• Title/Summary/Keyword: lytic phage

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Characterization of Endolysin LysECP26 Derived from rV5-Like Phage vB_EcoM-ECP26 for Inactivation of Escherichia coli O157:H7

  • Park, Do-Won;Park, Jong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.10
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    • pp.1552-1558
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    • 2020
  • With an increase in the consumption of non-heated fresh food, foodborne shiga toxin-producing Escherichia coli (STEC) has emerged as one of the most problematic pathogens worldwide. Endolysin, a bacteriophage-derived lysis protein, is able to lyse the target bacteria without any special resistance, and thus has been garnering interest as a powerful antimicrobial agent. In this study, rV5-like phage endolysin targeting E. coli O157:H7, named as LysECP26, was identified and purified. This endolysin had a lysozyme-like catalytic domain, but differed markedly from the sequence of lambda phage endolysin. LysECP26 exhibited strong activity with a broad lytic spectrum against various gram-negative strains (29/29) and was relatively stable at a broad temperature range (4℃-55℃). The optimum temperature and pH ranges of LysECP26 were identified at 37℃-42℃ and pH 7-8, respectively. NaCl supplementation did not affect the lytic activity. Although LysECP26 was limited in that it could not pass the outer membrane, E. coli O157: H7 could be effectively controlled by adding ethylenediaminetetraacetic acid (EDTA) and citric acid (1.44 and 1.14 log CFU/ml) within 30 min. Therefore, LysECP26 may serve as an effective biocontrol agent for gram-negative pathogens, including E. coli O157:H7.

A comparison of lytic activity of swine and chicken phages with Staphylococcus hyicus of swine, chicken and cattle origin (돼지, 닭 및 소유래 Staphylococcus hyicus에 대한 swine phage와 chicken phage의 용균능 비교)

  • Park, Cheong-Kyu;Jang, Han-Nae;Kwon, Young-Sam
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.87-94
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    • 2003
  • The lytic activity of the set of swine and chicken phages which were derived from lysogenic Staphylococcus hyicus strains of swine and chicken origin was compared by means of S. hyicus isolated from swine, chicken and cattle. Of the 80 strains each from swine and chicken, 71 (88.8%) of strains from swine and all the strains of chicken origin were found to be lysogenic. Swine phages showed wider range of lytic activity to the examined strains than that of chicken phages. Using chicken phages at $100{\times}routine$ test dilution (RTD), 25.0%, 85.6% and 50.0% of swine, chicken and bovine strains were lysed, respectively. However, when the set of swine phages was used at $100{\times}RTD$, higher frequency of the typable strains was found in strains of swine and chicken origin (73.8% and 90.2%). Phage F12 and L16 from chicken set were found to be highly active with chicken and bovine strains. On the contrary, all the swine strains were completely resistant to lysis by the two phages at $100{\times}RTD$. Thirteen (12.5%) of 104 S. aureus strains, 1 (1.8%) of 55 S. simudance strains, 31 (58.5%) of 53 S. chromo genes strains, and none of 31 strains of other coagulase-negative Staphylococcus species isolated from bovine mastitis were typable with the set of swine phages.

Biocontrol of Pectobacterium carotovorum subsp. carotovorum Using Bacteriophage PP1

  • Lim, Jeong-A;Jee, Samnyu;Lee, Dong Hwan;Roh, Eunjung;Jung, Kyusuk;Oh, Changsik;Heu, Sunggi
    • Journal of Microbiology and Biotechnology
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    • v.23 no.8
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    • pp.1147-1153
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    • 2013
  • Pectobacterium carotovorum subsp. carotovorum (formerly Erwinia carotovora subsp. carotovora) is a plant pathogen that causes soft rot and stem rot diseases in several crops, including Chinese cabbage, potato, and tomato. To control this bacterium, we isolated a bacteriophage, PP1, with lytic activity against P. carotovorum subsp. carotovorum. Transmission electron microscopy revealed that the PP1 phage belongs to the Podoviridae family of the order Caudovirales, which exhibit icosahedral heads and short non-contractile tails. PP1 phage showed high specificity for P. carotovorum subsp. carotovorum, and several bacteria belonging to different species and phyla were resistant to PP1. This phage showed rapid and strong lytic activity against its host bacteria in liquid medium and was stable over a broad range of pH values. Disease caused by P. carotovorum subsp. carotovorum was significantly reduced by PP1 treatment. Overall, PP1 bacteriophage effectively controls P. carotovorum subsp. carotovorum.

Characteristics of Structural Proteins of Synechococcus sp. Cyanophage (Synechoscoccus sp. cyanophage 구조단백질의 특성)

  • Kim, Seung-Won;Kim, Min;Leem, Mi-Hyea;Choi, Yong-Keel
    • Korean Journal of Microbiology
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    • v.33 no.4
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    • pp.242-246
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    • 1997
  • The protein profile of Synechococcus sp. cyanophage was investigated employing SDS-PAGE. The phage appears to be composed of two major proteins of 97 and 52 kDa and at least seven minor proteins of 70, 65, 60, 40, 35, 28, and 6 kDa. It seems that each subunit is combined to form a multimer although any disulfide bond does not exist in the phage structure. Lytic activity of the phage particle against cell wall was detected around the 52 kDa on renaturing SDS-PAGE using heat-killed Micrococcus luteus cells as substrate. The activity has the optimal pH between 9 and 10, and slightly inhibited by EDTA.

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Isolation, Physiological Characterization of Bacteriophages from Enhanced Biological Phosphorus Removal Activated Sludge and Their Putative Role

  • Lee, Sang-Hyon;Satoh, Hiroyasu;Katayama, Hiroyuki;Mino, Takashi
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.730-736
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    • 2004
  • This study aims at characterizing the bacteriophages isolated from activated sludge performing enhanced biological phosphorous removal (EBPR) to understand the interactions between the phage-host system and bacterial community. Sixteen bacterial isolates (E1-E16) were isolated as host bacterial strains from EBPR activated sludge for phage isolation. Forty bacteriophages based on their plaque sizes (2 plaques on E4, 4 on E8, 11 on E10, 5 on E14, 18 on E16) were obtained from filtered supernatant of the EBPR activated sludge. Each bacteriophage did not make any plaque on bacterial strains tested in this study except on its own host bacterial strain, respectively, indicating that the bacteriophages are with narrow host specificity. However, fourteen of the forty bacteriophages obtained in this study lost their virulent ability even on their own host bacteria. All of the lytic phages showed similar one-step growth patterns and had long latent period (about 9 hours) to reproduce their phage particles in their host bacterial cells. On the other hand, their probable burst sizes (6 to 48 per host cell) were large enough to actively lyse their host bacterial cells. Therefore, it could be implied that bacteriophages are also important members of the microbial community in EBPR activated sludge, and lytic phages directly decrease the population size of their host bacterial groups in EBPR activated sludge by lysis.

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 Characteristics of Bacteriophage-resistant Lactococcus lactis subsp. cremoris ATCC 11602-A1 (Lactococcus lactis subsp. cremoris ATCC 11602의 Bacteriophage 내성균주 A1의 특성에 관한 연구)

  • Lee, Chun-Hwa;Kang, Kuk-Hee;Bae, In-Hyu
    • Microbiology and Biotechnology Letters
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    • v.21 no.4
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    • pp.293-298
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    • 1993
  • The ppage resistance mechanism of Lactococcus lactis subsp. cremoris ATCC 11602-A1 was investigated. When parent and A1 were incubated at 30 and 40$^{\circ}C$, A1 grew well and multiplication of phage(MOI=1)on A1 slightly occurred at 40$^{\circ}C$ in contrast with parent. There was a great difference of proteolytic activity between parent and A1, irrespective of the temperature. As a result of ADS treatment oon culture broth, survival rate of A1 was 27% at the lethal concentration of parent and adsorption rate of phage was increased to 95~97%, which was considered to come from the exposure of phage receptor site masked by an unknown component. These results suggest that acridine orange (AO) treatment leads to the modification of cell wall, conferring resistance to high temperature and lytic phage. No change in plasmid profiles of A1 at 30 and 40$^{\circ}C$ were found, which suggests that plasmid is not relative to temperature-resistance of A1.

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The Characteristics, Detection and Control of Bacteriophage in Fermented Dairy Products (발효유제품에서 박테리오파지의 특성, 검출과 제어)

  • Ahn, Sung-Il;Azzouny, Rehab A.;Huyen, Tran Thi Thanh;Kwak, Hae-Soo
    • Food Science of Animal Resources
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    • v.29 no.1
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    • pp.1-14
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    • 2009
  • This study was to review the classification, detection and control of bacteriophage in fermented dairy products. Bacteriophage has lytic and/or lysogenic life cycles. Epidemiologically speaking, detected major phages are c2, 936 and p335. Among them p335 has been the largest concern in dairy industry. Traditionally, various analytical technologies, such as spot, starter activity, indicator test, ATP measurement and conductimetric analysis, have been used for the phage detection. In recent years, advanced methods such as flow cytometric method, petrifilm, enzyme linked immunosorbent assay (ELISA) and multiflex PCR diagnostic kit have been deveoloped. The phage contamination has been controlled by using heat, high-pressure treatment, and the combinations of heat and pressure, and/or chemical. Also some starter cultures with phage-resistant character have been developed to minimize the concentration of phages in dairy product. Bacteriophage inhibition media such as calcium medium was also mentioned. To prevent the contamination of bacteriophage in dairy industry, further researches on the detection and control of phage, and phage resistant starters are necessary in the future.

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

  • Younghee Kim
    • Journal of Environmental Science International
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    • v.1 no.2
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    • pp.99-103
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    • 1992
  • The bacteriophages lytic for Vibrio furnissi, Vibrio furniulis and Vibrio parahemolyticus were isolated from fish gills and shellfish. Nucleic acid of bacteriophage was prepared and restriction endonuclease profile was compared. All isolates contained deoxyribonucleic acid. V. fumissi bacteriophage from fish gills showed 2 bands with Bgl II, 1 with Pst, 3 with Hind III, 1 with Bm HI and 2 with EcoR I. V Puuialis phage represented 7 fragments with Bgl II, 1 with Pst, 4 with Hind III, and 2 with EcoR I. V parhemolyticn produced 13 sites with Hind III and 4 sites with EcoR I. 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. Key words: Bacteriophage, Vibrio furnissi, Vibrio fluvialis, Vibrio pnrahemolyticus, Deoxyribonucleic acid, Pst, Bam HI, Hind III, EcoR I, Bgl II.

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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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