Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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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 (Department of Food Science and Biotechnology, College of BioNano Technology, Gachon University) ;
  • Park, Do Won (Department of Food Science and Biotechnology, College of BioNano Technology, Gachon University) ;
  • Lee, Young-Duck (Department of Food Science and Engineering, Seowon University) ;
  • Park, Jong-Hyun (Department of Food Science and Biotechnology, College of BioNano Technology, Gachon University)
  • Received : 2018.08.28
  • Accepted : 2018.10.29
  • Published : 2018.12.31
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Abstract

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.

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.

Keywords

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Fig. 2. Stability of ECP33 and NOECP91 at (A) 65 and (B)70℃.

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Fig. 3. Stability of ECP33 and NOECP91 under various pH (A) and ethanol (B) conditions for 30 min and 1 h.

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Fig. 4. Stability of ECP33 and NOECP91 at (A) 50 ppm of NaClO and (B) 100 ppm of NaClO.

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Fig. 5. Growth inhibition using bacteriophage for E. coli NCCP 13930 (O157:H7) and E. coli NCCP 13937 (O103).

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Fig. 1. Transmission Electron Microscopy micrographs of (A) ECP33 and (B) NOECP91.

Table 1. Host spectrum of ECP33 and NOECP91 isolated for shiga-toxin producing E .coli

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