• Journal of Microbiology and Biotechnology
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• 제19권6호
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• pp.622-628
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• 2009

In the present study, the therapeutic potential of purified and well-characterized bacteriophages was evaluated in thermally injured mice infected with Klebsiella pneumoniae B5055. The efficacy of five Klebsiella phages (Kpn5, Kpn12, Kpn13, Kpn17, and Kpn22) was evaluated on the basis of survival rate, decrease in bacterial counts in different organs of phage-treated animals, and regeneration of skin cells as observed by histopathological examination of phage-treated skin. Toxicity studies performed with all the phages showed them to be non-toxic, as no signs of morbidity and mortality were observed in phage-treated mice. The results of the study indicate that a single dose of phages, intraperitoneally (i.p.) at an MOI of 1.0, resulted in significant decrease in mortality, and this dose was found to be sufficient to completely cure K. pneumoniae infection in the burn wound model. Maximum decrease in bacterial counts in different organs was observed at 72 h post infection. Histopathological examination of skin of phage-treated mice showed complete recovery of burn infection. Kpn5 phage was found to be highly effective among all the phages and equally effective when compared with a cocktail of all the phages. From these results, it can be concluded that phage therapy may have the potential to be used as stand-alone therapy for K. pneumoniae induced burn wound infection, especially in situations where multiple antibiotic-resistant organisms are encountered.

• Journal of Microbiology and Biotechnology
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• 제24권6호
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• pp.740-747
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• 2014

Bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae (Xoo) is a very serious disease in rice-growing regions of the world. In spite of their economic importance, there are no effective ways of protecting rice plants from this disease. Bacteriophages infecting Xoo affect the population dynamics of the pathogen and consequently the occurrence of the disease. In this study, we investigated the diversity, host range, and infectivity of Xoo phages, and their use as a bicontrol agent on BLB was tested. Among the 34 phages that were isolated from floodwater in paddy fields, 29 belonged to the Myoviridae family, which suggests that the dominant phage in the ecosystem was Myoviridae. The isolated phages were classified into two groups based on plaque size produced on the lawn of Xoo. In general, there was a negative relationship between plaque size and host range, and interestingly the phages having a narrow host range had low efficiency of infectivity. The deduced protein sequence analysis of htf genes indicated that the gene was not a determinant of host specificity. Although the difference in host range and infectivity depending on morphotype needs to be addressed, the results revealed deeper understanding of the interaction between the phages and Xoo strains in floodwater and damp soil environments. The phage mixtures reduced the occurrence of BLB when they were treated with skim milk. The results indicate that the Xoo phages could be used as an alternative control method to increase the control efficacy and reduce the use of agrochemicals.

• 식물병연구
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• 제17권1호
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• pp.90-94
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• 2011

Xanthomonas oryzae pv. oryzae에 의해서 발생하는 벼 흰잎마름병은 세계적으로 벼 재배지역에서 큰 피해를 주고 있다. 그러나, 이 병의 효과적인 방제 수단은 적은 실정이다. 본 연구에서는 국내 벼 재배지의 관개수에서 분리한 Xoo 파지를 이용하여 흰잎마름병을 생물적으로 방제하고자 하였다. 먼저 포장에서의 사용가능성을 확인하기 위하여 pH, 온도, 자외선에 대한 Xoo 파지의 안정성을 조사하였다. Xoo 파지는 pH 5-pH 10 사이에서는 안정적이었고, $50^{\circ}C$에서 1시간 이상 처리시 대부분 불활성화 되었다. 자외선을 조사하였을 때는 약 2분간의 노출에서도 불활성화 되었다. 벼의 유묘에 살포한 파지는 세균이 없이도 7일간 벼 잎에서 활성을 유지하였고, 스킴 밀크에 혼합하여 살포하면 PBS 완충액에 넣어 살포한 것 보다 10배 이상의 생존율을 보였다. 그러나 이러한 생존율의 증가에도 불구하고 포장에서의 방제효과는 저조하였다. 하지만 파지 혼합액을 Tecloftalam WP, ASM 등과 혼합하여 동시에 살포하면 방제효과가 월등히 증가하였다. 이상의 결과는 Xoo 파지가 흰잎마름병의 방제에 사용될 수 있으며, 약제의 사용 빈도나 농도를 줄여 줄 수 있을 것임을 의미한다.

• 한국식품위생안전성학회지
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• 제34권1호
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• pp.106-114
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• 2019

본 연구는 황색포도상구균 박테리오파지를 물리화학적 방법을 통해서 제어하기 위해 황색포도상구균을 대상으로 하는 SAP 84 와 SAP 89를 신규 분리하였다. 그리고, 황색포도상구균 박테리오파지에 대해 물리적 방법으로 $55^{\circ}C$ 및 $60^{\circ}C$ 열처리와 pH 4, pH 7, pH 10 처리하였다. 그 결과 SAP 84는 $60^{\circ}C$에서 급격히 감소하였고 SAP 89는 $60^{\circ}C$에서 25 분 만에 모두 활성을 잃었다. SAP 84는 pH 4 ~ 10에서 매우 안정적이었고 SAP 89는 pH 10 이상에서 비교적 불안정해지는 것을 보였다. 화학적 방법으로는 에탄올과 차아염소산나트륨 및 FAS 처리를 하였다. SAP 84가 50%, 70% 에탄올에서 급격히 감소하였다. 살균력이 좋은 100 ppm의 차아염소산 나트륨에서도 SAP 84와 SAP 89는 상당히 안정적인 상태를 유지하고 있다. Virucidal agent로 사용되는 FAS를 10% 농도로 처리하였을 때 SAP 84와 SAP 89 모두 처리 직후부터 제거되는 것을 확인할 수 있었다. 따라서 본 연구를 통해 SAP 84와 SAP 89 모두 화학적 처리 방법인 FAS 처리를 통해서 제어 가능함을 보여주었다. 이외에도 SAP 84는 물리적 방법인 열처리의 저감화 효과가 좋았으며 SAP 89는 화학적 방법인 에탄올 처리의 저감화 효과가 비교적 좋았다. 이렇듯 물리화학적 처리방법으로 황색포도상구균 박테리오파지를 제어 가능할 것으로 사료된다.

• Interdisciplinary Bio Central
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• 제4권3호
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• pp.9.1-9.7
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• 2012

Filamentous phages have been in the limelight as a new type of nanomaterial. In this study, genetically and chemically modified fd phage was used to generate a biomimetic phage self-assembly product. Positively charged fd phage (p8-SSG) was engineered by conjugating 3-aminopropyltriethoxysilane (APTES) to hydroxyl groups of two serine amino acid residues introduced at the N-terminus of major coat protein, p8. In particular, formation of a phage network was controlled by changing mixed ratios between wild type fd phage and APTES conjugated fd-SSG phage. Assembled phages showed unique bundle and network like structures. The bacteriophage based self-assembly approach illustrated in this study might contribute to the design of three dimensional microporous structures. In this work, we demonstrated that the positively charged APTES conjugated fd-SSG phages can assemble into microstructures when they are exposed to negatively charged wild-type fd phages through electrostatic interaction. In summary, since we can control the phage self-assembly process in order to obtain bundle or network like structures and since they can be functionalized by means of chemical or genetic modifications, bacteriophages are good candidates for use as bio-compatible scaffolds. Such new type of phage-based artificial 3D architectures can be applied in tuning of cellular structures and functions for tissue engineering studies.

• The Plant Pathology Journal
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• 제38권5호
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• pp.472-481
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• 2022

Brown blotch disease, caused by Pseudomonas tolaasii, is one of the most serious diseases in mushroom cultivation, and its control remains an important issue. This study isolated and evaluated pathogen-specific bacteriophages for the biological control of the disease. In previous studies, 23 varieties of P. tolaasii were isolated from infected mushrooms with disease symptoms and classified into three subtypes, Ptα, Ptβ, and Ptγ, based on their 16S rRNA gene sequences analysis and pathogenic characters. In this study, 42 virulent bacteriophages were isolated against these pathogens and tested for their host range. Some phages could lyse more than two pathogens only within the corresponding subtype, and no phage exhibited a wide host range across different pathogen subtypes. To eliminate all pathogens of the Ptα, Ptβ, and Ptγ subtype, corresponding phages of one, six, and one strains were required, respectively. These phages were able to suppress the disease completely, as confirmed by the field-scale on-farm cultivation experiments. These results suggested that a cocktail of these eight phages is sufficient to control the disease induced by all 23 P. tolaasii pathogens. Additionally, the antibacterial effect of this phage cocktail persisted in the second cycle of mushroom growth on the cultivation bed.

• The Plant Pathology Journal
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• 제38권3호
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• pp.248-253
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• 2022

Erwinia amylovora is a devastating bacterial plant pathogen that infects Rosaceae including apple and pear and causes fire blight. Bacteriophages have been considered as a biological control agent for preventing bacterial infections of plants. In this study, nine bacteriophages (ΦFifi011, ΦFifi044, ΦFifi051, ΦFifi067, ΦFifi106, ΦFifi287, ΦFifi318, ΦFifi450, and ΦFifi451) were isolated from soil and water samples in seven orchards with fire blight in Korea. The genetic diversity of bacteriophage isolates was confirmed through restriction fragment length polymorphism pattern analysis. Host range of the nine phages was tested against 45 E. amylovora strains and 14 E. pyrifoliae strains and nine other bacterial strains. Among the nine phages, ΦFifi044 and ΦFifi451 infected and lysed E. amylovora only. And the remaining seven phages infected both E. amylovora and E. pyrifoliae. The results suggest that the isolated phages were different from each other and effective to control E. amylovora, providing a basis to develop biological agents and utilizing phage cocktails.

• 한국미생물·생명공학회지
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• 제11권3호
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• pp.155-161
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• 1983

Bacteriophage f2에 대한 자외광선의 살균효과와 외투막 단백질의 구조에 미치는 영향을 Ray-onet photoreactor PPR-208을 사용하여 300nm의 광선으로 연구하였다. 처음 20분간의 조사에서는 약 4 log의 phage가 감소되고 그후 완만한 살균효과를 보이다가 90분 이상의 조사에서는 생존 바이러스가 발견되지 않았다. Tryptophan residue의 fluorescenve quenching, 자외선으로 조신한 phage에 부착시킨 ANS (8-anilino-1-napht-halene sulfonate)의 fluorescence emission의 감소, tryptophan에서 ANS로의 energy transfer 의 감소 등 spectroscopic technique에 의한 결과와 자외선 조사에 의하여 단백질 외투만이 파손되는 전자현미경 관찰의 결과에 의하여 자외광선은 phage f2의 외투막 단백질의 구조에 변화를 일으킨다는 것이 밝혀졌다.

• The Plant Pathology Journal
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• 제40권2호
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• pp.160-170
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• 2024

Erwinia amylovora is a plant pathogen that causes fire blight on apples and pears. Bacteriophages, which are viruses that selectively infect specific species of bacteria and are harmless to animal cells, have been considered as biological control agents for the prevention of bacterial pathogens. In this study, we aimed to use bacteriophages that infect E. amylovora as biocontrol agents against fire blight. We isolated bacteriophages Fifi044 and Fifi318 infecting E. amylovora, and characterized their morphology, plaque form, and genetic diversity to use as cocktails for disease control. The stabilities of the two phages were investigated at various temperatures and pH values and under sunlight, and long-term storage experiment was conducted for a year. To evaluate whether the two phages were suitable for use in cocktail form, growth curves of E. amylovora were prepared after treating the bacterial cells with single phages and a phage cocktail. In addition, a disease control test was conducted using immature apples and in vitro cultured apple plantlets to determine the biocontrol effects of the phage cocktail. The two phages were morphologically and genetically different, and highly stable up to 50℃ and pH value from 4 to 10. The phages showed synergistic effect when used as a cocktail in the inhibition of host bacterial growth and the disease control. This study demonstrated that the potential of the phage cocktail as a biocontrol agent for commercial use.

• 한국식품저장유통학회지
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• 제15권5호
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• pp.617-621
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• 2008

Sequential passes through $Sephadex^{TM}$ columns were used to select phages that displays ligands for dextran ($\alpha$-1,6 linked linear chains) from a phage antibody library. Those phages that bound to the $Sephadex^{TM}$ in each iteration were replicated in E. coli. A phage preparation isolated on the third round selection produced 5.4 nephelos turbidity units (NTU) in a dextran specific immunonephelometric assay, a 2.2 fold higher value than the phage preparation from the first round selection. This phage gave $72\;{\pm}\;10$ normalized intensity (N.I.) in a dip-stick assay against high molecular size dextran (T2000, $2\;{\times}\;10^6) and significantly lower color ($30\;{\pm}\;6$ N.I.) against low molecular size dextran (T10, $10^4$). The presence of an Fab insert in each of these phages was confirmed using a $\beta-galactosidase linked assay and polymerase chain reaction.