• Title/Summary/Keyword: Phage

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Mixture of Edwardsiella tarda specific Bacteriophage and Bacillus subtilis KM-1enhanced bactericidal activity against Edwardsiella tarda (Edwardsiella tarda의 특이 Bacteriophage와 Bacillus subtilis KM-1혼합액이 Edwardsiella tarda 에 미치는 항균효과)

  • Baek, Min Suk;Hwang, Yo Sep;Choi, Sanghoon
    • Journal of fish pathology
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    • v.26 no.3
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    • pp.185-191
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    • 2013
  • The present study was performed to investigate an antibacterial activity of specific bacteriophage (phage) and Bacillus subtilis KM-1 (B. subtilis) mixture against Edwardsiella tarda (E. tarda). An appropriate number of phage showing the most effective antibacterial activity was $2{\times}10^5$ PFU/ml with $1{\times}10^7$ CFU/ml of B. subtilis 36 h post incubation. On the other hand, B. subtilis showed a dose dependant manner in inducing antibacterial activity in the presence of phage ($2{\times}10^5$ PFU/ml). The phage and B. subtilis mixture showed higher antibacterial activity against E. tarda than phage or B. subtilis only. These results suggest that the phage and B. subtilis mixture could be utilized as an alternative to antibiotics in the control of fish diseases caused by E. tarda.

Cloning and Expression of K11 Phage RNA Polymerase (K11 RNA 중합효소의 Cloning 및 발현)

  • Lee, Sang-Soo
    • The Journal of Natural Sciences
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    • v.9 no.1
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    • pp.19-24
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    • 1997
  • Using the PCR(polymerase chain reaction method), gone 1 of phage K11 coding for K11 phage RNA polymerase has been cloned and expressed under the control of lac promoter. K11 phage RNA polymerase was conventionally purified through the DEAE-sephacel and Affigel blue column chromatographies. The 0.2-0.3 M $NH_4Cl$ fractions of DAEA-sephacel column chromatography showed K11 phage RNA polymerase activity and further purification with Affigel blue column chromatography showed nearly single protein band on SDS-polyacryl amide gel. K11 phage RNA polymerase, which is one of the T7 group phage RNA polymerase (E. coil phage T7, T3 and Salmonella tyhimurium phage SP6 RNA polymerase), shares high degrees of homology with the other T7 group phage RNA polymerase. Previously we constructed T7 and SP6 promoter variants and revealed promoter specificity of T7 and SP6 RNA polymerase (Lee and Kang, 1993). To investigate the promoter specificity of K11 RNA polymerase in vitro K11 promoter activity was measured with SP6 promoter variants. The SP6 promoter variant share highest degrees of sequence homology with K11 promoter sequence show strongest promoter activity.

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New Temperate Bacteriophages of Lactococcus garvieae (Lactococcus garvieae의 새로운 용원성파아지)

  • Park, Kyun-Hyun;Muroga, Kiyokuni;Jeong, Hyun-Do
    • Journal of fish pathology
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    • v.11 no.2
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    • pp.137-141
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    • 1998
  • Temperate phages were effectively induced from presumptive lysogenized cells of 96 strains out of 111 strains of L. garvieae No. 44 strains (phage type B) as the host cell. Similar cultures in distilled water-based TSB did not induce lytic infection in these cells. These temperate phages were also effectively induced by ultraviolet irradiation. All phages isolated were lytic only to L. garvieae No. 44 strain and the lytic nature was different from those of PLgY, PLgW, and PLgS. The virions appeared extracellularly after 1h of induction culture and increased in number until reaching the maximum of $10^6$ PFU/ml after 12h. This phage production was lower than that ($10^{10}$ PFU/ml) of the virulent phage.

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Evidence to Support the Therapeutic Potential of Bacteriophage Kpn5 in Burn Wound Infection Caused by Klebsiella pneumoniae in BALB/c Mice

  • Kumar, Seema;Harja, Kusum;Chhibber, Sanjay
    • Journal of Microbiology and Biotechnology
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    • v.20 no.5
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    • pp.935-941
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    • 2010
  • The emergence of antibiotic-resistant bacterial strains is one of the most critical problems of modern medicine. Bacteriophages have been suggested as an alternative therapeutic agent for such bacterial infections. In the present study, we examined the therapeutic potential of phage Kpn5 in the treatment of Klebsiella pneumoniae B5055-induced burn wound infection in a mouse model. An experimental model of contact burn wound infection was established in mice employing K. pneumoniae B5055 to assess the efficacy of phage Kpn5 in vivo. Survival and stability of phage Kpn5 were evaluated in mice and the maximum phage count in various organs was obtained at 6 h and persisted until 36 h. The Kpn5 phage was found to be effective in the treatment of Klebsiella-induced burn wound infection in mice when phage was administered immediately after bacterial challange. Even when treatment was delayed up to 18 h post infection, when all animals were moribund, approximately 26.66% of the mice could be rescued by a single injection of this phage preparation. The ability of this phage to protect bacteremic mice was demonstrated to be due to the functional capabilities of the phage and not due to a nonspecific immune effect. The levels of pro-inflammatory cytokines (IL-$1{\beta}$ and TNF-${\alpha}$) and anti-inflammatory cytokines (IL-10) were significantly lower in sera and lungs of phage-treated mice than phage untreated control mice. The results of the present study bring out the potential of bacteriophage therapy as an alternate preventive approach to treat K. pneumoniae B5055-induced burn wound infections. This approach not only helps in the clearance of bacteria from the host but also protects against the ensuing inflammatory damage due to the exaggerated response seen in any infectious process.

Dynamics Behavior of Phage-Host System Related to Microlunatus phosphovorus in Activated Sludge with Host Inoculation

  • Lee, Sang-Hyon;Otawa, Kenichi;Onuki, Motoharu;Satoh, Hiroyasu;Mino, Takashi
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1518-1522
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    • 2006
  • In the present study, it was observed how the phage-host system that is naturally reproduced in activated sludge is affected by the host inoculation. The system of Microlunatus phosphovorus and its phages was selected as the phage-host system native to an activated sludge system operated for 19 days under sequencing anaerobic-aerobic conditions with glutamate as the main carbon source. The phage-host system related to M. phosphovorus was monitored by plaque assay for the phages and by fluorescent in situ hybridization (FISH) for the bacterial host. In addition, the whole phage structure was also monitored by pulsed-field gel electrophoresis (PFGE). During the first 9 days, the phage-host system was more or less steady at approx. 9% (FISH/ DAPI) for M. phosphovorus and approx. 10,000 PFU/ml for its lytic phages. Microlunatus phosphovorus JCM9379 was inoculated into the activated sludge on day 10. Right after the inoculation, M. phosphovorus was approx. 24% (FISH/DAPI) whereas its lytic phages dropped down to approx. 500 PFU/ ml. After the host inoculation (within 9 days), however, the phage-host system eventually reverted to its original level in each population. On the other hand, the whole phage structure was not significantly changed by M. phosphovorus inoculation but stable throughout the process operation. Only the minor change that four phage groups gradually became abundant after the host inoculation was observed.

Biocontrol Potential of a Lytic Bacteriophage PE204 against Bacterial Wilt of Tomato

  • Bae, Ju Young;Wu, Jing;Lee, Hyoung Ju;Jo, Eun Jeong;Murugaiyan, Senthilkumar;Chung, Eunsook;Lee, Seon-Woo
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1613-1620
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    • 2012
  • Bacterial wilt caused by Ralstonia solanacearum is a devastating disease of many economically important crops. Since there is no promising control strategy for bacterial wilt, phage therapy could be adopted using virulent phages. We used phage PE204 as a model lytic bacteriophage to investigate its biocontrol potential for bacterial wilt on tomato plants. The phage PE204 has a short-tailed icosahedral structure and double-stranded DNA genome similar to that of the members of Podoviridae. PE204 is stable under a wide range of temperature and pH, and is also stable in the presence of the surfactant Silwet L-77. An artificial soil microcosm (ASM) to study phage stability in soil was adopted to investigate phage viability under a controlled system. Whereas phage showed less stability under elevated temperature in the ASM, the presence of host bacteria helped to maintain a stable phage population. Simultaneous treatment of phage PE204 at $10^8$ PFU/ml with R. solanacearum on tomato rhizosphere completely inhibited bacterial wilt occurrence, and amendment of Silwet L-77 at 0.1% to the phage suspension did not impair the disease control activity of PE204. The biocontrol activities of phage PE204 application onto tomato rhizosphere before or after R. solanacearum inoculation were also investigated. Whereas pretreatment with the phage was not effective in the control of bacterial wilt, post-treatment of PE204 delayed bacterial wilt development. Our results suggested that appropriate application of lytic phages to the plant root system with a surfactant such as Silwet L-77 could be used to control the bacterial wilt of crops.

Characterization of Phage Behaviors Against Antibiotic-Resistant Salmonella Typhimurium

  • Easwaran, Maheswaran;Ahn, Juhee
    • Journal of Food Hygiene and Safety
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    • v.35 no.6
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    • pp.602-606
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    • 2020
  • This study was designed to investigate the dynamic behaviors of phages against Salmonella enterica subsp. enterica serovar Typhimurium ATCC 19585 (STWT), S. Typhimurium KCCM 40253 (STKCCM), ciprofloxacin-induced S. Typhimurium ATCC 19585 strains (STCIP), and S. Typhimurium CCARM 8009 (STCCARM). Phages, including PBST-10, PBST-13, PBST-32, PBST-35, P-22, and P-22 B1 had narrow host ranges. The adsorption rates of all phages ranged from 47 to 85%, 58 to 95%, and 61 to 93%, respectively, against STWT, STKCCM, and STCIP, while the lowest adsorption rates ranged from 14 to 36% against STCCARM. The phage burst sizes were from 43 to 350, 37 to 530, 66 to 500, and 24 to 500 plaque-forming units (PFUs) per infected STWT, STKCCM, STCIP, and STCCARM, respectively. The STCIP strain was effectively inhibited by all phages at the early of incubation period. These results provide useful information for better understanding the phage behaviors against antibiotic-resistant and antibiotic-sensitive pathogens.

The distribution and Morphology of Bacillus thuringiensis Phages in Korea (Bacillus thuringiensis phage의 분포와 형태에 관한 연구)

  • Rhee, Tai-Woo;Ahn, Kyung-Joon
    • Applied Microscopy
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    • v.12 no.1
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    • pp.33-40
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    • 1982
  • Several phages of Bacillus thuringiensis distributed in Korea were isolated. The distribution and morphological characteristics of phages were studied. The results are as follows; 1. The isolated phages were highly specific for Bacillus thuringiensis var. thuringiensis. They were classified as YM series phages and designated as phage YM-1, YM-2 and YM-3 according to their morphological characteristics. 2. Most of these YM series phages were isolated from compost including domestic animal dung and soil under sewage. 3. The YM-1 phage was similar to Bacillus subtilis ${\phi}25$ in morphology. It has 94nm x 86nm head, contractile tail sheath and base plate with four cornered structure. 4. The YM-2 phage was similar to Bacillus subtilis GA-1 phage in morphology. It had 70nm x 56nm head and tail without contractile tail sheath. 5. The YM-3 phage was similar to Bacillus subtilis ${\phi}29$ phage. It had 56nm x 43nm head and tail with distal enlargement.

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P22-Based Challenge Phage Constructs to Study DNA-Protein Interactions between the $\sigma$54-Dependent Promoter, dctA, and Its Transcriptional Regulators

  • Kim, Euhgbin;Kim, Daeyou;Lee, Joon-Haeng
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.176-179
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    • 2000
  • A challenge phage system was used to study the DNA-protein interaction between C4-dicarboxylic acid transport protein D(DCTD) or $\sigma$54, and a $\sigma$54 -dependent promoter, dctAp. R. meliloti dctA promoter regulatory region replaced the Omnt site on the phage. S. typhimurium strains overproducing either DCTD or $\sigma$54 directed this challenge phage towards lysogency, indicating that DCTD or E$\sigma$54 recognized the dctA promoter on the phage and repressed transcription of the ant gene. These challenge phage constructs will be useful for examining interactions between DCTD(or $\sigma$54) and the dctA promoter region.

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Genomic analysis of WCP30 Phage of Weissella cibaria for Dairy Fermented Foods

  • Lee, Young-Duck;Park, Jong-Hyun
    • Food Science of Animal Resources
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    • v.37 no.6
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    • pp.884-888
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    • 2017
  • In this study, we report the morphogenetic analysis and genome sequence of a new WCP30 phage of Weissella cibaria, isolated from a fermented food. Based on its morphology, as observed by transmission electron microscopy, WCP30 phage belongs to the family Siphoviridae. Genomic analysis of WCP30 phage showed that it had a 33,697-bp double-stranded DNA genome with 41.2% G+C content. Bioinformatics analysis of the genome revealed 35 open reading frames. A BLASTN search showed that WCP30 phage had low sequence similarity compared to other phages infecting lactic acid bacteria. This is the first report of the morphological features and complete genome sequence of WCP30 phage, which may be useful for controlling the fermentation of dairy foods.