• Title/Summary/Keyword: phage inactivation

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Ozone Inactivation of Bacteriophage f2 (오존에 의한 bacteriophage f2의 살균작용(殺菌作用))

  • Kim, Chi-Kyung
    • Applied Microscopy
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    • v.11 no.1
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    • pp.29-38
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    • 1981
  • Bacteriophage f2 were treated with ozone at various concentrations for 20 minutes. The inactivation kinetics of f2 phage were examined during ozonation. In order to study the mode of action of ozone on the phage f2, absorption of the phage to the host pili was meassured by utilyzing radioactivity of tritium incorporated into the phage RNA. Sucrose density gradient analysis and electron microscopy were also used to prove the mechanism of ozone inactivation of the phage. Strucural proteins of the phage were broken by ozonation into many protein subunits. The extent of phage breakage was proportional to ozone concentration and reaction time. Percent decrease of the phage absorption to the host pili was coincident with the rate of ozone inactivation of the phage. Ozone inactivation of bacteriophage f2 was shown to be caused by the breakage of the structural protein and blockage of the phage absorption to the host pili.

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Inactivation of Bacteriophage f2 with Chlorine (염소에 의한 bacteriophage f2의 살균작용)

  • Chi Kyung KIM;Kyung Hee MIN
    • Korean Journal of Microbiology
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    • v.16 no.2
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    • pp.62-70
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    • 1978
  • Chlorine was used for inactivation of bacteriophage f2 at pH 5.5, 7.5, and 10.0 at $10^{\circ}C$. The inactivation rate phage with chlorine varied depending on the pH value and reaction time. Hypochlorous acid appeared to be the major species of free chlorine for the inactivation. Suevival of the phage treated with chlorine and infectivity of the RNA extracted from the chlorinated phage were examined. The RNA extracted from untreatd phage was chlorinated and its infectivity was assayed. All three samples showed similar rates of inactivation at pH 5.5 and 7.5, but the naked RNA was more susceptible to chlorine at pH 10.0. The rate of inactivation was compared naked RNA was more susceptible to chlorine at pH 10.0. The rate of inactivation was compared with specific and non-specific attachment of the phasge f2. The specific attachment of the phage increased after the phage had been inactivated by extended chlorination. Chlorine may penetrate to the becteriophage f2 by altering the structural integrity of the protein coat, but the main target of free chlorine for inactivation of the phage appeared to be the phage RNA.

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The Inactivation Effects of UV Light on Bacteriophage f2 (박테리오파지 f2에 대한 자외광선의 살균효과)

  • Kim, Chi-Kyung;Quae Chae
    • Microbiology and Biotechnology Letters
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    • v.11 no.3
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    • pp.155-161
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    • 1983
  • The effects of ultraviolet light on bacteriophage f2 were investigated to determine the inactivation kinetics and its mechanism. The 260nm light showed a little higher inactivation rate than the one of 300 nm. In this work, our main concern was whether structural and/or conformational changes in the protein capsid could occur by UV irradiation. The inactivation for the first 20 minutes irradiation was rapid with a loss of about 4 logs and followed by a slower rate during the next 40 minutes with no survival noted in the samples irradiated for 90 minutes or longer. The structural change of the protein capsid was examined by optical spectroscopic techniques and electron microscopy. The absorption spectra of the UV irradiated phages showed no detectable differences in terms of the spectral shape and intensity from the control phage. However, the fluorescence emission spectroscopic data, i.e. 1) fluorescence quenching of tryptophan residues upon irradiation of 300 nm light, 2) enhancement of fluorescence emission of ANS (8-aniline-1-naphthalene sulfonate) bound to the intact phages compared to the one in the UV-treated phages, and 3) decrease of energy transfer efficiency from tryptophan to ANS in the UV-treated samples, presented remarkable differences between the intact and UV-treated phages. Such a structural alteration was also observed by electron microscopy The UV-treated phages appeared to be broken and empty capsids. Therefore, the inactivation of the bacteriophage f2 by UV irradiation is thought to be attributed to the structural change in the protein capsid as well as damage in the viral RNA by UV irradiation.

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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.

Applicability Investigation of E.coli, RNA and DNA Bacteriophages for Possible Indicator Microorganisms Based on the Inactivation Effectiveness by UV (UV 불활성화 효과에 의거한 E.coli, RNA 및 DNA 박테리오파지의 대체 지표 미생물로서의 적용성 검토)

  • Kim, Il-Ho;Wahid, Marfiah AB;Tanaka, Hiroaki
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.11
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    • pp.1063-1068
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    • 2010
  • This study compared UV and UV/$H_2O_2$ inactivation of E.coli, a possible indicator microorganism for fecal contamination of water, and $Q{\ss}$ phage, an indicator for pathogenic viruses. UV inactivation of $Q{\ss}$, T4 and lambda phages in actual secondary effluent was investigated, too. As a result, similar inactivation efficiency between $Q{\ss}$ phage and E.coli was observed during UV treatment, while $Q{\ss}$ phage showed higher resistance to UV/$H_2O_2$ than E.coli. $Q{\ss}$ phage resistance to UV or UV/$H_2O_2$ does not reflect those of all pathogenic viruses. However, the result tells that the use of E.coli inactivation efficiency in evaluating microbiological safety of water could not always ensure the sufficient safety from pathogenic viruses. Meanwhile, $Q{\ss}$ phage showed less resistance to UV than T4 and lambda phages, indicating that the use of $Q{\ss}$ phage as an indicator virus may bring insufficient disinfection effectiveness by causing the introduction of lower UV dose than required. Consequently, it can be thought that T4 or lambda phages would be more desirable indicators in ensuring the sufficient disinfection effectiveness for various pathogenic viruses.

Studies on the Phage of Vibrio parahaemolyticus (Vibrio parahaemolyticus의 Phage에 관한 연구)

  • Ju, Jin-Woo;Lee, Ghee-Hee;Kim, Il
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.1
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    • pp.61-70
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    • 1987
  • Authors have isolated phages of V. parahaemolyticus from shellfish and investigated some of their characteristics. The results obtained were as follows: Twenty-three phage strains(9.2%) out of 250 specimens were isolated. Plaques of phages were small, clear or turbid and $0.5{\sim}1.5mm$ in diameter. The electron micrographs of K3 phages showed two morphology; one was a hexagonal head about 105nm with a tail about 12nm, the other was a hexagonnal head about 60nm with a tail about 25nm. The host ranges of pahges were limited to V. parahaemolyticus strains and there appeared to be no relationship between the K serotypes of V. parahaemolyticus strains and the host ranges of the phage isolates. The adsorption rate of phages were more than 80% for $10{\sim}15$ minutes, the inactivation rate at $60^{\circ}C$ was more than 99% for $40{\sim}45$ minutes. The pH stability range was between 6.0 and 8.0. The inactivation rate of phages by UV irradiation was more than 99% for $45{\sim}75$ seconds.

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Classification of Bacteriophage of Lactobacillus Casei Strain S-1 (Lactobacillus casei S-1 균주의 Bacteriophage 분류)

  • Kim, Young-Ki;Baek, Young-Jin;Bae, Hyung-Seok;Yoo Min
    • Microbiology and Biotechnology Letters
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    • v.13 no.3
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    • pp.265-271
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    • 1985
  • The classification of bacteriophage could be followed by several criteria. In this study three criteria were used for classification of Lactobacillus casei bacteriophag. In serological classification. antiserum was prepared by rabbit and used for classification. The inactivation effect of phage by antiserum was exponential and L. casei phage was classified in to three serological groups by inactivation rate (K-values). The Lac Y group was proved as a new serological group but the Lac J and Lac S group were shown the same results as previous reports. From the comparison of restriction enzyme pattern of phage DNA, Lac J group was divided into four sub-groups. According to the difference of host range, Lac J-II group was further subdivided into three groups. These results were shown that L. casei strains S-1 bacteriophage was classified into 8 sub-groups. The phage YK of Lac Y group was shown to consist of a icosahedral head about 95nm in diameter, a contractile tail about 150nm in length and 20nm in width. The tail of YK phage is composed of stacked disks(4nm repeat)and a hexagonal baseplate. The molecular weight of YK phage DNA was approximately 85.6 Mdalton.

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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.

Effect of Iron(II)-ascorbate Complex on Protein and DNA of Phages (파아지 단백질 및 DNA에 대한 2가철-아스코르빈산착체의 영향)

  • Lho, Il-Hwan;Murata, Akira
    • Korean Journal of Food Science and Technology
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    • v.25 no.1
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    • pp.46-51
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    • 1993
  • The inactivating effect of iron(II)-ascorbate complex (Fe-Asc) on various phages was previously reported. This paper describes the molecular target in the phage virion attacked by Fe-Asc. The effect of Fe-Asc on protein was investigated with bovine serum albumin and the structural protein of phage J1. There were no differences in the SDS-polyacrylamide gel electrophoresis (patterns of these two proteins when either they were treated) with Fe-Asc or not. Also, there were no changes in the amino acid composition and ultraviolet spectrum of the proteins. The effects of Fe-Asc on DNA was investigated with pUC18 DNA, M13mpB DNA and ${\lambda}$ DNA as well as DNA from phage J1. Fe-Asc caused initially nicking of the subsequently form of pUC18 DNA to yield the open circular form and then subsequently the linear form. Strand breaks were also confirmed with M13mp8 DNA and ${\lambda}$ DNA as well as J1 DNA. The results indicate that the strand breaks in phage DNA could be responsible for the inactivation of phages by Fe-Asc.

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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.