• Title/Summary/Keyword: Bacteriophages (phages)

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Recent Insights into Aeromonas salmonicida and Its Bacteriophages in Aquaculture: A Comprehensive Review

  • Park, Seon Young;Han, Jee Eun;Kwon, Hyemin;Park, Se Chang;Kim, Ji Hyung
    • Journal of Microbiology and Biotechnology
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    • v.30 no.10
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    • pp.1443-1457
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    • 2020
  • The emergence and spread of antimicrobial resistance in pathogenic bacteria of fish and shellfish have caused serious concerns in the aquaculture industry, owing to the potential health risks to humans and animals. Among these bacteria, Aeromonas salmonicida, which is one of the most important primary pathogens in salmonids, is responsible for significant economic losses in the global aquaculture industry, especially in salmonid farming because of its severe infectivity and acquisition of antimicrobial resistance. Therefore, interest in the use of alternative approaches to prevent and control A. salmonicida infections has increased in recent years, and several applications of bacteriophages (phages) have provided promising results. For several decades, A. salmonicida and phages infecting this fish pathogen have been thoroughly investigated in various research areas including aquaculture. The general overview of phage usage to control bacterial diseases in aquaculture, including the general advantages of this strategy, has been clearly described in previous reviews. Therefore, this review specifically focuses on providing insights into the phages infecting A. salmonicida, from basic research to biotechnological application in aquaculture, as well as recent advances in the study of A. salmonicida.

Isolation and Characterization of Temperate Phages in Enterococcus faecium from Sprouts (새싹채소 유래 Enterococcus faecium으로부터 Temperate Phage의 분리와 특성)

  • Lee, Young-Duck;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.46 no.3
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    • pp.323-327
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    • 2014
  • To analyze the characteristics of bacteriophages in Enterococcus faecium, D-19 and F6 phages were induced from five E. faecium isolated from sprouts by the treatment with mitomycin C. The bacteriophages of D-19 and F-6 had long, non-contractile tails and icosahedral heads, and were members of Siphoviridae family. As the host spectrum, D-19 phage lysed five out of 55 strains of E. faecium, whereas F6 phage lysed only three strains. Both D-19 and F6 phages displayed similar and high stabilities against ethanol and pH capable of resisting the exposure to 100% ethanol and pH 4.

Classification and Characterization of Bacteriophages of Lectobacillus casei (Lactobacillus casei Bacperiophage의 분류 및 특성에 관한 연구)

  • 김영창;박민철;강국희;윤영호;이광웅
    • Korean Journal of Microbiology
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    • v.17 no.4
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    • pp.165-178
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    • 1979
  • Phages of Lactobaciilus casei (PLC) isolated from plant drainage were classified and characterized. The results are as follows : 1. On the basis of host range pattern, phages could be divided into 2 groups (PLC-B and PLC-C). PLC-B group phages could be further divided into 5 sub-groups $(B_1, \;B_2, \;B_3, \;B_4, \;and\;B_5)$. Although PLC-C group phages had the same host range, they could be also divided into 2 sub-groups $(C_1\;and\;C_2)$ by morphlogical type. 2. It was $B_3$ group phages that represented a major proportion (44.4%) of phages tested. However, $B_1$ group phages were shown to have the widest host range. 3. Electron micrographs revealed that the phages fell into three different morphological types. $(B_1, \;B_2, \;and\;B_3)$ group phages hd a hexagonal head (52nm in diameter) and a sheathless noncontractile (245 nm in length). $B_4\;and\;C_2$ group phages had a hexagonal head (56 nm) and a short flexible tail (169nm) having no sheath. $B_5\;and\;C_1$ group phages were shown to have a hexagonal head (81 nm) and a contractile tail (140 nm) having a sheath, a base plate and tail fibers. 4. The inactivation of the phages by antisera indicated that serological relationships correlated completely with morphological types. 5. $B_1, \;C_1\;and\;C_2$ group phages produced a large (1, 2 mm in diameter) plaque with a clear ring. The morphology of plaques of $B_3\;and\;B_5$ group phages was the same as those produced by the above, but the average plaque sizes for $B_3\;and\;B_5$ were 0.8 mm abd 0.5 mm, respectively. $B_2\;and\;B_4$ group phages produced a small (0.5 mm) turbid plaque with an irregular edge. 6. The latent period and the average burst size of $B_1\;and\;B_3$ group phages were 90 min and 100, respectively. These phages reuqired calcium ions for their miltiplication. 7. $B_3$ group phages could not be absrobed to R-variant $KC_1$. 8. The order of resistance of phages to heat was $B_2\;>\;B_1, B_4\;and\;B_5\;>\;B_3\;and\;C_2, \;B_5$ group phages were more stable than $B_3$ in various pH values. $C_2$ group phages were more sensitive to UV irradiation than $B_1\;and\;B_3$ group phages. 9. Strains YIT9018 and IAM 1043 were induced by mitomycin C treatment. Phage particles detected in the lysates had a hexagonal head (38 and 49 nm, respectively), but no tail. Any sensitive indicator strain could not be isolated in spite of repaeated trials.

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Safety evaluation of bacteriophages for application as sanitizers (박테리오파지의 살균소독제 응용을 위한 안전성 평가)

  • Park, Do-Won;Lee, Young-Duck;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.52 no.1
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    • pp.109-112
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    • 2020
  • To evaluate the safety of bacteriophages for application of sanitizer, endotoxin content and cell cytotoxicity of two Escherichia coli and four Staphylococcus aureus phages were determined. Endotoxin ratio was determined by the Limulus amebocyte lysate (LAL) assay as a test for representative biological endotoxin content. The average endotoxin average content of the 9 log PFU/mL lysate was 18.6 EU/mL and that of the 10 log PFU/mL lysate was 5.9 EU/mL, suggesting that the phage lysate was not suitable for clinical applications, but suitable for food pathogen control applications. To confirm the cell cytotoxicity of the phage lysates, MTT assay was performed using Raw 264.7 cells treated with 9 log PFU/mL phages. Results of the assay indicated that the phage lysates did not significantly decrease the cell viability (p>0.05). These results indicated that bacteriophages would be suitable as a food safety sanitizer.

Phage Assembly Using APTES-Conjugation of Major Coat p8 Protein for Possible Scaffolds

  • Kim, Young Jun;Korkmaz, Nuriye;Nam, Chang Hoon
    • Interdisciplinary Bio Central
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    • v.4 no.3
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    • pp.9.1-9.7
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    • 2012
  • Filamentous phages have been in the limelight as a new type of nanomaterial. In this study, genetically and chemically modified fd phage was used to generate a biomimetic phage self-assembly product. Positively charged fd phage (p8-SSG) was engineered by conjugating 3-aminopropyltriethoxysilane (APTES) to hydroxyl groups of two serine amino acid residues introduced at the N-terminus of major coat protein, p8. In particular, formation of a phage network was controlled by changing mixed ratios between wild type fd phage and APTES conjugated fd-SSG phage. Assembled phages showed unique bundle and network like structures. The bacteriophage based self-assembly approach illustrated in this study might contribute to the design of three dimensional microporous structures. In this work, we demonstrated that the positively charged APTES conjugated fd-SSG phages can assemble into microstructures when they are exposed to negatively charged wild-type fd phages through electrostatic interaction. In summary, since we can control the phage self-assembly process in order to obtain bundle or network like structures and since they can be functionalized by means of chemical or genetic modifications, bacteriophages are good candidates for use as bio-compatible scaffolds. Such new type of phage-based artificial 3D architectures can be applied in tuning of cellular structures and functions for tissue engineering studies.

Pseudomonas tolaasii bacteriophage-specific polyclonal antibody formation and its cross reactivity to various phages (Pseudomonas tolaasii 박테리오파지에 특이적인 다클론항체 형성 및 이를 이용한 파지 교차 반응성)

  • Yun, Yeong-Bae;Park, Soo-Jin;Kim, Young-Kee
    • Journal of Applied Biological Chemistry
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    • v.62 no.3
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    • pp.287-292
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    • 2019
  • Pseudomonas tolaasii causes brown blotch disease on the oyster mushroom (Pleurotus ostreatus). Various pathogenic strains of P. tolaasii were isolated and divided into three subtypes, $P1{\alpha}$, $P1{\beta}$, and $P1{\gamma}$. For phage therapy, bacteriophages against to these subtype strains were applied to mushroom cultivation and very successful to prevent from the disease. In this study, bacteriophages were isolated against the representative strains of subtype pathogens and their polyclonal antibodies were synthesized to investigate structural relationship among capsid proteins of phages. Phage preparations over $10^{10}pfu/mL$ were injected to rabbit thigh muscle and polyclonal antibodies were obtained after three times of boost injection. Titers of the antibodies obtained were over $2{\times}10^7Ab/mL$ for the phage ${\phi}6264$, $1{\times}10^6Ab/mL$ for the phage ${\phi}HK2$, and $1{\times}10^7Ab/mL$ for the phage ${\phi}HK19$ and phage ${\phi}HK23$. High specific activities were observed between antibodies and the corresponding bacteriophages. Some cross-reactivities between the antibodies and non-corresponding bacteriophages were also measured. Antibody $Ab{\phi}6264$ inactivated all phages of $P1{\alpha}$ subtype and only phage ${\phi}HK16$ among $P1{\beta}$ subtype phages. Antibody $Ab{\phi}HK23$ of $P1{\gamma}$ subtype neutralized all phages of $P1{\beta}$ subtype as well as the phage ${\phi}HK23$, showing the widest phage-inactivation range. When the structural-similarity studies of phages were investigated by using phage antibodies, closeness obtained by phylogenetic analysis of 16S rRNA genes of pathogenic strains were quite different from that of polyclonal antibody-specific structural similarity of phage capsid proteins. In conclusion, there is weak correlation between the host strain specificity of bacteriophage and its capsid structural similarity measured by phage antibodies.

Comparison in Restriction Profile Analysis of Vibrio furnissi, Vibrio fluvialis, and Vibrio parahaemolyticus Bacteriophage from Sea Product

  • Kim, Young-Hee
    • Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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    • v.1 no.2
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    • pp.99-103
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    • 1997
  • The bacteriophages lytic for Vibrio furnissi, Vibrio fluvialis and Vibrio parahaemolyticus were isolated from fish gills and shellfish. Nucleic acid of bacteriophages was prepared and restriction endonuclease profile was compared. All isolates contained deoxyribonucleic acid. V. furnissi bacteriophage from fish gills showed 2 bands with Bgl II, 1 with Pst, 3 with Hind III, I with Bam HI and 2 with EcoR 1. V fluvialis phage represented 7 fragments with Bgl II, 1 with Pst, 4 with Hind III, and 2 with EcoR 1. V. parahamolyticus produced 13 sites with Hind III and 4 sites with EcoR 1. The fragment types were varied depending on the phage isolation. All three phages were digested with Hind III and EcoR I with different sizes. V. furnissi phage were digested with 5 different restriction enzymes.

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The Effect of Environmental Factors on Phage Stability and Infectivity on Their Host Bacteria: a Case Study for an Escherichia coli Phage (T7), a Listeria Phage (A511), and a Salmonella Phage (Felix O1)

  • Kim, Kwang-Pyo
    • Food Science and Biotechnology
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    • v.16 no.3
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    • pp.398-403
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    • 2007
  • The effectiveness of phage biocontrol depends on the activity of bacteriophage in a given environment. In order to investigate the infectivity and the stability of bacteriophages in representative environments, three virulent phages, Listeria phage A511, Salmonella phage Felix O1, and Escherichia coli phage T7, were subjected to different temperatures, pHs and salt concentrations (NaCl). Phage infectivity was also determined in the presence of divalent cations ($Mg^{2+}$ or $Ca^{2+}$). As a result, three phages exhibited a wide range of survival rates under various environments. Phage infectivity was directly correlated with bacterial growth under the applied conditions. One exception was Felix O1 that did not kill Salmonella grown in low pH (4.5). The failure was attributed to defective adsorption of Felix O1. This finding is significant as it provides an explanation for the inefficient phage biocontrol. Therefore, such information is crucial to improve phage biocontrol of pathogens.

Inhibition of Clostridium perfringens using Bacteriophages and Bacteriocin Producing Strains

  • Heo, Sunhak;Kim, Min Gon;Kwon, Mirae;Lee, Hee Soo;Kim, Geun-Bae
    • Food Science of Animal Resources
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    • v.38 no.1
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    • pp.88-98
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    • 2018
  • In this study, we isolated and characterized a bacteriocin-producing strain and two bacteriophages (P4, A3), showing antimicrobial effects against Clostridium perfringens, from chicken and swine feces by the spot-on-the lawn antagonism method. The selected strain was identified as Streptococcus hyointestinalis by 16S rRNA gene sequencing. The bacteriocin from the isolated strain exhibited strong inhibitory activity against four strains of C. perfringens and all the tested strains of Listeria monocytogenes, and the bacteriocin were highly heat- and pH-stable even at pH 2, pH 10 and $121^{\circ}C$ for 15 min. We also evaluated the combined effects of the isolated bacteriocin and phages. Combining the phage treatments and bacteriocin resulted in a synergetic effect compared with the phage or the bacteriocin alone. In addition, during the probiotic test, the bacteriocin-producing S. hyointestinalis B19 strain reduced the population of C. perfringens significantly. Treatment with S. hyointestinalis B19 and a cocktail of lytic bacteriophages eradicated the C. perfringens KCTC $3269^T$, completely. Consequently, the isolated bacteriocin and bacteriophages represent candidates for effective biocontrol of C. perfringens, and bacteriocin-producing S. hyointestinalis B19 is a potential probiotic candidate for use in domestic animals.

Ecological Characteristics of Bacteriophages Infecting Xanthomonas oryzae pv. oryzae and Their Use as Biocontrol Agents (벼 흰잎마름병균 파지의 생태학적 특성 및 이를 이용한 생물방제)

  • Yu, Sang-Mi;Noh, Tae-Hwan;Kim, Dong-Min;Jeon, Tae-Woog;Lee, Young-Kee;Lee, Se-Won;You, Oh-Jong;Kim, Byung-Seok;Lee, Yong-Hoon
    • Research in Plant Disease
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    • v.17 no.1
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    • pp.90-94
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    • 2011
  • Bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae (Xoo) is a very serious disease in rice growing regions of the world. There are no effective ways of protecting rice from the disease. In this study, the bacteriophage (phage) mixtures infecting Xoo were investigated as biological control agent on BLB. The effects of pH, heat and ultraviolet on the stability of phages were investigated to check and increase the possibility of practical use in the field. Phages were rather stable between pH 5 and pH 10. The infectivity dropped sharply when the phages were incubated at $50^{\circ}C$ and more than 90% of the phages were inactivated after two minutes of ultraviolet treatment. The phages were stable for 7 days at the rice plant leaves, and the phages survived 10 times more than other treatments when mixed with skim milk. Although the skim milk increased the stability of the phages, the control efficacy was not effective. However, the phage mixtures reduced the occurrence of BLB when they were treated with Tecloftalam WP or Acibenzolar-S-methyl simultaneously. The results indicated that the Xoo phages could be used as an alternative control method to increase the control efficacy and reduce the use of agrochemicals.