• 제목/요약/키워드: coliphages

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Virulence Factors and Stability of Coliphages Specific to Escherichia coli O157:H7 and to Various E. coli Infection

  • Kim, Eun-Jin;Chang, Hyun-Joo;Kwak, Soojin;Park, Jong-Hyun
    • Journal of Microbiology and Biotechnology
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    • 제26권12호
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    • pp.2060-2065
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    • 2016
  • Characteristics of E. coli O157:H7-specific infection bacteriophages (O157 coliphages) and broad-host-range bacteriophages for other E. coli serotypes (broad-host coliphages) were compared. The burst sizes of the two groups ranged from 40 to 176 PFU/infected cell. Distributions of the virulence factors stx1, stx2, ehxA, and saa between the two groups were not differentiated. Broad-host-range coliphages showed lower stability at $70^{\circ}C$, in relation to O157 coliphages. However, O157 coliphages showed high acid and ethanol tolerance by reduction of only 22% and 11% phages, respectively, under pH 3 and 70% ethanol for 1 h exposure. Therefore, these results revealed that the O157 coliphages might be more stable under harsh environments, which might explain their effective infection of the acid-tolerant E. coli O157:H7.

Fast Genetic Variation among Coliphage Quasispecies Revealed by a Random Amplified Polymorphic DNA (RAPD) Analysis

  • Kwon, Oh-Sik;Lee, Jae-Yung
    • Journal of Microbiology
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    • 제34권2호
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    • pp.166-171
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    • 1996
  • Genetic analysis was conducted on newly isolated coliphages form soil by using a RAPD assay. From the initial result, the coliphages were turned out to be different form one another but were closely related to .psi..lambda. due to the fact that they shared the samed RAPD maker in which other T phage testings failed to show. By using the primers EC01 or EC02, a fast genetic mutation of .psi.C1 was found by producing specific RAPD markers on the phages from the first filial progeny to the second filial progeny. When we made a RAPD assay with combined primers (EC01, EC05 and EC08), the genetic mutation was again confirmed in .psi.C1. The assay detection showed mutations in other coliphages such as .psi.C2 and .psi.C3 by revealing specific RAPD bands among different progeny phages, where genetic instability of the coliphages in implied.

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

  • 권오식
    • 대한의생명과학회지
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    • 제6권2호
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    • pp.131-139
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    • 2000
  • 분리한 한국형 대장균파아지군($\phi$C1, $\phi$C2, $\phi$C3 및 $\phi$C4)과 잘 알려진 미국형 대장균파아지군($\phi$T2, $\phi$T4, $\phi$T5, $\phi$T7 및 ${\phi}{\lambda}$)의 유전적 유연관계를 조사하기 위하여 분자적 계통분류를 위한 방법인 RAPD-PCR을 실시하고 컴퓨터분석을 하였다. 그 결과, 9개의 대장균파아지들은 5개의 그룹으로 나위어지면서 한국형 대장균파아지들만이 그들간의 유전적 유사도가 매우 높으면서 하나의 클러스터를 형성하였다. 반면 이국형 대장균파아지들은 오직 하나의 서브클러스터를 가지며 나누어졌다. 즉, 미국형 대장균파아지 중 $\phi$T2와 $\phi$T4($T_{even}$ 파아지)만이 하나의 서브클러스터를 형성하면서 $\phi$T5, $\phi$T7 및 ${\phi}{\lambda}$들과 뚜렷히 구분되고 있었다. 그리고 한국형 대장균파아지들은 미국형 대장균차이지 중 오직 ${\phi}{\lambda}$와 유전적 유연관계를 갖고 있음을 확인하였다. 한편 한국형 대장균파아지의 게놈의 크기는 25,000 bp~35,000 bp 정도 였으며, 이 중 $\phi$C2가 그 크기가 가장 작고 $\phi$C1이 가장 컸다. 그리고 $\phi$C3과 $\phi$C4의 게놈은 중간 크기로 비슷하였다.

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한강에서 분리한 이종 coliphage의 동정과 점토질에 대한 흡착 및 용출효과 (Identification of two coliphages from Han-river and its adsorption-elution effect on soil materials)

  • 홍순우;하영칠;안태석;이영숙
    • 미생물학회지
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    • 제20권4호
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    • pp.210-222
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    • 1982
  • Coliphages isolated from Han-River from September 1980 to August 1981 were classified by morphological and physiological characteristics. Effects of soil metrial on the fate of coliphage in nature were investigated. 1. The correlation coefficient between coliphage and E.coli which was host of coliphages in nature was 0.7173 (p=0.004). 2. Coliphage I isolated from Han-River of which DNA molecular weight was $27{\times}10^6$ daltons was identified as $T_1$ phage and coliphage II of which DNA molecular weight $72{\times}10^6$ daltons was classified as $T_5$ phage. 3. Soil material SW was composed of 63.65% silt and 21.92% clay. Clay was consisted of illite, kaolinite and chlorite evenly. Soil material J was composed of 68.92% silt and 11.67% clay. Clay consisted of smectite only. 4. Coliphage was absorbed to soil material J more than soil material SW, and $T_1$ coliphage was absorbed to soil material more than $T_5$ coliphage was. 5. The phage adsorption efficiency to soil material was enhanced at lower pH : the phage adsorption efficiency at pH 4 was 27 time higher than at pH 7. 6. Divalent $(Ca^{2+})\;and\;trivalention\;(Al^{3+})$ enhanced the phage adsorption efficiency to soil material from 4 to 39 and from 17 to 91 times higher than monovalent $ion(Na^+)$, respectively. 7. The concentration of organic compound was inversely related to the phage adsorption efficiency to soil. 8. Adsorption of phage onto soil material, and elution efficiency of elutants was in the order of D.D.W>tap water>river water>seawater. 9. The higher the concentration of organic compound was, the more were adsorbed phages to soil eluted. 10. Coliphages survived longer in sterile soil suspension than in nonsterile soil material suspension.

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지표미생물을 이용한 시화호 유입수의 수질평가 (Evaluation of Influent Water Quality Using Indicator Microorganisms in Lake Shiwha)

  • 이희태;김희연;박현진;조영은;유소영;이경진;정종선;고광표
    • 한국환경보건학회지
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    • 제34권1호
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    • pp.86-94
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    • 2008
  • Lake Shiwha, an artificial lake located near metropolitan Seoul, offers a unique water environment and has been suspected to have high levels of chemical and microbiological contaminations. Lake Shiwha was originally connected to the sea but currently has four major surface water inputs from agricultural, municipal, industrial areas and in addition an occasional inflow from the sea. The objectives of this study are to investigate the relative contribution of microbial contaminants from each of the inflowing surface waters and to identify appropriate microbial indicator organisms in this unique water environment. We measured the levels of microbial contaminations in the four inflowing surface waters. A number of microbial indicator organisms including total coliform (TC), fecal coliform (FC), E. coli, Enterococci, somatic and male-specific coliphages were analyzed. Bacterial indicator microorganisms were detected and quantified by the $Colilert^{(R)},\;Enterolert^{(R)}$ kit. Surface water (50 l) was sampled by $ViroCap^{TM}\;5"$ cartridge filters and analyzed by the single agar layer method for detecting coliphages. The concentrations of TC, FC, E. coli, and Enterococci were 1543 CFU/100 ml${\sim}1.99{\times}10^6$ CFU/100 ml, 0 CFU/100 ml${\sim}202$ CFU/100ml, 0 CFU/100 ml${\sim}1.80{\sim}10^5$ CFU/100ml, 74 CFU/100 ml${\sim}3408$ CFU/100 ml, respectively. The male-specific and somatic coliphages were detected in three different inflowing surface waters. Isolated E. coli and Enterococci strains were further analyzed by 16s rDNA amplification and subsequent phylogenetic analysis from Jungwang-chun, Ansan-chun, Banwol-chun and penstock of inflowing surface water. Our results indicated that the concentrations of different fecal indicator microorganisms might not be highly correlated with each other. Multiple microbial indicator organisms should be used for monitoring microbial contamination and microbial source tracking methods.

Exogenous Lytic Activity of SPN9CC Endolysin Against Gram-Negative Bacteria

  • Lim, Jeong-A;Shin, Hakdong;Heu, Sunggi;Ryu, Sangryeol
    • Journal of Microbiology and Biotechnology
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    • 제24권6호
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    • pp.803-811
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    • 2014
  • Concerns over drug-resistant bacteria have stimulated interest in developing alternative methods to control bacterial infections. Endolysin, a phage-encoded enzyme that breaks down bacterial peptidoglycan at the terminal stage of the phage reproduction cycle, is reported to be effective for the control of bacterial pathogenic bacteria. Bioinformatic analysis of the SPN9CC bacteriophage genome revealed a gene that encodes an endolysin with a domain structure similar to those of the endolysins produced by the P1 and P22 coliphages. The SPN9CC endolysin was purified with a C-terminal oligo-histidine tag. The endolysin was relatively stable and active over a broad temperature range (from $24^{\circ}C$ to $65^{\circ}C$). It showed maximal activity at $50^{\circ}C$, and its optimum pH range was from pH 7.5 to 8.5. The SPN9CC endolysin showed antimicrobial activity against only gram-negative bacteria and functioned by cutting the glycosidic bond of peptidoglycan. Interestingly, the SPN9CC endolysin could lyse intact gram-negative bacteria in the absence of EDTA as an outer membrane permeabilizer. The exogenous lytic activity of the SPN9CC endolysin makes it a potential therapeutic agent against gram-negative bacteria.

Genetic Analysis and Characterization of a Bacteriophage ØCJ19 Active against Enterotoxigenic Escherichia coli

  • Kim, Gyeong-Hwuii;Kim, Jae-Won;Kim, Jaegon;Chae, Jong Pyo;Lee, Jin-Sun;Yoon, Sung-Sik
    • 한국축산식품학회지
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    • 제40권5호
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    • pp.746-757
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    • 2020
  • Enterotoxigenic Escherichia coli (ETEC) is the major pathogenic E. coli that causes diarrhea and edema in post-weaning piglets. In this study, we describe the morphology and characteristics of ØCJ19, a bacteriophage that infects ETEC, and performed genetic analysis. Phage ØCJ19 belongs to the family Myoviridae. One-step growth curve showed a latent phase of 5 min and burst size of approximately 20 phage particles/infected cell. Phage infectivity was stable for 2 h between 4℃ and 55℃, and the phage was stable between pH 3 and 11. Genetic analysis revealed that phage ØCJ19 has a total of 49,567 bases and 79 open reading frames (ORFs). The full genomic sequence of phage ØCJ19 showed the most similarity to an Escherichia phage, vB_EcoS_ESCO41. There were no genes encoding lysogeny, toxins, virulence factors, or antibiotic resistance in this phage, suggesting that this phage can be used safely as a biological agent to control ETEC. Comparative genomic analysis in terms of the tail fiber proteins could provide genetic insight into host recognition and the relationship with other coliphages. These results showed the possibility to improve food safety by applying phage ØCJ19 to foods of animal origin contaminated with ETEC and suggests that it could be the basis for establishing a safety management system in the animal husbandry.

Applicability Evaluation of Male-Specific Coliphage-Based Detection Methods for Microbial Contamination Tracking

  • Kim, Gyungcheon;Park, Gwoncheol;Kang, Seohyun;Lee, Sanghee;Park, Jiyoung;Ha, Jina;Park, Kunbawui;Kang, Minseok;Cho, Min;Shin, Hakdong
    • Journal of Microbiology and Biotechnology
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    • 제31권12호
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    • pp.1709-1715
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    • 2021
  • Outbreaks of food poisoning due to the consumption of norovirus-contaminated shellfish continue to occur. Male-specific (F+) coliphage has been suggested as an indicator of viral species due to the association with animal and human wastes. Here, we compared two methods, the double agar overlay and the quantitative real-time PCR (RT-PCR)-based method, for evaluating the applicability of F+ coliphage-based detection technique in microbial contamination tracking of shellfish samples. The RT-PCR-based method showed 1.6-39 times higher coliphage PFU values from spiked shellfish samples, in relation to the double agar overlay method. These differences indicated that the RT-PCR-based technique can detect both intact viruses and non-particle-protected viral DNA/RNA, suggesting that the RT-PCR based method could be a more efficient tool for tracking microbial contamination in shellfish. However, the virome information on F+ coliphage-contaminated oyster samples revealed that the high specificity of the RT-PCR- based method has a limitation in microbial contamination tracking due to the genomic diversity of F+ coliphages. Further research on the development of appropriate primer sets for microbial contamination tracking is therefore necessary. This study provides preliminary insight that should be examined in the search for suitable microbial contamination tracking methods to control the sanitation of shellfish and related seawater.

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

  • 임가연;박도원;이영덕;박종현
    • 한국식품과학회지
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    • 제50권6호
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    • pp.594-600
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    • 2018
  • 본 연구는 시가독소 생성 대장균(STEC)를 제어하기 위하여 하천수 시료로부터 넓은 숙주저해범위를 갖는 ECP33 파지와 NOECP91 파지를 분리, 선별하였다. 총 44주의 STEC균주에 대해 ECP33 파지는 약 45.5%와 NOECP91 파지는 약 65.9%로 숙주저해특성을 가지는 것으로 나타났다. 형태학적 특성을 확인한 결과 모두 Myoviridae family에 속하였다. 환경에 대한 안정성 분석을 진행한 결과, 두 파지 모두 50 ppm 농도의 차아염소산 및 pH 4-10의 pH환경에 대해서는 최소 30분까지 비교적 안정한 것으로 나타났다. 반면, $70^{\circ}C$의 고온에서는 NOECP91 파지는 15분 내에, ECP33 파지는 30분 처리 시에 대부분 사멸하고, 유기용매(EtOH) 환경에서 NOECP91 파지는 30% 에탄올 1시간 처리에 모두 사멸하였으나, ECP33파지의 경우 70% EtOH/에탄올 1시간 처리에도 2 log PFU/mL 미만의 감소율을 보여 온도와 유기용매(EtOH) 환경에 대해 ECP33 파지가 NOECP91 파지보다 안정적인 것으로 나타났다. 아울러 분리된 ECP33 파지와 NOECP91 파지를 MOI 0.1 조건에 따른 STEC 균주 생육억제 정도를 분석한 결과, 배양 10시간대까지 생육억제가 이루어지는 것을 확인하였다. 따라서 본 연구를 통해 분리된 ECP33 파지와 NOECP91 파지는 시가독소 생성 대장균의 생물학적 제어제로서 적용이 가능할 것으로 사료된다.