• Title/Summary/Keyword: bacteriophage test

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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.

Isolation and Characterization of the Smallest Bacteriophage P4 Derivatives Packaged into P4-Size Head in Bacteriophage P2-P4 System

  • Kim, Kyoung-Jin;Song, Jae-Ho
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.530-536
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    • 2006
  • Bacteriophage P4, a satellite phage of coliphage P2, is a very useful experimental tool for the study of viral capsid assembly and cos-cleavage. For an in vitro cos-cleavage reaction study of the P2-P4 system, new shortened and selectable markers containing P4 derivative plasm ids were designed as a substrate molecules. They were constructed by swapping the non-essential segment of P4 DNA for either the kanamycin resistance (kmr) gene or the ampicillin resistance (apr) gene. The size of the genomes of the resulting markers were 82% (P4 ash8 delRI:: kmr) and 79% (P4 ash8 delRI:: apr) of the wild type P4 genome. To determine the lower limit of genome size that could be packaged into the small P4-size bead, these shortened P4 plasmids were converted to phage particles with infection of the helper phage P2. The conversion of plasmid P4 derivatives to bacteriophage particles was verified by the heat stability test and the burst size determination experiment. CsCl buoyant equilibrium density gradient experiments confirmed not only the genome size of the viable phage form of shortened P4 derivatives, but also their packaging into the small P4-size head. P4 ash8 delRI:: apr turned out to be the smallest P4 genome that can be packaged into P4-sized head.

Identification of K1 Polysaccharide Antigen of Escherichia coli Isolates from Urine Specimens of Urinary Tract Infections in Children (요로감염소아의 오줌에서 분리한 대장균 K1 다당류 항원의 동정)

  • 정희곤
    • The Korean Journal of Food And Nutrition
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    • v.11 no.4
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    • pp.416-419
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    • 1998
  • Identification of escherchia coli K1 polysaccharide antigen isolated from urine specimens of urinary tract infections in children were performed from of 1992 to 1993 in Kyoto, Japan. The serotypes of E. coli were categorized that O1:H7, O2:H6, O2:H7, O16:H6, O18:H7, O18:H ̄, and O135:H44 among 14 strains isolated from urine specimens of urinary tract infections in children by the serological test. And, one strain (O18:H ̄, isolation rate: 7.1%) of E. coli K1 polysaccharide antigen among 14 strains were isolated from urine specimens of urinary tract infections in children by the bacteriophage test.

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Method development for efficacy testing of veterinary disinfectants using bacteriophage MS2 (Bacteriophage MS2를 이용한 소독제 효력시험 확립에 관한 연구)

  • Rhee, Chae Hong;Kim, Soohee;Han, Bokhee;Kim, Young-Wook;Her, Moon;Jeong, Wooseog
    • Korean Journal of Veterinary Service
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    • v.44 no.3
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    • pp.149-155
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    • 2021
  • In virucidal efficacy testing, the chemical inactivation cannot be determined for all viruses due to the difficulties or the inability to culture sufficiently or the risk of exposure to the viruses. Therefore, disinfectants against these viruses could be evaluated by different methods and surrogate viruses are used as alternative. In this study we developed a method for efficacy testing of veterinary disinfectants using one of the candidate surrogate viruses, bacteriophage MS2, as part of the research on the selection of surrogate viruses for efficiency of efficacy testing of veterinary disinfectants. This method is based on the Animal and Plant Quarantine Agency (APQA) guidelines for efficacy testing of veterinary disinfectants. Bacteriophage and disinfectant are reacted in suspension in accordance with the APQA guidelines and then a newly established double agar layer method is applied for the efficacy test. The double agar layer method is summarized as follows: 1) The bottom agar with 1.5% agar is boiled and cooled before poured into petri dishes at volume of 20 mL, and dried under biological safety cabinet. 2) The top agar with 0.7% agar is boiled and kept at 50℃ before E. coli culture was seeded. 3) The serially diluted bacteriophage MS2-disinfectant mixtures 0.05 mL and E. coli host 0.01 mL (OD600 0.2~0.3) are mixed with 5 mL of top agar and incubate them at 50℃ for 5 min for reaction. 4) The resulting mixture is poured over top of a bottom agar plate and rocked sufficiently to ensure that the top agar covers the entire surface of the bottom agar. 5) The double agar layer is then placed under biological safety cabinet to allow the agar layer to solidify and subsequently incubated at 37℃ for 24 hr. 6) Following incubation, the plates may be inspected for plaques and record results.

Indicator Microorganisms Used as Fecal Contamination in Aquatic Environments (수계환경에서 분변성 오염의 지표로 사용되는 미생물들)

  • 이건형
    • Korean Journal of Environmental Biology
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    • v.20 no.3
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    • pp.189-196
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    • 2002
  • The direct detection of intestinal pathogens and viruses often requires costly, tedious, and time-consuming procedures. These requirements developed a test to show that the water was contaminated with sewage-borne pathogens by assessing the hygienic quality of water based on indicator microorganisms whose presence indicates that pathogenic microorganisms may also be present. Various groups of microorganisms have been suggested and used as indicator microorganisms. Proposed and commonly used microbial indicators are total coliforms, fecal coliforms, fecal streptococci, Clostridium perfringens, heterotrophic plate count, bacteriophage, and so on. Unfortunately, most, if not all, of these indicators are not ideal because of the sensitivity and resistance to environment stresses and disinfection. However, the development of gene probes and PCR technology may give hope for the discovery of rapid and simple methods toy detecting small number of fecal pathogens in various environments.

Binary Segmentation Procedure for Detecting Change Points in a DNA Sequence

  • Yang Tae Young;Kim Jeongjin
    • Communications for Statistical Applications and Methods
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    • v.12 no.1
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    • pp.139-147
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    • 2005
  • It is interesting to locate homogeneous segments within a DNA sequence. Suppose that the DNA sequence has segments within which the observations follow the same residue frequency distribution, and between which observations have different distributions. In this setting, change points correspond to the end points of these segments. This article explores the use of a binary segmentation procedure in detecting the change points in the DNA sequence. The change points are determined using a sequence of nested hypothesis tests of whether a change point exists. At each test, we compare no change-point model with a single change-point model by using the Bayesian information criterion. Thus, the method circumvents the computational complexity one would normally face in problems with an unknown number of change points. We illustrate the procedure by analyzing the genome of the bacteriophage lambda.

In Vitro Construction and Characterization of the Bacteriophage P4 Derivative, P4 sid71 cosP2, Containing the Bacteriophage P2 cos Region (박테리오파지 P2의 cos 지역을 함유하는 박테리오파지 P4 유도체인 P4 sid71 cosP2의 In vitro 조성과 정성 연구)

  • Kim, Kyoung-Jin
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.99-104
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    • 2013
  • Bacteriophage P2 sir mutants are inefficient helpers for their satellite bacteriophage P4. The term, "P2 sir-associated helper inefficiency" has been used to define this phenomenon and it has been suggested that the DNA sequence difference between the cos region of P2 and that of P4 is responsible. To test this hypothesis, P4 derivative phage, P4 sid71 cosP2, containing the cos region of P2 and sid71 allele was constructed through several in vitro DNA manipulation steps. Its burst size was determined using a one-step growth experiment. The results showed that the substitution of the cos region of P2 for the cos region of P4 in P4 sid71 cosP2 overcame "P2 sir-associated helper inefficiency". P4 sid71 cosP2 stock phages prepared with P2 wild type helper and P2 sir helper were analyzed using a CsCl buoyant equilibrium density gradient experiment. The results revealed that the phage particles containing three copies of the P4 genome were the predominant particles in both cases.

Epidemiological Studies on the Methicillin Resistant Staphylococcus aureus Isolated from Clinical Samples (임상가검물에서 분리한 Methicillin내성 Staphylococcus aureus의 분자역학적 연구)

  • Yang-Hyo Oh;Min-Jung Kim
    • Biomedical Science Letters
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    • v.5 no.2
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    • pp.135-145
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    • 1999
  • A total of 45 Staphylococcus aureus strains from clinical samples were tested for the biochemical test and antibiotic susceptibility test. Forty-five S. aureus strains were subjected to the molecular epidemiological study by susceptiblity test, antibiogram, bacteriophage typing, polymerase chain reaction and mec-associated hypervariable region gene in order to detect of mecA gene which was one of the structural gene related to antibiotic resistant expression factors. Three of 15 mecA-negative S. aureus isolates were classified as oxacillin resistant despite borderline minimal inhibitory concentration values. Methicillin susceptiblities were completely consistent with PCR results for these strains. On the other hand, 4 of 30 mecA-positive isolates yielded results in the oxacillin and methicillin susceptibility tests which were discrepant from those of PCR analysis. Except for SA6, the methicillin resistant S. aureus strains tested were highly resistant to penicillin, oxacillin, gentamicin, and chloramphenicol. In the phage typing, 27 strains were typable. The Iytic group III was as many as 12 strains, and 7 of 12 were 75/83A/84 type. In the PCR of specific mecA gene probe with chromosomal DNA of 30 methicillin resistant S. aureus, the amplified DNA band of 533 bp was confirmed in 30 strains and not in methicillin sensitive S. aureus. The single amplified band of hypervariable region related to mec was investigated in all of 30 methicillin resistant S. aureus, but in methicillin sensitive S. aureus it was amplified. The size of PCR products was between 200 bp and 600 Up. Four units was directly repeated.

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Isolation of Escherichia coli O157:H7 from animal feces and biochemical characteristics of Verotoxin-2 produced by these strains I. Study on the phages related to production of Verotoxin-2, and Isolation of Escherichia coli O157:H7 from feces of cattle and pigs (동물분변에서 Escherichia coli O157:H7의 분리 및 이들 균이 생산하는 Verotoxin-2의 생물화학적 특성 I. 소와 돼지의 분변에서 E coli O157:H7의 분리 및 Verotoxin-2 생산에 관여하여 파아지의 분리에 관하여)

  • Cha, In-ho;Kim, Yong-hwan
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.371-378
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    • 1996
  • Three and 2 strains of E coli O157 were isolated from fecal materials of cattle (390) and pigs (420) in Korea, respectively. One strain of O157:H7 isolated from cattle and 2 strains of O157:H7 isolated from pigs were identified as verotoxin-1 (VT-1) produing strains and 2 strains (O157:H7 and O157:H-) isolated from cattle were identified as verotoxin-2 (VT-2) producing strains by neutralization test on HeLa and Vero cells. Culture supernatants of the isolates were cytotoxic to HeLa and Vero cells. The levels of cytotoxin produced by isolates were $10^2{\sim}10^4$ cytotoxic dose($CD_{50}$)/ml. Also, VT-2-converting bacteriophage was isolated from KSC109 strain which had been isolated from cattle. Molecular weight of the phage DNA was determined as approximately 45 Kb in 0.8% agarose gel electrophoresis, and morphology of the phage stained with phosphotungstic acid was observed by transmissible electron microscopy.

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Removal of Bacteriophage T7 from Artificial groundwater by Layered Double Hydroxide (층상이중수산화물에 의한 인공지하수내의 박테리오파지 T7 제거)

  • Park, Jeong-Ann;Lee, Chang-Gu;Kang, Jin-Kyu;Kim, Song-Bae
    • Journal of Korean Society of Environmental Engineers
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    • v.33 no.6
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    • pp.426-431
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    • 2011
  • The objective of this study was to investigate the virus removal from artificial groundwater using Mg-Fe layered double hydroxide (LDH). Batch experiments were conducted under various experimental conditions to examine bacteriophage T7 removal with Mg-Fe LDH. Results showed that the removal of T7 by Mg-Fe LDH was a fast process, reaching equilibrium within 2~3 hrs. Mg-Fe LDH had the virus removal capacity of $1.57{\times}10^8pfu/g$ with a removal percent of 96%. Results also showed that the effect of solution pH on T7 removal was minimal between pH 6.2 and 9.1. The influence of anions ($SO_4^{2-}$, $CO_3^{2-}$, $HPO_4^{2-}$) on T7 removal was significant due to their competition with bacteriophage at the sorption sites on LDH, while the effect of $NO_3^-$ was negligible. This study demonstrated that Mg-Fe LDH could be applied as adsorbents for virus removal in water treatment.