Phage Therapy in Aquaculture: Current Developments and Future Challenges

A special issue of Pathogens (ISSN 2076-0817).

Deadline for manuscript submissions: closed (1 March 2022) | Viewed by 20870

Special Issue Editors


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Guest Editor
Institute of Marine Biology, Biotechnology and Aquaculture, Hellenic Centre for Marine Research, Former American Base of Gournes, Heraklion 71500, Crete, Greece
Interests: fish diseases; phage therapy; microbiology; marine aquaculture; biotechnology; vaccine development
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Guest Editor
Pontificia Universidad Católica de Valparaíso, Chile
Interests: Marine Microbiology,Bacteriophages

Special Issue Information

Dear Colleagues,

Aquaculture is the fastest growing sector of animal production worldwide. Very soon, most fishery products will come from aquaculture than from traditional fisheries. Intensification of aquaculture leads to an increase of disease outbreaks. When it comes to bacterial infections, administration of antibiotics is the first line of defense for the fish producer. Rational use of antibiotics is common practice in all animal production sectors; nevertheless, the emergence of antimicrobial resistance as one of the most significant threats for humanity has created an urgent need to replace antibiotics with more sustainable, alternative methods. Phage therapy is one of the most promising alternatives, and there is a steadily increasing volume of scientific publications towards this direction.

Phage research for aquaculture applications is currently being practiced in several parts of the world, including Europe, Asia, America, and Oceania. New phages with potency against fish and shellfish pathogens are continuously described, and new information is added to the scientific literature every day. However, there are many gaps in significant aspects before we can see the successful transition of phage therapy from the lab to the actual aquaculture production. These include stability of phages in the aquaculture environment, resistance development, upscaling of production, quality of the product, integration of modern approaches such as phage engineering, and of course regulation of phage therapy.

These are the main issues that we aspire to cover through this Special Issue of the journal Pathogens. The scope of the issue also includes current developments in phage research in relation to aquaculture (e.g., the role of prophages in virulence of aquaculture pathogens), discovery of new phages against aquaculture pathogens, and trials and applications in live feeds, fish, and invertebrates. We would like to encourage the submission of papers dealing with the use of phage lytic enzymes, the use of phages as probiotics or in combination with probiotics, and the role of phages in the intestinal environment of fish. 

Both original research and review articles are welcomed.

Dr. Pantelis Katharios
Prof. Roberto Bastías
Guest Editors

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Keywords

  • phage microbiology
  • aquaculture
  • phage therapy
  • antibiotic resistance
  • phage genomics

Published Papers (5 papers)

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Research

15 pages, 6929 KiB  
Article
Vibrio Phage Artemius, a Novel Phage Infecting Vibrio alginolyticus
by Stavros Droubogiannis, Lydia Pavlidi, Maria Ioanna Tsertou, Constantina Kokkari, Dimitrios Skliros, Emmanouil Flemetakis and Pantelis Katharios
Pathogens 2022, 11(8), 848; https://doi.org/10.3390/pathogens11080848 - 28 Jul 2022
Cited by 7 | Viewed by 2325
Abstract
Vibrio alginolyticus is an important pathogen of marine animals and has been the target of phage therapy applications in marine aquaculture for many years. Here, we report the isolation and partial characterization of a novel species of the Siphoviridae family, the Vibrio phage [...] Read more.
Vibrio alginolyticus is an important pathogen of marine animals and has been the target of phage therapy applications in marine aquaculture for many years. Here, we report the isolation and partial characterization of a novel species of the Siphoviridae family, the Vibrio phage Artemius. The novel phage was species-specific and could only infect strains of V. alginolyticus. It could efficiently reduce the growth of the host bacterium at various multiplicities of infection as assessed by an in vitro lysis assay. It had a genome length of 43,349 base pairs. The complete genome has double-stranded DNA with a G + C content of 43.61%. In total, 57 ORFs were identified, of which 19 were assigned a predicted function. A genomic analysis indicated that Vibrio phage Artemius is lytic and does not harbor genes encoding toxins and antibiotic resistance determinants. Full article
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25 pages, 7899 KiB  
Article
Genomic and Biological Profile of a Novel Bacteriophage, Vibrio phage Virtus, Which Improves Survival of Sparus aurata Larvae Challenged with Vibrio harveyi
by Stavros Droubogiannis and Pantelis Katharios
Pathogens 2022, 11(6), 630; https://doi.org/10.3390/pathogens11060630 - 30 May 2022
Cited by 16 | Viewed by 3259
Abstract
Due to the emergence of multidrug-resistant bacteria, commonly known as “superbugs”, phage therapy for the control of bacterial diseases rose in popularity. In this context, the use of phages for the management of many important bacterial diseases in the aquaculture environment is auspicious. [...] Read more.
Due to the emergence of multidrug-resistant bacteria, commonly known as “superbugs”, phage therapy for the control of bacterial diseases rose in popularity. In this context, the use of phages for the management of many important bacterial diseases in the aquaculture environment is auspicious. Vibrio harveyi, a well-known and serious bacterial pathogen, is responsible for many disease outbreaks in aquaculture, resulting in huge economic and production losses. We isolated and fully characterized a novel bacteriophage, Vibrio phage Virtus, infecting V. harveyi strain VH2. Vibrio phage Virtus can infect a wide spectrum of Vibrio spp., including strains of V. harveyi, V. owensii, V. campbellii, V. parahaemolyticus, and V. mediterranei. It has a latent period of 40 min with an unusually high burst size of 3200 PFU/cell. Vibrio phage Virtus has a double-stranded DNA of 82,960 base pairs with 127 predicted open reading frames (ORFs). No virulence, antibiotic resistance, or integrase-encoding genes were detected. In vivo phage therapy trials in gilthead seabream, Sparus aurata, larvae demonstrated that Vibrio phage Virtus was able to significantly improve the survival of larvae for five days at a multiplicity of infection (MOI) of 10, which suggests that it can be an excellent candidate for phage therapy. Full article
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38 pages, 7555 KiB  
Article
Biological and Genomic Characterization of a Novel Jumbo Bacteriophage, vB_VhaM_pir03 with Broad Host Lytic Activity against Vibrio harveyi
by Gerald N. Misol, Jr., Constantina Kokkari and Pantelis Katharios
Pathogens 2020, 9(12), 1051; https://doi.org/10.3390/pathogens9121051 - 15 Dec 2020
Cited by 22 | Viewed by 4396
Abstract
Vibrio harveyi is a Gram-negative marine bacterium that causes major disease outbreaks and economic losses in aquaculture. Phage therapy has been considered as a potential alternative to antibiotics however, candidate bacteriophages require comprehensive characterization for a safe and practical phage therapy. In this [...] Read more.
Vibrio harveyi is a Gram-negative marine bacterium that causes major disease outbreaks and economic losses in aquaculture. Phage therapy has been considered as a potential alternative to antibiotics however, candidate bacteriophages require comprehensive characterization for a safe and practical phage therapy. In this work, a lytic novel jumbo bacteriophage, vB_VhaM_pir03 belonging to the Myoviridae family was isolated and characterized against V. harveyi type strain DSM19623. It had broad host lytic activity against 31 antibiotic-resistant strains of V. harveyi, V. alginolyticus, V. campbellii and V. owensii. Adsorption time of vB_VhaM_pir03 was determined at 6 min while the latent-phase was at 40 min and burst-size at 75 pfu/mL. vB_VhaM_pir03 was able to lyse several host strains at multiplicity-of-infections (MOI) 0.1 to 10. The genome of vB_VhaM_pir03 consists of 286,284 base pairs with 334 predicted open reading frames (ORFs). No virulence, antibiotic resistance, integrase encoding genes and transducing potential were detected. Phylogenetic and phylogenomic analysis showed that vB_VhaM_pir03 is a novel bacteriophage displaying the highest similarity to another jumbo phage, vB_BONAISHI infecting Vibrio coralliilyticus. Experimental phage therapy trial using brine shrimp, Artemia salina infected with V. harveyi demonstrated that vB_VhaM_pir03 was able to significantly reduce mortality 24 h post infection when administered at MOI 0.1 which suggests that it can be an excellent candidate for phage therapy. Full article
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10 pages, 12123 KiB  
Article
Bacteriophage Cocktail for the Prevention of Multiple-Antibiotic-Resistant and Mono-Phage-Resistant Vibrio coralliilyticus Infection in Pacific Oyster (Crassostrea gigas) Larvae
by Hyoun Joong Kim, Jin Woo Jun, Sib Sankar Giri, Sang Guen Kim, Sang Wha Kim, Jun Kwon, Sung Bin Lee, Cheng Chi and Se Chang Park
Pathogens 2020, 9(10), 831; https://doi.org/10.3390/pathogens9100831 - 11 Oct 2020
Cited by 16 | Viewed by 2963
Abstract
Vibrio coralliilyticus (V. coralliilyticus) is a pathogen that causes mass mortality in marine bivalve hatcheries worldwide. In this study, we used a bacteriophage (phage) cocktail to prevent multiple-antibiotic-resistant (MAR) and phage-resistant (PR) V. coralliilyticus infection in Pacific oyster (Crassostreagigas [...] Read more.
Vibrio coralliilyticus (V. coralliilyticus) is a pathogen that causes mass mortality in marine bivalve hatcheries worldwide. In this study, we used a bacteriophage (phage) cocktail to prevent multiple-antibiotic-resistant (MAR) and phage-resistant (PR) V. coralliilyticus infection in Pacific oyster (Crassostreagigas) larvae. To prevent the occurrence of phage-resistant strains and decrease the effect of mono-phage treatment, we prepared a phage cocktail containing three types of V. coralliilyticus-specific phages and tested its prophylactic efficacy against MAR and PR V. coralliilyticus infection. The results of the cell lysis test showed that the phage cocktail showed an excellent bactericidal effect against the MAR and PR variants in contrast to the experimental group treated with two mono phages (pVco-5 and pVco-7). An in vivo test using Pacific oyster larvae also confirmed the preventive effect against MAR and PR variants. In conclusion, the application of the phage cocktail effectively prevented V. coralliilyticus infection in marine bivalve seedling production. Furthermore, it is expected to reduce damage to the aquaculture industry caused by the occurrence of MAR and PR V. coralliilyticus. Therefore, phage cocktails may be used for the control of various bacterial diseases. Full article
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13 pages, 3359 KiB  
Article
Isolation, Characterization, and Application of a Bacteriophage Infecting the Fish Pathogen Aeromonas hydrophila
by Muhammad Akmal, Aryan Rahimi-Midani, Muhammad Hafeez-ur-Rehman, Ali Hussain and Tae-Jin Choi
Pathogens 2020, 9(3), 215; https://doi.org/10.3390/pathogens9030215 - 13 Mar 2020
Cited by 54 | Viewed by 6603
Abstract
Bacteriophages are increasingly being used as biological control agents against pathogenic bacteria. In the present study, we isolate and characterize bacteriophage Akh-2 from Geoje Island, South Korea, to evaluate its utility in controlling motile Aeromonas septicemia. Akh-2 lysed four of the seven Aeromonas [...] Read more.
Bacteriophages are increasingly being used as biological control agents against pathogenic bacteria. In the present study, we isolate and characterize bacteriophage Akh-2 from Geoje Island, South Korea, to evaluate its utility in controlling motile Aeromonas septicemia. Akh-2 lysed four of the seven Aeromonas hydrophila strains tested. Transmission electron microscopy analysis showed that Akh-2 belongs to the Siphoviridae family, with head and tail sizes of 50 ± 5 and 170 ± 5 nm, respectively. One-step growth curve analysis revealed that the phage has a latent period of 50 ± 5 min and a burst size of 139 ± 5 plaque-forming units per infected cell. The phage appeared stable in a pH range of 6–8 and a temperature range of −80 to 46 °C. Based on next-generation sequencing analysis, its genome is 114,901 bp in size, with a 44.22% G + C content and 254 open reading frames. During an artificial induction of the disease, loach (Misgurnus anguillicaudatus) treated with Akh-2 showed an increased survival rate and time compared with the non-treated control. Our results suggest that Akh-2 is a potential biological agent for the treatment of Aeromonas infections in fish. Full article
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