대표적인 상업화된 박테리오신인 나이신은 Listeria monocytogenes 및 Staphylococcus aureus와 같은 병원성 세균에 대해 강력한 항균 활성을 보인다. 본 연구에서는 시판되는 나이신 제품을 박테리오파지 SAP84와 함께 병용 처리했을 때 S. aureus 억제에 대한 상승효과에 대하여 평가했다. S. aureus KCTC 3881 균주에 대해 나이신은 농도의존적으로 생균수를 감소시켰으며 18 IU/mL의 나이신은 대조구와 비교하여 6시간째에 4.03 Log CFU/mL의 균수가 감소된 반면, 동일 용량의 나이신이 박테리오파지 SAP84 (0.1 MOI)와 병용처리 되었을 때 5.54 Log CFU/mL의 생균수 감소가 관찰되었다. 또한 나이신과 SAP84 의 조합은 양상추에서 S. aureus 균주를 효과적으로 제어하는데 성공적으로 적용되었다.
일부 대장균 균주는 독성을 가지고 있으며 이들을 제어 하기 위해 박테리오파지와 같은 대체 항균물질에 대한 연구가 활발히 진행되고 있다. 본 연구에서는 E. coli DH5α 균주를 모델로 이 균주의 생육을 억제하는 박테리오파지 ECP27과 병용처리했을 때 상승효과를 나타낼 수 있는 항균물질을 탐색하였다. 후보물질로는 CaCl2, 젖산, 구연산 이 사용되었다. CaCl2의 경우 6시간째 농도 의존적으로 생육억제 상승효과가 나타났으나 12시간째 E. coli DH5α의 생균수가 회복되는 추세를 보였다. 그러나 30mM 농도에 서 젖산과 구연산은 박테리오파지 ECP27과 병용처리 했을 때 E. coli DH5α의 생육저해에 대하여 6시간째 상승 효과를 보였으며 12시간째 생균수가 검출되지 않았다. 또한 젖산과 구연산을 단독으로 처리했을 때 12시간째 E. coli DH5α의 생균수가 확인되지 않아 독자적으로도 항균력이 우수하였다. 따라서 박테리오파지와 유기산을 병용 처리하는 것은 대장균의 생육을 효과적으로 억제하는 좋은 전략이 될 수 있을 것이다.
The effect of physical and chemical treatments to reduce staphylococcal phages was investigated. To determine impact of physical treatment on viability of phages, two staphylococcal phages (SAP84 and SAP89) were treated with multiple heat (55oC and 60oC) and pH (pH4, 7, 10) conditions. Viability of SAP 84 was dramatically reduced at 60C and SAP 89 was completely inactivated at 60C within 25 min. Overall, the two phages were stable under all the pH conditions tested except for the SAP 89 at pH 10. Treatments, a 10% FAS (Ferrous Ammonium Sulfate) solution and various density of ethanol and sodium hypochlorite were used to reduce the two phages. SAP 84 was unstable in 50% and 70% ethanol. However, SAP 84 and SAP 89 showed high tolerance after exposure to 100 ppm of sodium hypochlorite which is known as an effective sterilizer. As soon as the two phages were treated with 10% FAS, which is used as a virucidal agent, they were inactivated and did not form any plaque. The result of this study provides additional evidence that staphylococcal phages can be controlled by various physicochemical treatments.
There has been an accelerating increase in water reuse due to growing world population, rapid urbanization, and increasing scarcity of water resources. However, it is well recognized that water reuse practice is associated with many human health and ecological risks due to numerous chemicals and pathogenic microorganisms. Especially, the potential transmission of infectious disease by hundreds of pathogenic viruses in wastewater is one of the most serious human health risks associated with water reuse. In this study, we determined the response of different bacteriophages representing various bacteriophage groups to chlorination in real wastewater in order to identify a more reliable bacteriophage indicator system for chlorination in wastewater. Different bacteriophages were spiked into secondary effluents from wastewater plants from three different geographic areas, and then subjected to various doses of free chlorine and contact time at 5˚C in a bench-scale batch disinfection system. The inactivation of φX174 was relatively rapid and reached ∼4 log10 with a CT value of 5 mg/L*min. On the other hand, the inactivation of bacteriophage PRD1 and MS2 were much slower than the one for φX174 and only ~1 log10 inactivation was achieved by a CT value of 10 mg/L*min. Overall, the results of this study suggest that bacteriophage both MS2 and PRD1 could be a reliable indicator for human pathogenic viruses for chlorination in wastewater treatment processes and water reuse practice.
Cronobactersakazakii is a newly emerging high hazard pathogen, which causes encephalomeningitis and necrotic colitis. Recently, successful biocontrol of harmful microorganisms in several foods through the use of bacteriophages has been reported. In this study, bacteriophages were isolated from kimchi and sewages. Morphological analysis by TEM indicated that phages belonged to the Myoviridae family. In case of heat stability, KCES2 and ESP 2949-2 phages were susceptible to temperatures above 70oC. KCES2 and ESP 2949-2 phages inhibited the growth of C. sakazakii in culture broth. When KCES2 and ESP 2949-2 phages were applied to biofilm-formed C. sakazakii, C. sakazakii was efficiently reduced. Therefore, newly isolated KCES2 and ESP 2949-2 phage for C. sakazakii might effectively reduce C. sakazakii in various foods.
The aim of study was to investigate the virulence profile of Escherichia coli O157:H7 bacteriophages isolated from sewage and livestock stools. Among 23 E. coli O157:H7 bacteriophages, 14 strains were isolated from sewage and 9 were from animal stools collected from 10 livestock farms in Korea. For each bacteriophage DNA sample, the presence of stx1, stx2, eae, aafII, ial, elt, estI, estII, astA, afa, and cnf was examined by polymerase chain reaction. The detection rate of eae, stx2, estI, astA, and ial was 100%, 69.6%, 13.0%, 13.0%, 8.7%, respectively. While all E. coli O157:H7 bacteriophages isolated from stools carried eae+stx2, stx2+eae, eae+astA, eae, stx2+eae+estI, eae+estI, stx2+eae+ial, and eae+ial were observed in bacteriophages isolated from sewage. As several plasmid-carrying virulence factors (estI, astA, and ial) were found in E. coli O157:H7 bacteriophages obtained from sewage and stools, the microbial safety of bacteriophages should be investigated in further study.
Continuous outbreaks of Shigella spp. have raised concerns about the lack of rapid and on-site applicable biosensor method for Shigella detection. Since a bacteriophage has recently been employed as an emerging bio-recognition element in biosensor method, Shigella sonnei-specific bacteriophage was isolated and purified from a slaughterhouse with the final concentration of 2.0×1012 PFU/mL in this study. Analysis of purified S. sonnei-specific bacteriophage using transmission electron microscopy indicated that it possessed an icosahedral head with a relatively long non-contractile tail. It was therefore classified as a member of the Siphoviridae family. Head width, head length, and tail length were 69.9±11.2 nm, 77.5±8.8 nm, and 264.4±33.9 nm, respectively. The genomic DNA size of the S. sonnei-specific bacteriophage was determined to be approximately 25 kb by using 0.4% agarose gel electrophoresis. In specificity test with 43 food-associated microorganisms, the S. sonnei-specific bacteriophage exhibited a clear plaque against S. sonnei only. In addition, the S. sonnei-specific bacteriophage was stable within a wide range of pH values (pH 3-11) and temperatures (4-37 ). Thus, the present study demonstrated the excellent specificity and stability of the S. sonnei-specific bacteriophage as a novel bio-recognition element for S. sonnei detection in foods.
최근 유산균을 이용한 다양한 발효 식품들이 연구, 개발되고 있으며, 유산균 발효 식품은 건강 증진 측면에서 효용성을 인정받아 전세계 인구가 섭취하고 있다. 이러한 유산균 발효 식품에 있어서 성공적인 제품 생산을 위해 박테리오파지의 중요도가 지속적으로 증대되고 있다. 특히, 발효 식품들 중 유제품에 대한 박테리오파지의 여러 가지 제어 연구가 다수 수행되었으며, 물리적, 화학적 위생처리를하거나 각각의 방법을 혼합하여 유제품 발효를 보다 성공적으로 수행하도록 하여 왔다. 또한, 최근에는 국내에서도GMP(Good Manufacturing Practice), CIP, SSOP(SanitationStandard Operating Procedures) 등을 유제품 생산 공장에적용하여 보다 효과적인 유제품 생산을 도모하고 있다.