vikas Yojana (RKVY) Grant RKVY-11:I3 “Development of biotechnological intervention strategies to enhance the safety and shelf life of fishery products” and the Board of Research in Nuclear Sciences (BRNS) Grant 55/14/02/2021-BRNS “Development of phage-based strategies for biocontrol of antibiotic resistant Aeromonas species in fishery products” to Anuj Tyagi.
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Sumeet Rai: Methodology, Investigation, Data Curation, Data Analysis, Visualization, Writing - Original Draft. Anuj Tyagi: Conceptualization, Methodology, Data Analysis, Writing - Review & Editing, Supervision, Project administration, Funding acquisition. Naveen Kumar B. T.: Investigation, Data Curation, Data Analysis, Writing - Review & Editing.
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All fish experiments in the present study were performed according to the Fish Experimentation Guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Government of India (protocol approval no. GADVASU/2022/IAEC/65/01).
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Communicated by Yusuf Akhter.
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Supplementary Table 1
: Mortalities in L. rohita fingerlings after intraperitoneal injection with A. hydrophila. Supplementary Table 2: Replicate-wise data of protection offered by phage cocktail against LD50 of A. hydrophila. Supplementary Table 3: Replicate-wise data of protection offered by phage cocktail against LD90 of A. hydrophila
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Rai, S., Tyagi, A. & B. T., N. Oral feed-based administration of phage cocktail protects rohu fish (Labeo rohita) against Aeromonas hydrophila infection. Arch Microbiol 206, 219 (2024). https://doi.org/10.1007/s00203-024-03951-3
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DOI: https://doi.org/10.1007/s00203-024-03951-3