• Title/Summary/Keyword: Phages

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CTX Prophages in Vibrio cholerae O1 Strains

  • Kim, Eun Jin;Lee, Dokyung;Moon, Se Hoon;Lee, Chan Hee;Kim, Dong Wook
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.725-731
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    • 2014
  • The classical biotype strains of the Vibrio cholerae O1 serogroup harbor the biotype-specific cholera-toxin encoding phage (CTX) $CTX^{cla}$, and the El Tor biotype strains contain CTX-1. Although the classical biotype strains have become extinct, a remnant of classical CTX phage is transferred to the El Tor biotype strains. The prototype El Tor strains, which produce the biotype-specific cholera toxin, are now being replaced by atypical El Tor variant strains producing classical biotype cholera toxin. The genome sequences of the CTX phages in atypical El Tor strains indicate that the CTX phages in atypical El Tor strains are a mosaic of $CTX^{cla}$ and CTX-1. Before the emergence of atypical El Tor stains in the early 1990s, unusual pre-seventh pandemic strains were isolated in the US Gulf Coast between 1973 and 1986. These strains have characteristics of atypical El Tor strains since they are El Tor biotype strains containing $CTX^{cla}$, yet the genome sequence of this CTX phage indicates that it is different from $CTX^{cla}$ and is therefore classified separately as $CTX^{US\;Gulf}$.

Therapeutic Effects of Bacteriophages Against Salmonella gallinarum Infection in Chickens

  • Hong, Sung Sik;Jeong, Jipseol;Lee, Jinju;Kim, Suk;Min, Wongi;Myung, Heejoon
    • Journal of Microbiology and Biotechnology
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    • v.23 no.10
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    • pp.1478-1483
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    • 2013
  • In this study the isolation and characterization of three bacteriophages (ST4, L13, and SG3) infecting Salmonella gallinarum were carried out. They were further tested for their in vivo efficacy in phage therapy. All three phages belong to the Siphoviridae family with isometric heads and non-contractile tails. They have a broad host range among serovars of Salmonella enterica. The burst sizes were observed to be 1670, 80, and 28 for ST4, L13, and SG3, respectively. The in vivo efficacy of the phages was tested in chickens. Layer chickens were challenged with S. gallinarum, whereas contact chickens were cohabited without direct challenge. Each bacteriophage was orally inoculated in the form of feed additives. Mortality was observed and S. gallinarum was periodically re-isolated from the livers, spleens, and cecums of the chickens. Bacterial re-isolation from the organs and mortality decreased significantly in both challenged and contact chickens treated with the bacteriophages compared with untreated chickens serving as the control. The three bacteriophages may be effective alternatives to antibiotics for the control of fowl typhoid disease in chickens.

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.

Identification of Dinitrotoluene Selective Peptides by Phage Display Cloning

  • Jang, Hyeon-Jun;Na, Jung-Hyun;Jin, Bong-Suk;Lee, Won-Kyu;Lee, Woong-Hee;Jung, Hyun-Jin;Kim, Seok-Chan;Lim, Si-Hyung;Yu, Yeon-Gyu
    • Bulletin of the Korean Chemical Society
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    • v.31 no.12
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    • pp.3703-3706
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    • 2010
  • Biomolecules specific to explosives can be exploited as chemical sensors. Peptides specific to immobilized dinitrotoluene (DNT) were identified using a phage display library. A derivative of DNT that contained an extended amine group, 4-(2,4-dinitrophenyl)butan-1-amine, was synthesized and immobilized using a self-assembled monolayer surface on gold. Filamentous M13 phages displaying random sequences of 12-mer peptides specific to the immobilized DNT-derivate were isolated from the M13 phage library by biopanning. A common peptide sequence was identified from the isolated phages and the synthesized peptides showed selective binding to DNT. When the peptide was immobilized on a quartz crystal microbalance (QCM) chip, it showed a binding signal to DNT, while toluene barely showed significant binding to the QCM chip. These results demonstrate that peptides screened by biopanning against immobilized DNT can be useful for quick and accurate detection of DNT.

Development of a Single Chain Antibody Using a Phage Display Cloning Method for the Detection of 2,4-Dinitrotoluene

  • Na, Jung-Hyun;Joo, Man-Seok;Lee, Won-Kyu;Shim, Hyunbo;Lim, Si-Hyung;Jung, Sang Taek;Yu, Yeon Gyu
    • Bulletin of the Korean Chemical Society
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    • v.34 no.2
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    • pp.460-464
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    • 2013
  • Single-chain variable fragments of antibodies (scFv) specific to 2,4-dinitrotoluene (DNT) were isolated from a phage library displaying synthetic human scFv fragments with 6 diversified complementary determining regions (CDRs). A DNT derivative that contained an extended amine group was synthesized and conjugated to the NHS-group that was linked to magnetic beads. Phages specific to the immobilized DNT derivatives were isolated from the library after 4 rounds of sequential binding and elution processes. The displayed scFv fragments from the isolated phages showed consensus CDR sequences. One DNT-specific scFv was expressed in E. coli and purified using Ni-affinity chromatography. The purified DNT-specific scFv binds specifically to the immobilized DNT-derivative with $K_D$ value of $6.0{\times}10^{-7}$ M. The scFv and DNT interaction was not disrupted by the addition of 4-nitrotoluene or benzoic acid. These data demonstrate that the screened scFv from the phage displayed library could be used for selective and sensitive detection of explosives such as TNT.

Characterization of Phage-Resistant Strains Derived from Pseudomonas tolaasii 6264, which Causes Brown Blotch Disease

  • Yun, Yeong-Bae;Han, Ji-Hye;Kim, Young-Kee
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2064-2070
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    • 2018
  • Pseudomonas tolaasii 6264 is a representative strain that causes bacterial blotch disease on the cultivated oyster mushroom, Pleurotus ostreatus. Bacteriophages are able to sterilize the pathogenic P. tolaasii strains, and therefore, they can be applied in creating disease-free mushroom cultivation farms, through a method known as "phage therapy". For successful phage therapy, the characterization of phage-resistant strains is necessary, since they are frequently induced from the original pathogenic bacteria in the presence of phages. When 10 different phages were incubated with P. tolaasii 6264, their corresponding phage-resistant strains were obtained. In this study, changes in pathogenic, genetic, and biochemical characteristics as well as the acquired phage resistance of these strains were investigated. In the phylogenetic analyses, all phage-resistant strains were identical to the original parent strain based on the sequence comparison of 16S rRNA genes. When various phage-resistant strains were examined by three different methods, pitting test, white line test, and hemolytic activity, they were divided into three groups: strains showing all positive results in three tests, two positive in the first two tests, and all negative. Nevertheless, all phage-resistant strains showed that their pathogenic activities were reduced or completely lost.

Development of an Antimicrobial Edible Coating to Improve Microbial Safety of Wild Vegetables (산채류의 미생물 안전성 증진을 위한 가식성 항균 코팅 개발 및 적용)

  • Jieun Choi;Bong Sun Kim;Yoonjee Chang;Jaewoo Bai
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.30 no.1
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    • pp.15-21
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    • 2024
  • The edible coating system, consisting of gelatin and bacteriophages, has been developed to enhance the microbial safety of wild vegetables. Newly isolated Escherichia coli phage EP and Staphylococcus aureus phage SP were loaded into the gelatin coating solutions. The phages remained significantly stable for up to 3 days, respectively, and exhibited rapid antibacterial capacity within 2 h of coating application (p < 0.05). The developed coating was applied to bracken and exhibited antibacterial efficacy against E. coli and S. aureus within 6 h (1.9-log CFU/mL and 1.5-log CFU/mL). Furthermore, the coated bracken significantly prevented weight loss and maintained firmness for 10 days (p < 0.05). Consequently, gelatin-based edible coatings containing phages have the potential as an antibacterial packaging strategy.

Molecular Phylogeny of Korean-type Coliphages and American-type Coliphages Determined by a RAPD Analysis (RAPD 분석법에 의한 한국형 대장균파아지와 미국형 대장균파아지의 분자적 계통분류)

  • 권오식
    • Biomedical Science Letters
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    • v.6 no.2
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    • pp.131-139
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    • 2000
  • RAPD-PCR was applied to identify the phylogenetic relationship between isolated Korean-type coliphages ($\phi$C1, $\phi$C2, $\phi$C3 and $\phi$C4) and well-known American coliphages ($\phi$T2, $\phi$T4, $\phi$T5, $\phi$T7 and ${\phi}{\lambda}$). Subsequently, a computer analysis was carried out with the results of RAPD-PCR. As a result, 9 individuals were divided into five groups. The Korean-type coliphages formed a single cluster which showed very high genetic similarity but the American-type coliphages revealed very low genetic similarity among them. In particular, the $\phi$T2와 $\phi$T4 (T$_{even}$ phages) made one sub-cluster among American coliphages, and they were very distant from $\phi$T5, $\phi$T7 and ${\phi}{\lambda}$. However, ${\phi}{\lambda}$ made a cluster with the Korean-type coliphages that we isolated. The genome size of Korean-type coliphages was ranged from 25,000 bp to 35,000 bp. Among them, the genome of $\phi$C2 was the smallest and that of $\phi$C1 was the biggest, while others were in the middle of the size.

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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.

Isolation and characterization of bacteriophages for the control of Shiga Toxin-producing E. coli (시가 독소 생성 대장균의 제어를 위한 박테리오파지의 분리와 특성 분석)

  • Lim, Ga-Yeon;Park, Do Won;Lee, Young-Duck;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.50 no.6
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    • pp.594-600
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    • 2018
  • Shiga toxin-producing Escherichia coli (STEC) is an important pathogenic bacterium. To control STEC, the characteristics of the ECP33 and NOECP91 coliphages, which belong to the Myoviridae family, were analyzed. The host inhibition range for a total of 44 STEC strains was 45.5% for ECP33 and 65.9% for NOECP91. ECP33 and NOECP91 were relatively stable at $65^{\circ}C$, 50 ppm of sodium hyperchlorite, and a pH value of 4-10. However, the two phages were susceptible to a temperature of $70^{\circ}C$. NOECP91 was killed within 1 h after exposure to 30% ethanol, but ECP33 showed high tolerance even after exposure to 70% ethanol for 1 h. Interestingly, the inhibition of STEC growth according to the multiplicity of infection of 0.1 was confirmed until no growth was observed after 10 hours of culture with the phages. Therefore, the ECP33 and NOECP91 phages may be applied as a biological control agent for Shiga toxin-producing E. coli.